LACTATE DEHYDROGENASE GENE GENOTYPIZATION FOR SPECIES IDENTIFICATION IN A FISH FARM ON THE RIVER NERETVA

There are severale Salmonid species, found in the river Neretva basin, among which S. trutta and S. obtusirostris. Also, natural hybrids such as S. obtusirostris x S. trutta have been observed. In one fish farm on the river Neretva, S. trutta and S. obtusirostris were decided to breed separately. Parental fishes were separated phenotypicaly on the basis of the morphological signs. PCR-RFLP analysis of the exon 3 to exon 4 part of the lactate dehydrogenase (LDH) C1* gene with restriction endonuclease RsaI was employed to identify the presence of other species representatives or intercrosses in two groups of juvenille fishes. Using this method, we were able to identify two S. trutta representatives in the S. obtusirostris group

Results of zeolite application in fattening pigs on deep litter

Istraživanje je provedeno na 120 svinja četveropasminskih križanaca oba spola, držanih na dubokoj stelji. Svinje su bile podijeljene u 4 skupine. Prva skupina bila je kontrolna, 2. i 3. skupina dobivale su u obroku prirodni zeolit Anivital (2,5% i 5%), dok je kod 4. skupine po stelji posipan zeolit Aromavital u količini od 3,5 kg/m2 obora. Tijekom provedbe istraživanja obavljena su mikroklimatska mjerenja, kao i mjerenja proizvodnih svojstava tovnih svinja. Po završetku tova svinje su zaklane, a na liniji klanja utvrđena su svojstva kakvoće svinjskih trupova i mesa. Svinje koje su dobivale Anivital u obroku imale su statistički značajno veće (P < 0,05) završne tjelesne mase, veće priraste, nižu konverziju i veću konzumaciju hrane u odnosu na kontrolnu skupinu svinja. Svinje kontrolne skupine imale su najveći udjel mišićnog tkiva u polovicama. Rezultati mikroklimatskih mjerenja pokazuju da su najpovoljniji mikroklimatski uvjeti utvrđeni kod 4. skupine svinja, kod koje je dodavan Aromavital, dok su najlošiji uvjeti bili kod kontrolne skupine. Može se zaključiti da dodatak Anivitala u obroke svinja povoljno utječe na poboljšanje nekih proizvodnih svojstava tovnih svinja. Isto tako, posipanjem Aromavitala po stelji u značajnoj se mjeri poboljšavaju mikroklimatski uvjeti u tovilištu.The research was carried out on 160 four-way crossbred pigs of both sexes, all kept on deep litter. Pigs were divided into 4 groups. The first group was the control, the 2nd and the 3rd were given natural zeolite Anivital (supplemented to diets in the amount of 2.5% and 5%, respectively), while in the 4th group zeolite Aromavital was spread on deep litter in the amount of 3.5 kg/m2 of the pen. During the research period, microclimatic indicators and productive traits of pigs were constantly measured. After slaughtering of pigs in a slaughterhouse, quality traits of their carcasses and meat were determined. Pigs that were given Anivital supplemented to diets had statistically significantly higher (P<0.05) end weights, better gains, lower conversion and better food consumption than the control group. Pigs in the control group had the highest portion of muscle tissue in carcass. Results of microclimatic measurements proved that the most favorable microclimatic conditions occurred in the 4th group of pigs that were given Aromavital, and the worst conditions were obtained in the control group. It is concluded that supplementation of Anivital to pig diets had positive effects on the improvement of some productive traits. Moreover, spreading of Aromavital on deep litter resulted in improved microclimatic conditions within pens

PRELIMINARNO ISTRAŽIVANJE ODNOSA POLIMORFIZAMA NA CAST GENU I KVALITETE SVINJSKOG MESA

The present study was performed in order to investigate a relationship between polymorphisms on the calpastatin gene (CAST) and pig meat quality traits. The investigation was carried out on 29 gilts and barrows, crosses of Large White x German Landrace randomly selected at slaughter line. Pigs were slaughtered at 130-150 kg of live weight and blood samples were taken for genomic DNA analysis. The following indicators of meat quality and meat chemical composition were evaluated: pH and electric conductivity measured 45 minutes post mortem in m. Longissimus dorsi (LD muscle) and in M. Semimembranosus (SM muscle); electric conductivity measured at the same locations after 24h of cooling; drip loss (determined by “bag method”); colour (measured with Minolta chromameter and expressed as Hunter L, a, b values); instrumental tenderness assessed as Warner-Bratzler (WB) shear force and moisture, fat, protein and collagen content (%) determined on cooked LD muscle after 24h of thawing. The amplification products of the CAST gene were digested with HinfI restriction endonuclease and three genotypes (AA, BB and AB) were revealed. Statistical analysis showed that meat originated from pigs of AB genotype had the lowest WB shear force and the highest protein content of cooked LD muscle. As for the indicators of technological meat quality, statistically significant differences (p<0.05) were found between genotype AA and both BB and AB genotype for electric conductivity measured in LD muscle after 24h of cooling, as well as between BB and both AA and AB genotypes for drip loss.Istraživanje je provedeno na 29 križanaca velikog jorkšira i njemačkog landrasa nasumično odabranih na liniji klanja u cilju utvrđivanja veze između polimorfizama na kalpastatin genu (CAST) i svojstava kvalitete mesa. Svinje su zaklane pri 130-150 kg tjelesne mase, kada su izuzeti uzorci krvi za izolaciju genomske DNK. Na liniji klanja izmjerena su slijedeća svojstva kvalitete mesa: početna pH vrijednost i početna vrijednost električne provodljivosti (mjereni na musculus Longissimus dorsi i musculus semimembranosus 45 minuta nakon klanja); završne vrijednosti električne provodljivosti (izmjerene na istim mjestima 24 h nakon hlađenja polovica); otpuštanje mesnog soka (“metodom vrećice”); boja (izmjerena Minolta kolorimetrom i izražena kao Hunter L, a i b vrijednosti); tvrdoća/nježnost mesa te udio vlage, masti, proteina i kolagena (na skuhanom odsječku MLD). Umnoženi produkti CAST gena podvrgnuti su restrikciji HinfI endonukleazom te su utvrđena tri genotipa (AA, AB i BB). Statističkom analizom utvrđeno je da su svinje AB genotipa imale najpoželjnije vrijednosti tvrdoće/nježnosti mesa te najviši udio proteina u MLD-u, dok su za tehnološka svojstva kvalitete mesa statistički značajne razlike utvrđene između genotipa AA i BB te AB za završne vrijednosti električne provodljivosti u MLD-u, kao i između BB te AA i AB za vrijednosti otpuštanja mesnog soka

