As a manuscript

4 LEK PP "Nikolaeva Ekaterina Valerievna

Researching taste preferences in fish

Moscow - 2000

The work was carried out at the Severtsov Institute of Ecology and Evolution of the Russian Academy of Sciences and at the Faculty of Biology of the Lomonosov Moscow State University.

Supervisor:

Scientific consultant:

RAS Academician D.S. Pavlov

Professor, Doctor of Biological Sciences A.O. Kasumyan

Official opponents: Professor, Doctor

biological auks Yu.B. Manteifel

Candidate

biological sciences V.M. Stygar

Lead organization: Moscow Agricultural

Academy named after K.A. Timiryazev

The defense of the dissertation will take place on December 19, 2000 at 10 o'clock at a meeting of the dissertation council D 002.48.01 on the defense of dissertations for the degree of candidate of biological sciences at the Severtsov Institute of Ecology and Evolution of the Russian Academy of Sciences, 117071 Moscow, Leninsky Prospekt , 33.

The dissertation can be found in the library of the General Biology Department of the Russian Academy of Sciences.

Scientific Secretary of the Council, L.T. Kapralova

: PhD in Biological Sciences

£ w. М - / ¿А Ш о к ML 3. NS

Relevance of the topic. Taste reception plays a leading role in the sensory support of the final phases of feeding behavior associated with the assessment of the taste properties of prey and its compliance with the food needs of fish, their use of adequate food items for food (Atema, 1980;

Pavlov, Kasumyan, 1990). Numerous studies of the gustatory system of fish, carried out to date, are mainly devoted to elucidating the morphological characteristics and topography of taste buds, their ultrastructure and innervation. the study of the morphological organization of the central section (Reutter, 1986; Jakubowski, Whiter, 1990). The study of the functional properties of the gustatory system is carried out in most cases using electrophysiological methods (Marui, Caprio, 1992). Methods for assessing the taste preferences of fish remained poorly developed and, as a result, information on the attitude of fish to the taste of various types of substances was practically absent until recently. The volume of experimental data obtained using behavioral "test reactions and revealing the taste preferences of fish is limited and does not give a clear idea of ​​the general patterns and specific features of the attitude of fish to taste stimuli, the similarity and differences in the spectra of effective flavoring substances in fish of different lifestyles and systematic provisions (Kasumyan, 1997).

The influence on the taste sensitivity of fish of various biotic and abiotic factors, the habitat of fish, and the characteristics of their nutrition remains extremely poorly studied. The question of the presence and nature of relationships between taste preferences and the breadth of the food spectrum of fish, the composition of consumed food organisms, and the individual food experience of an individual remains completely open. There is no information on the severity of sexual dimorphism in fish in gustatory

preferences, about the influence of the lifestyle of fish on the manifestation of various elements of the behavioral response to gustatory stimuli.

Study of these and others topical issues gustatory reception of fish is of great theoretical importance because it allows us to elucidate the role of this sensory system in the choice of adequate food objects by fish. Knowledge of the patterns of gustatory sensitivity of fish, the specific features of their response to various types of flavoring substances is of undoubted practical interest and can be used to solve various problems of fisheries and aquaculture. The purpose and objectives of the study. The purpose of this work was to study the characteristics of taste preferences in fish. The tasks of the work included:

Comparison of taste preferences in fish living in waters with different salinity;

Research, the relationship between the nature of nutrition, food spectrum and taste preferences of fish;

Study of the influence of food experience on the taste preferences of fish;

Determination of the nature of the inheritance of taste preferences in fish;

Study of sexual dimorphism of taste preferences in fish (by the example of guppies);

Elucidation of the features of the manifestation of the main elements of the gustatory behavioral response in fish of different ecology.

Scientific novelty. The scientific novelty of this dissertation work lies in the expansion of ideas about the functional features of the gustatory system of fish. In this work, for the first time, the taste preferences of 7 fish species to classical flavoring substances and 4 fish species to free amino acids have been clarified. For the first time, a statement was formulated about the absence of correlations between the taste preferences of fish and their

relation to water salinity, between the level of fish euryphagia and the breadth of the spectrum of stimulating substances. It has been shown that fish with a well-expressed plant component in their diet are characterized by a positive attitude to sugars (sucrose).

For the first time on isogenic individuals raised on different types of food, it was shown that the taste preferences of "fish are not modified by food experience, are under strict genetic control and their inheritance is patroclinous. On the example of sexually mature guppies, the absence of sexual dimorphism in taste preferences in fish has been established. It has been shown that sex differences can be expressed in the intensity of the manifestation of a number of fish - parameters of the behavioral gustatory response.It was found that in fish with different lifestyles the manifestation of some elements of the behavioral gustatory response (the number of grasping of the granule, the duration of the retention of the granule in the oral cavity) differs. that different food experiences have no effect on the overall pattern of fish gustatory response.

Practical significance. The results of this study can "serve as a basis for the development and introduction into aquaculture of gustatory chemical stimulants designed to optimize formulations of artificial feeds in order to increase their palatability for farmed fish. The results obtained can be used in the aquarium industry (guppies, cichlazoma severum) and sport fishing. (roach, goldfish).

The research results are used in the course of lectures "Physiology of Fish", read to students of the Department of Ichthyology, Faculty of Biology, Moscow State University.

Approbation of work. Thesis materials were presented at the International Symposium on the Ichthyofauna of the Northern Regions (Bergen, Norway, 1993), conference

"Problems of Study, Rational Use and Protection of Natural Resources of the White Sea" (St. Petersburg, 1995), the 2nd All-Russian Meeting on Fish Behavior (Borok, 1996), the XXVII Conference of Young Scientists of the Biological Faculty of Moscow State University (Moscow, 1997), colloquia of the laboratory Behavior of lower vertebrates of the Institute of Ecology and Evolution of N.N. Severtsov, Russian Academy of Sciences and the Laboratory of Chemoreception and Chemocommunication of Fish, Department of Ichthyology, Moscow State University.

Publications. The main provisions of the dissertation are presented in 6 publications.

Structure and scope of work. The thesis consists of an introduction, 4 chapters, conclusions and a list of references. The manuscript includes

Pages, of which ..... typewritten pages, 7 figures

and 33 tables. The list of references contains 133 works by domestic and 151 works by foreign authors.

MATERIAL AND METHOD

Experimental work was carried out at the Department of Ichthyology of the Faculty of Biology and the White Sea Biological Station of Moscow State University, as well as in the laboratory of the behavior of lower vertebrates IPEE named after I. A.N. Severtsov RAS in the period from 1989 to 1998. The objects of this study were 7 fish species:

Roach Rutilus rutilus, two-year-olds, length (L) 6.5 cm;

Goldfish Carassius auratus gibelio, fingerlings, length (L) 6.5 cm;

Guppy Poeciiia reticulata, 6-8 months, length (L) 2.5 cm;

Cichlazoma severum Heros (Cichlasoma) severus, 4-5 months, length (L) b cm;

Nine-spined stickleback Pungitius pungitius, yearlings, length (L) 4.8 cm;

Polar flounder Liopsetta glacialis, 6 months old, length (L) 6.3 cm;

Banded catfish Anarhichas lupus, 5-6 months, length (L)

For experimental work, gynogenetic individuals (yearlings, 4-5 cm) of a hybrid of goldfish (female) and carp Cyprinus carpió (male), obtained from the All-Russian Scientific Society for Fish Farming (Dmitrov, Moscow Region), were also used.

The fish were acclimated to the conditions of the experiment from 2 weeks to several months. At this time, they were kept in common aquariums, at the optimum temperature for each species and fed with live or freshly frozen bloodworms (Chironomídae mosquito larvae) or fish meat (marine species). Experiments to elucidate the influence of the food experiment on the taste preferences of fish were carried out on caracecarp hybrids. After being caught from the pond, the hybrids (underyearlings) were divided into three groups and over the next 6 months were kept in different aquariums when fed with artificial food for aquarium fish "Tetramine" (manufactured by "Tetra", Germany; group No. 1), a living tubule ( oligochaetes of the R. Tubifex; group No. 2) and live or freshly frozen bloodworms (group No. 3).

1–2 weeks before the start of the experiments, the fish were placed singly in small aquariums (5–12 L). The back and side walls of the aquariums were made of opaque vinyl plastic to visually isolate the fish from each other. The fish were observed through a transparent front wall. Experiments with polar "flounder and striped catfish were carried out in a pilot plant from communicating compartments (25 l each) of the flow-through type, into which sea ​​water... To assess the taste preferences of fish to chemical stimuli, the method proposed by A.O. Kasumyan and S.S. Sidorov was used (Kasumyan, Sidorov, 1992, 1993). It consists in recording the behavioral response of single fish individuals to

artificial agar-agar gel beads containing one of the test chemical substances... In this work, free amino acids (L-stereoisomers) and classical flavoring substances causing the main types taste sensations in humans. Substances with a high degree of chemical purity were used as irritants. The list of used flavorings and their concentrations are shown in table 1.

Before the start of the experiments, the experimental individuals were trained to grasp the pellets containing the feed extract (75 g / l). During the experiments, the following was recorded: 1) the eatability of the granules, i.e. the grabbed pellet was eaten or discarded; 2) the number of acts of setting the granule; 3) the duration of the retention of the granule after the first setting; 4) the total duration of the retention of the granule for the entire time of the experiment. Registration began from the moment of the first setting of the granule. The moment the fish swallowed the granules was determined by the completion of the characteristic chewing movements of the jaws and the restoration of rhythmic movements by the gill covers. The duration of the retention of the pellet by the fish in the oral cavity was recorded using Agat summing stopwatches. The pellets with different substances were fed to the fish in a random sequence. The supply of pellets containing one of the test substances was alternated with the supply of pellets containing the feed extract. In control experiments, dye-only beads were used. Experiments in which there was no grasping of the pellet within 1 minute from the moment of its presentation to the fish were not taken into account. In experiments on the striped catfish, the experimental fish were simultaneously served 10 agar-agar plates (4x4x2 mm) containing the test flavors. After the expiration of one minute, the number of uneaten fish was counted.

plates, which were then removed from the aquariums. The interval between separate experiments on the same experimental individual in all series of experiments was at least 10-15 minutes.

Pellets were prepared immediately before the start of the experiment from agar-agar gel (Reanal, 2%), into which one of the test substances or feed extract was added during preparation. To make the granules more visible, into a gel. added dyes to give it a bright red (Ponceau 4R, 0.0005 M) or green color(Cr203.0.3% - in experiments with cichlazoma severum and striped catfish). The size of the granules corresponded to the size of the experimental fish and could be easily swallowed by them. For roach, guppy, and nine-spined stickleback, the pellet diameter was 1.5 mm and the pellet length was 2.5 mm; for the goldfish, polar flounder, and crucian carp hybrids, the pellet diameter was 2.0 mm and the pellet length was 3.0 mm. The gel containing amino acids or classic flavoring agents was stored at + 5 ° C for no more than 7 days. The gel containing the feed extract was stored under the same conditions for no more than 3-4 days.

The primary material was processed using the Statgraphics (Version 3.0), MatCad, and Microsoft Excel (7.0) 97 software package. Statistical analysis of the results was carried out using the Stuodent t-test and Spearman's rank correlation coefficient (r5). A total of 17372 experiments were performed.

1. Taste responses to classic flavors and

free amino acids in fish with different attitudes to water salinity and different nutritional habits

One of the objectives of this work was to clarify the characteristics of taste preferences in fish living in waters with different salinity. The research was carried out

freshwater fish - roach, goldfish, guppies and cichlazoma severum, euryhaline species - nine-spined stickleback and sea fish - polar flounder and striped catfish. Of greatest interest was the comparison of the listed fish species according to their taste responses to sodium chloride and calcium chloride - substances that are essential components natural waters. Experiments have shown that for roach and polar flounder sodium chloride is a taste stimulant that increases the consumption of granules, for goldfish and cichlazoma severum is a deterrent that significantly reduces the consumption of granules. In guppies, nine-spined sticklebacks, and striped catfish, an indifferent attitude towards granules containing this substance was found. Calcium chloride only for cichlazoma severum was a taste stimulant, for other freshwater and sea ​​fish studied by us, it acted as an indifferent flavoring agent (Table 1). Literature data have confirmed this position. Freshwater or marine fish species studied by other authors more often exhibit different or even opposite taste responses to the same taste stimuli, and much less often the same type of responses. Freshwater fish such as Leuciscus leuciscus dace, grass carp Ctenopharyngodon idella, cristivomer char Salvelinus namaycush, European grayling Thymallus thymallus, and anadromous Caspian trout Salmo trutta caspius show a taste preference for sodium chloride. In freshwater Siberian sturgeon Acipenser baerii, anadromous stellate sturgeon Acipenser stellatus, and sea cod Eieginus navaga, the inclusion of sodium chloride in the granules caused a significant decrease in the consumption of granules (Kasumyan and Sidorov, 1993; Kasumyan et al., 1993a; Kasumyan and Kazhlayev, 1993; Kasumyan , 1995a; Kasumyan, 1997; Kasumyan, Morey, 1997). From the same literature, it is known that calcium chloride causes an increase in consumption

Table 1. Taste attractiveness of classic flavors and free amino acids in fish living in waters with different salinity

("+" - stimulating effect, "-" - deterrent effect, 0 - indifferent taste properties)

p / n Irritant. Concentration, M Roach Goldfish Guppy Tsichlazoma severum Nine-spined stickleback Arctic flounder Banded catfish

1 Sodium chloride 1.73 (10%) ttu - 0 - 0 0

2 Calcium chloride 0.9 (10%) 0 0 0 0 0 0

3 Citric acid 0.26 (5%) - -.

4 Sucrose 0.29 (10%) 0 DOESN'T! 0 0 0 0

5 Glycine 0.1 0 0

6 Alanya 0.1 0 0 0

7 Serin 0.1 vzhsha shashsh 0 0

8 Proline 0.1 0. 0 0

9 Cysteine ​​0.1 0 0 0

10 Glutamic acid 0.01 0 0 0

11 Glutamine 0.1 gdot 0 W8SHYA 0

12 Aslaragic acid 0.01 0 0 0 0

13 Asparagine 0.1 0 0 0 0

14 Threonine 0.1 ¡SED 0 0

15 Tryptophan 0.01 0 0 0

16 Tyrosine 0.001 Myad-yayish 0 0

17 Norvalin 0.1 adayazya 0 0 0

18 Valine 0.1 0 EEZKZZZZ 0 0

19 Phenylalanine 0.1 0 - 0 0

20 Methionine 0.1 0 0 0 0

21 Leucine 0.01 0 shsh + tt 0 0

22 Lysine 0.1 0 0 886 * 355 $ 0

23 Isoleucine 0.01 0 0 0

24 Arginine 0.1 VJ + ZhZ 0 0

25 Histidine 0.1 0 0 0 0

pellets in common carp, chub Leodcum serba1us, christiwomer char, European grayling, rainbow trout Parabaitho tu ^ B and Caspian trout, and in Siberian sturgeon it causes a decrease in consumption of pellets (Kasumyan and Sidorov, 1993; Kasumyan and Kazhlaapuyev, 1993; Hoy, 1995a; Kasumyan, Morey, 1996; Kasumyan, 1997).

Comparison of taste preferences in fish living under conditions of different salinity was continued with the example of other classic flavoring substances - citric acid and sucrose. Unlike salts, citric acid and sucrose are not found in marine or fresh water natural reservoirs. It was found that for all the fish species we used, regardless of their relation to salinity, citric acid is a highly effective gustatory stimulus. For roach, goldfish and severum cichlazoma, as well as for polar flounder, citric acid has a repulsive taste properties... At the same time, it served as a gustatory stimulant for the striped catfish, guppy, and nine-spined stickleback, increasing the consumption of granules (Table 1). Literature data on other fish species support our results that citric acid is a highly effective gustatory irritant. In common carp, lacustrine char, chrystivomer, davatchan loach BajenpiB alpiru $ eryllusb and European grayling, citric acid caused a sharp increase in the consumption of pellets, and in grass carp, Siberian sturgeon and chum salmon Opsorphyns kela, a sharp decrease in consumption of pellets (1992 Kasumyan et al., 1993; Kasumyan, Kazhlaev, 1993; KaBituap, Sikogou, 1995a, 19956; Kasumyan, Morey, 1996, 1997; Kasumyan, 1997). Sucrose was an indifferent flavoring agent for polar flounder, catfish, severum cichlazoma, goldfish, nine-spined stickleback and served as a flavoring stimulant for roach and guppy. Analysis of own results and data of others

The fish species studied by us differ not only in their attitude to water salinity, but also in the nature of their nutrition, composition and breadth of food spectra. Of particular interest from the point of view of searching for connections between the above-mentioned nutritional characteristics of fish and their taste preferences are fish, in the diet of which a significant proportion is plant food... According to data concerning other groups of vertebrates, animals eating plant or mixed food usually show positive taste responses to mono- and disacchara (Bronstein, 1956; Harborne, 1985), as well as some other substances that have a sweet taste for humans (Kassil , 1972).

