The identification and classification of endogenous retroviruses in the horse genome

Endogenous retroviruses (ERVs) are sequences that derived from ancient retroviral infections of germ cells and integrated in humans, mammals and other vertebrates millions years ago. These ERVs are inherited according to Mendelian expectations in the same way as all other genes in the genome. Size of complete endogenous retrovirus is between 8-12 kb long in average and contains gag, pro, pol and env genes that always occur in the same order. Coding sequences are flanked by two LTRs (Long Terminal Repeat sequences). Most ERVs are defective that are carrying multitude of inactivating mutations. However some ERVs still have open reading frames in their genome. These ERVs settle close to functional genes or within the genes and can influence or control functions of the host genes using their LTRs. Most integration has deleterious effects. However some integration could be example of positive co-adaptation as syncitin which is involved to form the syncytial layer of the placenta. The first equine endogenous beta retrovirus which is EcERV-Beta1 has been found in 2011 by Antoinette C. van der Kuyl1. The first known beta retrovirus and few pol gene similar to foamy retrovirus were only known endogenous retroviruses fixed in the domestic horse (Equus caballus) genome. Our aim of the study was to identify other endogenous retrovirus sequences in an equine genome and classify them into groups. Based on the high number of SINEs (Equine Repetitive Element) in the horse genome we hypothesized that certain ERVs will be located sufficiently close to SINEs that they will be amplified using an unbiased SINE-PCR approach with degenerate primers. The nearest SINE element was located 5.5 k bp upstream at the 5’of the EcERV-Beta1. Pan-pol PCR was also used to find novel ERVs based on 640 bp long region of pol gene which is the most conserved region of ERVs. 27 complete and novel ERVs that are 13 beta, 13 gamma, 1 spuma and 249 candidate endogenous retroviruses have been revealed using LTR_STRUC tool and double checked by Retrotector© online tool and NCBI-BLAST tool. It was proven that EcERV-Beta1 which has 2 LTRs with 1% divergence between LTRs has a polymorphism among 13 different breeds

The prevalence of headache disorders in children and adolescents in Mongolia: a nationwide schools-based study

Background The Global Campaign against Headache collects data from children (7–11 years) and adolescents (12–17 years) both to inform health and education policies and to contribute to the Global Burden of Disease (GBD) study. This survey in Mongolia was part of this global enquiry. Methods Following the generic protocol for the global enquiry, this was a schools-based cross-sectional survey. Self-completed structured questionnaires were administered, within classes, in seven schools in four districts of the Capital city and three rural areas of Mongolia, selected to represent the country’s diversities. Headache diagnostic questions were based on ICHD-3 criteria but for the inclusion of undifferentiated headache (UdH). Results Of 4515 potential participants, 4266 completed the questionnaire (children 2241 [52.5%], adolescents 2025 [47.5%]; males 2107 [49.4%], females 2159 [50.6%]). Children were therefore slightly over-represented, although overall mean age was 11.3 ± 3.3 years (range: 6–17; median 11). The non-participation proportion was 4.5%. Observed lifetime prevalence of headache was 81.0%. Gender- and age-adjusted 1-year prevalence was 59.4% (migraine: 27.3%; tension-type headache [TTH]: 16.1%; UdH: 6.6%; all headache on ≥15 days/month: 4.2%; probable medication-overuse headache: 0.7%). All headache types except UdH were more prevalent among females than males, and all were more prevalent among adolescents than children, although UdH represented a higher proportion of all headache in children (13.0%) than in adolescents (10.0%). Headache yesterday was reported by 15.9% of the sample, 26.0% of those with headache. Conclusions At least in adolescents, headache in Mongolia is no less common than in adults. The clear difference from similar studies in other countries was a lower prevalence of UdH, perhaps a consequence of reporting bias in a non-troublesome headache (mild and short-lasting by definition). This study informs policy in Mongolia and, with no similar study yet from elsewhere in Western Pacific Region, makes an important contribution to the global enquiry

International interlaboratory validation of a nested PCR for molecular detection of Babesia bovis and Babesia bigemina, causative agents of bovine babesiosis

