Two cases of hydrophobia in the Republic of Tatarstan: In vivo and postmortem laboratory diagnosis

The results of rabies in vivo and postmortem laboratory detection in two cases registered in the Republic of Tatarstan are reported: a victim bitten by a wolf in 2002 and another one bitten by a stray dog on Goa Island, India, in 2013. In the patient bitten by a wolf cornea imprints studies using the method of fluorescent antibodies (MFA) showed rabies-positive result 6 days before the patient's death. The results were confirmed by postmortem examination of different parts of the brain and salivary glands using the MFA, enzyme-linked immunosorbent assay (ELISA), optical microscopy, and bioassay methods. In the patient bitten by a stray dog the rabies virus specific antigen was detected by eye cornea studies using the MFA method and saliva studies using the ELISA. The rabies virus genome was also isolated from saliva and tear fluid using nested reverse-transcription polymerase chain reaction (RT-PCR) 9 days before the patient's death. The in vivo studies results were consistent with the postmortem study of different parts of the brain using the MFA, enzyme-linked immunosorbent assay (ELISA), optical microscopy, and bioassay methods. All the infection-positive results of both in vivo and postmortem studies were consistent with the clinical studies, i.e. rabies diagnosis was confirmed. The analysis of the rabies virus gene G fragment nucleotide sequence of 238 nd length showed a slight difference between the studied isolates (2 rabies) and the RABV AY956319 (1.68%), difference by 10.5% from the Vnukovo-32 vaccine strains and by 10.9 % from the SAD B19 rabies strain, respectively (rabies viruses of 1st genotype). It was also significantly different from the lissaviruses of 2,4,5, and 6 genotypes (21.0-32.7%). The obtained results indicate phylogenetic closeness of the studied isolates (2 rabies) with the RABV AY956319 rabies virus strain belonging to the 1st genotype

Persistent form of bovine viral diarrhea

The review provides an analysis of literature data on the persistent form of Bovine Viral diarrhea/Mucosal disease (BVD) and is focused on virus and host factors, including those related to immune response, that contribute the persistence of the virus. BVD is a cattle disease widespread throughout the world that causes significant economic damage to dairy and beef cattle. The disease is characterized by a variety of clinical signs, including damage to the digestive and respiratory organs, abortions, stillbirths and other failures of reproductive functions

Проба с диаскинтестом при диагностике туберкулеза животных

The objective of the study: to compare the diagnostic value of intracutaneous tests with tuberculin and diaskintest for detection of tuberculosis in cattle.Subjects and methods. In this study, PPD tuberculin for mammals was used, it was made from M. bovis strain AN-5, manufactured by BIOK, and Diaskintest (recombinant tuberculous allergen, solution forintracutaneous administration) manufactured by ZAO FF Lecco, series 030307.Laboratory animals (guinea pigs, rabbits, chickens) and farm animals (pigs and cattle) were used in the study.Results. It has been established on laboratory animals that diaskintest does not cause any adverse events. When assessing sows sensitized with M. avium, it was found that the intracutaneous administration of PPD tuberculin to mammals resulted in up to 8.5% of positive reactions, while diaskintest was negative in all of them. Among cattle sensitized to non-tuberculous mycobacteria, intracutaneous administration of tuberculin for mammals revealed up to 4.6% of positive reactions, and diaskintest – up to 0.6%. In the farm with the unfavorable TB situation where animals infected with M. bovis were previously detected, when examining 177 cows, tuberculin test was positive in 102 (57.6%) of them, and diaskintest in 88 (49.7%). Diaskintest can be used for differential diagnosis of tuberculosis and sensitization by non-tuberculous mycobacteria in pigs and cattle.Цель исследования: сравнение диагностической ценности внутрикожных проб с туберкулином и диаскинтестом для выявления туберкулеза крупного рогатого скота.Материалы и методы. В работе использовали ППД туберкулин для млекопитающих, изготовленный из M. bovis штамма AN-5, производства фирмы «БИОК» и препарат диаскинтест (аллерген туберкулезный рекомбинантный, раствор для внутрикожного введения) производства ЗАО ФФ «Лекко», серия 030307. Исследования проводили на лабораторных животных (морских свинках, кроликах, курах) и сельскохозяйственных животных (свиньях и крупном рогатом скоте).Результаты. На лабораторных животных установлено, что проба с диаскинтестом не вызывает нежелательных реакций. При исследовании сенсибилизированных M. avium свиноматок установлено, что внутрикожное введение ППД туберкулина для млекопитающих выявляет до 8,5% положительных реакций, при этом проба с диаскинтестом была у всех отрицательной. Среди крупного рогатого скота с сенсибилизацией к нетуберкулезным микобактериям внутрикожное введение туберкулина для млекопитающих выявляло до 4,6% положительных реакций, а проба с диаскинтестом – до 0,6%. В неблагополучном по туберкулезу хозяйстве, где ранее выявлялись животные, инфицированные M. bovis, при исследовании 177 коров проба с туберкулином была положительной у 102 (57,6%), а проба с диаскинтестом – у 88 (49,7%). Проба с диаскинтестом может использоваться при дифференциальной диагностике туберкулеза и сенсибилизации нетуберкулезными микобактериями у свиней и крупного рогатого скота

