Black Pied Research Articles

Методы ГЖХ, ДСК и ИК-спектроскопии в изучении коровьего молочного жира

Для целей надежной идентификации натурального коровьего молочного жира получены газохроматографические данные с применением ионизационно-пламенного детектора (ГЖХ-ПИД) о жирнокислотном составе молока 4 пород – красно-пестрой, черно-пестрой, голштинской и джерсейской в переходный период от пастбищного к стойловому содержанию, по методике, рекомендованной ГОСТ 32261-2013; получены также газохроматографические данные (ГЖХ-ПИД) о триглицеридном составе по ГОСТ Р 70238-2022. В комплексе с хроматографическими методиками изучены термограммы молочного жира методом дифференциальной сканирующей калориметрии (ДСК), а также проанализированы ИК-спектры образцов молочного жира. При статистическом анализе данных о суммарном влиянии генетических и фенотипических факторов на химический состав жиров и масел, на теплофизические параметры (температуры и тепловые эффекты плавления, полиморфных превращений) выявлены тесные природные корреляции между некоторыми отдельными жирными кислотами, триглицеридами и теплофизическими параметрами ДСК, характерные для конкретных липидов. Нарушение выявленных корреляций может указывать на внесение в продукт жиров-заменителей, фракционирование, переэтерификацию или иное физико-химического воздействие на образец масложирового сырья. Применение комплекса методик ГЖХ-ПИД, ДСК и ИК-спектроскопии позволяет не только надежно идентифицировать натуральное сырье, но и зачастую его происхождение, т.е. породу скота и условия, в которых произведена сельхозпродукция.

Comparative assessment of beef characteristics from young bulls of different breeds and the influence of storage conditions on meat quality indicators

Comparative assessment results for quality indicators of meat samples obtained from Black Pied, Simmental and Aberdeen Angus young bulls and the influence of different temperature conditions on the quality of vacuum-packed beef during refrigerated storage are presented. The general chemical composition, physicochemical properties, and biological value of the samples based on the content of nonessential and essential amino acids were determined, as well as the protein quality index (PQI) and amino acid score. Analysis of the general chemical composition revealed higher moisture and protein content and the lowest fat content in Black Pied beef compared to other breeds. The amino acid composition of the protein showed a higher content of essential amino acids and the highest PQI value in Simmental beef. According to the calculation results, higher amino acid scores for lysine (149.1% and 129.1%) and tryptophane (200.0% and 240.0%) were noted in meat from Simmental and Aberdeen Angus young bulls, respectively. For the process of storing vacuum-packed meat in a cooled (at a temperature of 2.0 ± 0.5 °C) and superchilled state (minus 2.0 ± 0.5 °C) at a relative air humidity of 85%, a comparative analysis of changes in free amino acids and dynamics of hydrolytic and oxidative spoil- age of meat samples from various breeds was conducted. After 21 days of storage in a superchilled state, the content of free amino acids in Black Pied, Simmental and Aberdeen Angus beef was lower by 13.1% (P > 0.05), 24.1% (P ≤ 0.05) and 17.0% (P ≤ 0.01) compared to storage in a cooled state, respectively. For all studied samples stored in a cooled state, the acid number values were 40% to 41% (P ≤ 0.01) higher than in a superchilled state and peroxide number values were 20% to 23% (P ≤ 0.05) higher than in a superchilled state. It has been established that lowering the storage temperature of vacuum-packed beef helps to better preserve quality and ensure safety for meat from all breeds studied.

Effect of an enzymatic-probiotic preparation on the microfauna of the rumen of the Russian Black Pied and Ayrshire cows.

Effect of an enzymatic-probiotic preparation on the microfauna of the rumen of the Russian Black Pied and Ayrshire cows.

