scholarly journals TECHNOLOGY OF BOVINE LEUKEMIA VIRUS GENODIAGNOSTICS IN CATTLE, IN PRODUCED RAW MATERIALS AND PRODUCTS

2021 ◽  
pp. 119-125
Author(s):  
R. R. Vafin ◽  
Kh. Kh. Gilmanov ◽  
A. G. Galstyan ◽  
N. S. Pryanichnikova ◽  
A. V. Bigaeva ◽  
...  

The most important task of the dairy cattle industry is to obtain high quality raw milk. To achieve it, a set of measures is required, including aimed at increasing the biological safety of produced raw materials. The aim of the study was to create a scientific and methodological basis for the Bovine leukemia virus (BLV) gene diagnostics in a combined format of pathogen indication and identification. This required updating the strategy of BLV PCR-RFLP genotyping, consistent with its phylogenetic classification, taking into account the growing knowledge about the genetic diversity of 11 genotypes of the studied viral pathogen. When staging nested PCR, oligonucleotide primers were used, which initiate at the final stage of the reaction the production of a 444 bp env-gene fragment of the pathogen. Five restriction endonucleases were used in PCR-RFLP BLV genotyping of: PvuII, SspI, AsuHPI, HaeIII, and BstX2I. As a result of verification of the developed Bovine leukemia virus method for gene identification with an updated genotyping strategy, a technical result was obtained, expressed in the ability to identify all 11 BLV genotypes discovered to date by interpreting the generated 58 genotype-associated combinations of PCR-RFLP profiles.

2018 ◽  
Vol 6 (2) ◽  
pp. 314-324 ◽  
Author(s):  
Irina Donnik ◽  
Irina Donnik ◽  
Ramil Vafin ◽  
Ramil Vafin ◽  
Aram Galstyan ◽  
...  

Molecular genetic research methods make it possible to evaluate the genetic diversity of bovine leukemia virus (BLV) and are the most informative approaches to its genetic identification. Molecular genetic research methods work well for the phylogenetic analysis of sequenced nucleotide DNA sequences of the provirus, as well as for the polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) according to the phylogenetic classification of the pathogen. The purpose of the research was to study the scientific and methodological approaches to the genetic identification of bovine leukemia virus, integrated into the molecular monitoring of infection of cattle with BLV genotypes. The authors used PCR-RFLP-genotyping and comparative phylogenetic analysis of aligned nucleotide sequences of the env gene fragment of the BLV provirus isolates to detect the genotypic affiliation of the cattle from twenty-one livestock farms of the Republic of Tatarstan. As a result, isolates of four out of ten BLV genotypes were found in the Tatarstani cattle, namely genotypes 1, 4, 7, and 8. The research involved a comparative analysis of 505 nucleotide sequences of a fragment of the BLV env gene, including those deposited in GenBank NCBI. The analysis confirms the inconsistency of several earlier PCR-RFLP typing strategies with the current approach in assessing the genotypic diversity by phylogenetic analysis. The improved strategy of PCR-RFLP genotyping of BLV corresponds with its modern phylogenetic classification. The strategy makes it possible to identify all the known genotypes of the viral pathogen. Its validity has been proved by in silico modelling of restrictogrammes and a phylogenetic analysis of the env gene fragment of 57 reference isolates of ten BLV genotypes that generate 57 genotype-associated combinations of diagnostically significant PCR-RFLP profiles.


2020 ◽  
Vol 50 (3) ◽  
pp. 525-535
Author(s):  
Khamid Gilmanov ◽  
Vladislav Semipyatniy ◽  
Alana Bigaeva ◽  
Ramil Vafin ◽  
Svetlana Turovskaya