Among the species used by us, the plant component in the diet is most pronounced in roach (Domrachev, Pravdin, 1962; Klyuchareva, I960; Grandilevskaya-Deksbakh, 1961; Poddubny, 1966; Rodionova, 1969; Wetherley, 1987; Giles et al., 1990; Horppila, 1994) and guppies (Dussault, Kramer, 1981). It was these two fish species out of seven studied that showed a taste preference for sucrose - in roach, the consumption of granules with this substance was twice that of the control granules, and in guppies - 9 times and 22 times (for groups 1 and 2). In goldfish, algae and macrophytes are also found in the diet (Aristovskaya, 1935; Borutsky; 1950; Kharitonova, 1963, etc.), but sucrose was an indifferent irritant for him. In the rest of the fish studied by us, plant objects in the diet are poorly expressed or absent altogether, and all of them are characterized by an indifferent attitude to sucrose (Table 1). It is known from the literature that for dace and grass carp they are fish in the diet of which plants are present (Verigin, 1961; Lupacheva, 1967; Bobrova, 1968;

Stuge, 1973; Popov, 1975; Weatherley, 1987), sucrose had the properties of a taste stimulant that increased the consumption of granules (Kasumyan and Morey, 1997; Kasumyan, 1997). For fish such as carp, chub, minnow Phoxinus phoxinus, rainbow trout, char-

Davatchan, char-christivomer, Siberian sturgeon, stellate sturgeon, cod

Gadus morhua, navaga, Caspian trout, and chum sucrose were an indifferent flavoring agent (Kasumyan and Sidorov, 1992, 1993; Kasumyan and Kazhlaev, 1993; Kastimyan, Sidorov, 1995b; Kasumyan and Morey, 1996; Kasumyan, 1997). Of the above listed species, only carp and minnow have plants in their diet, as well as in the studied goldfish, but their share in the diet of these fish is not large (Stepanova, 1953; Dmitrieva, 1957; Cheremisova, 1958; Kharitonova, 1963; Lebedev , Spanovskaya, 1983). The rest of the species do not use plants for food, or feed on them sporadically, they are characterized by zoophagy.

The fish species studied by us also differ in the breadth of the range of food items consumed. So, according to the data of many * researchers, roach belongs to euryphages (Zheltenkova, I960; Shmidtov, 1962; Nebolsina, 1965; Grigorash et al., 1973; Ermolin, 1977, etc.). The goldfish also belongs to this group of fish (Aristovskaya, 1935; Dmitrieva, 1957; Stepanova, 1953; Cheremisova, 1958; Kharitonova, 1963). Polar flounder can be classified as benthophages with a wide range of food (Voevodin, 1996; Shubnikov et al., 1970; Yu.S. Reshtnikov, oral communication), planktophages with a wide range of food include guppies (Dassault and Kramer, 1981) , nine-spined stickleback (Wootton, 1976; Vvedenskaya, 1992, 1993; Maksimenkov and Tokranov, 1994) and severum cichlazoma (Axelrod and Vorderwinkler, 1993; Pausan, 1984; D.D. Zvorykin, oral communication). The striped catfish is characterized by pronounced stenophagy (Barsukov, 1983; Karamushko, Shatunovsky, 1994).

Analysis of the taste responses of the studied fish to classical flavoring substances shows that the breadth of the nutritional spectra does not affect the amount of flavoring substances with attractive properties. So, only one substance out of 4 classical taste stimuli had a stimulating effect for the polar flounder, severum cichlazoma and nine-spined stickleback - fish with a wide range of nutrition, as well as for the stenophage of the striped catfish. For the goldfish euryphage, none of the classic flavoring substances had a stimulating effect, while the roach and guppy had two such substances each.

Similar results were obtained when free amino acids were used as flavoring agents. Thus, in roach and goldfish there were 8 stimulating amino acids, in guppies - 5, in polar flounder - 0 (Table 1). Thus, the relative number of attractive amino acids for the four fish studied by us, which have broad food spectra, varies from 38 % (roach and goldfish) to 0% (polar flounder). The involvement of other researchers in the analysis of the results confirms the absence of an obvious connection between the level of euryphagia in fish and the number of amino acids with an attractive taste. euryphages, the amount of stimulating amino acids reaches 90% and 66%, while in carp, minnow and chub, which have a no less wide range of nutrition, this number is only 23%, 19% and 5% (Kasumyan, 1997). , only for goldfish were detergent amino acids (9.5%) found. properties makes the role of compounds of this class in the regulation of fish nutrition, their determination of taste properties more understandable.

captured prey, making a decision by fish to reject or swallow food. It was of interest to find out whether there is a relationship between the level of euryphagia in fish and the number of amino acids that have a repulsive taste for fish. Analysis of our own and published data shows that, as in the case of stimulating amino acids, such a relationship cannot be identified. Nevertheless, a certain tendency towards its manifestation, in our opinion, may nevertheless exist. Thus, amino acids with a deterrent effect were absent or were present in insignificant amounts, not only in the fish studied by us with a wide food spectrum (roach - 0%, guppy - 0%, polar flounder -0%, goldfish 9.5%), but also in dace (0%), minnow (19%), Siberian sturgeon (5%), cod (14%) (Kasumyan and Sidorov, 1994a; Kasumyan, 1997) are fish that consume a variety of food organisms. In fish with a limited food spectrum, the proportion of detergent amino acids may be higher (char-cristivomer - 57%). On the other hand, a large number of amino acids causes deterrent taste responses in the European grayling euryphage - (48%).

Correlation analysis of taste preferences for the classic flavoring substances of the fish studied by us from 21 possible option pairwise comparison of 7 species revealed 2 cases of significant negative correlation and no cases of significant positive correlation. The use of data from the literature on gustatory behavioral responses to classical gustatory stimuli of other fish species (22 species in total, including our own results) for analysis showed that out of 253 possible options for pairwise comparison, only in 25 cases (10%) a significant correlation was found, and in This relationship was positive in 12 cases and negative in 13 cases. Of the 12 pairs of species for which a positive correlation was found, only 7 pairs were combined freshwater species, 2 pairs consisted of

freshwater and anadromous species, 2 pairs - from anadromous and marine species of fish, and only one pair included freshwater and marine species (guppies, group No. 1 - navaga). A significant negative correlation was found for 13 pairs of species, of which 6 pairs consisted of freshwater fish, three pairs - from freshwater and anadromous species, two pairs - from freshwater and marine species, one pair - from anadromous and marine fish and one pair - from marine fish. Statistical analysis of taste preferences for free amino acids out of 21 possible variants of pairwise comparison of 7 fish species revealed 2 cases of significant correlation. A positive correlation was found for a pair of guppies - carp, and negative - for a pair of guppies - chum salmon.

These results show that the taste preferences of fish are characterized by a strongly expressed species specificity. In most cases, the taste spectra of the studied species do not correlate with each other. In cases of positive correlation, pairs usually unite fish that are distant in their lifestyle, diet, or taxonomy. Undoubtedly, for more rigorous conclusions about the nature of the relationships under consideration, an increase in the number of studied fish species is required, but even now it can be assumed that a direct relationship between the level of food specialization of fish, or their relation to water salinity and the breadth of the spectrum of stimulating flavoring substances, seems to be , absent. The only exception is fish with a well-pronounced plant component in their diet, which are characterized by a positive attitude to sucrose, while for zoophagous fish, sucrose has indifferent taste properties. The lack of correlation between taste preferences and the lifestyle of fish, in our opinion, can be explained by the strongly pronounced specificity of taste sensitivity in fish, which is manifested even in closely related species or in fish with a similar ecology (Kdsumyan, 1997; our data). Received

The results show that the species specificity of taste preferences is manifested in fish not only at the chemosensory level, as it was established earlier using electrophysiological methods (Oob and Tatiga, 1980; Mags et al., 1983b; Ishida, Lshaka, 1987), but also at the organismic level. Apparently, it is largely due to this particular feature of taste reception - the high specificity of taste preferences that the specific originality of the nutritional spectra, the divergence of jointly living fish different types by the composition of consumed organisms (Nikolsky, 1974).

2. Influence of food experience on taste responses.

One of the objectives of this work was to identify the modifying effect of the food consumed by fish, their individual nutritional experience on taste preferences. This problem was solved by us on the example of gynogenetic individuals of a hybrid between goldfish (female) and carp (male), characterized by a high level of genetic homogeneity.

It was found that hybrids grown for 6 months on different feeds (artificial feed "Tetramine", tubifex or bloodworms) are characterized by a high similarity of taste responses to classic flavoring substances and amino acids (Fig. 1). It was found that all groups of hybrids equally intensively consume granules with sodium chloride, and the consumption of granules with calcium chloride and citric acid slightly different among representatives different groups... The indices are characterized by a high degree of similarity in fish of different groups taste preference amino acids. The performed correlation analysis revealed significant positive correlations between the consumption of granules with the substances used by the caracecarp hybrids of the three experimental groups.

Consumption of pellets. %

<23456786 группа №1

group Кз2

123456780 groups »KsJ

Rice. 1. Flavoring. attractiveness of classic flavoring substances and selective free amino acids for caracecarp hybrids grown on artificial feed "Tetramine" (group No. 1), oligochaetes Tubifex (group No. 2), "larvae of chironomids (group Jfe 3).

1 - sodium chloride 1.73M (10%), 2 - calcium chloride 0.9M (10%), 3 - citric acid 0.26M (5%), 4 - sucrose 0.29M (10%), 5 - proline 0.1M, 6 - 0.1M series, 7 - 0.1M alanine, 8 - 0.1M glycine, 9 - control.

*, *** - reliability of differences in relation to control, p< 0.05 и 0.001."

The differences between the compared groups of fish are small, but they nevertheless may indicate some shifts in taste preferences in fish during prolonged feeding on different foods. Another feature is the higher palatability of the extract of the usual food for fish (except for group No. 2 - tubifex extract was less attractive than bloodworm extract). This indicates that the resulting diet leads not only to some shifts in taste preferences, but also to an increase, apparently, on

conditioned reflex basis, taste sensitivity to substances that are part of the consumed feed.

Thus, using the example of classical gustatory stimuli and free amino acids, it has been shown that the nature of the consumed food has a weak effect on the gustatory preferences of fish. This indicates a relatively low plasticity of gustatory chemoreception of fish in relation to such an important trophic factor as the composition of the diet, and also demonstrates a high genetic determination of gustatory preferences. Our results on the insignificant effect of the food experience on the taste preferences of fish are confirmed by the literature data (Kasumyan and Morey, 1997; Kasumyan, Morsy, 1998), according to which juvenile grass carp reared for 6 months on an animal (bloodworm) or vegetable (Lemna duckweed) minor, lettuce Lactuca sativa) feed, is characterized by a significant similarity in taste preferences. For other groups of vertebrates, it was also found that taste preferences are under strict genetic control (Ramirez and Sprott, 1978; Lush, 1981; Shingai and Beidler, 1985; Sinclair et al., 1992).

Comparison of the gustatory spectra of caracecarpic hybrids and parental species makes it possible to estimate the character of inheritance of gustatory preferences in fish. The results show that, in terms of "taste responses, the hybrids are much closer to carp than to silver carp (data on carp were taken from Kasumyan and Morey, 1996). This similarity is especially pronounced when comparing the taste responses of fish to citric acid, calcium chloride and some others. Thus, among the classic flavoring substances, the level of consumption of granules with citric acid was the lowest in goldfish, while in hybrids and carp citric acid is one of the most attractive substances in taste.

taste responses between the original species and hybrids allows us to speak about the patroclinous type of inheritance of taste preferences of substances during interspecific crosses. This conclusion is confirmed by the performed statistical calculations, which made it possible to reveal the existence of a positive correlation of taste responses to amino acids and classical taste substances between carp and hybrids (r5 = 0.51, p< 0.01). Достоверной связи между ответами серебряного карася и гибридов не установлено (rs = - 0.08, р > 0.05).

"3. Study of sexual dimorphism of taste preferences in fish

One of the previously not developed areas of research on taste reception is the question of the presence of sexual dimorphism of taste preferences in fish. The number of studies investigating sex differences in fish nutrition is extremely small (Baird, 1965; Dussault and Kramer, 1981; Sakashita, 1992; Aburto-Oropeza et al., 2000; Laland, Reader, 1999). According to these data, the nutrition of females and males of fish inhabiting the same biotopes may differ, which in turn indicates a possible discrepancy in taste preferences in fish of different sexes.

One of the species for which sex differences in nutrition have been found is the guppy (Dussault and Kramer, 1981). The objectives of this work were to carry out a comparative study of taste preferences and features of the manifestation of a behavioral response to various types of flavoring substances in individuals of the opposite sex using this fish species as an example. For the experiments, various types of flavoring substances were used - stimulating (sucrose, citric acid, glycine, L-glutamic acid), detergent (L-histidine) and

Consumption of pellets,%

1 00 90 8 070 -6 05 0403 020 -1 0 -

■ Males 1; S a m k and

1-G-T-1-G "T-I-g-t-g-

Chemical incentives

Rice. 2. Ranked ranks of chemical stimuli, compiled according to the magnitude of their palatability for male and female guppies.

1 - control, 2 - calcium chloride 0.9M (10%), 3 - sodium chloride 1.73M (10%), A - citric acid 0.26M (5%), 5 - b-histidine 0.1M, 6 - b-glutamic acid acid 0.01M, 7 - sucrose 0.29M (10%), 8 - glycine 0.1M, 9 - chironomid extract 75g / l.

**. *** - reliability of differences p< 0.05, 0.01, 0.001 соответственно, до косой черты - по отношению к контролю, после косой черты - между самцами и самками.

indifferent (sodium chloride, calcium chloride). Our results showed that there are no pronounced sex differences in taste preferences in guppies. For males and females guppies, the same substances from the number of used substances have taste efficiency, and

sequence of substances in ranked ranks constructed

separately for individuals of both sexes, almost completely the same (Fig. 2). The high value of the correlation coefficient between

consumption by males and females of guppies of granules containing substances with different levels of palatability (r5 = 0.97; p< 0.01). Поскольку у большинства видов рыб, в отличие от гуппи, отсутствует ярко выраженный половой диморфизм в морфологии, размерах и поведении, в частности, пищевом, вполне справедливым представляется предположение о том, что для этих рыб в целом, по-видимому, не характерен половой диморфизм по их вкусовым предпочтениям.

The differences between females and males of guppies relate only to the intensity of the manifestation of individual elements of the behavioral gustatory response. Thus, significant differences were revealed between males and females, according to such characteristics as the number of grasps and the duration of the retention of the granule after the first grasp and during the entire experiment. These differences affect only the intensity of the manifestation of these elements of the behavioral gustatory response, which in fish such as guppies, belonging to viviparous fish and characterized by pronounced sexual dimorphism in body size, may be associated with the peculiarities of the biology of this species (Dussault and Kramer, 1981; Laland , Reader, 1999).

4. Features of the behavioral response to food pellets in fish of different lifestyles

The behavioral response of fish to the presented granules is outwardly quite simple and includes several basic elements: grasping and rejecting the granule, its repeated grasping, retention of the granule in the oral cavity, in some cases chewing the granule, like in brown trout (Kasumyan and Sidorov, 1993, 19946). The frequency or intensity of the manifestation of these elements by fish varies and depends on the lifestyle of the fish and on the taste properties of the food item (Kasumyan, 1997).

In roach, goldfish, and cichlazoma severum, a significant negative correlation was found between the number of grasping acts and the attractiveness of the granule with classical flavoring substances and free amino acids from all the fish studied by us. The higher the attractiveness of the pellet for these fish, the fewer the number of grasping acts the fish performed in relation to the pellets with this irritant. This confirms the earlier conclusion that “attractive-tasting granules are swallowed by fish predominantly from the first try (Kasumyan, 1997). other species show that many river fish (minnow, dace, chub, European grayling, brown trout, rainbow trout, etc.) during experiments relatively rarely reject and then re-seize pellets (Kasumyan, Sidorov, 1993, 19946; Kavituap, B Clowy, 1995a; Kasumyan, 1997) Such a feature of the behavioral gustatory response of these fish is undoubtedly associated with habitation in sections of rivers and streams with high flow rates. In such conditions, rejected food items will be quickly demolished downstream and will become inaccessible, especially for fish leading a territorial lifestyle and not making long hunting dashes for prey. In fish (carp, etc.) that live mainly in slow-flowing or stagnant waters, as well as in marine fish such as cod and navaga, repeated grasping of the granule occurs much more often (Kasumyan and Morey, 1996;

Kasumyan, 1997). However, unlike cod and navaga,

the polar flounder studied by us, an inverse relationship was observed, which is possibly associated with its habitation in the littoral zone of the northern seas, where tidal flows of water are especially pronounced,

In the gustatory responses of roach, goldfish, guppy, polar flounder, and nine-spined stickleback to classic gustatory stimuli and amino acids, a positive correlation was found between the palatability of the granule and the duration of its retention in the oral cavity. These fish are characterized by longer testing of pellets with attractive substances in taste. Earlier (Kasumyan, Morey, 1996), it was suggested that such a feature of the gustatory response may be associated with the benthic type of fish feeding, which requires a long time for separation of food from a large amount of detritus captured along with the food. 8 "Linen £, 1991). Our results demonstrate that a similar dependence can be characteristic not only of benthophages, but also of planktophages, as is the case in nine-spined stickleback and guppies. Apparently, confident and unerring identification of substances attractive in taste, accompanied in some cases by multiple repeated taste tests of the granule, and the triggering of the reflex act of swallowing food on the basis of such information requires a longer time. Faster rejection of tasting food can be attributed to the minimization of unproductive feeding time. The assumptions made, however, are not universal, since, for example, in chum salmon, the relationship between the duration of taste testing of food and its attractiveness has, unlike many other species, the opposite character (Kasumyan and Sidorov, 1992). A number of authors have suggested that> long-term retention of a food object in the oral cavity can contribute to a more accurate assessment of its taste (Tamar, 1976; Kasumyan, Sidorov, 1993, 19946).

It was found that the taste responses of caracecarp hybrids grown on different feeds are characterized by high

the similarity of not only the level of consumption of the same type of granules, but also the coincidence of the general pattern of the taste response - an insignificant number of rejections and re-grasping of the granule, a pronounced positive correlation between the taste attractiveness of the granule and the duration of its retention in the oral cavity. This indicates that different food experiences have no effect on the manifestation of the main elements of the gustatory response by fish.