Composition and functions of rumen and endometrial microorganisms associated with endometritis in dairy cows

Despite the attention given in the recent years to the microbiological causes of endometritis and the potential sources of uterine microbiota infestation, more questions than answers remain in this research field. This paper describes an experiment carried out at the livestock farm of the Leningrad region on 6 dairy cows of the Holsteinized Russian Black Pied breed of the second lactation in the period after calving. The animals were divided into two groups (n = 3): Group 1 included clinically healthy animals, Group 2 included animals diagnosed with purulent-catarrhal post-calving endometritis. Metagenomic sequencing was performed using the MiSeq genomic sequencer (Illumina, Inc., USA) with the MiSeq Reagent Kit v3 (Illumina, Inc., USA). Based on the next-generation sequencing of microbiota of endometrial scrapings, 7 phyla of microorganisms were detected in clinically healthy cows and only 4 phyla of microorganisms in cows diagnosed with endometritis. The increase in the proportion of Fusobacteriota taxon bacteria permanently present in endometrial scrapings and the decrease in Bacteroidota phylum bacteria in the group of animals with endometritis could be related to the occurrence of this disease. Bacteria Alloprevotella, Campylobacter, Caviibacter, Falsiporphyromonas, Veillonella present only in the endometrial tissue of sick cows may be the etiological origin of endometritis. In the rumen of animals with endometritis, there was an increase in Bacteroidota phylum microorganisms (p≤0.05) against a decrease in Firmicutes phylum bacteria (p≤0.05) compared to the clinically healthy group. Using the PICRUSt2 software package (v.2.3.0), it was shown that the microbiome of cows diagnosed with endometritis showed inhibition (p≤0.05) of the potential of 9 metabolic pathways compared to healthy animals. This could have negative consequences for the body of animals and be a consequence of metabolic disorders.

Comparative evaluation of russian black pied and holstein cattle breeds of foreign breeding

A related Holstein breed has recently been used to improve domestic dairy cattle and create highly productive herds by purchasing semen from breeding bulls, as well as a large number of heifers and calves of foreign breeding. The purpose of the work is a comparative assessment of the productive qualities of Black Pied and Holstein cows of foreign breeding. Purebred Holstein cows differ from animals of Black Pied breed in milk yield per lactation by 1064 kg or 15.2% (P≤0.01, in favor of Holstein cows). In terms of quality indicators of milk, that are the content of fat and protein in milk, the superiority remained with Black Pied cows. The difference was significant at P≤0.05 for fat content in milk and at P≤0.001 for protein content. Both groups of cows exceeded the breed standard for milk yield per lactation, mass fraction of fat and mass fraction of protein in milk. The milk of the Russian Black Pied cattle contains more essential amino acids. The milk of cows of both breeds can be considered complete, since the amount of limited amino acids in the milk of purebred Holstein cows of foreign breeding is over 95% of their content in the ideal protein.

The interrelation of the Russian Black Pied breed productive qualities of the Ural type depending on live weight

Holsteinized Russian Black Pied cattle of the Ural type are bred in the Sverdlovsk region, which is represented by large, highly productive animals with a high genetic potential for productivity. The purpose of the work was to study the effect of first heifers’ live weight on their productivity and the interrelation between milk yield and milk quality indicators. It was found that there was a positive tendency for an increase in milk yield with an increase in the cows’ live weight from less than 550 kg to 624 kg. The most significant increase in milk yield in cows was in the range from 574 to 599 kg. The difference in milk yield between these groups was 692 kg or 8.1% in favor of animals with live weight of 575-599 kg. An increase in cows’ live weight over 625 kg leads to a decrease in milk yield, which amounted to 2.3%. Changes in correlation are observed from a negative average value in cows with a live weight of less than 550 kg to a positive average value in cows with a live weight of more than 625 kg. In first heifers with live weight from 575 to 599 kg, a very low positive interrelation between live weight and milk yield per lactation was recorded (0.04).