ГЕМОСПОРИДИОЗЫ СЕЛЬСКОХОЗЯЙСТВЕННЫХ, ДОМАШНИХ И ДИКИХ ЖИВОТНЫХ НА ТЕРРИТОРИИ РОССИЙСКОЙ ФЕДЕРАЦИИ

Objective of research:  analysis of epizootic situation on protozoan blood parasitic diseases in domestic and wild animals on the territory of Russian Federation Materials and methods:  Veterinary reporting forms, sources of literature. Results and discussion:  The data on epizootic situation related to protozoan blood parasitic diseases in animals on the territory of  RF have been obtained and analyzed in this article. It was determined that for a proper evaluation of epizootic situation it is necessary to combine the activities of scientific-research institutes and veterinary services for the purpose of development of monitoring based on recommendations of the International Epizootic Bureau (IEB).  Цель работы – анализ  эпизоотической ситуации по протозойным кровепаразитарным болезням домашних и диких  животных в Российской Федерации. Материалы и методы. Формы ветеринарной отчётности, литературные источники. Результаты и обсуждение.  В статье обобщены и проанализированы данные об эпизоотической ситуации по протозойным кровепаразитарных болезням животных в субъектах РФ. Показано, что для достоверной  оценки эпизоотической ситуации необходимо объединение научно-исследовательских институтов и ветеринарной службы с целью разработки системы мониторинга с учётом рекомендаций МЭБ (Международное Эпизоотическое бюро).

ДНК-ДИАГНОСТИКА АНАПЛАЗМОЗА КРУПНОГО РОГАТОГО СКОТА

Objective of research: The purpose of our research was to develop a DNA diagnostic method for anaplasmosis in cattle. Materials and methods: Blood samples were obtained from the tail vein with the use of ethylenediaminetetraacetic acid as an anticoagulant. The extraction of DNA was performed with the kit Sorb-M. To analyze the gene msp4 the following sequences belonging to different isolates Anaplasma marginale were used. To select primers the conserved elements of sequences were detected with the server СlustalW2. The specificity of primers was checked by a BLASTN search. The results of polymerase chain reaction (PCR) were estimated using 2% agarose gel electrophoresis. Electrophoresis was performed within 40 minutes at the field intensity 5 V/cm. Fragments of gene msp4 obtained as a results of polymerase chain reaction (PCR) were purified, ligated and cloned in E. coli cells. Transformation was performed using the heat shock method. Search for E. coli colonies containing pGEM-msp4 plasmid was conducted by the PCR method using standard M13 primers with the following analysis of PCR results by electrophoresis. Target colonies of E. coli were cultured overnight at 37 °С in 2 ml LB medium containing ampicillin in a 100 mcg /ml concentration. Sequencing of received plasmids pGEM-msp4 was carried out by Sanger method and the genetic analyzer Applied Biosystems 3130.  Results and discussion: Development and approbation of primers on the basis of the MSP4 gene of Anaplasma marginale to perform DNA diagnostics of anaplasmosis in cattle by PCR method were described. Due to PCR sensitivity along with the use of primers, it is possible to identify 100 and more gene copies. TheЦель исследования – разработка метода ДНК-диагностики анаплазмоза крупного рогатого скота.  Материалы и методы. Пробы крови отбирали из хвостовой вены с использованием ЭДТА в качестве антикоагулянта. ДНК выделяли с помощью набора Sorb-M. Для анализа гена msp4 были использованы соответствующие последовательности, принадлежащие разным изолятам Anaplasma marginale. Выявление консервативных участков последовательностей для подбора праймеров проводили с помощью сервера СlustalW2. Видоспецифичность праймеров проверяли с использованием алгоритма BLASTN. Результаты полимеразной цепной реакции (ПЦР) оценивали методом электрофореза в 2%-ном агарозном геле. Электрофорез проводили в течение 40 минут при напряженности поля 5 В/см. Полученные в результате ПЦР фрагменты гена msp4 были очищены, лигированы и клонированы в клетках E. coli. Трансформацию проводили методом теплового шока. Поиск колоний E. coli DH5α, содержащих плазмиду pGEM-msp4, проводили методом ПЦР с использованием стандартных праймеров M13 с последующим анализом результатов ПЦР методом электрофореза. Целевые колонии наращивали в течение ночи при 37 °С в 2 мл среды LB, содержащей ампициллин в концентрации 100 мкг/мл. Секвенирование полученных плазмид pGEM-msp4 осуществляли по методу Сэнгера и генетического анализатора Applied Biosystems 3130. Результаты и обсуждение. Описаны разработка и апробация праймеров к гену msp4Anaplasma marginale для ДНК-диагностики анаплазмоза крупного рогатого скота методом ПЦР. Чувствительность ПЦР с использованием этих праймеров позволяет выявить 100 и больше копий гена. Проведенные испытания свидетельствуют о 100%-ной повторяемости и воспроизводимости данного метода