Conclusion

The results of the study show that fish of different ecology have a well-developed taste reception, which allows them to distinguish between the taste properties of substances of different types. Taste responses of fish are genetically strictly determined, - when interspecific crossing of goldfish and carp, they are inherited according to the patroclinic type. The relationship between the taste preferences of fish and the conditions of their habitat, or

the nature of nutrition is not detected, apparently due to the high level of species specificity of taste spectra, manifested not only at the chemosensory, but also at the organismal level. This feature clearly indicates the important and, of course, the determining role of gustatory reception in ensuring the selectivity of fish nutrition, their ability to selectively consume their characteristic food items.

The obtained data, along with the available data on the absence of noticeable population specificity of taste preferences in fish (Kasumyan and Sidorov, 1995) and on the absence of a noticeable influence of the individual feeding experience of fish (Kasumyan, Morey, 1997; KaBituap, Mogueu, 1998; our data), give grounds to assert that the taste spectra in fish can be considered as species suprapopulation traits, independent of sex or food consumed.

Many patterns of gustatory perception of fish, specific features of their response to various types of flavoring substances are of important practical interest and can be used to solve various problems of fisheries and aquaculture. The studies carried out show that it is promising to search for and create highly effective stimulants and detergents for fish, and serve as a biological basis for the development of methods for controlling the feeding behavior of fish using gustatory stimuli. The results obtained by us can be used to increase the palatability of feed, carry out work on the correction of their formulation by introducing special substances with a high stimulating effect, or by excluding components containing detergent compounds from the feed composition. This will not only reduce

direct losses of artificial feed during cultivation, but it will also provide a more efficient conversion of feed for fish growth, since it is known (Takeyeo and Takp, 1992) that the consumption of chemosensorically attractive feeds in fish is accompanied by a more intense secretion of digestive enzymes.

1. The investigated fish species of different ecology have well-expressed gustatory behavioral responses to chemical substances of various types - free amino acids and classic gustatory substances. A high level of species specificity of taste preferences in fish has been confirmed.

2. There is no correlation between the taste preferences of fish and their attitude to water salinity. Marine, freshwater and euryhaline fish do not differ in the latitude and composition of the spectra of substances with stimulating, indifferent or deterrent gustatory properties.

3. No relationship was found between the main characteristics of fish nutrition (the nature of nutrition, the breadth of the spectrum of consumed food organisms) and the taste preferences of fish (the number of stimulating flavoring substances, their composition).

4. It was found that fish with a well-expressed plant component in their diet are characterized by a positive attitude to sucrose. For fish - zoophages, sucrose has an indifferent taste.

5. The taste preferences of fish are strictly genetically determined. Individual food experience has little modifying effect on the taste preferences of fish. Inheritance of taste preferences in fish when

interspecific crossing (goldfish, female x carp, male) is patroclinous.

6. There are no gender differences in taste preferences in guppies. The differences between individuals of different sex relate only to the intensity of manifestation of individual elements of the gustatory behavioral response, which reflects the peculiarities of the biology of guppy reproduction.

7. It has been shown that the quantitative characteristics of the main elements of the behavioral gustatory response (the number of grasping of the granule, the duration of the retention of the granule in the oral cavity) differ in fish of different ecology. In most of the studied fish species, a negative correlation is observed between the number of repeated acts of grasping a food object and its palatability. The smallest number of seizure acts is observed in rheophilic fish. The majority of benthophagous fish are characterized by a longer retention of the food object in the oral cavity. No modifying effect of the food experience on the manifestation of the gustatory behavioral response by the fish was found.

1. Kasumyan A.O., Sidorov S.S., Platova-Nikolaeva E.V. 1993. The behavioural responses of juvenile wolffish to classical taste stimuli // In: Abstracts of scientific papers and posters for the Symposium on Mass Rearing of Juvenile Fish. Bergen. Norway 21-23 June 1993. Copenhagen. P. 37.

2. Kasumyan A.Ö., Platova-Nikolaeva E.B. 1995. Study of the gustatory sensitivity of some fish of the White Sea with different attitudes to water salinity // In collection: Problems

study, rational use and protection of natural resources of the White Sea. St. Petersburg. ZIN RAS. S. 122-123.

3. Kasumyan A.O., Marusov E.A., Nikolaeva E.V. 1996. Influence of food experience on gustatory and olfactory sensitivity of fish // In collection: Behavior of fish, abstracts. 2nd All-Russian. conference., Borok. S. 35-36.

4. Kasumyan A.O., Nikolaeva E.V. 1997. Taste preferences of guppies, Poecilia reticulata (Cyprinodontiformes, Pisces) // Questions of ichthyology. T. 37.No. 5.S. 696-703.

5. Kasumyan A.O., Marusov E.A., Morsy A.M. N., Nikolaeva E.V. 1998. Comparison of olfaction and taste behavioral responses to free amino acids in two cyprinid fishes // Chemical Senses. V. 23, No. 5, P. 642-643. "

6. Nikolaeva E.B., Kasumyan A.O. 2000. Comparative analysis of taste preferences and behavioral response to taste stimuli in female and male guppies, Poecilia reticulata // Questions of ichthyology. T. 40. No. 4. S. 560-565.

Chapter 1. Literature review.

Chapter 2. Material and method.

2.1. Research objects.

2.2. Keeping conditions and preparation of experimental fish for experiments.

2.3. Methodology for conducting behavioral experiments.

2.4. Technique for the preparation of experimental agar-agar granules.

2.5. General characteristics of the material.

Chapter 3. Results.

3.1. Study of the gustatory behavioral response of fish of different ecology to classic flavoring substances and free amino acids.

3.1.1. Roach.

3.1.2. Goldfish.

3.1.3. Guppy.

3.1.4. Tsikhlazoma severum.

3.1.5. Nine-spined stickleback.

3.1.6. Polar flounder.

3.1.7. Striped catfish.

3.2. Taste behavioral responses of karacecarp hybrids grown on different feeds.

3.3. Gustatory behavioral responses of male and female guppies.

Chapter 4. Discussion of results. 54 Conclusion. 90 List of cited literature. 92 Appendix

Introduction Thesis in biology on the topic "Research of taste preferences in fish"

Taste reception plays a leading role in the sensory support of the final phases of feeding behavior associated with the assessment of the taste properties of prey and its compliance with the food needs of fish, and their use of adequate food items for food. Numerous studies of the taste system of fish, carried out to date, are mainly devoted to elucidating the morphological characteristics and topography of taste buds, their ultrastructure and innervation, and the study of the morphological organization of the central section. The study of the functional properties of the gustatory system is carried out in most cases using electrophysiological methods. Methods for assessing the taste preferences of fish remained poorly developed and, as a result, information on the attitude of fish to the taste of various types of substances was practically absent until recently. The volume of experimental data obtained using behavioral test reactions and revealing the taste preferences of fish is limited and does not give a clear idea of ​​the general patterns and specific features of the attitude of fish to taste stimuli, the similarity and differences in the spectra of effective flavoring substances in fish of different lifestyles and systematic position. ...

The influence on the taste sensitivity of fish of various biotic and abiotic factors, the habitat of fish, and the characteristics of their nutrition remains extremely poorly studied. The question of the presence and nature of relationships between taste preferences and the breadth of the food spectrum of fish, the composition of consumed food organisms, and the individual food experience of an individual remains completely open. There is no information on the severity of sexual dimorphism in taste preferences in fish, on the influence of the lifestyle of fish on the manifestation of various elements of the behavioral response to taste stimuli.

The study of these and other topical issues of taste reception in fish is of great theoretical importance, since it makes it possible to elucidate the role of this sensory system in the choice of adequate food objects by fish. Knowledge of the patterns of gustatory sensitivity of fish, the specific features of their response to various types of flavoring substances is of undoubted practical interest and can be used to solve various problems of fisheries and aquaculture.

The purpose of this work was to study the characteristics of taste preferences in fish. The tasks of the work included: comparison of taste preferences in fish living in waters with different salinity; study of the relationship between the nature of nutrition, food spectra and taste preferences of fish; study of the influence of food experience on the taste preferences of fish; determination of the nature of the inheritance of taste preferences in fish; study of sexual dimorphism of taste preferences in fish (by the example of guppies); elucidation of the features of the manifestation of the main elements of the gustatory behavioral response in fish of different ecology.

The scientific novelty of this dissertation work lies in the expansion of ideas about the functional features of the gustatory system of fish. In this work, for the first time, the taste preferences of 7 fish species to classical flavoring substances and 4 fish species to free amino acids have been clarified. For the first time, a position was formulated on the absence of correlations between the taste preferences of fish and their relation to water salinity, between the level of fish euryphagia and the breadth of the spectrum of stimulating substances. It has been shown that fish with a well-expressed plant component in their diet are characterized by a positive attitude to sugars (sucrose).

For the first time on isogenic individuals raised on various types of food, it was shown that the taste preferences of fish are not modified by food experience, are under strict genetic control, and their inheritance is patroclinous. On the example of mature guppies, the absence of sexual dimorphism in taste preferences in fish was established. It has been shown that sex differences can be expressed in the intensity of the manifestation of a number of parameters of the behavioral gustatory response by fish. It was found that in fish with different lifestyles, the manifestation of some elements of the behavioral gustatory response (the number of grasping of the granule, the duration of the retention of the granule in the oral cavity) differs. It was found that different food experiences have no effect on the general pattern of fish gustatory response.

The results of this study can serve as a basis for the development and introduction into aquaculture of gustatory chemical stimulants designed to optimize the formulations of artificial feeds in order to increase their palatability for farmed fish. The results obtained can find application in the aquarium industry (guppies, cichlazoma severum) and sport fishing (roach, goldfish).

Conclusion Dissertation on the topic "Ichthyology", Nikolaeva, Ekaterina Valerevna

CONCLUSION

The main chemosensory systems of fish - olfactory and gustatory - can participate in the formation of the behavioral response of fish to granules containing chemical substances. A comparative analysis of the behavioral responses of anosmic and control (intact) fish (Kasumyan and Morey, 1996), evaluated using a method identical to ours, shows that depriving fish of their olfactory sensitivity does not lead to a change in their response to granules with substances. In anosmic and intact fish, the threshold concentrations of substances introduced into the pellets also coincide. These data give grounds to assert that the olfactory system is not involved in the sensory supply of the observed behavioral responses of experimental fish to granules with different substances and that the nature and intensity of these responses are controlled by intraoral taste reception.

The results of the study show that fish of different ecology have a well-developed taste reception, which allows them to distinguish between the taste properties of substances of different types. The gustatory responses of fish are strictly genetically determined; during interspecific crossing of goldfish and carp, they are inherited according to the patroclinic type. The relationship between the taste preferences of fish and the conditions of their habitat or the nature of nutrition is not found, apparently due to the high level of species specificity of taste spectra, which manifests itself not only at the chemosensory, but also at the organismal level. This feature clearly indicates the important and, of course, the determining role of gustatory reception in ensuring the selectivity of fish nutrition, their ability to selectively consume their characteristic food items.

The obtained data, along with the available data on the absence of a noticeable population specificity of taste preferences in fish (Kasumyan and Sidorov, 1995) and on the absence of a noticeable effect of the individual feeding experience of fish (Kasumyan and Morey, 1997; Kasumyan, Morsy, 1998; our data), give grounds to assert that the gustatory spectra in fish can be considered as species supra-dyopulation traits, independent of sex or food consumed.

Many patterns of gustatory perception of fish, specific features of their response to various types of flavoring substances are of important practical interest and can be used to solve various problems of fisheries and aquaculture. The studies carried out show that it is promising to search for and create highly effective stimulants and detergents for fish, and serve as a biological basis for the development of methods for controlling the feeding behavior of fish using gustatory stimuli. The results obtained by us can be used to increase the palatability of feed, to carry out work on the correction of their formulation by introducing special substances with a high stimulating effect, or by excluding components containing detergent compounds from the feed composition. This will not only reduce the direct loss of artificial feed during rearing, but also ensure a more efficient conversion of feed for fish growth, since it is known (Takeda, Takii, 1992) that the consumption of chemosensorically attractive feeds is accompanied by more intense secretion of digestive enzymes in fish.

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The concept of aesthetic taste was formed in European culture in a relatively late historical period on the basis of individualization of spiritual experience and became a condition for the diversity of the content of spiritual values. Aesthetic taste is defined as the ability of a person, depending on the feeling of pleasure or displeasure, to perceive and evaluate the degree of aesthetic completeness of the objective world and spiritual phenomena. Aesthetic taste objectifies itself in value judgments, as well as in all forms of formative activity, starting from everyday manifestations of style in clothing, in lifestyle, in relation to social, in particular artistic, values. The classic definition of aesthetic taste is contained in Kant's work "Anthropology in a pragmatic sense". The philosopher writes: "Taste is the ability of the aesthetic judgment to make choices that are universal." As the philosopher notes, we are talking about the correspondence of our judgments to the objective qualities of the subject of judgment, which is the guarantee of their universality. The objectivity of the judgment of taste is evidenced by the presence of a developed aesthetic experience in relation to the world. On this basis, the German philosopher I. Sulzer puts developed taste along with such intellectual abilities as reasonable knowledge and moral attitude: "Taste ... is nothing more than the ability to feel beauty, just as reason is the ability to cognize the true, perfect, true, and moral feeling is the ability to feel good. "

The problem of taste has been put forward in aesthetic theory at one of the leading places, starting from the Renaissance, as a reflection of the phenomenon of individualization of the spiritual experience of a person. In artistic formation, she carries out a departure from the canons, and in aesthetic assessments, she begins to move away from the well-established ideas about the meaning of perfection. Pushing the boundaries of the established in aesthetic experience, carriers of taste assert new facets of the value of phenomena or offer a new vision of them. Thus, medieval ideas about bodily beauty as sinful are changed by the assertion of bodily beauty, a hymn of harmony of the bodily and spiritual principles in a person is sung. We find interesting ideas about the essence of taste in the treatises of the epoch leaders L. Valla, M. Ficino, Pico de la Miran-dole, Leonardo da Vinci.

In the XVII century. the concept of "taste" begins to be used in a categorical sense, in particular, thanks to the works of the Spanish philosopher Gracian i-Morales ("Hero", "Clever", etc.). Let us recall that philosophers of the 17th - 18th centuries pay great attention to the development of the theory of taste. in France (Battier, La Rochefoucauld, Tremblay, Rousseau, Helvetius, Voltaire), in England (Shaftesbury, Hutcheson, Burke, Hume, Mandeville), in Germany (Winkelmann, Lessing, Herder, Sulzer, Kant, Schiller). The attention of researchers is focused on questions of the nature of taste: whether it is rational or irrational, based on reason or feelings, taste is acquired in education or is an innate ability. La Rochefoucauld raises the question of the individual definiteness of taste (treatise "Maxima"). Voltaire in his work "Smack" characterizes this phenomenon as a sensory response to good and bad, based on the ability of the intellect to distinguish between the objective qualities of the objective world. Voltaire singles out such a modification of aesthetic taste as artistic taste. The philosopher defines the concept of "taste" in the system of aesthetic knowledge as a kind of "metaphor" designed to indicate sensitivity to the beautiful and the ugly in the arts. Depending on this ability, he divides tastes into good, bad and distorted. “Taste in art is distorted,” writes Voltaire, “is expressed in love for subjects that disturb the educated mind, the preference for the burlesque over the noble, the pretentious and prim over the simple and natural beauty, this is a disease of the spirit (italics by V. M.) 11 Such a characteristic sounds very relevant now, in conditions of subordination of artistic culture to the demands of the market.The spread of bad taste is harmful to the general spiritual development of the individual.

Voltaire emphasizes that artistic taste is the result of a long and careful upbringing. A person must slowly learn to listen and peer into the natural world and master artistic values. Habit and reflection make her able to suddenly feel pleasure, discerning previously inaccessible to her. The philosopher singles out the phenomenon of individualization of taste not only at the level of the face, but also at the level of the nation as a whole: "Taste is slowly brought up in the nation, because it slowly perceives the spirit of its best artists."

Voltaire also gives an interpretation of one of the pressing problems of the theory of taste: can you argue about tastes? The philosopher clearly divides taste as a physiological property of the organism and an aesthetic taste. Of course, there is no dispute about tastes when it comes to preferences associated with bodily pleasures: what is pleasing to one person may be unpleasant to another. However, this does not apply to art. "Since there is real beauty in art, then there is a good taste that distinguishes it, and a bad taste that does not perceive it, and the defects of the mind - the source of spoiled taste - must be corrected." the practice of aesthetic education. First, the source of the formation of developed taste is beauty. The objective source of beauty is art, which means that it is "an active factor in the formation of developed taste. Secondly, the beauty that objectively exists in perfect works of art requires the development of sensory-intellectual structures in order to open up in its qualities to the subject. Thirdly, penetration into the world of beauty is possible only under the condition of coordinated interactions of the subject's spiritual structures: the ability of sensory perception and the activity of the mind, reveals the quality of an object of indifference. This is the correspondence of the object to the idea of ​​the expedient (inner vitality) and the perfection of its identification in the work as a spiritual whole.

The aesthetic theory differentiates the level of tastes. So, Helvetius (treatise "On the mind") divides them into two types: "the taste of habit" and "conscious taste". This, according to the now widespread classification, in accordance with the taste is limited and developed. For the theory of taste and the practice of educating developed taste, Helvetius's reasoning about the difference between these two levels of taste is important. The philosopher sees it in the difference between the ideas about the essence of beauty, despite the fact that both types are based on aesthetic experience. The first - "the taste of a little call11" - is distinguished by a certain skill of evaluating phenomena. The judgment of the taste of connoisseurs of this kind is characterized by the fact that they consider valuable only that which has already been consolidated in their experience. "- writes Helvetius. The second type -" conscious taste "- is based on a deep knowledge of the subject of assessment and the spiritual experience produced by culture. Its carriers are able to appreciate new artistic phenomena and their assessment will be objective. The formation of this type of taste is achieved by a long study of works of art and scientific ideas, reveals knowledge of the truly beautiful.

