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  • Application of semen evaluation techniques

    Laboratory methods of semen evaluation are used to select males for artificial insemination. The current review describes several techniques that have been recently used for sperm analysis. Conventional microscopic methods in combination with the objective computerassisted sperm motility and morphology analyzers and flow cytometry, allows to obtain more precise information about the membrane and functional status of spermatozoa. By using several methods to detect motility, viability, acrosomal and capacitation status besides DNA integrity sperm biology and some of the mechanism involved in sperm cry injury can be better understood. The number of possible targets related to sperm quality is increasing, and possible that some of them could enable sperm analysis for predicting freezability and fertility to be improved.

  • Comparing skull formation of the Hungarian (Hortobágy) Zackel sheep breed by geometrics morphometrics

    This work seeks to explore the morphological changes of the Hungarian (Hortobágy) Zackel sheep's skull, which occurred in the past 50–70 years. In this study, we compared individuals skull forms by geometric morphometric methods. The origin of the breed is not known, we do not know when entering the Carpathian Basin. Therefore, the comparison involved the only known early archaeological findings. We have shown that there is no difference between each period colour variations, but over time change has occurred in the skull formation of the breed.

  • Seasonal variations in somereproductive parameters of Dorper Rams in Hungary

    The purpose of the present study was to test the hypothesis that season has an affect on semen quality and scrotal circumference of Dorper rams. The experiment was carried out with six Dorper rams aged between 15 and 18 month. Semen samples were collected with artificial vagina and volume, concentration (x 109/ml), total sperm number/ejaculate (x 109), mass motility (0–5), progressive motility (%), scrotal circumference (cm) was observed. Significant differences (P<0.05) were observed in concentration, total sperm number/ejaculate, scrotal circumference in different seasons. Volume was the highest in autumn (1.4±0.5 ml) and the lowest in the spring (1.3±0.4 ml). Concentration of semen was lower in spring (2.6±1.5 x 109) and summer (3.3±1.5 x 109) as compared to fall (4.1±1.1 x 109) (P<0.05). Regarding total sperm number/ejaculate (x 109), scrotal circumference (cm) all the seasons differed significantly (P<0.05), although the season had no effect on mass motility and progressive individual motility (P<0.05). In conclusion the present study showed that semen quality parameters and scrotal circumference of Dorper rams were better in autumn than in the other sea- sons.

  • The results of the investigation of the β-casein gene polymorphism in sheep breed – Review

    β-casein is the most abundant protein fraction in sheep milk, and has at least six different alleles (A, B, C, G, X, Y). The alleles of the β-casein gene may influence on the quality and quantity of milk. Knowing the gene polymorphism has an important role in the process of milk production. The properties of milk could be positively influenced by themolecular genetic methods.

  • Fractionation of chicken egg proteome by isoelectric point in liquid phase

    The 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.

  • Investigation of the β-casein (CSN2) gene polymorphism in goat breed – Review

    Milk plays an important role in the human nutrition as an important protein and energy source. Β-casein is the most abundant casein fraction in the goat milk. The β-casein gene has at least 9 polymorphism in goat breed (A, A1, B, C, C1, D, E, 0 and 0’). The most important alleles are the zero (0) and zero’ (0’) alleles, because they decrease the quantity of β-casein protein in milk. The lower quantity of the β-casein changes the properties of the coagulation and the allergenin milk. This review is about the β-casein gene polymorphism in goat breed.

  • Coincidences between molecular genetic and studbook data of gidrán mare families on the basis of mtDnA

    The traditional Hungarian horse breed, Gidran has been close to the edge of extinction several times. Despite the multiple bottleneck effect, the breed has retained a part of its genetic variability, and performed prominently in carriage driving and show-jumping competitions. Maintaining of the Gidran breed is important in the point of view of world heritage; because besides Hungary, smaller Gidran populations exist only in Bulgaria and Romania. Taking advantage of the special inheritance features of mtDNA, our study focused on two mtDNA regions of Gidran mares. Altogether, 251 hair samples from various Hungarian studs were examined. The analysis was successfully made in case of 251 samples of the cytochrome b and in case of 246 samples of D-loop regions. Because of the distinct mutation rates of the two mtDNA markers, the number of the haplotypes and the way of grouping samples into haplotypes was different. Our key finding was that most haplotypes may be compatible with mare families of the stud book; however incidental mistakes in stud book have occurred only in a few cases. Our results indicate the importance of the preservation and breeding those mare families, which are molecular genetically more diverse than the others, and are in the edge of extinction.

