Institute of Veterinary Medicine

Belgrade, Serbia

Institute of Veterinary Medicine

Belgrade, Serbia
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Sivakumar T.,Obihiro University of Agriculture and Veterinary Medicine | Altangerel K.,Obihiro University of Agriculture and Veterinary Medicine | Battsetseg B.,Institute of Veterinary Medicine | Battur B.,Institute of Veterinary Medicine | And 6 more authors.
Veterinary Parasitology | Year: 2012

We developed a new nested PCR (nPCR) assay based on the Babesia bigemina apical membrane antigen-1 (AMA-1) gene sequence for parasite-specific detection. The primers were designed to amplify 738-bp and 211-bp fragments of the AMA-1 gene by primary and nested PCRs, respectively. The assay was proven to be specific for the B. bigemina, whereas the previously established SpeI-AvaI nPCR assay amplified not only the target fragment of B. bigemina but also a homologous one from Babesia ovata. The AMA-1 nPCR assay was also evaluated using field DNA samples extracted from 266 bovine blood samples collected from Mongolia in 2010. In a comparative evaluation, 90 (33.8%) and 25 (9.4%) of the blood samples showed positive reactions for B. bigemina by the SpeI-AvaI nPCR and AMA-1 nPCR assays, respectively. The sequencing analysis of the nPCR products confirmed that the AMA-1 nPCR method had specifically detected the target B. bigemina DNA. However, 4 different kinds of sequences were determined among the SpeI-AvaI nPCR amplicons. Two of them were derived from B. bigemina and B. ovata, while the origins of the others were unknown. In the current study, the presence of B. bigemina was clearly demonstrated among Mongolian cattle populations by the current nPCR assay for the first time. Furthermore, our findings also indicate that the AMA-1 nPCR assay may be a useful diagnostic tool for the specific detection of B. bigemina. © 2012 Elsevier B.V.

Munkhjargal T.,Obihiro University of Agriculture and Veterinary Medicine | Munkhjargal T.,Institute of Veterinary Medicine | Yokoyama N.,Obihiro University of Agriculture and Veterinary Medicine | Igarashi I.,Obihiro University of Agriculture and Veterinary Medicine
Parasitology Research | Year: 2016

Human babesiosis is the most important zoonotic protozoan infection in the world. This is the first report of the cloning, expression, purification, and immunobiochemical characterization of a methionine aminopeptidase 1 (MetAP1) protein from Babesia microti (B. microti). The gene encodes a MetAP1 protein of B. microti (BmMetAP1) of approximately 66.8 kDa that includes glutathione S-transferase (GST) tag and shows MetAP activity. BmMetAP1 was detected in a lysate of B. microti and further localized in cytoplasm of the B. microti merozoite. rBmMetAP1 was found to be immunogenic, eliciting a high antibody titer in mice. Moreover, rBmMetAP1 stimulated the production of IFN-γ and IL-12 but not IL-4. Finally, rBmMetAP1 was able to provide considerable protection to mice against a B. microti challenge infection based on a reduction in peak parasitemia levels and earlier clearance of the parasite as compared with control mice. Taken together, these results suggest that rBmMetAP1 confers significant protection against experimental B. microti infection and might be considered a potential vaccine target against human babesiosis. © 2016 Springer-Verlag Berlin Heidelberg

Brenig B.,Institute of Veterinary Medicine | Beck J.,Chronix Biomedical GmbH | Floren C.,Institute of Veterinary Medicine | Bornemann-Kolatzki K.,Chronix Biomedical GmbH | And 3 more authors.
Animal Genetics | Year: 2013