Educational function in the formation of developed tastes of society in the 18th-19th centuries. performed literary and art criticism, which at that time was a special type of aesthetic activity. She played a huge positive role in the formation of European culture (including Eastern European) by awakening interest in the highest, most aesthetically perfect treasures of national and world culture, orienting the public to such samples and producing in their aesthetic analysis the criteria of aesthetic perception and judgment of taste. Unfortunately, in the culture of postmodernity, this tradition has been lost, and therefore the public taste is spoiled.

The complex structure of aesthetic taste is explored in the aesthetics of English sensationalism. According to E. Burke, taste is formed by "primary pleasures of the senses from the perception of phenomena, secondary sweets of the imagination and conclusions of the mind about the various relationships between them, as well as about human passions, morals and actions." So, taste is not a revelation of immediate sensibility as such, not a sphere of the purely irrational, but also not a purely conceptual sphere. Taste is an organic interplay of sense gratification, imagination enjoyment and reasoning. It is also noted that the said interaction of intellectual and sensory structures is common for any type of aesthetic attitude, and tastes can be improved constantly thanks to the expansion of the experience of cognition, deepening as an object and constant exercises in aesthetic knowledge. Let's pay attention to the all-encompassing phenomenon as revealed by the philosopher. Taste is viewed as an intellectual ability and as a consequence of the conscious choice of objects and the creation of an individualized attitude towards them. It is also important that the sphere of individualization of aesthetic experience, objectifies itself in judgments of taste, does not define any phenomena, but perfect artistic phenomena, that is, carriers of the universal value in their content. The latter is important in the construction of a scientific theory of aesthetic education and self-education of the individual. The classical aesthetic theory sees a person as an active subject of an aesthetic attitude, and removes the idea of ​​relativism from the process of aesthetic education both in terms of the content of values ​​and in terms of purpose.

Let us return to German classical aesthetics, focusing on the theory of taste in Kant's aesthetics, developed in the following works: Observations of the Sense of Beauty and the Sublime (1764), Critique of Judgment (1790), Anthropology in a Pragmatic Ratio (1798). In developing the theory of taste, Kant largely relied on the ideas of E. Burke, D. Hume and other English sensationalist philosophers. the play of cognitive abilities. "The first point contains the idea of" non-cycling11 judgment of taste. It proves that taste is associated with aesthetic pleasure, due to the qualities of the object of indifference. The second and fourth points assert the beautiful as such, that in the judgment of taste it is cognized without concept, since it is "the subject of necessary pleasure." That is, taste is based on the sense of beauty. Art, embodies the sensual manifestations of beauty, was revealed by I. Kant as a source of a special way of cognition - cognition in images that give tremendous spiritual pleasure in the perfection of forms. Cognition based on sweetness is imperceptible: cognitive abilities seem to play rather than work. The third aspect of the judgment of taste asserts its intrinsic value - "expediency without a goal", since the object of the aesthetic judgment of taste is a goal for the senses due to the purposeful and perfect inner vitality. Taking into account its internal purposefulness, Kant compares a work of art with nature in its inherent purposefulness of forms, while realizing what is different between them. The philosopher notes such an inner perfection of a work of art when he acts as a spiritually formative principle, collects human spiritual structures into integrity, combining in the judgment of taste the coherence of intellect and feelings. The philosopher interprets taste as the ability to evaluate (natural phenomena, artistic phenomena), even allows "feeling ... to convey to everyone else." In other words: the artistic persuasiveness of a work is capable of awakening and shaping feeling and intellect, prompting the adoption of an aesthetic judgment of taste adequate to the qualities of the work.

An important problem in judgments of aesthetic taste, Kant sees the dialectics of the individual and everything in common. If individual judgment includes the principle of universality, then this principle must be inherent in the aesthetic feeling itself. The feeling of satisfaction is conditioned by the general purposefulness, it is subjectively defined as an a priori principle of consciousness, and objectively it appears as the "pure form" of the object. Kant substantiates the path to the acquisition of universal experience by consciousness, putting forward the concept of "three maxims of everyday consciousness." They can help in explaining "criticism of taste", namely: 1) have their own judgment; 2) mentally put yourself in the place of each other; 3) always think in harmony with yourself.

Clarifying the concept, the philosopher argues that the first of them means the maxim of a way of thinking free from prejudices; the second is a broad way of thinking, that is, the ability, in one's own way of judging, to get out of a universal point of view (which can be found only by sharing the views of others). Finally, the third maxim - the Poslead way of thinking - is achieved only through a combination of the first and the second and such interaction between them that it becomes a skill. This maxim is achieved most. The three named maxims, according to Kant, cover all spheres of the intellect, since the first of them is the maxim of reason, the second is the ability of judgment, and the third is of the mind. their dialectical connection is built as follows: imagination in its freedom awakens the activity of reason, which, without the mediation of concepts, provides the rightness to the play of the imagination: what is presented to others will not be seen as a thought, but "as an inner feeling of the expedient state of the soul (italics by V. M.)." Kant regards the appearance of "the feeling and judgment of taste as a need conditioned by the nature of man - a creature destined to live in society, and therefore to feel the need for communication."

In Kant's aesthetics, the idea of ​​the practically uninterested nature of the aesthetic judgment of taste is consistently traced. Interested judgments of taste are based both on their prerequisite, on an interest in a morally good and on the makings of a morally good way of thinking. The judgment of taste in relation to nature and art is a way of presenting "expedient without a purpose", because it is necessary to create a culture of the soul's abilities for communication between people. The general perception of pleasure provides that this pleasure of pleasure, based not only on sensations, but is the pleasure of reflection, that is, it is associated with the reflexive ability of judgment.

The history of the formation of the theory of taste is analyzed, which is evidenced by its open science connection with truth, the ability to adequately perceive sensory manifestations of perfection in natural phenomena and artistic phenomena, the ability to reflect on perceived phenomena and one's own experience of their qualities, and finally, the ability to convey in the judgment of taste one's feelings caused by the object. The qualitative definiteness of judgments of taste, developed by the aesthetic theory, is characterized - this is its truth (a desired sample, an ideal model). In the practice of value judgments, it manifests itself as a developed or aesthetic taste. Its bearer is a person with rich spiritual experience, capable not only of making objective judgments about value, but also of creating them. it is characterized by a sense of proportion in self-expression, the presence of a criterion in aesthetic judgments and in relations with the world (attitude towards other people, towards the moral and artistic values ​​of society and humanity, etc.). her experiences of phenomena and judgments about them are characterized by individual uniqueness with a pronounced correspondence to the universality of the content of judgments (evidence of the possession of truth about things).

Along with developed taste, modern aesthetic theory distinguishes a bad, or spoiled (distorted) type of tastes. The bearers of this taste get pleasure from the contemplation of ugly phenomena and are indifferent to beauty. The danger of such value ideas that their carriers spread negative in human manifestations, in artistic and practical formation, implanting it for granted, as the norm in understanding the nature of the valuable. In addition, the way pseudo-values ​​are asserted is aggressive in nature, which corresponds to the very subject of the assertion. The danger of such content of value ideas and value judgments that they are capable of spoiling public tastes, spreading interest in the ugly and immoral under the slogans of "fashionable", "original".

With regard to quantitative characteristics, then alongside the developed one, an undeveloped ("narrow") taste is rightfully distinguished. Its carriers are people of a low level of culture, limited experience of communication with artistic values. They lack the criterion of good and bad, beautiful and ugly. they are distinguished by non-argumentation of evaluations, helplessness in judging the quality of an object. They are not able to explain to themselves why and what attracted them to this or that subject of judgment. their assessments do not include the general content of value concepts. These judgments are based on empirical experience, therefore, they are arbitrary. It is worth noting that for communication they choose what is understandable for themselves and thus pleasant, and therefore are in a circle of limited experience. Standard objects neutralize tastes, cause monotony of judgments regarding their quality.

Note that undeveloped taste lends itself to education and, subject to systematic communication with aesthetic values ​​and bearers of developed taste, can deepen and improve. The formation of a developed aesthetic taste is the goal of the pedagogical process, because children are usually undeveloped tastes due to little experience of communication with artistic values. It is worth noting, however, that on an intuitive level, they are able to grasp aesthetically expressive phenomena more accurately than adults, although they are not able to evaluate their choice and determine the evaluation criteria.

Let us focus on the social value of a developed aesthetic taste, taking into account its formative capabilities both in relation to a person and in relation to society as a certain spiritual whole. Usually the meaningful level of taste is due to the "zeitgeist". Interest in the truth or, conversely, ignoring it is reflected in the nature of the response to the phenomena of a large circle of people (it is legitimate to talk about the tastes of an era, nation, class, etc.). Kant argues that good taste occurs only during the period of healthy, not just refined taste. Developed taste universalizes feelings, directing them towards understanding the universal in the spiritual experience of relationship, and at the same time individualizes them. An individualized manifestation of taste is valuable in that it demonstrates the nuances of the qualities of the object of care and allows you to convey to others the feeling experienced by the person. Consequently, the foundations are created for co-creation in evaluative-experiencing interactions. Therefore, aesthetic taste serves as an effective means of spiritual unification of people.

Note that the spiritually forming function is performed only by "good", developed taste, based on a rich experience of communication with aesthetic values. The most effective educational impact is artistic culture, first of all, the classical artistic heritage, arises as a perfect embodiment of a high level of aesthetic experience. The richness of its value content and the perfection of the artistic image of his being determine the effectiveness of the formative possibilities in the sphere of the spirit. Let us pay attention to the opinion of I. Kant once again, good taste distinguishes what corresponds to the "concept of a thing," and the meaning of choice is the ability to choose what everyone likes, that is, to make a choice that would be public with feelings. Let us also pay attention to the fact that the object of feelings and value judgments of taste is not an object as such (material-material reality), but its spiritual and value sense. The deeper the subject of evaluative activity penetrates as an object of judgment, the more complete and deep will be the judgment of taste, and his experience unfolds in judgment as individually unique, creative in character. The carriers of the developed taste are able to reveal what, in the words of I. Kant, "I like it for a long time."

The present invention relates to a method for preparing a palatability enhancer for use in pet foods with low, medium and high moisture content. The method includes: (i) reacting a substrate with at least one protease in the absence of any added lipase, wherein the substrate contains protein and fatty materials, (ii) heat inactivating said protease and filtering the resulting cleavage product; d) reacting the resulting emulsion with at least one lipase in the absence of any added protease to obtain a second stage reaction product: e) adding to said second stage reaction product at least one reducing sugar and at least at least one nitrogen compound and heating the resulting mixture. 7 n. and 7 p.p. f-crystals, 6 dwg., 12 tbl., 5 ex.

The technical field to which the invention relates

The present invention relates to a method of enhancing the palatability of compositions (including foods, additives, condiments, toys, and the like) for companion animals. In particular, the invention relates to a method for preparing a palatability enhancer for use in pet food with low, medium or high humidity.

The invention relates to the field of pet food, such as dogs and cats. All references herein to food of any kind are intended to refer only to food that is manufactured and sold for domestic animals such as dogs and cats. Although trials have so far been limited to dogs and cats only, the invention can also be adapted for use in rodents and other types of companion animals. If necessary, the invention disclosed herein can also be tested to determine its applicability for use in various classes of animals, including rodents (such as: hamsters, guinea pigs, rabbits and the like), birds, as well as horses and any type of livestock. However, since dogs and cats exhibit a higher degree of sensitivity to taste, taste and odor than rodents, horses, livestock, etc., and since generally dogs and cats receive food with an improved taste, the term “domesticated food” is used in this text. animals "will refer to all animals that appear to be receiving an improved palatable food as disclosed herein, and the term" animal food "will refer to all types of food offered to those animals.

It should be understood that the expression "animal feed" as used herein includes foods having a low, medium or high moisture content. There are three categories of animal feed: (1) dry foods or foods with a low moisture content (usually less than 15%), which are usually high in nutrients, cheaper packaging, easier to handle, but less palatable; (2) canned or moist foods or foods with a high moisture content (greater than about 50%), which usually have the greatest palatability to pets; (3) semi-wet or semi-dry foods or foods with intermediate or medium moisture content (usually 15% to 50%;), which usually have less palatability than canned foods, but more palatability than dry foods.

Prior art

Caring owners provide pets with a suitable choice of food. These foods may be a regular diet and may include their usual diet, supplements, additional treatments, and toys. Pets, like humans, prefer and more often and more willingly eat food that is palatable to them. Therefore, palatability enhancers are extremely important for animal consumption. Animal feeds — such as pet foods, typically contain flavoring compositions to enhance their palatability and appeal to the pet. To date, a large number of flavoring compositions (palatability enhancers or palatability factors) have been described. For example, US Pat. Nos. 3,857,968 and 3,968,255 to Haas and Lugay disclose a palatability-enhancing composition for use in dry animal foods, especially dry dog ​​foods, containing fat and proteins, which is prepared by a process comprising emulsifying fat, treating the composition with a mixture of enzymes, containing lipase and protease, and, optionally, enzyme inactivation. Another example is described in US Pat. No. 4,713,250, in which a palatability enhancing composition for dog food is prepared by a multi-step enzymatic reaction comprising firstly contacting an aqueous proteinaceous or mealy material with a protease and / or amylase, then preparing an emulsion containing fat and the product obtained in the first stage, and reacting said emulsion with lipase and protease. The following example is described in US patent No. 4089978, where Lugay et al. propose a composition with improved palatability for use in animal feed, which is prepared by a method comprising reacting at a moderate temperature an aqueous mixture of reducing sugar, animal blood, yeast and fat with an enzyme mixture containing lipase and protease, followed by an increase in temperature for more complete formation taste and enzyme inactivation.

However, the palatability of such compositions is generally different for different animal species. For example, a flavoring agent that is effective for cats is often not effective for dogs. Moreover, a flavoring agent that is effective in dry pet foods is generally ineffective in semi-moist or wet pet foods. Therefore, there is a continuing need for new palatability enhancers that provide a lasting mouthfeel and that can be used easily and effectively for pets, including dogs and cats, in a variety of feed types such as dry, intermediate and wet foods.

The essence of the invention

Thus, it is an object of the present invention to provide such a "generally applicable" palatability enhancer, ie palatability enhancer, which is effective regardless of animal species and feed type. The method of the present invention significantly enhances the palatability of low, medium or high moisture pet foods.

Thus, a method for producing a palatability enhancer is presented. This method involves staged enzymatic hydrolysis followed by a flavor-generating thermal reaction of raw materials (eg, poultry, pork, beef, lamb, fish products, and the like) to produce a highly effective palatability enhancer. The palatability enhancer can be added to pet food as a liquid product by coating spraying, as a dry powder in a coating spray process, or as a liquid or dry product by mixing it with pet food ingredients prior to packaging or canning. Alternatively, the palatability enhancer can be mixed with the fat and added at the same time.

The present invention also contemplates combining two or more palatability enhancers, including one as disclosed herein, to form compositions or mixtures useful for enhancing palatability of pet foods. The different types of palatability enhancers can be mixed together before being incorporated into the feed (palatability enhancer blends can be stored this way until they are used). Alternatively, the various palatability enhancers can be combined in situ, that is, directly in the pet food.

Surprisingly, it has been found that the palatability enhancer according to the present invention is the first one that is highly attractive to pets, especially dogs and cats, when added to pet food having a low, medium or high moisture content.

Brief description of the figures

Fig. 1 is a graph showing palatability results of Super Premium palatability enhancer SP1 versus SP2 (Example 1).

Figure 2: A set of graphs showing palatability results of palatability enhancers XLHM versus Super Premium SP1 and SP2 (Examples 1 and 2).

FIG. 3 is a graph showing palatability results of a palatability enhancer from Super Premium PRODUCT B versus PRODUCT C (Example 5).

Detailed description of the invention

The term "palatability" means the relative preference for an animal of one food composition over another. Palatability can be determined by a standard test procedure in which the animal has equal access to both compositions. This preference can arise from any of the animal's senses, but is usually associated with taste, smell, mouthfeel, texture, mouthfeel. It is determined here that the pet food having the increased palatability is the food that the animal prefers over the control composition.

The terms "palatability enhancers" or "flavors" or "flavors" (flavors, flavors), or "palatability factors" or "palatability factors" denote any material that enhances the palatability of a food composition for an animal. The palatability enhancer can be a single material or a mixture of materials, and it can be natural, processed or untreated, synthetic or partially natural and partially synthetic materials.

As used herein, the term "slice" ("kibble") refers to specific slices or nuggets obtained in a squeeze or extrusion process. Typically, the slices are made to produce dry and semi-moist pet food. The pieces can vary in size and shape, depending on the process or equipment. As used herein, the term "loaf" refers to edible foodstuffs obtained as moist products and includes terrines, pates, mousses, and the like. More generally, the term "pet food" encompasses all forms of food, including slices and breads as defined above, that are suitable for consumption by pets.

As is well known in the art, a "protease" is an enzyme that proteolizes, that is, triggers protein catabolism, by hydrolyzing the peptide bonds that link amino acids together in a polypeptide chain. Lipase is a water-soluble enzyme that catalyzes the hydrolysis of ester bonds in water-insoluble lipid substrates. Importantly, since lipases are proteins, they can be at least partially hydrolyzed by proteases when lipases and proteases are used as an enzyme mixture, as disclosed in previous studies. This is an adverse reaction that the present invention seeks to avoid.

The terms "fat" and "oil" as used herein are synonymous and also encompass mixtures of fats or oils. Animal fats as well as vegetable and / or sea oils can be used. Any commercially available sources of animal, vegetable, sea fat can be tested. Vegetable oils that are available in large quantities are typically rapeseed oil, soybean oil, corn oil, olive oil, sunflower oil, flax seed oil, palm oil, saffron oil, and the like, and their by-products. Typical animal fats are lard, lard, poultry fat, and the like, and their by-products. Sea oils are usually tuna oil, sardine oil, salmon oil, anchovy oil, fish oil, and the like, and their by-products. It also covers fats derived from animal, vegetable, marine sources, or produced (produced) by animals and plants.

