Michal Gábor

Identification of cryptic allele for merle patterning in dogs by molecular genetics methods.

Abstract Merle patterning in dogs, caused by the insertion of a short interspersed element (SINE) in the genetic structure of SILV gene, is characterized by patches of diluted pigment intermingled with normal melanin. Sequencing analyses of SINE element localized in the canine SILV gene discovered a variability of the poly (A)-tail length which is responsible for the different expression of merle pattern. The SINE element with the length of poly(A)-tail between 91 - 101 nucleotides is responsible for the merle phenotype with all characters of merle pattern. On the contrary the dogs which have SINE element with the shorter length of poly(A) tail between 54-65 nucleotides are referred as cryptic merles without expresion of Merle pattern. The aim of this study was to improve molecular genetics method for the detection of cryptic allele for merle patterning in dogs. A total of 40 dogs of four breeds - Border collie, Shetland sheepdog, Australian Shepherd dog, and Chihuahua were used in this study. Canine genomic DNA was isolated from samples of whole blood and buccal cells by commercial column kit. Detection of merle (M), cryptic merle (Mc) and non-merle (m) alleles was done using M13-tailed primer protocol and two different allele-sizing methods for the verification of the electrophoresis result. In the analyzed population of dogs were detected 20 dogs with non-merle genotype mm, 17 dogs with merle genotype Mm, 2 dogs with double merle genotype MM and one dog with merle phenotype but with the presence of cryptic merle allele Mc with the consequential genotype MMc. Mramorirana šara pasa, nastala umetanjem kratkih ponovaca elemenata (SINE) u genetsku strukturu SILV gena, karakteriše se prisustvom slabije pigmentisanih polja pomešanih sa poljima normalne pigmentacije. Analizom SINE elementa lokalizovanog u SILV genu pasa sekvencioniranjem otkrila je varijabilnost poli(A) repa koji je odgovoran za različitu ekspresiju mramorirane šare. SINE element sa poli(A) repom dužine od 91 do 101 nukleotida je odgovoran za mramorirani fenotip sa svim karakterima mramorirane šare. Nasuprot tome, psi koji poseduju SINE element sa kratkim poli(A) repom dužine od 54 do 65 nukleotida su opisani kao prikriveni mramorirani bez ekspresije mramorirane šare. Cilj ove studije je poboljšanje metoda molekularne genetike za detekciju skrivenog alela za mramoriranu šaru pasa. Ukupno 40 pasa četiri rase: border koli, šetlandski ovčar, australijski ovčar i čivava bilo je uključeno u studiju. Genom DNK pasa bio je izolovan iz uzoraka pune krvi i ćelija sluznice usne duplje bio je izolovan pomoću komercijalnih kitova. Detekcija mramoriranog (M), prikriveno mramoriranog (Mc) i ne-mramoriranog (m) alela je izvršena pomoću „M13-tailed“ prajmera protokola i dve različite metode određivanja veličine alela za verifi kaciju rezultata elektroforeze. U analiziranoj populaciji pasa otkriveno je 20 pasa sa ne - mramoriranim genotipom mm, 17 pasa sa mramoriranim genotipom Mm, 2 psa sa duplim mramoriranim genotipom MM i jedan pas sa mramoriranim fenotipom, ali sa prisustvom prikrivenog mramoriranog alela Mc sa posledičnim genotiom MMC.

GENETIC MARKERS AS ONE OF TOOLS FOR PRODUCTION OF TENDERNESS MEAT IN CATTLE

Meat tenderness is one of the major characteristic quality of beef not only for consumers but for breeders of beef cattle too. Selection of cattle focussed on an increment of meat tenderness is complicated because this trait has large variability not only between different breeds but between individuals of equal breed too. Similarly a measurement of meat tenderness is expensive because it make after slaughter of animal and ageing of meat post mortem. Therefore it was developed a several methods, by the help of which is possible increase tenderness of meat. However still exist variance in values of meat tenderness which are caused by distinctness genetic base of animal. By using molecular genetic methods was described the most significant candidate genes (CAPN1, CAST) coding formation of the calpains-calpastatin proteolytic system, which exercise an influence on tenderness. The single nucleotide polymorphisms (SNPs) in these genes were using to design commercially genetic marker panels GeneSTAR Tenderness and Igenity Tender-GENE. By help this commercially test is possible to make genotyping and selection of animals for production of tenderness beef meat in meat industry. doi:10.5219/102

Trace elements content in semen and their interactions with sperm quality and RedOx status in freshwater fish Cyprinus carpio: A correlation study

Objective of the present study was to investigate interactions between trace elements content and RedOx status, as well as sperm quality parameters (motility features, DNA fragmentation) in fish spermatozoa in natural conditions. Reproductively mature male freshwater fish (n = 16) of Cyprinus carpio breed were used in the study. Trace elements content was determined in fish milt samples by inductively-coupled plasma optical emission spectrometry (ICP-OES) and by cold-vapor atomic absorption spectroscopy (CV-AAS). Sperm quality evaluation was realized by computer-assisted sperm analysis (CASA) quantifying several parameters: concentration, total motility, progressive motility, distance average path, distance curved line, distance straight line, velocity average path, velocity curved line, velocity straight line, straightness, linearity, amplitude of lateral head displacement and beat cross frequency. The general scheme of descending concentrations of trace metals in semen samples was following: Zn > Fe > Cu > As > Sr > Ni > Mn > Se > Pb > Cr > Cd > Hg. Total motility of spermatozoa was relatively high (91.45%), however progressive motility was not even half of this value (39.47%). Sperm DNA fragmentation values were relatively low (4.00-6.29%). The percentage of immotile spermatozoa showed a significant correlation with all RedOx status parameters and also with DNA fragmentation. Positive statistically significant correlations were observed between trace elements (Mn, Se, Sr, and Zn) and some qualitative spermatozoa parameters (velocity and distance parameters). Cu and Hg content shows similar negative associations with progressive motility. Hg also interacted with production of malondialdehyde. Overall, the present study suggests application of multi-component mixtures of environmentally related trace elements concentrations when assessing the potential reproductive risk.