White Galloway cattle exhibit three different white coat colour phenotypes, that is, well marked, strongly marked and mismarked. However, mating of individuals with the preferred well or strongly marked phenotype also results in offspring with the undesired mismarked and/or even fully black coat colour. To elucidate the genetic background of the coat colour variations in White Galloway cattle, we analysed four coat colour relevant genes: mast/stem cell growth factor receptor (KIT), KIT ligand (KITLG), melanocortin 1 receptor (MC1R) and tyrosinase (TYR). Here, we show that the coat colour variations in White Galloway cattle and White Park cattle are caused by a KIT gene (chromosome 6) duplication and aberrant insertion on chromosome 29 (Cs29) as recently described for colour-sided Belgian Blue. Homozygous (Cs 29/Cs29) White Galloway cattle and White Park cattle exhibit the mismarked phenotype, whereas heterozygous (Cs29/wt 29) individuals are either well or strongly marked. In contrast, fully black individuals are characterised by the wild-type chromosome 29. As known for other cattle breeds, mutations in the MC1R gene determine the red colouring. Our data suggest that the white coat colour variations in White Galloway cattle and White Park cattle are caused by a dose-dependent effect based on the ploidy of aberrant insertions and inheritance of the KIT gene on chromosome 29. © 2013 The Authors, Animal Genetics © 2013 Stichting International Foundation for Animal Genetics.

Zang Y.-T.,Institute of Veterinary Medicine | Tan Y.-P.,Institute of Veterinary Medicine | Hu Y.-N.,Institute of Veterinary Medicine | Lu C.-H.,Institute of Veterinary Medicine
Procedia Engineering | Year: 2012

Animal health risk analysis technology on large-scale farm is becoming more important, but the assessment of relevant external risk factors of disease spreading into pig farm is an complex multi-dimensional process. The analytic hierarchy process (AHP) has been accepted as a robust and flexible multi-criteria decision-making tool for dealing with complex decision problems. On this study, The index system of external risk factors on large-scale farm is built based on AHP. The result shows that farm management practices, Biosecurity and site are major risk factors and reveals AHP can be used in animal risk analysis for disease control and prevention. © 2012 Published by Elsevier Ltd.

Schutz E.,Institute of Veterinary Medicine | Brenig B.,Institute of Veterinary Medicine
Genetics Selection Evolution | Year: 2015

Background: Parentage control is moving from short tandem repeats- to single nucleotide polymorphism (SNP) systems. For SNP-based parentage control in cattle, the ISAG-ICAR Committee proposes a set of 100/200 SNPs but quality criteria are lacking. Regarding German Holstein-Friesian cattle with only a limited number of evaluated individuals, the exclusion probability is not well-defined. We propose a statistical procedure for excluding single SNPs from parentage control, based on case-by-case evaluation of the GenCall score, to minimize parentage exclusion, based on miscalled genotypes. Exclusion power of the ISAG-ICAR SNPs used for the German Holstein-Friesian population was adjusted based on the results of more than 25 000 individuals. Results: Experimental data were derived from routine genomic selection analyses of the German Holstein-Friesian population using the Illumina BovineSNP50 v2 BeadChip (20 000 individuals) or the EuroG10K variant (7000 individuals). Averages and standard deviations of GenCall scores for the 200 SNPs of the ISAG-ICAR recommended panel were calculated and used to calculate the downward Z-value. Based on minor allelic frequencies in the Holstein-Friesian population, one minus exclusion probability was equal to 1.4×10-10 and 7.2×10-26, with one and two parents, respectively. Two monomorphic SNPs from the 100-SNP ISAG-ICAR core-panel did not contribute. Simulation of 10 000 parentage control combinations, using the GenCall score data from both BeadChips, showed that with a Z-value greater than 3.66 only about 2.5% parentages were excluded, based on the ISAG-ICAR recommendations (core-panel: ≥ 90 SNPs for one, ≥ 85 SNPs for two parents). When applied to real data from 1750 single parentage assessments, the optimal threshold was determined to be Z = 5.0, with only 34 censored cases and reduction to four (0.2%) doubtful parentages. About 70 parentage exclusions due to weak genotype calls were avoided, whereas true exclusions (n = 34) were unaffected. Conclusions: Using SNPs for parentage evaluation provides a high exclusion power also for parent identification. SNPs with a low GenCall score show a high tendency towards intra-molecular secondary structures and substantially contribute to false exclusion of parentages. We propose a method that controls this error without excluding too many parent combinations from the evaluation. © 2015 Schütz and Brenig; licensee BioMed Central.