A "thermal reaction" is, according to the present invention, a reaction obtained by combining at an elevated temperature at least one carbohydrate, preferably a reducing sugar, and at least one nitrogen compound. Such a reaction can actually include various concomitant and / or sequential reactions, including, for example, the Maylor reaction. It is possible for a complex combination of reactions to take place, depending on the conditions used. Preferably, the "reducing sugar" is selected from hexose, pentose, glucose, fructose, xylose, ribose, arabinose, starch hydrolysates and the like, and combinations thereof. The term "nitrogen compound" as used herein encompasses twenty known naturally occurring amino acids as well as aminoacyl sequences, that is, peptides, oligopeptides, and proteins or polypeptides. Also included are all compounds containing nitrogen of any origin that are acceptable for use in pet food. Suitable nitrogen compounds are selected from thiamine, methionine, cystine, cysteine, glutathione, hydrolyzed plant proteins (HPP), yeast autolysates, yeast extracts, and combinations thereof. Of course, the term "nitrogen compound" encompasses any sulfur-containing nitrogen compounds that are acceptable for use in pet food, such as sulfur-containing amino acids.

A first aspect of the present invention relates to a method for making a palatability enhancer for use in pet food, comprising at least:

a) providing a first-stage reaction product obtained by:

(i) reacting with at least one exogenous and / or endogenous protease in the absence of any added (or exogenous) lipase, wherein the substrate contains protein and fatty materials in amounts, at pH and temperature conditions and for a time effective to carry out proteolytic reaction,

(ii) heat inactivating said protease and filtering the resulting cleavage product;

b) optional fat addition;

c) emulsification of the specified reaction product of the first stage;

d) reacting said emulsion with at least one lipase in the absence of any added protease, in amounts, at pH and temperature conditions and for a time effective to effect the lipolytic reaction to obtain a second-stage reaction product.

After step (a) (ii), where the protease (s) have been (were) inactivated by heat, it is also useful to cool the resulting product, for example, to a temperature of from about 20 ° C to about 50 ° C (preferably from about 25 ° C to about 45 ° C) so that the mixture is at a temperature effective for the subsequent lipolytic reaction, as suggested in step (d). For example, this cooling is useful immediately after step (a) (ii) and the temperature will then be kept the same in steps (b), (c) and (d).

Desirably, the first stage reaction product is prepared and stored under suitable conditions until later used. This first-stage reaction product can also conveniently be obtained from a commercial source if the commercial digest was obtained after only one proteolytic reaction.

An essential feature of the present invention is a sequential reaction scheme consisting first of using a protease and then using a lipase. Indeed, this stepwise enzymatic treatment provides an increase not only in the efficiency and reaction rate, but also in the degree of enhancement of the palatability of the product. In addition, an unexpected advantage of the resulting palatability enhancer is that it can be used very widely in widely varying feeds such as dry, semi-dry and wet pet foods, including at least cats and dogs.

The addition of fat in step (b) is optional, but preferably done for best results.

Proteases can be present in the original substrate containing protein and fatty materials. Therefore, the addition of proteases in step (i) is optional. However, for best results, it is preferable to add at least one protease in step (a) (i).

A second aspect of the present invention relates to a palatability enhancer for use in pet food that can be prepared by the method described above.

The palatability enhancer of the present invention can be in the form of a liquid (eg solution) or dry substance (eg powder).

A third aspect of the present invention relates to a palatability enhancing composition for use in pet food, which comprises at least one palatability enhancer as described above.

Alternatively, said palatability enhancing composition comprises two or more palatability enhancers, at least one of which is a palatability enhancer according to the present invention.

A fourth aspect of the present invention relates to a method for preparing a palatable pet food comprising at least: administering at least one palatability enhancer or at least one palatability enhancing composition as disclosed above in an amount effective for increasing the palatability of said pet food.

The introduction of the palatability enhancer can be accomplished by coating (eg, spraying or spraying) or by adding to the bulk of the pet food.

A fifth aspect of the present invention relates to a pet food having an improved palatability obtained by the method described above.

The present invention also encompasses a palatable pet food comprising at least one palatability enhancer or at least one palatability-enhancing composition as described above.

Such pet food can be selected from the group consisting of dry, semi-dry and wet foods.

A sixth aspect of the present invention is directed to a method for feeding pets, comprising at least:

a) providing pet food as described above;

Preferably, the pets are selected from the group consisting of cats and dogs.

Thus, the invention relates to a method for improving the palatability of pet foods comprising lipolysis of the starting materials followed by a thermal reaction, such as the Maillor reaction. By starting materials is meant an animal and / or marine and / or vegetable digest obtained after hydrolysis with endogenous enzymes present in tissues or added proteases. Commercially available sources of starting materials include poultry, pork, beef, lamb, fish, and the like, and combinations thereof. Raw materials can be used as starting materials (for example, entrails or entrails and liver from poultry, pork, beef, lamb, fish and the like, and their combinations) and proteolysis before continuing the process in the form of lipolysis and thermal reactions.

The enzymes used in the present invention are proteases and lipases. Commercial proteases and lipases are isolated from plants, animals and microorganisms such as bacteria, yeast and fungi. In practice, commercially available proteases may not be completely pure in the sense that they may exhibit residual lipase activity. Accordingly, commercially available lipases can exhibit residual proteolytic activity. Of course, one skilled in the art will be able to select suitable enzymes to avoid or minimize possible unwanted side effects. For this reason, steps (a) (i) and (d) indicate that the proteolytic and lipolytic reactions are carried out respectively “in the absence of any added lipase” (step (a) (i)) and “in the absence of any added protease "(step (d)). This means that only protease (s) in step (a) (i) and lipase (lipases) in step (d) are present or added. Therefore, if any residual lipase or protease activity is present, respectively, in steps (a) (i) and (d), they are negligible. The only significant enzymatic activities of interest are proteolytic activity in step (a) (i) and lipolytic activity in step (d). The enzymes are generally used in amounts of from about 0.01% to 10%, preferably from 0.01% to 5%, more preferably from 0.01% to 2%, based on the weight of the final palatability enhancer.

To obtain an optimal hydrolysis rate, temperature and pH should be correlated with the enzymes used. This will be fairly obvious to those skilled in the art. The pH can be adjusted to the desired value with any suitable compound suitable for use in pet food, such as phosphoric acid, caustic soda, other common and suitable acid and alkaline regulators, and combinations thereof.

If raw tissues are used as starting material, after proteolysis and before lipolysis, a step of heat inactivation of enzymes (for example, pasteurization) is carried out, followed by filtration, at a temperature usually from about 70 ° C to 95 ° C, for a sufficient time - for example, about from 5 to 20 min. This allows proteases to be inactivated prior to lipolysis.

To ensure that the lipolysis step is carried out, it is important to first emulsify the mixture before adding lipases. Emulsification can be accomplished by adding at least one emulsifier suitable for use in pet food. Suitable emulsifiers are sodium stearoyl lactylate (SLN), succinylated monoglycerides, gum (gum arabic), sodium alginate, lecithin, and the like. Typically, emulsifiers are added in an amount of about 0.01% to 10%, preferably 0.01% to 8%, and more preferably 0.01% to 5%, based on the weight of the final palatability enhancer.

As already indicated above, you can try any commercially available source of animal fat and / or vegetable oil. Suitable sources of vegetable oils available in large quantities are rapeseed oil, soybean oil, corn oil, olive oil, sunflower oil, flax seed oil, palm oil, saffron oil, and the like, as well as by-products thereof. Suitable sources of animal fats include lard, lard, poultry fat, and the like, as well as their by-products. Suitable sources of marine oils are tuna oil, sardine oil, salmon oil, anchovy oil, fish oil, and the like, and their by-products. It also includes fats obtained from animal, vegetable, marine sources, or produced by animals and plants. Typically, the fat is present in an amount of from about 2% to about 30%, preferably from 5% to about 20%, based on the weight of the final palatability enhancer.

After lipolysis, a thermal reaction is carried out to complete the off-flavor of the product. Conveniently, the carbohydrate and nitrogen compound are added at a concentration of from about 0.01% to 30%, preferably from 0.1% to 20%, more preferably from 0.1% to 15% for the first, and from about 0.01% to 30% preferably from 0.01% to 20%, more preferably from 0.01% to 15% for the latter. A suitable temperature is selected in the range from about 70 ° C to 130 ° C, preferably from 80 ° C to 120 ° C, and the heat treatment is carried out for a time sufficient to further develop the flavor of the product, for example for at least 30 minutes.

To ensure long shelf life, preservatives such as natural or synthetic antioxidants can be added (suitable antioxidants include, but are not limited to: butyloxyanisole (BHA), butyloxytoluene (BHT), propyl gallate, octyl gallate, tocopherols, rosemary extracts, and the like), sorbitic acid or sorbitol salts; and other acids such as phosphoric acid and the like.

The palatability enhancer of the present invention can be used directly by itself, usually in an amount of about 0.01% to 20%, preferably 0.01% to 10%, more preferably 0.01% to 5% by weight relative to the weight of the pet food composition. Alternatively, it can be combined with other palatability enhancers, and all palatability enhancers can be administered simultaneously or sequentially.

In one embodiment of the present invention, a dry palatability enhancer formulation is prepared by combining the palatability enhancer in an appropriate ratio with carriers and mixing the components. The mixture is then dried by evaporation to form a dry palatability enhancer.

The palatability enhancers of the present invention are useful in pet foods such as dry pet foods, semi-moist pet foods, having a moisture content of about 50% or less by weight and which is a nutritionally balanced mixture containing proteins, fibers ( fiber), carbohydrates and / or starch. Such mixtures are well known to those skilled in the art and their composition depends on many factors, such as, for example, the required nutritional balance for a particular type of companion animal. In addition to these basic elements, the feed may contain vitamins, salts and other additives such as condiments, preservatives, emulsifiers and moisturizers. Nutritional balance, including the relative amounts of vitamins, salts, lipids, proteins and carbohydrates, is determined according to known veterinary nutritional standards - for example, as recommended by the National Research Council (NRC) or the American Association of Representatives for the Control of food quality (American Asociation of Feed Control Officials, AAFCO).

All common protein sources can be used, especially vegetable proteins such as soy or groundnuts, animal proteins such as casein or albumin, and raw animal tissues such as raw meat tissue and raw fish tissue, or even elements that are dry or dried, such as fish meal, bird meal, meat meal, and bone meal. Other types of suitable proteinaceous materials include wheat or corn gluten and proteins from microorganisms such as yeast. You can also use ingredients that contain a significant proportion of starches or carbohydrates, such as corn, milo, alfalfa, wheat, barley, rice, soybean hulls, and other low protein grains.

Other ingredients such as whey and milk by-products such as carbohydrates can be added to the feed. In addition, you can add known seasonings, including corn syrup or molasses.

By way of example, a typical dry cat food formulation into which a palatability enhancer according to the present invention can be incorporated consists of the following components (weight percent): about 0-70% of a bread base such as flour (corn, wheat, barley or rice); approximately 0-30% of animal by-products (from poultry or meat); about 0-25% corn gluten; approximately 0-25% of crude animal tissue - such as that of poultry or cattle; about 0-25% soy flour; approximately 0-10% animal fat; about 0-20% seafood base; approximately 0-25% raw fish tissue; about 0-10% high fructose corn syrup; about 0-10% dry molasses; about 0-1.5% phosphoric acid; and about 0-1.5% citric acid.

Vitamins and salts can be added, including calcium carbonate, potassium chloride, sodium chloride, choline chloride, taurine, zinc oxide, iron sulfate, vitamin E, vitamin A, vitamin B12, vitamin D3, riboflavin, niacin, calcium pantothenate, biotin, thiamine mononitrate, copper sulfate, folic acid, pyroxidine hydrochloride, calcium iodate and menadione complex with sodium bisulfite (source of vitamin K activity).

Dry pet foods are generally prepared in a variety of ways. One of these methods, which is widely used, is the cook-extrusion method. In a cook-extrusion process, the dry ingredients are first mixed together to form a mixture. This mixture is transferred to a steam conditioner where it is sufficiently humidified to be squeezed out. The mixture is then introduced into a cooking extruder, where it is cooked at elevated temperature and pressure, and then extruded from the apparatus through a press. This press gives the extrusion product a specific shape. Individual pieces of product are obtained by periodically cutting from the end of the strip of the extruded product. The individual pieces or slices are then dried in a hot air dryer. Typically, the product is dried until it contains less than 15% moisture, preferably about 5 to 10% moisture. The dried particles or pieces are then transferred by a loading conveyor to a coating drum and sprayed with fat. Other liquids, such as, for example, phosphoric acid, can alternatively be applied to the pieces or can be applied along with the fat. The resulting granules or slices constitute the base composition to which the palatability enhancer coating can be applied.

In one embodiment of the present invention, the palatability enhancers of the present invention may be administered by coating. The term "coating" as used herein refers to the surface application of a palatability enhancer or flavoring composition to the surface of a base composition, for example, by spraying, spraying, and the like. For example, slices of uncoated, squeezed out basic pet food can be placed in a container for mixing, such as a tube or drum for coating. Fat, such as lard or poultry fat, is heated and then sprayed onto the pet food to coat the slices. The coating does not require a continuous layer, but should preferably be uniform. After the fat, a palatability enhancer can be applied, either as a liquid or as a dry powder, during the mixing process. The liquid palatability enhancer is usually sprayed on, while the dry palatability enhancer is usually sprayed on. Alternatively, palatability enhancers can be mixed with fat and applied at the same time. In another alternative coating method, palatability enhancers are applied prior to application of the fat.

In another embodiment of the present invention, the palatability enhancer is contacted with the raw materials of the pet food composition prior to cooking. In this case, the palatability enhancer is combined with proteins, fibers, carbohydrates and / or starch of the base composition and cooked together with these materials in a cooking extruder.

The palatability enhancers of the present invention are also useful for wet pet foods having a moisture content of more than 50% and presenting a nutritionally balanced mixture. Wet food may contain one or more ingredients selected from mealy materials (such as grain and flour based materials), animal by-products, raw animal tissues, raw fish tissues, animal and vegetable fats, marine materials, vitamins, salts, preservatives, emulsifiers, surfactants, structuring agents, colorants and the like. Such ingredients are well known to those skilled in the art and may be suitably selected depending on the type of wet food.

Most types of meat gravy-based pet foods are prepared by pulverizing meat, simulated meat or meat by-products and then forming the mashed mixture by reduced pressure extrusion through a steam tunnel where the food is cooked. Then add starch and binders, after which the mixture is cut into pieces, mixed with water, starch and binders. After that, the mixture is packed and closed in cans and boiled in a hydrostat with continuous or rotary sterilization. Non-gravy wet pet foods are prepared by soaking meat, simulated meat or meat by-products and shaping the soaked materials with starch, water, and binders. After that, the mixture is packed and closed in cans and boiled in a hydrostat with continuous or rotary sterilization.

A liquid or dry palatability enhancer can be incorporated into a base such as a gravy or jelly while mixing along with the remaining ingredients (structuring agents, stabilizers, colorants, and nutritional supplements). A liquid or dry palatability enhancer can also be incorporated into meat by-product mixtures for making slices or sticks. In this case, it can be added to the raw material before or after the grinding process. The meat by-product mix can be steam-cooked or grilled for slices, or directly soldered into cans for chopsticks.

The palatability enhancers described above provide significant advantages over previous solutions. The effects of the present invention can be measured in what is commonly referred to as the "two bowl test" or "comparison test". Of course, one skilled in the art is free to use any other suitable test instead of the two bowl test described herein to determine the advantage. Such alternative tests are well known in the art.

Two Bowl Test Principle:

The test is based on the postulate that the more feed is eaten, the more palatability it has. Preference tests of individual animals were performed using the two-bowl method, based on a comparison of two foods. Tests were performed on either groups of 36 dogs or groups of 40 cats, depending on the purpose of the test.

Test method:

The same amounts of food A and food B were weighed and placed in the same bowls. The amount in each diet provides the daily food requirement.

Distribution of bowls:

Dog test: Bowls were placed in individual food trays available for dogs.

Cat test: Bowls were placed at the same time in front of each cat in individual locked boxes and their position was changed with each meal to eliminate the influence of the position of the bowls.

Test duration:

Dog test: 15 minutes maximum (if one of the two bowls was completely emptied in less than 15 minutes, both bowls were removed and the test was stopped).

Test for cats: minimum 15 minutes (if the contents of one of the bowls were completely eaten in less than 30 minutes, both bowls were removed and the test was stopped).

Investigated parameters:

Measured parameters: First food eaten and the amount of each food eaten at the end of the test.

Calculated parameters: Individual consumption ratio (CR).

SP A = feed intake A (in d) × 100 / intake A + B (in d)

SP B = consumption of feed B (in d) × 100 / consumption of A + B (in d).

Average intake ratio (SDR) is the average of all individual ratios (all animals are equally significant, regardless of their size and their respective food intake). If the consumption of animals is higher or lower than certain values, they are not taken into account in statistical processing.

Statistical analysis:

Statistical analysis was applied to determine if there is a significant difference between the two ERP ratios. The Student's t-test was used with three threshold error values, namely 5%, 1% and 0.1%.

The Chi test was used to determine if there is a significant difference between the number of dogs or cats with a preference for food A and the number of dogs or cats with a preference for food B.