Two cases of hydrophobia in the Republic of Tatarstan: In vivo and postmortem laboratory diagnosis

The results of rabies in vivo and postmortem laboratory detection in two cases registered in the Republic of Tatarstan are reported: a victim bitten by a wolf in 2002 and another one bitten by a stray dog on Goa Island, India, in 2013. In the patient bitten by a wolf cornea imprints studies using the method of fluorescent antibodies (MFA) showed rabies-positive result 6 days before the patient's death. The results were confirmed by postmortem examination of different parts of the brain and salivary glands using the MFA, enzyme-linked immunosorbent assay (ELISA), optical microscopy, and bioassay methods. In the patient bitten by a stray dog the rabies virus specific antigen was detected by eye cornea studies using the MFA method and saliva studies using the ELISA. The rabies virus genome was also isolated from saliva and tear fluid using nested reverse-transcription polymerase chain reaction (RT-PCR) 9 days before the patient's death. The in vivo studies results were consistent with the postmortem study of different parts of the brain using the MFA, enzyme-linked immunosorbent assay (ELISA), optical microscopy, and bioassay methods. All the infection-positive results of both in vivo and postmortem studies were consistent with the clinical studies, i.e. rabies diagnosis was confirmed. The analysis of the rabies virus gene G fragment nucleotide sequence of 238 nd length showed a slight difference between the studied isolates (2 rabies) and the RABV AY956319 (1.68%), difference by 10.5% from the Vnukovo-32 vaccine strains and by 10.9 % from the SAD B19 rabies strain, respectively (rabies viruses of 1st genotype). It was also significantly different from the lissaviruses of 2,4,5, and 6 genotypes (21.0-32.7%). The obtained results indicate phylogenetic closeness of the studied isolates (2 rabies) with the RABV AY956319 rabies virus strain belonging to the 1st genotype

Two cases of hydrophobia in the Republic of Tatarstan: In vivo and postmortem laboratory diagnosis

The results of rabies in vivo and postmortem laboratory detection in two cases registered in the Republic of Tatarstan are reported: a victim bitten by a wolf in 2002 and another one bitten by a stray dog on Goa Island, India, in 2013. In the patient bitten by a wolf cornea imprints studies using the method of fluorescent antibodies (MFA) showed rabies-positive result 6 days before the patient's death. The results were confirmed by postmortem examination of different parts of the brain and salivary glands using the MFA, enzyme-linked immunosorbent assay (ELISA), optical microscopy, and bioassay methods. In the patient bitten by a stray dog the rabies virus specific antigen was detected by eye cornea studies using the MFA method and saliva studies using the ELISA. The rabies virus genome was also isolated from saliva and tear fluid using nested reverse-transcription polymerase chain reaction (RT-PCR) 9 days before the patient's death. The in vivo studies results were consistent with the postmortem study of different parts of the brain using the MFA, enzyme-linked immunosorbent assay (ELISA), optical microscopy, and bioassay methods. All the infection-positive results of both in vivo and postmortem studies were consistent with the clinical studies, i.e. rabies diagnosis was confirmed. The analysis of the rabies virus gene G fragment nucleotide sequence of 238 nd length showed a slight difference between the studied isolates (2 rabies) and the RABV AY956319 (1.68%), difference by 10.5% from the Vnukovo-32 vaccine strains and by 10.9 % from the SAD B19 rabies strain, respectively (rabies viruses of 1st genotype). It was also significantly different from the lissaviruses of 2,4,5, and 6 genotypes (21.0-32.7%). The obtained results indicate phylogenetic closeness of the studied isolates (2 rabies) with the RABV AY956319 rabies virus strain belonging to the 1st genotype