Correlation between mitochondrial dysfunction of spermatozoa and their biological adequacy

One of the most significant indicators affecting male fertility is the sperm nuclear and mitochondrial DNA fragmentation index (DFI). DNA damage depends on biotic and abiotic factors, leading to oxidative stress (O.S.). This research aimed to investigate the relationship between mitochondrial dysfunction of spermatozoa and their biological adequacy. The research material was frozen-thawed sperm samples from the Ayrshire, Russian Black Pied Holstein, Russian Red Pied Holstein, Limousin, and Polled Russian breeding bulls. Assessments of mobility, morphology, and fragmentation index were performed using computer-assisted sperm analysis (CASA). It was found that there is a negative correlation between sperm activity and mitochondrial dysfunction with the correlation coefficient r = -0.24. The incidence of abnormal spermatozoa correlated with sperm dysfunction r = 0.77. The nDNA fragmentation index in chromatin varied from 0 to 25%.

Holsteinization results of Russian Black Pied cattle

Holsteinized Russian Black Pied cattle of the Ural type are bred in the Sverdlovsk region, which is represented by large, highly productive animals with a high genetic potential for productivity. The aim of the study was study holsteinization results of Russian Black Pied cattle in one of the pedigree reproducers of the Sverdlovsk region. Over the past 20 years, there have been significant changes in the rearing heifers’ raising intensity; their live weight at the first insemination increased and their age decreased. In terms of live weight at the first insemination, there was an increase from 2006 to 2013 to over 400 kg; then, when switching to early insemination terms, there was a decrease in live weight to 390 kg. The age of first insemination has decreased since 2000 from 20.2 months to 14.4 months in 2018. An inversely proportional dependence of the milk yield of first heifers on the age of the rearing heifers’ first insemination has been established. An increase in milk yield occurs with a decrease in the age of the first insemination and vice versa, an increase in the age of heifers’ first insemination is observed with a decrease in milk yield. Moreover, this also happened when raising both hybrid heifers and heifers of the new Ural type of Holsteinized Russian Black Pied cattle.

Single nucleotide polymorphism rs110861313 in the intergenic region of chromosome 23 is associated with the development of leukosis in the Russian Black Pied cattle

In recent years, using a genome-wide association study (GWAS), a number of single nucleotide polymorphisms (SNPs) have been suggested to be associated with susceptibility to leukemia in cattle. However, all studies have been done with purebred Holstein cows and their hybrids. In this regard, it is important to confirm the functional role of polymorphisms previously identified in a GWAS study in Russian cattle breeds. The aim of this study was to verify the association between rs110861313 in the intergenic region of bovine chromosome 23 and leukemia in the Russian Black Pied cattle. Based on the levels of bovine leukemia virus (BLV)-specific antibodies detected in serum using serodiagnostic techniques, animals were divided into three groups: healthy animals (n = 115), asymptomatic virus carriers (n = 145) and animals with leukemia (n = 107). Genotyping of rs110861313 was carried out using polymerase chain reaction followed by analysis of restriction fragment length polymorphisms. A significant decrease in the frequency of the A/A genotype (11.2 %) was revealed in animals with persistent lymphocytosis compared to virus carriers (27.6 %) (p < 0.002). At the same time, the frequency of animals with the C/C genotype in animals with persistent lymphocytosis (41.1 %) was significantly higher than that of virus carriers (21.4 %) (p < 0.001). In this case, asymptomatic virus carriers can be considered a more suitable control than healthy animals that have not been in contact with the virus. According to bioinformatics analysis, resistance to BLV can be due to the presence of the transcription factor FOXM1 binding site in the region of rs110861313. FOXM1 is expressed in immune cells and can potentially affect the expression of the neighboring genes (LY6G5B, GPANK1, ABHD16A, LY6G6F, LY6G6E, CSNK2B, ApoM). Thus, we found that SNP rs110861313 in the intergenic region of bovine chromosome 23 is associated with the development of leukemia following BLV infection in the Russian Black Pied cattle.