Significance levels are indicated as follows:

ND difference is insignificant (p> 0.05)

* reliably (p<0,05)

** high degree of reliability (p<0,01

*** very high confidence level (p<0<001).

In the following examples, various types of fats or fat blends are tested as defined above. These fats are hereinafter referred to as Fat 1, Fat 2, Fat 3. It is shown below that regardless of whether a fat or a fat blend is used, the palatability of the product according to the present invention is very high.

Example 1: XLHM product with starting raw material

Recipe:

The raw materials, exogenous and / or endogenous proteases, preservatives and antioxidants are mixed together and heated at about 60 ° C to 70 ° C for at least 30 minutes (step a) (i)).

The mixture is heated and pasteurized at about 85 ° C for at least 10 minutes, then cooled at about 25 ° C to 45 ° C, preferably with concomitant filtration, to obtain the first stage reaction product (step a) (ii)).

Optionally, a storage step may be added here under suitable conditions for a given period, preceded by an acidification process.

The pH is then adjusted to about 7 to 10 with caustic soda or an alkalinity regulator and emulsifiers, fat and lipase enzymes are added for lipolysis, lipolysis is carried out for at least 120 minutes, preferably about 120 to 420 minutes, to obtain the reaction product the second stage (stages b) to d) is carried out simultaneously).

Reducing sugars and nitrogen compounds are added and the resulting mixture is heated at about 90 ° C to 110 ° C for at least 30 minutes, resulting in a palatability enhancer (step e).

Finally the product is cooled and phosphoric acid, potassium sorbate, preservatives and antioxidants are added for long-term storage, with a final pH of 2.9, resulting in a ready-to-use palatability enhancer product (designated XLHM).

XLHM version A: base - raw material from poultry;

XLHM version B: base - raw material from poultry, used fat 1;

XLHM version C: poultry raw material base, fat 2 used;

XLHM version D: poultry raw material base, fat used 3.

Results of comparing palatability of SP1 and SP2 for dogs:

Consumption of SP1 and SP2 is significantly different, which demonstrates the best performance of the palatability enhancer SP2 from Super Premium. The test results are shown in the graph in Fig. 1.

Taste Test Results for Dogs XLHM Compared to SP1 and SP2

All four versions of XLHM exhibit increased palatability compared to the SP1 product. For all fat variants used, the palatability of XLHM is equal to or greater than the palatability of SP2.

Example 2: XLHM product with original cleavage

Recipe:

The starting material used in this example is the digest (cleavage product) obtained after step a) (i) and (ii), as illustrated in example 1, that is, it is the product of the first reaction.

The method begins with steps b), c) and d), where the pH is adjusted in the range from about 7 to 10 using caustic soda or alkalinity regulators, for lipolysis, emulsifiers, fat and lipase enzymes are added, lipolysis is carried out for at least 120 min, preferably for about 120 to 420 minutes to obtain the reaction product of the second stage.

Then, reducing sugars and nitrogen compounds are added and the resulting mixture is heated at a temperature of about 90 ° C to 110 ° C for at least 30 minutes, resulting in a palatability enhancer (step e).

Finally the product is cooled and phosphoric acid, potassium sorbate, preservatives and antioxidants are added for long-term storage, with a final pH of 2.9, resulting in a ready-to-use palatability enhancer product (designated XLHM version ...).

Evaluation of palatability for dogs:

SP1 and SP2 are Super Premium liquids from the existing range with different palatability levels, with SP2 being more palatable than SP1.

XLHM products are palatability enhancers according to the present invention:

XLHM version E: first liquid digest, used fat 1;

XLHM version F: liquid digest first, fat 2 used;

XLHM version G: liquid digest first, fat used 3.

All test results are presented in the graph in figure 2.

When using the original digest at the beginning of the process, the result is the same as before. That is: higher palatability than SP1, and at least equal to that of SP2, and often higher than SP2.

Comparative example 3: XLHM product (D ") with starting raw material and without related proteolysis step

Recipe:

Here the method begins with the stage of joint action of proteases and lipases.

First, the protease-containing raw materials, water, preservatives and antioxidants are mixed together, the pH is adjusted to the range from about 7 to 10 with caustic soda, emulsifiers, salts, fat 3 and lipase enzymes are added to carry out the hydrolysis step, the hydrolysis is carried out at a temperature of about 25 ° C to 45 ° C, for at least 120 minutes, preferably for about 120 to 420 minutes, to obtain product 1.

Then, reducing sugars and nitrogen compounds are added and the resulting mixture is heated at a temperature of about 90 ° C to 110 ° C for at least 30 minutes to obtain product 2.

Finally, product 2 is cooled and phosphoric acid, potassium sorbate, preservatives and antioxidants are added for long-term storage, with a final pH of 2.9 to obtain a ready-to-use product (product XLHM D ").

Evaluation of palatability for dogs:

Compared to the results obtained for the product XLHM version D (see example 1), XLHM version D "is worse compared to SP2, but retains a high palatability compared to SP1. In other words, the results obtained with the combination of proteolysis and lipolysis, not as good as the results obtained by separating proteolysis and lipolysis.

Comparative example 4: XLHM product (B ") with starting raw material and with reordering of enzymatic steps

Recipe:

In this example, the method begins with a lipolysis step followed by a proteolysis step.

First, the raw materials, water, preservatives and antioxidants are mixed together, the pH is adjusted to the range from about 7 to 10 with caustic soda, emulsifiers, salts, fat 1 and lipase enzymes are added to carry out the lipolysis step, lipolysis is carried out at a temperature of about 25 ° C. to 45 ° C, for at least 120 minutes, preferably for about 120 to 420 minutes, to obtain product 1.

Then, protease enzymes, reducing sugars and nitrogen compounds are introduced and heated at a temperature of about 60 ° C to 70 ° C for at least 30 minutes to obtain product 2.

The resulting mixture is heated at about 90 ° C to 110 ° C for at least 30 minutes to obtain product 3.

Finally, product 3 is cooled and phosphoric acid, potassium sorbate, preservatives and antioxidants are added for long-term storage, with a final pH of 2.9 to obtain a ready-to-use product (product XLHM B ").

Evaluation of palatability for dogs:

Compared to the results obtained for the product XLHM version B (see example 1), XLHM version B "is worse than SP1.

Conclusion: Better results are obtained when proteolysis is performed prior to lipolysis than when lipolysis precedes proteolysis.

Example 5: Adding XLHM version G to cat loaves

Product B recipe

Product recipe C

Definition: C "sens W9P is a commercial Super Premium SPF palatability enhancer for incorporation into wet pet food.

Raw materials (pork lungs, pork liver, chicken lungs and liver, chicken carcasses) were thawed overnight at room temperature. Then they were ground in a vertical grinder (Stephen, Germany) for 5 min at a frequency of 1500 strokes per min. Water was added to the glass. Powders (structuring agents, a mixture of vitamins and salts and wheat flour) and XLHM version G or C "sens W9P were dissolved in water using a homogenizer (Dynamic, France). The solution was added to the crushed meat products and stirred for another 5 min under vacuum ( -1 bar) in a grinder.The porridge was transferred to a vacuum dispenser (Handtmann, Germany) and filled into 400 g iron cans. The cans were closed and heated in a Microflow retort (Barriquand, France) using the following regime: heating to 127 13 min, the temperature is maintained at 127 ° C for 55 min, cooling to 20 ° C for 15 min.

Evaluation of palatability for cats

The results show that Product B (containing XLHM version G) has a higher palatability than Product C. The test results are plotted in FIG. 3.

Thus, there is shown and described a new and useful method for improving the palatability of pet food compositions. Although the present invention has been accompanied by examples for purposes of illustration and the description has been given with reference to specific embodiments, it should be apparent to those skilled in the art that various modifications, variations, and equivalents of the illustrative examples are possible. It is believed that any such changes which directly follow from the set forth herein and which do not depart from the spirit and scope of the present invention are covered by the present invention.

1. A method of obtaining a palatability enhancer for use in pet food, comprising at least:
a) providing a first-stage reaction product obtained by:
(i) reacting with at least one exogenous and / or endogenous protease in the absence of any exogenous or added lipase, wherein the substrate contains protein and fatty materials in amounts, at pH and temperature conditions and for a time effective to effect the proteolytic reaction , (ii) heat inactivating said protease and filtering the resulting cleavage product;
b) optional fat addition;
c) emulsification of the specified reaction product of the first stage;
d) reacting said emulsion with at least one lipase in the absence of any added protease in amounts, at pH and temperature conditions and for a time effective to effect the lipolytic reaction to obtain a second-stage reaction product;
e) adding to said second-stage reaction product of at least one reducing sugar and at least one nitrogen compound and heating the resulting mixture to a temperature and for a time effective to further form the palatability of the mixture, resulting in a palatability enhancer.

2. The method according to claim 1, further comprising step a) (iii) cooling the product obtained in step a) (ii) to a temperature effective for carrying out the subsequent lipolytic reaction in step d).

3. A method according to claim 1 or 2, further comprising the step of cooling the mixture obtained in step e).

4. A process according to claim 1 or 2, wherein the reaction product of the first stage is prepared and stored under suitable conditions until further use.

5. A palatability enhancer for use in pet food, which can be obtained by the method according to any one of claims 1 to 4.

6. The palatability enhancer of claim 5, wherein said palatability enhancer is a liquid or a powder.

7. A palatability enhancing composition for use in pet food, comprising at least one palatability enhancer according to claim 5 or 6.

8. A method of preparing pet food having increased palatability, comprising at least:
introducing into the pet food at least one palatability enhancer according to claim 5 or 6 or at least one palatability-enhancing composition according to claim 7 in an amount effective to increase the palatability of said pet food.

9. The method of claim 8, wherein said administration is by coating or by addition to the bulk of the pet food.

10. Pet food having an increased palatability, which can be obtained by the method according to claim 8 or 9.

11. A palatable pet food comprising at least one palatability enhancer according to claim 5 or 6, or at least one palatability-enhancing composition according to claim 7.

12. The pet food of claim 10 or 11, wherein said pet food is selected from the group consisting of dry, semi-dry, and wet pet foods.

13. A method of feeding pets, comprising at least:
a) providing pet food according to any one of claims 10-12;
b) feeding said pet food to pets.

14. The method of claim 13, wherein said companion animals are selected from the group consisting of cats and dogs.

Similar patents:

The invention relates to the feed industry, in particular to the production of feed additives and feed for farm animals and poultry with high enzymatic (cellulose) activity, improved probiotic properties and simultaneously having antimicrobial properties.

The invention relates to the field of food for pets, such as dogs and cats

The taste system is used to evaluate taste of products- a complex of sensations perceived during their testing, – tastings (evaluation of the food product in the mouth).

Taste – a feeling that occurs when the taste buds are excited and is determined both qualitatively and quantitatively.

Evaluation of taste comes down to determining the type of taste and its intensity. The standards of primary flavoring substances in commodity science are considered to be, respectively: sweet – sucrose; sour – hydrochloric acid; salty – salt; bitter – quinine (caffeine). All other types and shades of taste can be obtained by mixing three out of four possible primary tastes in the required proportions.

The qualitative definition of taste is caused by the effect of substances on the taste buds, which are located mainly on the tongue. In addition, they are found on the mucous surface of the oral cavity, pharyngeal walls, tonsils, and larynx. The total number of taste buds in the human oral cavity reaches 9 thousand. In addition, the definition of taste is associated with the sense of food in the oral cavity.

The gustatory apparatus of the human mouth is a chemical analyzer, and more sensitive than modern devices. All the richness of various shades, combinations of taste sensations arises as a result of irritation of special sense organs – taste buds (buds), each of which consists of several sensitive chemoreceptor cells connected to sensory neurons. Chemoreceptor cells respond to certain chemicals.

Taste bulbs are differentiated for the perception of the main types of taste: sweet, salty, sour and bitter. The bulbs on the tip of the tongue are most sensitive to sweet taste, at the edges of the front of the tongue to salty, at the edges of the back of the tongue – to sour, at the base – to bitter.

All substances that determine the taste of food are soluble in water. Only in dissolved form can they affect the chemical analyzers of the taste apparatus.

The sensation threshold also depends on the temperature of the solution, which is probably due to a change in the state of the taste bud protein molecule. The best perception of flavoring substances occurs at a solution temperature close to the temperature of the human body (36.5 ° C). Hot solutions of the same substances in the indicated concentrations seem tasteless, that is, they do not cause any sensations. When cooled to 30 ° C, the sweet taste appears rather than salty or bitter.

When evaluating taste, the speed of the taste is also important: the salty taste is perceived the fastest, and the sweet and sour taste is perceived slower. Bitter taste is perceived most slowly.

The following flavor characteristics are distinguished.

Sour taste-- characterizes the basic taste characteristic of aqueous solutions of most acids (for example, citric and tartaric acids); complex olfactory sensation, caused mainly by the presence of organic acids.

Acidity – an organoleptic property of individual substances or mixtures that produces a sour taste.

Sour taste- characterizes a lesser degree of intensity of the taste of a sour product.

Bitter taste - characterizes the main taste caused by aqueous solutions of chemicals such as quinine and caffeine, as well as some alkaloids.

Bitterness – organoleptic property of individual substances or mixtures that cause a bitter taste.

Salty taste - characterizes the main taste caused by aqueous solutions of chemicals such as sodium chloride.

Salinity – an organoleptic property of individual substances or mixtures that cause a salty taste.

Sweet taste - characterizes the main taste caused by aqueous solutions of chemicals such as sucrose.

Sweetness – organoleptic property of individual substances or mixtures that cause sweet taste.

Alkaline taste--characterizes the basic taste caused by aqueous solutions of chemicals such as sodium bicarbonate.

Alkalinity – organoleptic property of individual substances or mixtures that cause an alkaline taste.

Astringent, tart taste - characterizes the complex sensation caused by the contraction of the mucous surfaces of the oral cavity and arising from the effects of substances such as tannins. Astringency – organoleptic property of individual substances or mixtures that cause astringent taste.

Metallic taste - characterizes the main taste caused by aqueous solutions of chemicals such as ferrous sulfate.

Tasteless, insipid taste – a product that does not have a characteristic taste.

Aftertaste – a taste sensation that appears after swallowing or removing a product from the oral cavity, which is different from the sensations that were perceived while it was in the oral cavity.

Foods have either one taste (sugar – sweet, table salt – salty, acid – sour), or differ in a combination of basic types of taste. In this case, we can talk about a harmonious and inharmonious combination of taste . So, r harmoniously as a whole, sweet or salty tastes are combined with sour or bitter ones. An example is the sweet-sour taste of fruits, confectionery, the bittersweet taste of chocolate, sour-salty pickled vegetables, and salty-bitter olives. Inharmonious combinations such as salty-sweet, bitter-sour are considered. These combinations are perceived as two different tastes, they are unusual for food products, they are rare and usually arise as a result of spoilage (for example, the bitter-sour taste of pickled vegetables).

The perception of taste depends on the chemical composition, viscosity and amount of food; the nature of odorous and flavoring substances and the intensity of release, the rate of removal, the nature of their effect on the organ of taste; availability of these substances in a certain period; the conditions of food intake (in particular, respiration, the amount and speed of saliva flow, duration, temperature) and the quality of chewing of the product.

It was found that the assessment of the intensity of the main tastes can be influenced by the color of the product. Thus, yellow and light green colors increase the assessment of the intensity of the acidity of the product, and the red color enhances the assessment of the intensity of sweetness in comparison with colorless. You should also consider the moments of suppression of one taste by another. For example, sour taste is suppressed by sweet taste, and to a lesser extent – salty and bitter. Salty and bitter tastes are suppressed by certain concentrations of sucrose and citric acid; the sweet taste is slightly suppressed by small concentrations of citric acid.

The taste that persisted despite the termination of the impulse that caused it is called secondary taste ... It can be the same and contrasting. The same it is because after the cessation of the gustatory impulse, a gustatory sensation remains, which is identical in quality to that which was at the time of exposure to the gustatory impulse. Contrasting is called the secondary taste, which arises after the removal of the active gustatory impulse.

All food products evoke the sensation of a certain secondary taste, the same or contrasting. If the secondary flavor is the same and coincides with the main flavor profile of a product and quickly disappears when a piece of this product is swallowed, this proves that the quality of the product is high. But if, after swallowing, a secondary taste remains in the mouth, then the product is inferior to the first in consumer value. In the practice of organoleptic analysis, a secondary contrasting taste is very often encountered, for example, distilled water after rinsing the mouth with a solution of table salt seems sweetish. After a short tasting of the sweet taste, the sour taste is perceived more sharply, and the unpleasant sensation is intensified. Therefore, aged wines are not judged after sweet wines; do not evaluate lightly salted products after evaluating dry-salted products.

Delicacy (flavor– flavor) – a complex sensation in the mouth caused by the taste, smell and texture of the food product, assessed (assessed) both qualitatively and quantitatively.

The palatability can be influenced by tactile, thermal, pain and / or kinesthetic sensations.

Evaluation of the quality of a food product based on a comprehensive perception and analysis of optical, olfactory, tactile, gustatory, acoustic and other impulses (stimuli) is called the evaluation of tastyness or flavor as opposed to its taste.

Quantitative changes in the individual components of goodness at a certain moment lead to a qualitative leap and, as a result, a high-quality product is formed with a harmonious or complete gustatory merit. Over time, the balance between the individual components of the deliciousness is disturbed and this leads to a deterioration in the quality of the product. An example is the process of maturation, aging and withering away of wine.

When assessing taste, it is necessary to take into account such phenomena due to the physiological characteristics of the sensory organs as adaptation and fatigue.

Adaptation – this is a decrease in the impressionability of the organ of taste caused by prolonged exposure to a taste impulse of the same quality and constant intensity. When testing a large number of samples with the same taste, the same intensity, adaptation causes distorted results. The organs of taste, in contrast to sight and similar to the sense of smell, are inherent in rapid adaptation.

Fatigue- reduces the perception of taste due to fatigue of the taste buds under the influence of a repetitive impulse. It occurs after a different period of time, depending on the properties of the product, physiological and psychological state of the testers, training, working conditions.