Vukovic Z.,Vinča Institute of Nuclear Sciences | Radenkovic M.,Vinča Institute of Nuclear Sciences | Stankovic S.J.,Vinča Institute of Nuclear Sciences | Vukovic D.,Institute of Veterinary Medicine
Journal of the Serbian Chemical Society | Year: 2011

The distribution and accumulation of assorted heavy metals and a long-lived radionuclide (Cu, Zn, Pb, Cd, U, Th and 137Cs) in the water and sediment of the River Sava (in Serbia) were investigated at three locations in the vicinity of industrial and urban settlements (Sabac, Obrenovac, Belgrade). The concentrations of heavy metals in the sediment were found to be in the ranges (mg kg-1): 29.6-145.1 for Cu, 53.2-253.6 for Zn, 14.2-78.6 for Pb, 0.3- -24.6 for Cd, and 4.0-12.5 Bq l -1 for 137Cs. These values correlate to the concentrations in the river water if expressed by equilibrium distribution coefficients Kd (dm 3 g -1) between the solid and liquid phases. The degrees of accumulation and enrichment of tracer metals were determined.

Yondon M.,Institute of Veterinary Medicine | Zayat B.,Institute of Veterinary Medicine | Nelson M.I.,U.S. National Institutes of Health | Heil G.L.,University of Florida | And 7 more authors.
Emerging Infectious Diseases | Year: 2014

Because little is known about the ecology of influenza viruses in camels, 460 nasal swab specimens were collected from healthy (no overt illness) Bactrian camels in Mongolia during 2012. One specimen was positive for influenza A virus (A/camel/Mongolia/335/2012[H3N8]), which is phylogenetically related to equine influenza A(H3N8) viruses and probably represents natural horse-to-camel transmission. © 2014, Centers for Disease Control and Prevention (CDC). All rights reserved.

PubMed | Hue University, Veterinary Research Institute, Obihiro University of Agriculture and Veterinary Medicine and Institute of Veterinary Medicine
Type: | Journal: Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases | Year: 2015

Babesia bovis is the most virulent Babesia organism, resulting in a high mortality rate in cattle. The genetic diversity of B. bovis merozoite surface antigens (MSAs), such as MSA-1, MSA-2b, and MSA-2c, might be linked to altered immune profiles in the host animals. The present study aimed to develop type-specific PCR assays for Asian msa-1 genotypes, thereby re-analyzing the genetic diversity of msa-1 in Sri Lanka, Mongolia, and Vietnam. Specific primers were designed for nine Asian msa-1 genotypes, which had been detected based on the phylogeny constructed using msa-1 gene sequences retrieved from the GenBank database. Specificity of the type-specific PCR assays was confirmed using plasmids containing the inserts of msa-1 gene fragments that represent Asian genotypes. Furthermore, no amplicons were observed by these PCR assays when DNA samples of Babesia bigemina, Babesia ovata, Theileria annulata, Theileria orientalis, Trypanosoma evansi, Trypanosoma theileri, Anaplasma marginale, and Anaplasma bovis, and non-infected bovine blood were analyzed. In total, 109 B. bovis-positive blood DNA samples sourced from Sri Lanka (44 cattle), Mongolia (26 cattle), and Vietnam (23 cattle and 16 water buffaloes) were then screened by the type-specific PCR assays. The sequences derived from all of the PCR amplicons were phylogenetically analyzed. Out of 109 DNA samples, 23 (20 from cattle and 3 from water buffaloes) were positive for at least one genotype. In agreement with previous studies, five and four different genotypes were detected among the DNA samples from Sri Lanka and Vietnam, respectively. In contrast, four genotypes, including three novel genotypes, were detected from Mongolia. Five DNA samples were found to be co-infected with multiple genotypes. The sequences of the PCR amplicons clustered phylogenetically within the corresponding clades. These findings indicated that the type-specific PCR assays described herein are useful for the determination of genotypic diversity of the B. bovis msa-1 gene in Asia.