Association of haplotypes for SNPs in the LTR regions of bovine leukemia virus with hematological indices of cattle

Molecular typing of BLV samples isolated from Holsteinized Russian Black Pied cattle was carried out, and various cytofluorometric and morphological blood indices were examined. We performed the total count of white blood cells (WBC), lymphocyte (lymf), granulocyte (gran), monocyte (mon), red blood cell (RBC), hemoglobin (HGB), hematocrit (HTC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red blood cell distribution width (RDW), platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW), and platelet crit count (PCT). The LTR-region of BLV was haplotyped. Only viruses of haplotypes I (0.33±0.03) and III (0.67±0.03) of the eight possible were detected. The ratio of hematologically sick, healthy, and suspected carriers of BLV of haplotypes I and II was comparable with the results of other researchers. The numbers of leukocytes, erythrocytes and platelets in the blood of carriers of haplotype III exceeded the corresponding parameters of cattle affected by the virus of haplotype I. It is interesting to note that the difference in the hemolytic status of animals was manifested not only by the concentration of leukocytes as direct immune agents but also by the count of erythrocytes and platelets, which are not directly involved in the immune response. The number of particles of haplotype III of the BLV circulating in the blood of infected individuals exceeded that of the carriers of haplotype I. In this connection, an assumption was made about the evolutionary advantage of the more virulent haplotype III. However, the results of our own research in conjunction with the data of other scientists indicate that the high virulence of individual virus strains is a consequence of the tendency to implement the maximum possible intensity of the synthesis of virus particles but not of the high damaging effect alone. It is shown that high lethality is evolutionarily disadvantageous for viruses, since the extinction of the carrier as a biological species is fraught with the disappearance of the virus itself.

Genome-wide associations and functional gene analyses for endoparasite resistance in an endangered population of native German Black Pied cattle

BackgroundGastrointestinal nematodes (GIN), liver flukes (Fasciola hepatica) and bovine lungworms (Dictyocaulus viviparus) are the most important parasitic agents in pastured dairy cattle. Endoparasite infections are associated with reduced milk production and detrimental impacts on female fertility, contributing to economic losses in affected farms. In quantitative-genetic studies, the heritabilities for GIN and F. hepatica were moderate, encouraging studies on genomic scales. Genome-wide association studies (GWAS) based on dense single nucleotide polymorphism (SNP) marker panels allow exploration of the underlying genomic architecture of complex disease traits. The current GWAS combined the identification of potential candidate genes with pathway analyses to obtain deeper insights into bovine immune response and the mechanisms of resistance against endoparasite infections.ResultsA 2-step approach was applied to infer genome-wide associations in an endangered dual-purpose cattle subpopulation [Deutsches Schwarzbuntes Niederungsrind (DSN)] with a limited number of phenotypic records. First, endoparasite traits from a population of 1166 Black and White dairy cows [including Holstein Friesian (HF) and DSN] naturally infected with GIN, F. hepatica and D. viviparus were precorrected for fixed effects using linear mixed models. Afterwards, the precorrected phenotypes were the dependent traits (rFEC-GIN, rFEC-FH, and rFLC-DV) in GWAS based on 423,654 SNPs from 148 DSN cows. We identified 44 SNPs above the genome-wide significance threshold (pBonf = 4.47 × 10− 7), and 145 associations surpassed the chromosome-wide significance threshold (range: 7.47 × 10− 6 on BTA 1 to 2.18 × 10− 5 on BTA 28). The associated SNPs identified were annotated to 23 candidate genes. The DAVID analysis inferred four pathways as being related to immune response mechanisms or involved in host-parasite interactions. SNP effect correlations considering specific chromosome segments indicate that breeding for resistance to GIN or F. hepatica as measured by fecal egg counts is genetically associated with a higher risk for udder infections.ConclusionsWe detected a large number of loci with small to moderate effects for endoparasite resistance. The potential candidate genes regulating resistance identified were pathogen-specific. Genetic antagonistic associations between disease resistance and productivity were specific for specific chromosome segments. The 2-step approach was a valid methodological approach to infer genetic mechanisms in an endangered breed with a limited number of phenotypic records.