Effect of selected polymorphisms of genes lep, MTHFR and FTO to BMI level and gender-specificity

The aim of this study is to investigate the effect of selected polymorphisms LEP (G2548A), MTHFR (C677T) and FTO ( rs1121980 ) on body mass index in humans. In the study participated 79 people from Slovakia with some genetic relatedness. Genomic DNA was isolated from the buccal swabs using a commercial kit Qiagen DNA Mini Kit (Qiagen). The detection of SNP polymorphisms in human genes LEP, MTHFR and FTO was performed using molecular genetics methods such as PCR-RFLP and ARMS. The most common genotypes in all 3 polymorphism were found in heterozygous form (for LEP AG = 0.5190, for MTHFR CT = 0.519, for FTO CT =0.4051). The LEP gene had increased frequency of G allele (0.5506), the MTHFR gene T allele (0.6646) and FTO gene T allele (0.50635). The least frequent genotype in LEP was AA (0.1899), in MTHFR was TT (0.0759), in FTO it was CC (0.2911). According to the results we can assume that the genotypes AA (LEP G2548A), CC (FTO rs1121980 ) and TT (MTHFR C677T) in case of women have a protective effect on the prevalence of obesity, based on BMI, compared to the other genotypes and this polymorphism is gender-specific. Added anthropometric measurements, blood test and extension of the group in the future evaluation could increase the statistical relevance in relation to obesity and gender-specificity.

Effect Of Chromium Nicotinate On Oxidative Stability, Chemical Composition And Meat Quality Of Growing-Finishing Pigs

The effect of different organic sources of Cr on growth, feed efficiency and carcass value is known but there is a lack of information between chromium nicotinate (CrNic) and pork quality. Therefore, purpose of this research was to investigate the effects of CrNic on chemical composition, quality and oxidative stability of pork meat. In the study, pigs of Large White breed (40 pcs) were used. The pigs were divided into two groups, namely the control and the experimental of 20 pcs with equal number of barrows and gilts. The pigs were fed the same diet which consisted of three feed mixtures applied at the different growth phases, from 30 - 45 kg OS-03, 45 - 70 kg OS-04 and 70 - 100 kg OS-05. The pigs were allowed ad libitum access to feed and water. The diet of experimental group was supplemented with 0.75 mg.kg -1 CrNic in the form of chromium-inactivated yeast Saccharomyces cerevisiae . The fattening period in pigs lasted from 30 to 100 kg. The chromium supplementation led to a significantly higher content of chromium in longissimus thoracis muscle (LT) of experimental pigs. In addition, the results showed a statistically significant difference ( p ≤0.05) in retention of chromium in the LT, monounsaturated and omega-3 polyunsaturated fatty acids content in experimental group compared with control. Moreover, there was highly significant ( p £0.05) difference in essential fatty acids, as well as in oxidative stability in 7 days, among the groups. The highly significant differences were also observed among sexes, namely in total water, protein and intramuscular fat contents, colour CIE b* in both times, and oxidative stability. However, physical-technological parameters (pH, drip loss, shear force and meat colour) were not affected when pigs were fed the supplement. On the whole, the positive effect of chromium nicotinate in most of investigated parameters may be beneficial not only for pork industry but also for consumers. Normal 0 21 false false false CS JA X-NONE

THE TESTING OF SANITIZERS EFFICACY TO ENTEROCOCCI ADHERED ON GLASS SURFACES

The aim of this work was to test the ability of 6 strains of enterococci to adhere on glass surfaces in environment with different content of milk residues and then to evaluate efficacy of 2 commercial sanitizers (alkaline and acidic) used in milk production. Tested enterococci were isolated from milk, dairy products and from rinse water after sanitation milking machine. Suspension of enterococci (8 log CFU.mL -1 ) was prepared in phosphate buffered saline (PBS), PBS with content 0.1% and 1% of skimmed reconstituted milk. Glass plates were immersed into bacterial suspension for 1 h at 37 °C. The number of enterococci adhered on glass surface in PBS achieved an average value 3.47 log CFU.mm -2 , in PBS with 0.1% of milk 2.90 CFU.mm -2 , in PBS with 1% of milk 2.63 CFU.mm -2 . Differences between the tested files were not statistically significant (p >0.05). In the second part of work the glass plates with adhered enterococci were exposed to the effect of alkaline sanitizer (on basis of NaOH and NaClO), respectively acidic sanitizer (on basis of H3PO4). Sanitation solutions were prepared and tested according to manufacturer recommendations (concentration 0.25%, contact time 20 min, temperature 20 °C). Alkaline sanitation solution was 100% effective against all tested enterococci regardless to content of milk residues in environment. Acidic sanitation solution was 100% effective only against E. faecalisD (isolated from rinse water after sanitation). Average value of reduction of enterococci with acidic sanitation solution, which were on glass plates in environment PBS was 2.84 CFU.mm -2 , in PBS with 0.1% of milk was 2.45 CFU.mm -2 and in PBS with 1% of milk was 2.16 CFU.mm -2 . It can be concluded, that increase of milk residues in environment decrease the adhesion of enterococci on