  • Detection of DNA mutations by PCR-TTGE method

    In 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.

  • Ragweed components in honey

    The aim of this research was to prove that the syrup containing ragweed used for feeding the bees was transferred to the honey. To reach this goal we developed a method to analyse the volatile components of the samples. We applied two sample preparation methods. The first one was SPME (Solid Phase Micro Extraction) preparation from the headspace of all of the samples. In the second case only the raw leaves of the ragweed were extracted with n-hexane and dichloromethane to complete the SPME results. The measurements were performed on GC-MS equipment. Chromatograms and data evolution showed that the components of the ragweed were also present in honey so they were transferred by bees.

  • Role of hypothalamic neuropeptides in feed intake regulation of livestock species (literary review)

    Energy 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.

  • Genetic diversity of the Hungarian draft horse assessed by mitochondrial DNA

    Hungarian draft is a horse breed with a recent mixed ancestry. It was developed in the 1920s by crossing local mares with draught horses imported from France and Belgium. To genetically characterize the breed and to set up the basis for a conservation programme, we have employed a molecular marker: a 256-bp D-loop mitochondrial DNA fragment. We analyzed 124 horses representing Hungarian draft horses to assess the maternal phylogeography of the breed. Sequence analysis of a 256-bp segment revealed a total of 34 haplotypes with thirty-four polymorphic sites. High haplotype and nucleotide diversity values (Hd=0.953±0.001; π=0.024±0.001) were detected. The average number of pairwise differences were k=5.998. This breed counts 800 mares today, and only survive due to breeding programmes, this way each haplotype frequency depends on the extent to which mares are involved into the breeding. The reduced number of surviving maternal lineages emphasizes the importance of establishing a conservation plan for this endangered breed. Due to the revealed 34 polymorphic sites we could presuppose twelve maternal linages, which could be a first step for making a breeding programme.

  • Genetic diversity study in Hungarian coldblooded horses

    Because of the feeding technology innovation, accelerated transport and communication facilities breeds of high performance breeds replaced local autochone breeds worldwide. These latter species however have an important role in gene conservation. Hungarian cold-blooded horse breeding stock are lacking pedigree, the actual founder breed mares are not known. For this reason, it is an major priority defining the genetic backround of the existing flock, for that breeding could operate with purposeful using of origin maternal founders. In the present study 195 cold-blooded Hungarian mares tail and mane sample were analized. Our analysis was carried out between 15531–15752 base pairs in mithocrondial DNA D-loop region, which reported a total of 222 base pairs. Fourtyone polymorphic sites were determined, which resulted in 39 haplotypes (h=39). The average pairwise differences were k=6.825. High haplotype and nucleotide diversity values were observed (Hd=0.968±0.003, π=0.026±0.003). Based on the previously defined variable positions of haplotypes defined by Jansen et al (2002), we groupped our haplotypes into haplogroups. 23 percent of the studied population (45 mares) belonged to haplogroup F1. Nearly 97% of the analyzed population was classified into one of eight  haplogroups defined by al. (2002). This study gives genetic information nearly 25% of the Hungarian population. Another possibility would be patterning more mares or involving more genetic marker in the study which will assuming the possibility of a more comprehensive analysis.

  • Differences in content of beet root and beet root juice, and it’s suitable to develop functional foods from animal sources

    In Western medicine is beginning to increasingly focus on the key role of healthy nutrition and lifestyle in preventing certain diseases. Therefore, it is important to emphasize the importance of primary prevention in our country, and to promote it; not only the health care team work, but broaden the recommended foods, which have benefit for human health as well.

    Polyphenols are antioxidants, which are easily consumable with diet, however, food processing can influence the level of them.

    In our study, nutritional assessment was made of raw beet root and a juice, made from this beet root. We investigated in which special diet referenced to eat this foods and whether this is suitable to develop functional foods from animal sources as a row material.