PubMed | Korea University, Institute of Veterinary Medicine, Seoul National University and Animal and Plant Quarantine Agency
Type: | Journal: Comparative immunology, microbiology and infectious diseases | Year: 2017

Mongolia has a high incidence of brucellosis in human and animals due to livestock husbandry. To investigate the genetic characteristics of Mongolian B. melitensis, an MLVA (multi-locus variable-number tandem-repeat analysis)-16 assay was performed with 94 B. melitensis isolates. They were identified as B. melitensis biovar (bv.) 1 (67), 3 (10) and Rev. 1 vaccine strains (17) using a classical biotyping and multiplex PCR. In genotyping, three human isolates were grouped at 2 genotypes with sheep isolates, and it implies that B. melitensis are cross-infected between human and livestock. In the parsimony analysis, Mongolian B. melitensis isolates had high genetic similarity with Chinese strains, likely due to the geographical proximity, clustered distinctively as compared with other foreign isolates. B. melitensis Rev. 1 vaccine strains were divided into 4 genotypes with 92% similarity. In the analysis of Rev.1 strains, the risk of mutation of vaccine strain might not be overlooked. Animal quarantines should be strengthened to prevent the spread of Brucella species among adjacent countries.

News Article | November 2, 2016

In the back seat of the Cruiser, Duke master of science in global health students Laura Pulscher and Thomas Moore braced themselves during the ride as best they could, relaxing when Inka stopped the vehicle to ask a goat herder for directions. Pulscher, unfazed by hours of jouncing, said slowly with a matter-of-fact smile, "We're lost." It's a commonly uttered phrase for visitors and natives alike. A meager 10 percent of Mongolia's road network is paved. The vast majority of travel in this vast, storied land of Genghis Khan is done on dirt trails and grassy paths. "If you had asked me before beginning the global health master's program that I'd be coming to Mongolia, I would have laughed," Moore would say later at a campsite on the edge of a livestock pasture. But Mongolia was the opportunity presented to Moore and Pulscher to conduct their field research projects, and they both barely blinked before saying yes. "It's one thing to learn something in a classroom setting," Pulscher said, "but another to actually apply it to your field of study in a developing nation. I don't think it ever crossed my mind that I'd end up in Mongolia." Yet, there they were for three months. Early in their three-month stay in Mongolia, Moore and Pulscher spent three weeks traveling north from the capital of Ulaanbaatar toward the Siberian border. With them in a second vehicle were veterinarians Myagmarsuk "Myagaa" Yondon; Igori "Khatnaa" Khatanbaatar; and Purevdorj "Zula" Munkhzul from the Mongolian Institute of Veterinary Medicine, who collected blood samples from herders' horses, goats, and sheep. Moore and Pulscher captured small rodents—Mongolian gerbils, Daurian ground squirrels, Siberian chipmunks, hamsters and field mice—in humane, live animal traps, and injected them with a mild sedative. Pulscher collected blood and serum samples from the animals' tails and took small ear biopsies. Moore's work was far more tedious—combing the oft-matted fur of the rodents, in search of ticks. Their individual projects differed, but the students' goals were the same: to detect in rodents' blood and ticks the presence of dangerous pathogens—Borrelia (Lyme disease), Rickettsia (spotted fever), and anaplasma (blood disease seen mostly in livestock but can cause mild fever in humans) – and share lab results with local health officials and veterinarians who could take prophylactic measures, such as educating herders and their families about ways to protect themselves and their livestock. In Mongolia, animals have near royalty status. It is a nation of just 3 million people and more than 50 million horses, camels, sheep, goats, and yaks, which are serious players in the economic cycle, and critical for the more than one million herder families who feed off their livestock and make a living selling animals for meat, hides or wool. "Observing these diseases as they circulate throughout wildlife and livestock has a tremendous impact on the health and welfare of humans here," Moore said. Holed up for two days in the dome tent she shared with Sola as an atypical summer downpour drenched the steppe, Pulscher was confident the rain would subside and the rodent trapping continue. Moore entered the tent and announced, "My tent has turned into a swimming pool." By afternoon the sun reappeared and the drying-out process commenced. The quest for rodents resumed about a half-hour from