EPIZOOTOLOGICAL AND PHYLOGENETIC EVALUATION OF THE BOVINE LEUKEMIA VIRUS IN RUSSIAN FEDERATION

The genetic characterization of BLV is an important task in scientific research in many countries of the world. According to the sequenced gene region - env BLV isolates allocated in different geographical locations of the world, up to 10 different genetic groups of the virus were identified and classified. Acknowledgments: The research was carried out at the expense of the Russian Science Foundation grant (project No. 17-76-10051). As part of the research, we monitored the epizootic situation of the BLV in Russia. Groups of animals (n=50) Holstein-Frisian (imported breed) and Russian Black Pied (local breed) were selected, belonging to agricultural organizations of the Tyumen region. Serological screening methods (ELISA, AGID) were used in cattle to identify infected animals. Immunological evaluation of animals in all test groups is given. A primary A nested-PCR study was performed, which resulted in a fragment of the env 444 bp gene in the studied samples. RFLP analysis of this fragment allowed to establish that in 94 % of the samples there was a «Belgian type» of the leukemia virus, in 4 % of samples - «Australian» and in 2 % - a «mixed type». All samples were sent for NGS sequencing. By phylogenetic evaluation of the BLV genome env site in the isolated samples and the immunological evaluation of the infected animals, new data will be obtained that will allow updating information on the genetic groups of the BLV in the territory of the Russian Federation.

Development of a Bioinformatics Framework for the Detection of Gene Conversion and the Analysis of Combinatorial Diversity in Immunoglobulin Heavy Chains in Four Cattle Breeds.

We have developed a new bioinformatics framework for the analysis of rearranged bovine heavy chain immunoglobulin (Ig) variable regions by combining and refining widely used alignment algorithms. This bioinformatics framework allowed us to investigate alignments of heavy chain framework regions (FRHs) and the separate alignments of FRHs and heavy chain complementarity determining regions (CDRHs) to determine their germline origin in the four cattle breeds Aubrac, German Black Pied, German Simmental, and Holstein Friesian. Now it is also possible to specifically analyze Ig heavy chains possessing exceptionally long CDR3Hs. In order to gain more insight into breed specific differences in Ig combinatorial diversity, somatic hypermutations and putative gene conversions of IgG, we compared the dominantly transcribed variable (IGHV), diversity (IGHD), and joining (IGHJ) segments and their recombination in the four cattle breeds. The analysis revealed the use of 15 different IGHV segments, 21 IGHD segments, and two IGHJ segments with significant different transcription levels within the breeds. Furthermore, there are preferred rearrangements within the three groups of CDR3H lengths. In the sequences of group 2 (CDR3H lengths (L) of 11–47 amino acid residues (aa)) a higher number of recombination was observed than in sequences of group 1 (L≤10 aa) and 3 (L≥48 aa). The combinatorial diversity of germline IGHV, IGHD, and IGHJ-segments revealed 162 rearrangements that were significantly different. The few preferably rearranged gene segments within group 3 CDR3H regions may indicate specialized antibodies because this length is unique in cattle. The most important finding of this study, which was enabled by using the bioinformatics framework, is the discovery of strong evidence for gene conversion as a rare event using pseudogenes fulfilling all definitions for this particular diversification mechanism.

Design of a DNA panel for genomic studies in Russian cattle breeds

A panel of 96 DNA samples (Russian Cattle Genomic Diversity Panel 1.0 or RCGDP 1.0) characterizing the breadth of genetic diversity in popular Russian cattle breeds was designed. The panel contains from four to eight animals from each of 11 dairy and six dairy-meat and meat breeds. The main criterion for inclusion of individual animals in the panel was that they had, according to the pedigree data, the greatest number of unrelated haploid genomes. The resulting panel of 96 animals was estimated to contain 186.1 unshared haploid genomes and was expected to allow a 95% probability of detecting alleles with a frequency greater than 0.016 in the group. This is comparable to the USDA MARC Beef Cattle Diversity Panel version 2.1. Analysis of three SNPs in the Galloway, Hereford, Grey Ukrainian and Black Pied breeds showed no significant differences in the allele frequencies between the animals in the panel and in the herds. Thus, the designed DNA panel may be useful for SNP selection in the genetic analysis of economically important traits, for identification of purebred and hybrid animals, and, probably, for tentative estimation of allele frequencies in the populations.