Spread Dynamics of Leucosis in Cattle in Livestock Farms of the Russian Federation for 2000–2018

Leucosis occupies a leading position in the modern nosological structure of cattle infectious diseases. This is due to the high infection of livestock and large economic damage in affected farms [1–3]. Leucosis is a chronic tumor disease caused by an RNA-containing virus of the Retroviridae family. The disease is characterized by uncontrolled reproduction of immature hematopoietic cells [4, 5]. The objective is to assess in dynamics the indicators of cattle infection with the leucosis virus and the incidence of leucosis in farms of all categories of the Russian Federation in the period of 2000–2018. The data of the analysis of the epizootic situation for leucosis in cattle in farms of all categories of the Russian Federation for 2000-2018 are presented. Over this period of time, the number of diagnostic serological studies in the immunodiffusion reaction increased 1.88 times in all subjects. According to the results of epizootological monitoring in farms of all categories of the Russian Federation, the rates of infection of the bovine leucosis virus decreased 1.89 times, the incidence rates decreased 2.02 times. At present, the epizootic situation has improved markedly, but so far there are problems with incomplete release of farms from leucosis in cattle. It is necessary to continue work on the elimination of this dangerous chronic disease in farms of all categories in the territory of the Russian Federation

HEMOSPORIDOSIS OF FARM, DOMESTIC AND WILD ANIMALS ON THE TERRITORY OF RUSSIAN FEDERATION

Objective of research:  analysis of epizootic situation on protozoan blood parasitic diseases in domestic and wild animals on the territory of Russian Federation Materials and methods:  Veterinary reporting forms, sources of literature. Results and discussion:  The data on epizootic situation related to protozoan blood parasitic diseases in animals on the territory of  RF have been obtained and analyzed in this article. It was determined that for a proper evaluation of epizootic situation it is necessary to combine the activities of scientific-research institutes and veterinary services for the purpose of development of monitoring based on recommendations of the International Epizootic Bureau (IEB)

DNA DIAGNOSTICS OF ANAPLASMOSIS IN CATTLE

Objective of research: The purpose of our research was to develop a DNA diagnostic method for anaplasmosis in cattle. Materials and methods: Blood samples were obtained from the tail vein with the use of ethylenediaminetetraacetic acid as an anticoagulant. The extraction of DNA was performed with the kit Sorb-M. To analyze the gene msp4 the following sequences belonging to different isolates Anaplasma marginale were used. To select primers the conserved elements of sequences were detected with the server СlustalW2. The specificity of primers was checked by a BLASTN search. The results of polymerase chain reaction (PCR) were estimated using 2% agarose gel electrophoresis. Electrophoresis was performed within 40 minutes at the field intensity 5 V/cm. Fragments of gene msp4 obtained as a results of polymerase chain reaction (PCR) were purified, ligated and cloned in E. coli cells. Transformation was performed using the heat shock method. Search for E. coli colonies containing pGEM-msp4 plasmid was conducted by the PCR method using standard M13 primers with the following analysis of PCR results by electrophoresis. Target colonies of E. coli were cultured overnight at 37 °С in 2 ml LB medium containing ampicillin in a 100 mcg /ml concentration. Sequencing of received plasmids pGEM-msp4 was carried out by Sanger method and the genetic analyzer Applied Biosystems 3130.  Results and discussion: Development and approbation of primers on the basis of the MSP4 gene of Anaplasma marginale to perform DNA diagnostics of anaplasmosis in cattle by PCR method were described. Due to PCR sensitivity along with the use of primers, it is possible to identify 100 and more gene copies. Th