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Detection of DNA mutations by PCR-TTGE method
21-25Views:186In our study PCR-temporal temperature gelelectrophoresis (TTGE) and MeltINGENY bioinformatic program were used to analyse the mutations in the genes of melanocortin-1 receptor (MC1R) and pituitary adenylate-cyclase activating polypeptide (PACAP) in cattle. Amplification of target DNA by PCR was performed with GC-clamp primers and non-GC-clamp primers in simplex PCR reactions. The fragments were separated by denaturing polyacrylamide gelelectrophoresis (denaturing agents: high temperature, urea) after PCR reactions.MC1R homozygous individuals were used for the reaction.
We concluded that MeltINGENY program makes the decision and detection system easier, and more simple as the melting profile of target sequence is determined by the software. In case of MC1R gene, PCR-TTGE method is appropriate for SNP detection, however PACAP gene polymorphism can not be identified by the method, because PACAP mutations are not included in melting domains, therefore PCR-TTGE cannot detect them.
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Search for polymorphism in exon 5 of cattle pituitary adenylate cyclase activating polypeptide gene
17-20Views:193Pituitary adenylate cyclase-activating polypeptide is a neuropeptide expressed primarily in the hypothalamus and in other tissues, which divergent and extensive physiological functions are proved. Large number of SNPs in PACAP are published involved SNP that is associated with phenotypic trait of cattle as well. Hungarian Grey, Hungarian Simmental, Holstein, Charolais and Angus cattle breeds were involved in this study to search for polymorphism in exon 5 1–391 bp and G/A transition. Our results of PCR RFLP and PCR SSCP did not prove the occurrence neither G/A transition, nor any other SNP in cattle breeds involved.
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Fractionation of chicken egg proteome by isoelectric point in liquid phase
39-42Views:190The application of proteomics is relevant to physiology, reproduction, immunology, muscle and lactational biology in animal science, altough its use is still limited. One of the greatest challenges of proteome analysis is the reproducible fractionation of the complex protein mixtures. The fractionation methods can increase the probability of biomarker protein discovery. The fractionation by liquid-phase isoelectric focusing is one of the prefractionation methods. As a result, protein fractions can be easily collected, pooled and refractionated. There is a lack in the knowledge of gel-based proteomic methods of egg as only a limited number of protocols can be found in the literature, thus sample purification and fractionation require a time consuming optimisation procedure. The aim of this study was to fractionate egg yolk and white proteins by isoelectric point in liquid phase.
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The effect of different genotype cattle grazing on the nutrient content of saline grasslands vegetation
46-50Views:195Maintaining saline grasslands in good condition, preserving their yield and diversity is important not only for the purpose of nature conservation, but also for farming. Therefore, the primary purpose of our study is to analyse the effect of pasture use of the smaller weight extensive and the larger weight intensive beef cattle on the grassland vegetation and nutrient content. In this way, we can answer the question whether grazing for nature conservation can be achieved with more profitable, more economical and more intense varieties.
The tests were carried out in May 2016 and May 2017, in the Hortobágy National Park (Pap-ere and Zám puszta), where a total of 16 sample areas were analysed. These areas are grazed with extensive beef cattle (Hungarian grey) and mixed genotype of intensive cattle. The associations were selected along a moisture gradient, such as wet salt marsh (Bolboschoenetum maritimi) and drier saline meadow (Beckmannion eruciformis). All the vegetative material collected both years in May was analysed for the following parameters: dry matter, crude protein, crude fibre and life-sustaining net energy content. We compared the effects of medium grazing (0.46 livestock/ha) and abandonment on vegetation and nutrient content.
We examined the effect of (i) grazing, (ii) different grasslands (salt marsh, saline meadow) and (iii) grazing of different cattle breeds (Hungarian grey, intensive beef) on the nutrient content of the vegetation of grasslands Based on our results, it was found that grazing had an impact on crude protein and life-sustaining net energy content. The highest crude protein content (12.75 m/m%) was obtained in the year 2017 in the area where higher density had been grazed for two years. For the lifesustaining net energy, the highest value (5.05 MJ/kg d.m.) was also obtained in 2017 and the lowest in 2016. Furthermore, it was found that there was no significant difference between the effect of the two cattle breeds on the parameters examined. Significant effects were observed only in the case of life-sustaining net energy: in the area of intensive beef cattle we received a higher value (5.15 MJ/kg body weight) than in the area with extensive beef cattle (4.96 MJ/kg body weight).
Our results have also shown that cattle grazing is of the utmost importance for the maintenance of both wet and mesophilous habitats. Based on our three-year study, we can say that grazing by both extensive and intensive cattle breeds is suitable for the management of saline habitats.
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Influence of 17-alpha methyl testosterone on the production parameters of common carp (Cyprinus carpio L.) fry
37-43Views:257During our research we aimed at finding an answer as to what extent the different concentrations of 17-alpha methyl testosterone incorporated in the diet of common carp fries can influence the production parameters of the species, as well as how efficient their sexreversal can be with the use of this method. To this end, an aquarium experiment was conducted in the course of which four different hormone treatments were set and monitored. The fish feed was enriched with 17-alpha methyl testosterone in 50 ppm, 75 ppm, 100 ppm, 500 ppm dosages.
The obtained figures revealed that the hormone treatments had no influence on the production parameters and conservation of the common carp fries. Further on, our team is to determine the sex of the fish through the examination of gonads during autopsy when they reach the 500 g average weight.
Furthermore, a male specific test method which was supposed to be of great help in our attempt to select the sex-reversed specimens in the subsequent processes was also put to the trial. During the experiment the DNA-isolation of different sample types (muscle tissue, fin, mucus) of common carp with identified sex was successfully carried out. The extracted PCR product was examined with agarose gel. Our results indicated that the ccmf2 marker was applicable, however, the obtained figures were not reliable.
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Reference genes for livestock gene expression profiling – Literature review
81-89Views:176Quantitative real-time polymerase chain reaction (qPCR) is an essential tool for understanding animal cell’s response to developmental progression or to different experimental conditions at gene expression level. However the reliability of this method heavily lies on proper normalization (measuring a target and a reference gene’s expression from the same sample to correct for technical related variations).
Our literature review aimed to summarize the articles addressing the most important livestock species in regards of reference gene stability used as normalizers for quantitative real-time polymerase chain reaction experiments. Stably expressing reference genes were categorized into 14 distinct groups according to gene function. The number of reference genes tested and the publication numbers according to years and the ranking algorithms were also noted.
Counting showed that genes encoding ribosomal protein components are ranked as most stable in majority of cases and therefore should be taking into account for qPCR stable normalizer gene finding experiments. -
Role of hypothalamic neuropeptides in feed intake regulation of livestock species (literary review)
63-68Views:203Energy balance is the net result of the energy intake (nutrition) and expenditure (basic metabolic rate). The purpose of the daily feed intake is to provide energy and nutrients for maintenance, production and fill and maintain energy storages in form of glycogen and fat. Animals can adjust their feed intake to ensure their energy demand. Food intake regulation in animals and human is a very complex process, in which the digestive system, the central nervous system, the joining hormonal and non-hormonal factors, and the integrating hypothalamus take part. This review primarily focuses on the action mechanism of some important appetite regulating neuropeptides, and their impacts on the performance traits of the economically significant animal species.