The present invention relates to a method for preparing a palatability enhancer for use in pet foods with low, medium and high moisture content. The method includes: (i) reacting a substrate with at least one protease in the absence of any added lipase, wherein the substrate contains protein and fatty materials, (ii) heat inactivating said protease and filtering the resulting cleavage product; d) reacting the resulting emulsion with at least one lipase in the absence of any added protease to obtain a second stage reaction product: e) adding to said second stage reaction product at least one reducing sugar and at least at least one nitrogen compound and heating the resulting mixture. 7 n. and 7 p.p. f-crystals, 6 dwg., 12 tbl., 5 ex.

Drawings to the RF patent 2476082

The technical field to which the invention relates

The present invention relates to a method of enhancing the palatability of compositions (including foods, additives, condiments, toys, and the like) for companion animals. In particular, the invention relates to a method for preparing a palatability enhancer for use in pet food with low, medium or high humidity.

The invention relates to the field of pet food, such as dogs and cats. All references herein to food of any kind are intended to refer only to food that is manufactured and sold for domestic animals such as dogs and cats. Although trials have so far been limited to dogs and cats only, the invention can also be adapted for use in rodents and other types of companion animals. If necessary, the invention disclosed herein can also be tested to determine its applicability for use in various classes of animals, including rodents (such as: hamsters, guinea pigs, rabbits and the like), birds, as well as horses and any type of livestock. However, since dogs and cats exhibit a higher degree of sensitivity to taste, taste and odor than rodents, horses, livestock, etc., and since generally dogs and cats receive food with an improved taste, the term “domesticated food” is used in this text. animals "will refer to all animals that appear to be receiving an improved palatable food as disclosed herein, and the term" animal food "will refer to all types of food offered to those animals.

It should be understood that the expression "animal feed" as used herein includes foods having a low, medium or high moisture content. There are three categories of animal feed: (1) dry foods or foods with a low moisture content (usually less than 15%), which are usually high in nutrients, cheaper packaging, easier to handle, but less palatable; (2) canned or moist foods or foods with a high moisture content (greater than about 50%), which usually have the greatest palatability to pets; (3) semi-wet or semi-dry foods or foods with intermediate or medium moisture content (usually 15% to 50%;), which usually have less palatability than canned foods, but more palatability than dry foods.

Prior art

Caring owners provide pets with a suitable choice of food. These foods may be a regular diet and may include their usual diet, supplements, additional treatments, and toys. Pets, like humans, prefer and more often and more willingly eat food that is palatable to them. Therefore, palatability enhancers are extremely important for animal consumption. Animal feeds — such as pet foods, typically contain flavoring compositions to enhance their palatability and appeal to the pet. To date, a large number of flavoring compositions (palatability enhancers or palatability factors) have been described. For example, US Pat. Nos. 3,857,968 and 3,968,255 to Haas and Lugay disclose a palatability-enhancing composition for use in dry animal food, especially dry dog ​​food, containing fat and proteins, which is prepared by a process comprising emulsifying fat, treating the composition with a mixture of enzymes, containing lipase and protease, and, optionally, enzyme inactivation. Another example is described in US Pat. No. 4,713,250, in which a palatability enhancing composition for dog food is prepared by a multi-step enzymatic reaction comprising first contacting an aqueous proteinaceous or mealy material with a protease and / or amylase, then preparing an emulsion containing fat and the product obtained in the first stage, and reacting said emulsion with lipase and protease. The following example is described in US patent No. 4089978, where Lugay et al. propose a composition with improved palatability for use in animal feed, which is prepared by a method comprising reacting at a moderate temperature an aqueous mixture of reducing sugar, animal blood, yeast and fat with an enzyme mixture containing lipase and protease, followed by an increase in temperature for more complete formation taste and enzyme inactivation.

However, the palatability of such compositions is generally different for different animal species. For example, a flavoring agent that is effective for cats is often not effective for dogs. Moreover, a flavoring agent that is effective in dry pet foods is generally ineffective in semi-moist or wet pet foods. Therefore, there is a continuing need for new palatability enhancers that provide a lasting mouthfeel and that can be used easily and effectively for pets, including dogs and cats, in a variety of feed types such as dry, intermediate and wet foods.

The essence of the invention

Thus, it is an object of the present invention to provide such a "generally applicable" palatability enhancer, ie palatability enhancer, which is effective regardless of animal species and feed type. The method of the present invention significantly enhances the palatability of low, medium or high moisture pet foods.

Thus, a method for producing a palatability enhancer is presented. This method involves staged enzymatic hydrolysis followed by a flavor-generating thermal reaction of raw materials (eg, poultry, pork, beef, lamb, fish products, and the like) to produce a highly effective palatability enhancer. The palatability enhancer can be added to pet food as a liquid product by coating spraying, as a dry powder in a coating spray process, or as a liquid or dry product by mixing it with pet food ingredients prior to packaging or canning. Alternatively, the palatability enhancer can be mixed with the fat and added at the same time.

The present invention also contemplates combining two or more palatability enhancers, including one as disclosed herein, to form compositions or mixtures useful for enhancing palatability of pet foods. The different types of palatability enhancers can be mixed together before being incorporated into the feed (palatability enhancer blends can be stored this way until they are used). Alternatively, the various palatability enhancers can be combined in situ, that is, directly in the pet food.

Surprisingly, it has been found that the palatability enhancer according to the present invention is the first one that is highly attractive to pets, especially dogs and cats, when added to pet food having a low, medium or high moisture content.

Brief description of the figures

Fig. 1 is a graph showing palatability results of Super Premium palatability enhancer SP1 versus SP2 (Example 1).

Figure 2: A set of graphs showing palatability results of palatability enhancers XLHM versus Super Premium SP1 and SP2 (Examples 1 and 2).

FIG. 3 is a graph showing palatability results of a palatability enhancer from Super Premium PRODUCT B versus PRODUCT C (Example 5).

Detailed description of the invention

The term "palatability" means the relative preference for an animal of one food composition over another. Palatability can be determined by a standard test procedure in which the animal has equal access to both compositions. This preference can arise from any of the animal's senses, but is usually associated with taste, smell, mouthfeel, texture, mouthfeel. It is determined here that the pet food having the increased palatability is the food that the animal prefers over the control composition.

The terms "palatability enhancers" or "flavors" or "flavors" (flavors, flavors), or "palatability factors" or "palatability factors" denote any material that enhances the palatability of a food composition for an animal. The palatability enhancer can be a single material or a mixture of materials, and it can be natural, processed or untreated, synthetic or partially natural and partially synthetic materials.

As used herein, the term "slice" ("kibble") refers to specific slices or nuggets obtained in a squeeze or extrusion process. Typically, the slices are made to produce dry and semi-moist pet food. The pieces can vary in size and shape, depending on the process or equipment. As used herein, the term "loaf" refers to edible foodstuffs obtained as moist products and includes terrines, pates, mousses, and the like. More generally, the term "pet food" encompasses all forms of food, including slices and breads as defined above, that are suitable for consumption by pets.

As is well known in the art, a "protease" is an enzyme that proteolizes, that is, triggers protein catabolism, by hydrolyzing the peptide bonds that link amino acids together in a polypeptide chain. Lipase is a water-soluble enzyme that catalyzes the hydrolysis of ester bonds in water-insoluble lipid substrates. Importantly, since lipases are proteins, they can be at least partially hydrolyzed by proteases when lipases and proteases are used as an enzyme mixture, as disclosed in previous studies. This is an adverse reaction that the present invention seeks to avoid.

The terms "fat" and "oil" as used herein are synonymous and also encompass mixtures of fats or oils. Animal fats as well as vegetable and / or sea oils can be used. Any commercially available sources of animal, vegetable, sea fat can be tested. Vegetable oils that are available in large quantities are typically rapeseed oil, soybean oil, corn oil, olive oil, sunflower oil, flax seed oil, palm oil, saffron oil, and the like, and their by-products. Typical animal fats are lard, lard, poultry fat, and the like, and their by-products. Sea oils are usually tuna oil, sardine oil, salmon oil, anchovy oil, fish oil, and the like, and their by-products. It also covers fats derived from animal, vegetable, marine sources, or produced (produced) by animals and plants.

A "thermal reaction" is, according to the present invention, a reaction obtained by combining at an elevated temperature at least one carbohydrate, preferably a reducing sugar, and at least one nitrogen compound. Such a reaction can actually include various concomitant and / or sequential reactions, including, for example, the Maylor reaction. It is possible for a complex combination of reactions to take place, depending on the conditions used. Preferably, the "reducing sugar" is selected from hexose, pentose, glucose, fructose, xylose, ribose, arabinose, starch hydrolysates and the like, and combinations thereof. The term "nitrogen compound" as used herein encompasses twenty known naturally occurring amino acids as well as aminoacyl sequences, that is, peptides, oligopeptides, and proteins or polypeptides. Also included are all compounds containing nitrogen of any origin that are acceptable for use in pet food. Suitable nitrogen compounds are selected from thiamine, methionine, cystine, cysteine, glutathione, hydrolyzed plant proteins (HPP), yeast autolysates, yeast extracts, and combinations thereof. Of course, the term "nitrogen compound" encompasses any sulfur-containing nitrogen compounds that are acceptable for use in pet food, such as sulfur-containing amino acids.

A first aspect of the present invention relates to a method for making a palatability enhancer for use in pet food, comprising at least:

a) providing a first-stage reaction product obtained by:

(i) reacting with at least one exogenous and / or endogenous protease in the absence of any added (or exogenous) lipase, wherein the substrate contains protein and fatty materials in amounts, at pH and temperature conditions and for a time effective to carry out proteolytic reaction,

(ii) heat inactivating said protease and filtering the resulting cleavage product;

b) optional fat addition;

c) emulsification of the specified reaction product of the first stage;

d) reacting said emulsion with at least one lipase in the absence of any added protease, in amounts, at pH and temperature conditions and for a time effective to effect the lipolytic reaction to obtain a second-stage reaction product.

After step (a) (ii), where the protease (s) have been (were) inactivated by heat, it is also useful to cool the resulting product, for example, to a temperature of from about 20 ° C to about 50 ° C (preferably from about 25 ° C to about 45 ° C) so that the mixture is at a temperature effective for the subsequent lipolytic reaction, as suggested in step (d). For example, this cooling is useful immediately after step (a) (ii) and the temperature will then be kept the same in steps (b), (c) and (d).

Desirably, the first stage reaction product is prepared and stored under suitable conditions until later used. This first-stage reaction product can also conveniently be obtained from a commercial source if the commercial digest was obtained after only one proteolytic reaction.

An essential feature of the present invention is a sequential reaction scheme consisting first of using a protease and then using a lipase. Indeed, this stepwise enzymatic treatment provides an increase not only in the efficiency and reaction rate, but also in the degree of enhancement of the palatability of the product. In addition, an unexpected advantage of the resulting palatability enhancer is that it can be used very widely in widely varying feeds such as dry, semi-dry and wet pet foods, including at least cats and dogs.

The addition of fat in step (b) is optional, but preferably done for best results.

Proteases can be present in the original substrate containing protein and fatty materials. Therefore, the addition of proteases in step (i) is optional. However, for best results, it is preferable to add at least one protease in step (a) (i).

A second aspect of the present invention relates to a palatability enhancer for use in pet food that can be prepared by the method described above.

The palatability enhancer of the present invention can be in the form of a liquid (eg solution) or dry substance (eg powder).

A third aspect of the present invention relates to a palatability enhancing composition for use in pet food, which comprises at least one palatability enhancer as described above.

Alternatively, said palatability enhancing composition comprises two or more palatability enhancers, at least one of which is a palatability enhancer according to the present invention.

A fourth aspect of the present invention relates to a method for preparing a palatable pet food comprising at least: administering at least one palatability enhancer or at least one palatability enhancing composition as disclosed above in an amount effective for increasing the palatability of said pet food.

The introduction of the palatability enhancer can be accomplished by coating (eg, spraying or spraying) or by adding to the bulk of the pet food.

A fifth aspect of the present invention relates to a pet food having an improved palatability obtained by the method described above.

The present invention also encompasses a palatable pet food comprising at least one palatability enhancer or at least one palatability-enhancing composition as described above.

Such pet food can be selected from the group consisting of dry, semi-dry and wet foods.

A sixth aspect of the present invention is directed to a method for feeding pets, comprising at least:

a) providing pet food as described above;

Preferably, the pets are selected from the group consisting of cats and dogs.

Thus, the invention relates to a method for improving the palatability of pet foods comprising lipolysis of the starting materials followed by a thermal reaction, such as the Maillor reaction. By starting materials is meant an animal and / or marine and / or vegetable digest obtained after hydrolysis with endogenous enzymes present in tissues or added proteases. Commercially available sources of starting materials include poultry, pork, beef, lamb, fish, and the like, and combinations thereof. Raw materials can be used as starting materials (for example, entrails or entrails and liver from poultry, pork, beef, lamb, fish and the like, and their combinations) and proteolysis before continuing the process in the form of lipolysis and thermal reactions.

The enzymes used in the present invention are proteases and lipases. Commercial proteases and lipases are isolated from plants, animals and microorganisms such as bacteria, yeast and fungi. In practice, commercially available proteases may not be completely pure in the sense that they may exhibit residual lipase activity. Accordingly, commercially available lipases can exhibit residual proteolytic activity. Of course, one skilled in the art will be able to select suitable enzymes to avoid or minimize possible unwanted side effects. For this reason, steps (a) (i) and (d) indicate that the proteolytic and lipolytic reactions are carried out respectively “in the absence of any added lipase” (step (a) (i)) and “in the absence of any added protease "(step (d)). This means that only protease (s) in step (a) (i) and lipase (lipases) in step (d) are present or added. Therefore, if any residual lipase or protease activity is present, respectively, in steps (a) (i) and (d), they are negligible. The only significant enzymatic activities of interest are proteolytic activity in step (a) (i) and lipolytic activity in step (d). The enzymes are generally used in amounts of from about 0.01% to 10%, preferably from 0.01% to 5%, more preferably from 0.01% to 2%, based on the weight of the final palatability enhancer.

To obtain an optimal hydrolysis rate, temperature and pH should be correlated with the enzymes used. This will be fairly obvious to those skilled in the art. The pH can be adjusted to the desired value with any suitable compound suitable for use in pet food, such as phosphoric acid, caustic soda, other common and suitable acid and alkaline regulators, and combinations thereof.

If raw tissues are used as starting material, after proteolysis and before lipolysis, a step of heat inactivation of enzymes (for example, pasteurization) is carried out, followed by filtration, at a temperature usually from about 70 ° C to 95 ° C, for a sufficient time - for example, about from 5 to 20 min. This allows proteases to be inactivated prior to lipolysis.

To ensure that the lipolysis step is carried out, it is important to first emulsify the mixture before adding lipases. Emulsification can be accomplished by adding at least one emulsifier suitable for use in pet food. Suitable emulsifiers are sodium stearoyl lactylate (SLN), succinylated monoglycerides, gum (gum arabic), sodium alginate, lecithin, and the like. Typically, emulsifiers are added in an amount of about 0.01% to 10%, preferably 0.01% to 8%, and more preferably 0.01% to 5%, based on the weight of the final palatability enhancer.

As already indicated above, you can try any commercially available source of animal fat and / or vegetable oil. Suitable sources of vegetable oils available in large quantities are rapeseed oil, soybean oil, corn oil, olive oil, sunflower oil, flax seed oil, palm oil, saffron oil, and the like, as well as by-products thereof. Suitable sources of animal fats include lard, lard, poultry fat, and the like, as well as their by-products. Suitable sources of marine oils are tuna oil, sardine oil, salmon oil, anchovy oil, fish oil, and the like, and their by-products. It also includes fats obtained from animal, vegetable, marine sources, or produced by animals and plants. Typically, the fat is present in an amount of from about 2% to about 30%, preferably from 5% to about 20%, based on the weight of the final palatability enhancer.

After lipolysis, a thermal reaction is carried out to complete the off-flavor of the product. Conveniently, the carbohydrate and nitrogen compound are added at a concentration of from about 0.01% to 30%, preferably from 0.1% to 20%, more preferably from 0.1% to 15% for the first, and from about 0.01% to 30% preferably from 0.01% to 20%, more preferably from 0.01% to 15% for the latter. A suitable temperature is selected in the range from about 70 ° C to 130 ° C, preferably from 80 ° C to 120 ° C, and the heat treatment is carried out for a time sufficient to further develop the flavor of the product, for example for at least 30 minutes.

To ensure long shelf life, preservatives such as natural or synthetic antioxidants can be added (suitable antioxidants include, but are not limited to: butyloxyanisole (BHA), butyloxytoluene (BHT), propyl gallate, octyl gallate, tocopherols, rosemary extracts, and the like), sorbitic acid or sorbitol salts; and other acids such as phosphoric acid and the like.

The palatability enhancer of the present invention can be used directly by itself, usually in an amount of about 0.01% to 20%, preferably 0.01% to 10%, more preferably 0.01% to 5% by weight relative to the weight of the pet food composition. Alternatively, it can be combined with other palatability enhancers, and all palatability enhancers can be administered simultaneously or sequentially.

In one embodiment of the present invention, a dry palatability enhancer formulation is prepared by combining the palatability enhancer in an appropriate ratio with carriers and mixing the components. The mixture is then dried by evaporation to form a dry palatability enhancer.