camp on a tract of land where a herd of horses grazed nearby. Small, rectangular, aluminum traps were set. When the researchers returned early that evening (the summer sun sets around 10 p.m.) the sound of scratching was heard coming from within several of them. "It's great that we still have a project," Pulscher jested as she suited up in a full-body Tyvek suit to protect her from any pathogens the rodents might harbor. "I'm glad it stopped raining." The veterinarian Khatnaa, with heavily gloved hands, cradled a Mongolian gerbil taken from one of the traps. Pulscher—with Khatnaa's guidance—injected it with the mild sedative. It was Khatnaa, with his well-spoken English, who had baptized the students to rodent testing in the field. He taught them how to safely handle the animals, determine the correct dose of sedative, clip the tail and ear so as not to cause lasting injury, and collect the blood and tick samples. After four gerbils were tested and gear repacked inside the Land Cruiser, Moore considered the importance that the team of Mongolian vets played in the success of his and Pulscher's respective projects. "They are a tremendous help to us," Moore said. "They not only teach us different tricks of the trade to catch rodents here, but they act as interpreters when we meet a herder family and provide us with local knowledge of the different species of animals." This partnership between Mongolian vets and Duke Global Health students was forged by Gregory Gray, MD, MPH, a professor of global health and infectious diseases, who established a research relationship with Mongolia nearly 10 years ago. Gray, who also holds an appointment in the Nicholas School of the Environment, is a passionate disciple of the One Health concept of health care that champions the conviction that human health is intimately tied to veterinary and environmental health. Grey, Moore, and Pulsher were part of a roughly 10-member Duke contingent that participated in the 5th Annual International Symposium on One Health in Ulaanbaatar prior to the students' embarking on their three-week fieldwork. Pulscher and Moore were presenters at an infectious disease workshop. It is rare, they said, for master's students to have the opportunity to present at an international symposium. "It was a tremendous opportunity to collaborate with international leaders and share our upcoming research," Moore said. The Institute of Veterinary Medicine in Ulaanbaatar is a 1980s-era Soviet-looking building that houses multiple labs with Mongolian scientists exploring various aspects of zoonotic diseases—those that can or might some day jump from animals to humans. This was home base for Moore and Pulscher for the bulk of their three-month Mongolian stay. It was here that the blood and tick samples they collected in the field were analyzed for signs of the dangerous pathogens. The Duke students extracted DNA from the samples using magnetic beads inside small test tubes. Because DNA has a charge, it is attracted to the beads, which are then separated from the rest of the sample and the DNA extracted. Performing field and lab work is usually an either/or proposition for students interested in global health. Rarely do master's students have the opportunity to do both. "It's unique that we're able to collect our own specimens in the environment and bring them back and test them," Moore said. "This takes our education to the next level." "It makes us more well-rounded individuals," Pulscher added. "A lot of people don't necessarily have both lab and field skills." They worked in the lab with Myagaa, who accompanied them on their fieldwork. His own project involved collecting mosquitoes and testing them for West Nile virus. Learning about his work, and watching the other Mongolian vets in the field lasso livestock and take blood samples was an unexpected, welcomed educational component, they said. The fresh yogurt was great, the dried, fermented milk curds and mare's milk not so much, the students concluded. But that is the way one gets to know the culture of a people – through their food. The hospitality of the Mongolian herder families is a memory that won't soon fade, they said. "It is amazing how welcoming they were to let us into their homes, feed us, let us get to know them, and let us camp on their land," Pulscher said. On a cool summer night in one of those camps as he watched the sun slowly set over a verdant hillside, lighting the horizon in an indigo purple haze, Moore reflected on the allure of the landscape surrounding him. "This place is beautiful," he said. "Being here has been such a privilege." Explore further: Saving lives with better flu diagnosis in Mongolia

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