  • HPLC-MS analysis of the active ingredients of chamomile (Matricaria chamomilla) bee feeding syrup

    Apiarists let prepare by bee honey products containing medicinal drugs. Our aim was to prove that the active ingredients originated from the herbs are also present in the bee products. This publication focuses only to the chamomile (Matricaria chamomilla) containing feeding syrup and the difference between the various syrup phases. To fulfil this task, we developed a method analysing the non-volatile components of the syrup using the flowers of chamomile. The method involves a filtration followed by HPLC-MS analysis. The analyses and the data evaluations proved that the non-volatile components of the herb were transferred to syrup. As the characteristic components of certain herbs could be identified also in the syrup, it is obvious that the components responsible for the medicinal effects are also transferred.

  • Examination of microsatellite markers of Dorper sheep breed

    Number of not woolly and molty sheep exceeds 60 million throughout the world. Their numbers and their importance is growing, still they have appeared in the past two decades all over in North-America, Australia, New-Zealand and also in Europe. The South African Dorper has been a pioneer among them in Hungary. It was introduced in 2006 in the country. The Dorper sheep is the second largest breed in South Africa, which was developed from the crossing of Dorset Horn and the Blackhead Persian. The aim of the EU Member States in terms of this specific breed is increasing the small populations, improving the productive qualities, in addition to this avoiding inbreeding. However, finding appropriate breeding stock is difficult due to the small size of available populations and also to the suspected common of origin. With the help of various molecular genetic methods we could get a total view of the genetic background of these flocks. Nowadays the most commonly known and used genetic markers are microsatellites, because their applications give fast, accurate and easily reproducible results. There is no specific descriptive information on the genetic background of Dorper populations in the various EU countries , also regarding diversity between populations. Therefore in our work we want to optimize the conditions of applicability of 31 selected microsatellite reactions as a first step of mapping the entire genetic background of the different EU Dorper populations.

  • Identification of Hucul mare families by mtDNA markers

    Hundred animal species have disappeared during the last century. By this time, approximately one-third of domestic animals have been in the endangered category. Hucul horses are also in this category; furthermore saving the genetic diversity beside the race preservation is an important challenge as well. The number of mares and stallions is only one of the expressive elements of genetic diversity; together with their quality determine the genetic variability of this breed. Beyond that, if an exact breed can originates from more founders, it can be more renewed genetically. Stud book documents these data by registering the mare families and stallions’ genealogical lineage. Molecular genetics, especially mitochondrial DNA analysis can make the precise identification of mare families possible. As a result of these molecular based methods, protection of genetic diversity, as well as breed preservation became more reliable. After the primer designing, the optimal primer pair was chosen which targets a 1092 bp length DNA sequence in the cytochrome b region. After the successful PCR optimalisation, we determined 170 Hucul mares’ sequences. According to our results, the samples compose ten haplotypes, which are much less, than the registered number of mare families in the stud book. Further investigations are needed to reach more representative results, and drawn the further consequences.

  • Search for polymorphism in exon 5 of cattle pituitary adenylate cyclase activating polypeptide gene

    Pituitary 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.

  • Morpometric study of Hungarian honey bee (Apis mellifera L.) colonies

    The honey bees are essential for the pollination of agricultural plants. The Pannonian honey bee, Apis mellifera pannonica, is native to Hungary, only these subspecies are being bread in our country. The parameters have been separated the pannon and italian honey bee subspecies, the colour of tergit, the cubital index and proboscis length. The morphometric analisys is of special importance because this, on the one hand, shows correlation with honey bee production and on the other hand, the pure morphometric charactersitics is the basis of any potential honey bee export. Mitochondrial DNA and microsatellites are the common methods to define genetic diversity and the separation of subspecies.

  • Reference genes for livestock gene expression profiling – Literature review

    Quantitative 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.

  • Study the correlation of morphological and production traits of honey bee (Apis mellifera L.) (Review

    There are several morphological differences among the honey bee subspecies, which proofs that they adapted different way to the environmental factors of the given area. Morphological observations are not the only way to separate subspecies, it is possible on their genetic speciality as well. The authors’s aim in this present study was to examine morphological characteristics of subspecies, furthermore looking for correlation between morphological parameters and honey yield. According to their results, among the most important parameters considered by Hungarian breeders, in the case of the native Apismelliferacarnica subspecies, the proboscis length could be the indirect tool for honey-productivity focused selection.