Introduction. High-quality dairy products depend on the properties of raw materials. Milk is the basic raw material for all dairy products. Its quality is subject to requirements of biological safety, and its physicochemical and technological properties comply with strict indicators. ϰ-casein gene is a basic milk protein. Its allelic variants affect milk productivity, as well as its composition and technological properties, e.g. cheeseability and heat resistance. The research objective was to develop a new method for determining the ratio of the relative proportions of selection-significant kappa-casein gene alleles in bulk cow’s milk powder by means of molecular genetics and bioinformation system. The method can help to test raw materials for further processing rationality. Study objects and methods. The research included a genetic analysis of samples of bulk whole milk powder, bulk powdered skimmed milk, and bulk raw milk. The developed method involved DNA extraction, combined PCR-RFLP technique, electrophoretic results, and analysis detection of information data using new mathematical algorithms and software. Results and discussion. The ratio of the relative proportions of the kappa-casein gene alleles in milk powder was determined using primer sets JK5 and JK3, as well as restriction endonuclease HinfI for PCR-RFLP analysis. The experiment showed satisfactory reproducibility and interpretation of the obtained data. The program provided a ratio calculation of the kappa-casein gene alleles relative proportions in the studied milk powder samples, expressed as a percentage of the A allele proportion with additional indication of the absolute and relative errors. It also placed the information block of the generated numerical indicators into the percentage scale. Conclusion. The developed methodology was implemented by a set of laboratory procedures. The new specialized program “Calculation of the relative proportions ratio of ϰ-casein alleles in bulk milk” is on open access. It provides a correct and prompt data interpretation, generated during the analysis of dry milk raw materials of bulk origin.


2020 ◽  
Vol 165 (12) ◽  
pp. 2961-2966
Author(s):  
Chihiro Ochiai ◽  
Yuzuru Katagiri ◽  
Sota Kobayashi ◽  
Ikunori Naitoh ◽  
Syuji Yoneyama ◽  
...  

2019 ◽  
Vol 50 (1) ◽  
Author(s):  
Sonoko Watanuki ◽  
Shin-nosuke Takeshima ◽  
Liushiqi Borjigin ◽  
Hirotaka Sato ◽  
Lanlan Bai ◽  
...  

AbstractBovine leukemia virus (BLV) infects cattle and causes serious problems for the cattle industry, worldwide. Vertical transmission of BLV occurs via in utero infection and ingestion of infected milk and colostrum. The aim of this study was to clarify whether milk is a risk factor in BLV transmission by quantifying proviral loads in milk and visualizing the infectivity of milk. We collected blood and milk from 48 dams (46 BLV seropositive dams and 2 seronegative dams) from seven farms in Japan and detected the BLV provirus in 43 blood samples (89.6%) but only 22 milk samples (45.8%) using BLV-CoCoMo-qPCR-2. Although the proviral loads in the milk tended to be lower, a positive correlation was firstly found between the proviral loads with blood and milk. Furthermore, the infectivity of milk cells with BLV was visualized ex vivo using a luminescence syncytium induction assay (LuSIA) based on CC81-GREMG cells, which form syncytia expressing enhanced green fluorescent protein (EGFP) in response to BLV Tax and Env expressions when co-cultured with BLV-infected cells. Interestingly, in addition to one BLV-infected dam with lymphoma, syncytia with EGFP fluorescence were observed in milk cells from six BLV-infected, but healthy, dams by an improved LuSIA, which was optimized for milk cells. This is the first report demonstrating the infectious capacity of cells in milk from BLV-infected dams by visualization of BLV infection ex vivo. Thus, our results suggest that milk is a potential risk factor for BLV vertical spread through cell to cell transmission.


Author(s):  
N.V. Bateneva ◽  
◽  
P.N. Smirnov ◽  
I.V. Mikhnovich ◽  
◽  
...  