The palatability enhancers of the present invention are useful in pet foods such as dry pet foods, semi-moist pet foods, having a moisture content of about 50% or less by weight and which is a nutritionally balanced mixture containing proteins, fibers ( fiber), carbohydrates and / or starch. Such mixtures are well known to those skilled in the art and their composition depends on many factors, such as, for example, the required nutritional balance for a particular type of companion animal. In addition to these basic elements, the feed may contain vitamins, salts and other additives such as condiments, preservatives, emulsifiers and moisturizers. Nutritional balance, including the relative amounts of vitamins, salts, lipids, proteins and carbohydrates, is determined according to known veterinary nutritional standards - for example, as recommended by the National Research Council (NRC) or the American Association of Representatives for the Control of food quality (American Asociation of Feed Control Officials, AAFCO).

All common protein sources can be used, especially vegetable proteins such as soy or groundnuts, animal proteins such as casein or albumin, and raw animal tissues such as raw meat tissue and raw fish tissue, or even elements that are dry or dried, such as fish meal, bird meal, meat meal, and bone meal. Other types of suitable proteinaceous materials include wheat or corn gluten and proteins from microorganisms such as yeast. You can also use ingredients that contain a significant proportion of starches or carbohydrates, such as corn, milo, alfalfa, wheat, barley, rice, soybean hulls, and other low protein grains.

Other ingredients such as whey and milk by-products such as carbohydrates can be added to the feed. In addition, you can add known seasonings, including corn syrup or molasses.

By way of example, a typical dry cat food formulation into which a palatability enhancer according to the present invention can be incorporated consists of the following components (weight percent): about 0-70% of a bread base such as flour (corn, wheat, barley or rice); approximately 0-30% of animal by-products (from poultry or meat); about 0-25% corn gluten; approximately 0-25% of crude animal tissue - such as that of poultry or cattle; about 0-25% soy flour; approximately 0-10% animal fat; about 0-20% seafood base; approximately 0-25% raw fish tissue; about 0-10% high fructose corn syrup; about 0-10% dry molasses; about 0-1.5% phosphoric acid; and about 0-1.5% citric acid.

Vitamins and salts can be added, including calcium carbonate, potassium chloride, sodium chloride, choline chloride, taurine, zinc oxide, iron sulfate, vitamin E, vitamin A, vitamin B12, vitamin D3, riboflavin, niacin, calcium pantothenate, biotin, thiamine mononitrate, copper sulfate, folic acid, pyroxidine hydrochloride, calcium iodate and menadione complex with sodium bisulfite (source of vitamin K activity).

Dry pet foods are generally prepared in a variety of ways. One of these methods, which is widely used, is the cook-extrusion method. In a cook-extrusion process, the dry ingredients are first mixed together to form a mixture. This mixture is transferred to a steam conditioner where it is sufficiently humidified to be squeezed out. The mixture is then introduced into a cooking extruder, where it is cooked at elevated temperature and pressure, and then extruded from the apparatus through a press. This press gives the extrusion product a specific shape. Individual pieces of product are obtained by periodically cutting from the end of the strip of the extruded product. The individual pieces or slices are then dried in a hot air dryer. Typically, the product is dried until it contains less than 15% moisture, preferably about 5 to 10% moisture. The dried particles or pieces are then transferred by a loading conveyor to a coating drum and sprayed with fat. Other liquids, such as, for example, phosphoric acid, can alternatively be applied to the pieces or can be applied along with the fat. The resulting granules or slices constitute the base composition to which the palatability enhancer coating can be applied.

In one embodiment of the present invention, the palatability enhancers of the present invention may be administered by coating. The term "coating" as used herein refers to the surface application of a palatability enhancer or flavoring composition to the surface of a base composition, for example, by spraying, spraying, and the like. For example, slices of uncoated, squeezed out basic pet food can be placed in a container for mixing, such as a tube or drum for coating. Fat, such as lard or poultry fat, is heated and then sprayed onto the pet food to coat the slices. The coating does not require a continuous layer, but should preferably be uniform. After the fat, a palatability enhancer can be applied, either as a liquid or as a dry powder, during the mixing process. The liquid palatability enhancer is usually sprayed on, while the dry palatability enhancer is usually sprayed on. Alternatively, palatability enhancers can be mixed with fat and applied at the same time. In another alternative coating method, palatability enhancers are applied prior to application of the fat.

In another embodiment of the present invention, the palatability enhancer is contacted with the raw materials of the pet food composition prior to cooking. In this case, the palatability enhancer is combined with proteins, fibers, carbohydrates and / or starch of the base composition and cooked together with these materials in a cooking extruder.

The palatability enhancers of the present invention are also useful for wet pet foods having a moisture content of more than 50% and presenting a nutritionally balanced mixture. Wet food may contain one or more ingredients selected from mealy materials (such as grain and flour based materials), animal by-products, raw animal tissues, raw fish tissues, animal and vegetable fats, marine materials, vitamins, salts, preservatives, emulsifiers, surfactants, structuring agents, colorants and the like. Such ingredients are well known to those skilled in the art and may be suitably selected depending on the type of wet food.

Most types of meat gravy-based pet foods are prepared by pulverizing meat, simulated meat or meat by-products and then forming the mashed mixture by reduced pressure extrusion through a steam tunnel where the food is cooked. Then add starch and binders, after which the mixture is cut into pieces, mixed with water, starch and binders. After that, the mixture is packed and closed in cans and boiled in a hydrostat with continuous or rotary sterilization. Non-gravy wet pet foods are prepared by soaking meat, simulated meat or meat by-products and shaping the soaked materials with starch, water, and binders. After that, the mixture is packed and closed in cans and boiled in a hydrostat with continuous or rotary sterilization.

A liquid or dry palatability enhancer can be incorporated into a base such as a gravy or jelly while mixing along with the remaining ingredients (structuring agents, stabilizers, colorants, and nutritional supplements). A liquid or dry palatability enhancer can also be incorporated into meat by-product mixtures for making slices or sticks. In this case, it can be added to the raw material before or after the grinding process. The meat by-product mix can be steam-cooked or grilled for slices, or directly soldered into cans for chopsticks.

The palatability enhancers described above provide significant advantages over previous solutions. The effects of the present invention can be measured in what is commonly referred to as the "two bowl test" or "comparison test". Of course, one skilled in the art is free to use any other suitable test instead of the two bowl test described herein to determine the advantage. Such alternative tests are well known in the art.

Two Bowl Test Principle:

The test is based on the postulate that the more feed is eaten, the more palatability it has. Preference tests of individual animals were performed using the two-bowl method, based on a comparison of two foods. Tests were performed on either groups of 36 dogs or groups of 40 cats, depending on the purpose of the test.

Test method:

The same amounts of food A and food B were weighed and placed in the same bowls. The amount in each diet provides the daily food requirement.

Distribution of bowls:

Dog test: Bowls were placed in individual food trays available for dogs.

Cat test: Bowls were placed at the same time in front of each cat in individual locked boxes and their position was changed with each meal to eliminate the influence of the position of the bowls.

Test duration:

Dog test: 15 minutes maximum (if one of the two bowls was completely emptied in less than 15 minutes, both bowls were removed and the test was stopped).

Test for cats: minimum 15 minutes (if the contents of one of the bowls were completely eaten in less than 30 minutes, both bowls were removed and the test was stopped).

Investigated parameters:

Measured parameters: First food eaten and the amount of each food eaten at the end of the test.

Calculated parameters: Individual consumption ratio (CR).

SP A = feed intake A (in d) × 100 / intake A + B (in d)

SP B = consumption of feed B (in d) × 100 / consumption of A + B (in d).

Average intake ratio (SDR) is the average of all individual ratios (all animals are equally significant, regardless of their size and their respective food intake). If the consumption of animals is higher or lower than certain values, they are not taken into account in statistical processing.

Statistical analysis:

Statistical analysis was applied to determine if there is a significant difference between the two ERP ratios. The Student's t-test was used with three threshold error values, namely 5%, 1% and 0.1%.

The Chi test was used to determine if there is a significant difference between the number of dogs or cats with a preference for food A and the number of dogs or cats with a preference for food B.

Significance levels are indicated as follows:

ND difference is insignificant (p> 0.05)

* reliably (p<0,05)

** high degree of reliability (p<0,01

*** very high confidence level (p<0<001).

In the following examples, various types of fats or fat blends are tested as defined above. These fats are hereinafter referred to as Fat 1, Fat 2, Fat 3. It is shown below that regardless of whether a fat or a fat blend is used, the palatability of the product according to the present invention is very high.

Example 1: XLHM product with starting raw material

Recipe:

The raw materials, exogenous and / or endogenous proteases, preservatives and antioxidants are mixed together and heated at about 60 ° C to 70 ° C for at least 30 minutes (step a) (i)).

The mixture is heated and pasteurized at about 85 ° C for at least 10 minutes, then cooled at about 25 ° C to 45 ° C, preferably with concomitant filtration, to obtain the first stage reaction product (step a) (ii)).

Optionally, a storage step may be added here under suitable conditions for a given period, preceded by an acidification process.

The pH is then adjusted to about 7 to 10 with caustic soda or an alkalinity regulator and emulsifiers, fat and lipase enzymes are added for lipolysis, lipolysis is carried out for at least 120 minutes, preferably about 120 to 420 minutes, to obtain the reaction product the second stage (stages b) to d) is carried out simultaneously).

Reducing sugars and nitrogen compounds are added and the resulting mixture is heated at about 90 ° C to 110 ° C for at least 30 minutes, resulting in a palatability enhancer (step e).

Finally the product is cooled and phosphoric acid, potassium sorbate, preservatives and antioxidants are added for long-term storage, with a final pH of 2.9, resulting in a ready-to-use palatability enhancer product (designated XLHM).

XLHM version A: base - raw material from poultry;

XLHM version B: base - raw material from poultry, used fat 1;

XLHM version C: poultry raw material base, fat 2 used;

XLHM version D: poultry raw material base, fat used 3.

Results of comparing palatability of SP1 and SP2 for dogs:

Consumption of SP1 and SP2 is significantly different, which demonstrates the best performance of the palatability enhancer SP2 from Super Premium. The test results are shown in the graph in Fig. 1.

Taste Test Results for Dogs XLHM Compared to SP1 and SP2

All four versions of XLHM exhibit increased palatability compared to the SP1 product. For all fat variants used, the palatability of XLHM is equal to or greater than the palatability of SP2.

Example 2: XLHM product with original cleavage

Recipe:

The starting material used in this example is the digest (cleavage product) obtained after step a) (i) and (ii), as illustrated in example 1, that is, it is the product of the first reaction.

The method begins with steps b), c) and d), where the pH is adjusted in the range from about 7 to 10 using caustic soda or alkalinity regulators, for lipolysis, emulsifiers, fat and lipase enzymes are added, lipolysis is carried out for at least 120 min, preferably for about 120 to 420 minutes to obtain the reaction product of the second stage.

Then, reducing sugars and nitrogen compounds are added and the resulting mixture is heated at a temperature of about 90 ° C to 110 ° C for at least 30 minutes, resulting in a palatability enhancer (step e).

Finally the product is cooled and phosphoric acid, potassium sorbate, preservatives and antioxidants are added for long-term storage, with a final pH of 2.9, resulting in a ready-to-use palatability enhancer product (designated XLHM version).

Evaluation of palatability for dogs:

SP1 and SP2 are Super Premium liquids from the existing range with different palatability levels, with SP2 being more palatable than SP1.

XLHM products are palatability enhancers according to the present invention:

XLHM version E: first liquid digest, used fat 1;

XLHM version F: liquid digest first, fat 2 used;

XLHM version G: liquid digest first, fat used 3.

All test results are presented in the graph in figure 2.

When using the original digest at the beginning of the process, the result is the same as before. That is: higher palatability than SP1, and at least equal to that of SP2, and often higher than SP2.

Comparative example 3: XLHM product (D ") with starting raw material and without related proteolysis step

Recipe:

Here the method begins with the stage of joint action of proteases and lipases.

First, the protease-containing raw materials, water, preservatives and antioxidants are mixed together, the pH is adjusted to the range from about 7 to 10 with caustic soda, emulsifiers, salts, fat 3 and lipase enzymes are added to carry out the hydrolysis step, the hydrolysis is carried out at a temperature of about 25 ° C to 45 ° C, for at least 120 minutes, preferably for about 120 to 420 minutes, to obtain product 1.

Then, reducing sugars and nitrogen compounds are added and the resulting mixture is heated at a temperature of about 90 ° C to 110 ° C for at least 30 minutes to obtain product 2.

Finally, product 2 is cooled and phosphoric acid, potassium sorbate, preservatives and antioxidants are added for long-term storage, with a final pH of 2.9 to obtain a ready-to-use product (product XLHM D ").

Evaluation of palatability for dogs:

Compared to the results obtained for the product XLHM version D (see example 1), XLHM version D "is worse compared to SP2, but retains a high palatability compared to SP1. In other words, the results obtained with the combination of proteolysis and lipolysis, not as good as the results obtained by separating proteolysis and lipolysis.

Comparative example 4: XLHM product (B ") with starting raw material and with reordering of enzymatic steps

Recipe:

In this example, the method begins with a lipolysis step followed by a proteolysis step.

First, the raw materials, water, preservatives and antioxidants are mixed together, the pH is adjusted to the range from about 7 to 10 with caustic soda, emulsifiers, salts, fat 1 and lipase enzymes are added to carry out the lipolysis step, lipolysis is carried out at a temperature of about 25 ° C. to 45 ° C, for at least 120 minutes, preferably for about 120 to 420 minutes, to obtain product 1.

Then, protease enzymes, reducing sugars and nitrogen compounds are introduced and heated at a temperature of about 60 ° C to 70 ° C for at least 30 minutes to obtain product 2.

The resulting mixture is heated at about 90 ° C to 110 ° C for at least 30 minutes to obtain product 3.

Finally, product 3 is cooled and phosphoric acid, potassium sorbate, preservatives and antioxidants are added for long-term storage, with a final pH of 2.9 to obtain a ready-to-use product (product XLHM B ").

Evaluation of palatability for dogs:

Compared to the results obtained for the product XLHM version B (see example 1), XLHM version B "is worse than SP1.

Conclusion: Better results are obtained when proteolysis is performed prior to lipolysis than when lipolysis precedes proteolysis.

Example 5: Adding XLHM version G to cat loaves

Product B recipe

Product recipe C

Definition: C "sens W9P is a commercial Super Premium SPF palatability enhancer for incorporation into wet pet food.

Raw materials (pork lungs, pork liver, chicken lungs and liver, chicken carcasses) were thawed overnight at room temperature. Then they were ground in a vertical grinder (Stephen, Germany) for 5 min at a frequency of 1500 strokes per min. Water was added to the glass. Powders (structuring agents, a mixture of vitamins and salts and wheat flour) and XLHM version G or C "sens W9P were dissolved in water using a homogenizer (Dynamic, France). The solution was added to the crushed meat products and stirred for another 5 min under vacuum ( -1 bar) in a grinder.The porridge was transferred to a vacuum dispenser (Handtmann, Germany) and filled into 400 g iron cans. The cans were closed and heated in a Microflow retort (Barriquand, France) using the following regime: heating to 127 13 min, the temperature is maintained at 127 ° C for 55 min, cooling to 20 ° C for 15 min.

Evaluation of palatability for cats

Thus, there is shown and described a new and useful method for improving the palatability of pet food compositions. Although the present invention has been accompanied by examples for purposes of illustration and the description has been given with reference to specific embodiments, it should be apparent to those skilled in the art that various modifications, variations, and equivalents of the illustrative examples are possible. It is believed that any such changes which directly follow from the set forth herein and which do not depart from the spirit and scope of the present invention are covered by the present invention.

CLAIM

1. A method of obtaining a palatability enhancer for use in pet food, comprising at least:

a) providing a first-stage reaction product obtained by:

(i) reacting with at least one exogenous and / or endogenous protease in the absence of any exogenous or added lipase, wherein the substrate contains protein and fatty materials in amounts, at pH and temperature conditions and for a time effective to effect the proteolytic reaction , (ii) heat inactivating said protease and filtering the resulting cleavage product;

b) optional fat addition;

c) emulsification of the specified reaction product of the first stage;

d) reacting said emulsion with at least one lipase in the absence of any added protease in amounts, at pH and temperature conditions and for a time effective to effect the lipolytic reaction to obtain a second-stage reaction product;

e) adding to said second-stage reaction product of at least one reducing sugar and at least one nitrogen compound and heating the resulting mixture to a temperature and for a time effective to further form the palatability of the mixture, resulting in a palatability enhancer.

2. The method according to claim 1, further comprising step a) (iii) cooling the product obtained in step a) (ii) to a temperature effective for carrying out the subsequent lipolytic reaction in step d).

3. A method according to claim 1 or 2, further comprising the step of cooling the mixture obtained in step e).

4. A process according to claim 1 or 2, wherein the reaction product of the first stage is prepared and stored under suitable conditions until further use.

5. A palatability enhancer for use in pet food, which can be obtained by the method according to any one of claims 1 to 4.

6. The palatability enhancer of claim 5, wherein said palatability enhancer is a liquid or a powder.

7. A palatability enhancing composition for use in pet food, comprising at least one palatability enhancer according to claim 5 or 6.

8. A method of preparing pet food having increased palatability, comprising at least:

introducing into the pet food at least one palatability enhancer according to claim 5 or 6 or at least one palatability-enhancing composition according to claim 7 in an amount effective to increase the palatability of said pet food.

9. The method of claim 8, wherein said administration is by coating or by addition to the bulk of the pet food.

10. Pet food having an increased palatability, which can be obtained by the method according to claim 8 or 9.

11. A palatable pet food comprising at least one palatability enhancer according to claim 5 or 6, or at least one palatability-enhancing composition according to claim 7.

12. The pet food of claim 10 or 11, wherein said pet food is selected from the group consisting of dry, semi-dry, and wet pet foods.

13. A method of feeding pets, comprising at least:

a) providing pet food according to any one of claims 10-12;

b) feeding said pet food to pets.

14. The method of claim 13, wherein said companion animals are selected from the group consisting of cats and dogs.