  • Strategic Steps in the Sheep and Goat Branches

    The balance of the sheep and goat branches can be summarised as follows:
    If the increase in stock size, specific yields, genetic improvement and modernisation are not realised, several thousand shepherds will lose their jobs and possibilities to work, and their families will lose their only source of livelihood.
    Several hundred hectares of grassland will become weedy and the costs of crop protection will increase in these regions. Pollen pollution, the number of allergic ailments and medical costs will increase, mentioning only the main disadvantages. Its financial requirement is nearly equal to the one required for development without any essential compensation or production.
    In order to develop a competitive branch, a subsidisation of 10-20 billion HUF is needed until the accession of Hungary to the EU, depending on the scale and progressivity of development. The subsidisation and realisation of the strategic steps would result in:
    – the establishment of a registered livestock with 1.5 million ewes, which would ensure better market position for the branch in the EU (5%),
    – the establishment of a competitive and marketable branch with a production value of 15-25 billion HUF, calculated using the present price level (after accession, the branch would be self-operating and profitable, with the 6-8 billion HUF/year normative subsidisation by the EU),
    – the establishment of a genetic potential which would ensure our participation in profitable international breeding animal market, which provides extra profit,
    – the establishment of a sheep husbandry with better genetic potential and phenotypic performance, producing a quality of international level,
    – the maintenance of the workplace of more than 10.000 people (saving 3-5 billion HUF/year of the budget),
    – the maintenance of the condition of growing areas of grasslands with their advantages (crop protection and public health) whose financial effect is considerable.

  • Investigation of the heat shock gene Hsp70 polymorphism in different sheep genotypes

    Nowadays the climate change has an increasing effect on the animals. The warming climate brings up several problems on the area of the animal husbandry, which ones are really important. From the first time the living beings have defensive mechanisms against the heat shock. In current examination we use– from Hungary and from other countries collected –samples of sheep breeds, which are living on different climate. Our fundamental assumption was, that the animals living on other climate adapted to the changes of the environment and there are differeces in their genetic background. These fixed mutations we are looking for in the HSP70 heat shock gene, but we haven’t found any polymorphism yet. We are going to involve further breeds and more individuals in the investigations.

  • Age of puberty and conception rates of Hungarian Merino, (Merino x Somali) F1 and (Merino x Barbados Blackbelly) F1 ewes

    The aim of our examination was to detect the puberty of the Hungarian Merino and its hairsheep crossbred ewes, and also to determine their average conception rates after having ram exposed in May and June. Hormon diagnostation was used to evaluate the exact date of the very first ovulation. Ultrasound technique was used for checking the conception rates of ewes. The pregnancy detection was applied after a month of the date of taking off the ram.
    We evaluated the ages of ewes at puberty. Statistical deviation and standard error were calculated. The results proved the Hungarian Merino x Somali (N=10), ((x ± s) = 173 ± 43) and also the Hungarian Merino x Barbados Blackbelly crossbred genotypes (N=7), ((x ± s) = 186 ± 19) have their puberty much earlier as compared to the fullbred Hungarian Merino ewes (N=10), ((x ± s) = 231 ± 95). We experienced the highest conception rate in the group of the (Hungarian Merino x Somali) F1 ewes after exposing the ram in the „out-of season” period of time.

  • Effect of the storage temperature onto the quality of the semen of rams of different breeds

    Semen of 10 Tsigai,3 Prolific Merino and 7 Barbados Blackbelly rams was taken in January, out of the season. Split samples of the diluted semen samples were kept at 23, v.s. 8oC and their motilitywas evaluated daily for 3 days by subjective microscopic investigation. The ratio of motile spermatozoa was strongly decreased during the first day, later the motility rate was sinking more slowly. It could be stated by multivariance analysis that the motility value of diluted semen of Prolific Merino rams was higher and showed a slower reduction as compared to the other two breeds. The decrease of the motility rates was slower at 8oC than at 23oC.

  • Tenability of ram semen

    It could be stated that the diluted semen of Awassi rams taken in the breeding season preserved its fertilizing abilityat different temperatures for different periods of time. The motility of spermatozoa kept at 23 vs. 8oC was checked daily. The largest spread of data was observed 24 hours after taking the semen, then the motility rate of cells showed a linear decrease. Motility results of fresh and frozen-thawed samples were compared also after heat resistance test and significant differences were found between these groups. Significant individual differences were observedin the sperm motility after heat resistance test.