Author(s):  
В.Н. СУРОВЦЕВ ◽  
Е.Н. ПАЮРОВА

Проанализированы проблемы на рынке молока и молочных продуктов России в условиях глобального экономического кризиса: перепроизводство молока в основных странах-экспортерах, снижение закупочных цен на сырое молоко, снижение цен на биржевые товары в мире и России в 2020 году, тренд на уменьшение спроса на молочные продукты на мировом рынке в среднесрочном периоде, снижение общего спроса на молочные продукты на внутрироссийском рынке при падении реальных доходов населения, изменение структуры потребления. Проведена оценка новых возможностей и угроз для развития отрасли: со стороны потребителей — рост цен на продовольствие, снижение доходов; с позиции производителей молока — снижение закупочных цен, рост требований к сырью для производства продукции с увеличенными сроками годности, дефицит рабочей силы, вероятное сокращение господдержки в результате снижения цен на углеводороды, попытки регулирования цен; со стороны перерабатывающих предприятий — сокращение спроса, снижение цен на готовую продукцию вслед за мировыми ценами. Обоснованы приоритеты инвестирования в молочном скотоводстве и основные формы совершенствования государственной поддержки отрасли, обеспечивающие эффективную адаптацию производителей молока к новым экономическим условиям, повышение устойчивости отрасли при усилении макроэкономических рисков. The article analyzes the problems in the Russian milk and dairy products market in the context of the global economic crisis: overproduction of milk in the main exporting countries, lower purchase prices for raw milk, lower prices for commodities in the world and in Russia in 2020, trend to reduce demand for dairy products on the world market in medium term, a decrease in total demand for dairy products in the Russian market with a decrease in the purchasing power of the population, a change in the structure of consumption. An assessment of new opportunities and threats to the development of the industry was carried out: on the part of consumers — rising food prices, lower incomes; from the perspective of dairy producers — reduction in purchase prices, increased requirements for raw materials for the production of products with extended periods, labor shortages, the likely reduction in state support as a result of lower prices for hydrocarbons, attempts to regulate prices; on the part of processing enterprises — reduction of demand, reduction of prices for finished goods following world prices. The investment priorities in dairy cattle breeding and the main forms of improving state support for the industry, ensuring the effective adaptation of milk producers to new economic conditions, increasing the sustainability of the industry with increasing macroeconomic risks, are substantiated.


2019 ◽  
Vol 07 (01) ◽  
Author(s):  
Kelly Cristina Santos Montanari ◽  
Marcia Mayumi Fusuma ◽  
Alessandra Maria Dias Lacerda ◽  
Líria Hiromi Okuda ◽  
Edviges Maristela Pituco ◽  
...  

Domiati cheese is the most popular brand of cheese ripened in brine in the Middle East in terms of consumed quantities. This study was performed to investigate the impact of the microbiological quality of the used raw materials, the applied traditional processing techniques and ripening period on the quality and safety of the produced cheese. Three hundred random composite samples were collected from three factories at Fayoum Governorate, Egypt. Collected samples represent twenty-five each of: raw milk, table salt, calf rennet, microbial rennet, water, environmental air, whey, fresh cheese, ripened cheese & swabs from: worker hands; cheese molds and utensils; tanks. All samples were examined microbiologically for Standard Plate Count (SPC), coliforms count, Staphylococcus aureus (S. aureus) count, total yeast & mould count, presence of E. coli, Salmonellae and Listeria monocytogenes (L. monocytogenes). The mean value of SPC, coliforms, S. aureus and total yeast & mould counts ranged from (79×102 CFU/m3 for air to 13×108 CFU/g for fresh cheese), (7×102 MPN/ cm2 for tank swabs to 80×106 MPN/ml for raw milk), (9×102 CFU/g for salt to 69×106 CFU/g for fresh cheese) and (2×102 CFU/cm2 for hand swabs to 60×104 CFU/g for fresh cheese), respectively. Whereas, E. coli, Salmonella and L. monocytogenes failed to be detected in all examined samples. There were significant differences in all determined microbiological parameters (p ≤0.05) between fresh and ripened cheese which may be attributed to different adverse conditions such as water activity, pH, salt content and temperature carried out to improve the quality of the product.


2012 ◽  
Vol 6 (3) ◽  
pp. 1-7 ◽  
Author(s):  
M Kazemimanesh ◽  
O Madadgar ◽  
MR Mahzoonieh ◽  
T Zahraei-Salehi ◽  
F Steinbach ◽  
...  

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