Biological value of meat of bulls with different genotypes

The quality of meat obtained from young bulls of Azerbaijan zebu, Cuban zebu × Azerbaijan zebu crosses, and hybrids of Azerbaijan zebu with Latvian Brown, Black Pied, and Caucasian Brown breeds is investigated. The meat of bulls of all groups is characterized by high biological value; however, the highest protein quality index is noted in crosses and Azerbaijan zebu × Latvian Brown hybrids. The cutting resistance of meat is the lowest and color intensity and water binding capacity the highest in crosses of Cuban zebu with the Azerbaijan zebu; however, these indices were within the norm in all animals. During tasting, the meat and its products received a high evaluation.

Polymorphism of intron 2 of the SDF1 gene in Galloway, Hereford, and Russian Black Pied cattle

The region of intron 2 of the SDF1 gene encoding a chemokine of the CXC subfamily has been resequenced in Galloway, Hereford, and Black Pied cattle. Five of the single-nucleotide polymorphisms (SNPs) that were earlier detected by other authors in various breeds of cattle in North America (99C/G, 128T/C, 206C/T, 267C/G and 313C/T) have been found. The 270insC polymorphic marker has proved to be monomorphic in Russian cattle breeds. Hereford cattle significantly differ from Galloway and Black Pied cattle in the frequencies of some SNP variants and their combinations. The number of SNP combinations in Hereford and Galloway cattle exceeds that in Black Pied cattle.

Quality of hides obtained from cattle-zebu hybrids

Three-breed hybrids Cuban zebu x (Azerbaijan zebu x Aberdeen Angus) surpassed the Azerbaijan zebu and two-breed Azerbaijan zebu with Brown Latvian, Black Pied, and Aberdeen Angus breeds in weight, length, width, area, and thickness of hides. Minimum indices were obtained or purebred animals.

Qualitative characteristics and processing properties of milk from various cow breeds

Kappa-casein gene (CSN3) polymorphism has been established in cows of the Belarus Black Pied breed and Belarus Red breed group. The frequency of genotypes and alleles across gene CSN3 has been calculated. More cheese and curd can be made from milk of cows of the Belarus Red breed group with genotype CSN3 BB than from milk of animals with genotype CSN3 AB and CSN3 AA.

Ventilation and Haemodynamic Indicators in Spontaneously Breathing Pigs under General Anaesthesia

The aim of this study was to obtain ventilation and haemodynamic data of healthy piglets under general anaesthesia for future patho-physiological experimental studies. A total of 34 domestic piglets of the Czech Black Pied (Přeštice) breed were used in the study. The animals (male to female ratio 8 : 9) were six weeks old and their average body mass was 22 kg. A general anaesthetic (fentanyl and azaperon) was introduced via a pulmonary artery catheter and the spontaneously breathing animals were monitored for 60 min. Cardiac output and haemodynamic indicators were established using intermittent pulmonary artery thermodilution. Blood gas data were deduced using fan dynamic parameters of ventilation and ventilation indices. The study yielded reliable data of dynamic lung indicators (p < 0.05); e.g. the tidal volume 6.00 ± 0.82 ml/kg, hypoxemic index 350.47 ± 42.50 mmHg, lung compliance 0.63 ± 0.12 ml/cmH2O/kg, and haemodynamic indicators (p < 0.01) such as cardiac output 2.12 ± 0.75 l/min, pulmonary vascular resistance 3.92 ± 0.52 and systemic vascular resistance 15.8 ± 6.81 Woods units. Reliable data regarding lung dynamics, cardiac output, preload and afterload of both heart ventricles in spontaneously breathing healthy piglets under general anaesthesia were achieved.

Creation of a new population of black pied dairy cattle of the Vyatskii type

A complex reproductive crossing has been used in the Volga-Vyatsk region to breed a new competitive dairy cattle type Vyatskii which combines the gene pool of four breeds: Istoben, Dutch, Estonia Black Pied, and Holstein. Animals of the new type are distinguished by good acclimatization abilities and have the high milk productivity of Holsteins while retaining the milk fat qualities of the Istoben and Dutch breeds (patent No. 3778 of November 26, 2007).