GENETIC ANALYSIS OF BLACK SLAVONIAN PIG

Pairs (18) of microsatelite primers were used in this study to detect the genetic relationship within Black Slavonian Pig and between Turopolje Pig, Mangalitsa breed and Croatian Wild Pigs. The second goal of this study was to determine phylogenetic relationships among these breeds and some Asian and European pigs using the mtDNA D-loop sequence polymorphism. The third goal was to determine the MC1R genotype of Black Slavonian pigs and to find an efficient and simple PCR-RFLP method, based on differences in MC1R genotype, to distinguish between purebred Black Slavonian pigs and their crossings with commercial pig breeds and Wild Boars. Aiming to conduct microsatellite analysis each animal was genotyped for 18 microsatelite markers, chosen based on their quality, size, polymorphism and location on the porcine genome as proposed by the FAO. Two pairs of primers amplified a 511-bp fragment of control region between sites 15 390 and 15 900 (Mit1.F and Mit1.R) and a 810-bp fragment between sites 15 825 and 16 634 (Mit2.F and Mi2.R) were genotyped for mtDNA. Two primer pairs were used to amplify the majority of the single exon of MC1R gene aiming to determinate MC1R genotype of Black Slavonian pig. The first pair of primers, MERL1 and EPIG2, was used to amplify a 428-bp product from the 5’ half of the exon, whereas EPIG1 and EPIG3 amplified a 405-bp product from the 3’ half. Our results showed that the 18 microsatellites used in this study were useful markers to study genetic diversity among Croatian autochthonous pig breeds. This set of microsatellites may be used for identifying individuals and for genetic diversity studies for selection and conservation of the Black Slavonian pig, Turopolje pig and Mangalitsa breed. Genetic distances between populations made with Principal Component Analysis (PCA) method noticed that studied populations are mostly clearly geneticaly defined. mtDNA analysis suggested that Black Slavonian and Turopolje pig showed genetic distances from the other European and Chinese pig breeds. Black Slavonian and Turopolje pig clustered together. Mangalitza was used in the development of Black Slavonian pig and this can explain their presence in clustering. Also, with this work, we were able to identify a simple PCR-RFLP method, based on different coat color MC1R gene genotypes, by which it is possible to detect potential crossings of autochthonous Black Slavonian pig with commercial pig breeds and including wild boars

APPLICATION OF ANALYTICAL METHODS FOR FOOD AUTHENTICATION

Autentifikacija proizvoda služi za dokazivanje sastava i podrijetla poljoprivrednih i prehrambenih proizvoda. Republika Hrvatska ima veliki broj poljoprivrednih i prehrambenih proizvoda koji su zbog specifičnih organoleptičkih, tehnoloških i prehrambenih specifičnosti deklarirani kao proizvodi visoke vrijednosti te zaštićeni oznakom izvornosti, zemljopisnog podrijetla ili zajamčeno tradicionalnog specijaliteta. Provođenje postupka autentifikacije predstavlja multidisciplinarni postupak koji uključuje analitičke metode kombinirane s informatikom, matematikom i statistikom, a sve s ciljem dobivanja što točnijih i pouzdanijih podataka kako bi se mogao donijeti zaključak o podrijetlu i sastavu proizvoda. Zanimanje javnosti za konzumiranjem kvalitetne i sigurne hrane dovelo je do brzog razvoja metoda za autentifikaciju proizvoda. U analitičke metode analize ubrajaju se molekularne metode analize, kromatografske metode, spektrometrijske metode i imunološke metode. U radu su opisane analitičke metode koje se primjenjuju u postupcima dokazivanja izvornosti, zemljopisnog podrijetla ili tradicijskog specijaliteta proizvoda u svijetu.Food Authentication is primarily used for testing the composition and origin of agricultural and food products. Croatia has a large number of agricultural and food products, which are due to the specific organoleptic, technological and nutritional specifics declared as high-value and they are protected by designation of origin, geographical indications and traditional specialties guaranteed. The implementation of the authentication procedure is multidisciplinary process which includes analytical methods combined with computer science, mathematics and statistics, all with the goal of achieving a more accurate and reliable information in order to make a conclusion about the origin and composition of the product. Public interest in the consumption of high-quality and safe food has led to a rapid development of methods for products authentication. The analytical approach to the analysis is divided in several categories related to the molecular analysis methods, chromatographic methods, immunological methods and spectroscopic methods. This paper describes the analytical methods used in the procedures of proving the origin, geographical indications or traditional specialties product