Babesia bovis and B. bigemina are tick-transmitted parasites causing bovine babesiosis, characterized by significant morbidity and mortality leading to economic losses to the livestock industry in tropical and subtropical regions worldwide. Animals that recover from acute infection remain carriers with low parasitemia acting as a source of transmission, and often escape detection. An improved diagnosis of a B. bovis and/or B. bigemina infection of carrier animals is enabled by the availability of detection methods with high sensitivity. To this end, two nested PCR assays targeting the cytochrome b (cytb) genes of B. bovis and B. bigemina (cytb-nPCR), have been recently developed and an increased sensitivity with respect to reference protocols has been shown (Romero-Salas et al., 2016). In this study, the specificity against a panel of hemoparasites that potentially co-occur with B. bovis and B. bigemina was demonstrated to ensure applicability of the cytb-nPCR assays in a wide range of regions where bovine babesiosis is endemic. Furthermore, we compared both reported cytb-nPCR assays with reference nPCR and qPCR protocols for (i) their capability to detect carrier animals in the field, and (ii) their reproducibility when performed in different laboratories by independent operators. We show that, in a panel of bovine field samples (n = 100), the cytb-nPCR assays detected a considerably higher number of 25% B. bovis and 61% B. bigemina-positive animals compared to 7% and 20% B. bovis and 55% and 49% B. bigemina-positive animals when tested by reference nPCR and qPCR protocols, respectively. Cytb-nPCRs were also found superior in the detection of carrier animals when field samples from Africa were analyzed. In addition, both the B. bovis and B. bigemina cytb-nPCR assays were independently validated in a single blinded study in three laboratories. Importantly, no significant differences in the number/percentage of infected animals was observed using cytb-nPCR assays. In summary, the cytb-nPCR assays detected a considerably higher number of chronically infected B. bovis and B. bigemina carrier animals compared to reference nPCR and qPCR protocols, when applied in different epidemiological field situations. Furthermore, a high reproducibility between laboratories could be demonstrated.Instituto de PatobiologíaFil: Ganzinelli Sabrina Belen. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; ArgentinaFil: Ganzinelli Sabrina Belen. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Byaruhanga, Charles. University of Pretoria. Faculty of Veterinary Science. Department of Veterinary Tropical Diseases. Vectors and Vector-borne Diseases Research Programme; SudáfricaFil: Primo, María Evangelina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea; ArgentinaFil: Primo, María Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Lukanji, Zinathi. University of Pretoria. Faculty of Veterinary Science. Department of Veterinary Tropical Diseases. Vectors and Vector-borne Diseases Research Programme; SudáfricaFil: Sibeko, Kgomotso. University of Pretoria. Faculty of Veterinary Science. Department of Veterinary Tropical Diseases. Vectors and Vector-borne Diseases Research Programme; SudáfricaFil: Matjila, Tshepo. University of Pretoria. Faculty of Veterinary Science. Department of Veterinary Tropical Diseases. Vectors and Vector-borne Diseases Research Programme; SudáfricaFil: Neves, Luis. University of Pretoria. Faculty of Veterinary Science. Department of Veterinary Tropical Diseases. Vectors and Vector-borne Diseases Research Programme; SudáfricaFil: Benitez, Daniel Francisco. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mercedes; ArgentinaFil: Enkhbaatar, Batmagnai. Obihiro University of Agriculture and Veterinary Medicine. National Research Center for Protozoan Diseases; JapónFil: Enkhbaatar, Batmagnai. Mongolian University of Life Sciences. Institute of Veterinary Medicine. Laboratory of Molecular Genetics; MongoliaFil: Nugraha, Arifin Budiman. Obihiro University of Agriculture and Veterinary Medicine. National Research Center for Protozoan Diseases; JapónFil: Nugraha, Arifin Budiman. IPB University. Faculty of Veterinary Medicine. Department of Animal Infectious Diseases and Veterinary Public Health; IndonesiaFil: Igarashi, Ikuo. Obihiro University of Agriculture and Veterinary Medicine. National Research Center for Protozoan Diseases; JapónFil: Florin-Christensen, Mónica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; ArgentinaFil: Florin-Christensen, Mónica. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; ArgentinaFil: Schnittger, Leonhard. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Detection and molecular characterization of rabies virus in Mongolia during 2008-2010

Aim: We aimed to investigate the prevalence and molecular characterization of rabies virus (RABV) from wild and domestic animals in Mongolia during 2008-2010. Materials and Methods: Brain tissue samples were collected from 24 rabid animals in Zavkhan, Omnogovi, Tov, Dundgovi, Govi-Altai, Selenge, Ovorkhangai, and Khentii provinces in Mongolia. Herein, samples were included from 13 domestic animals (dogs, cattle, camels, sheep, and goat) and 11 wild animals (wolves and foxes) in this study. Direct fluorescent antibody (DFA) test and reverse transcriptase polymerase chain reaction (RT-PCR) were performed for detection of RABV, and positive samples were further processed for molecular characterization of the virus using nucleoprotein gene. Subsequently, the molecular characterization was determined based on the nucleoprotein gene. Results: Out of 24 samples, 22 samples were detected positive for RABV by DFA test, and its nucleoprotein gene was amplified in all of the 24 samples by RT-PCR. These Mongolian RABVs were classified within steppe-type virus clade by phylogenetic analysis of nucleoprotein gene sequences. This steppe-type virus clade was clearly divided by two Sub-clades (A and B). The most of Mongolian RABVs belongs to the Sub-clade A in the phylogenetic tree. Conclusion: These findings have clearly confirmed RABV in domestic and wild animals of Mongolia. Further molecular characterization indicated that this Mongolian strain is steppe-type virus clade consisting of two sub-clades; the Subclade A might be prevalent in Altai, Khangai, Khentii Mountains as a major genotype, whereas the Subclade B seems to be cosmopolitan in the steppe-type virus clade, is spread in northern central Eurasia

Molecular identification and risk factor analysis of the first Lumpy skin disease outbreak in cattle in Mongolia

Lumpy skin disease (LSD) is a transboundary viral infectious disease in cattle caused by a Capripoxvirus. LSD has been recently introduced in some Asian countries. However, in Mongolia, no report of LSD is publicly available. We clinically examined LSD symptoms in 1,034 cattle from 4 soum (district) in Dornod province in Mongolia. Sixty-one cattle of them were confirmed with symptoms of LSD and then viral P32 gene was detected by a PCR. The overall prevalence of LSD in cattle was 5.9%. Females odds ratios (OR)=2.27 than males, adults (>2.5-years-old, OR=3.68) than young (1-2.5-years-old) and calves (<1-year-old) were at higher risks for LSD cases in Mongolia, while locations near the tube well and pond water are major risk areas for viral transmission due to density of insects often is high. For virus isolation, skin nodule tissue samples of 4 cattle located in four distinct soums were used for viral propagation using the MDBK cell line. Internal terminal repeat region and RPO30 gene of 4 Mongolian isolates were amplified and sequenced. In the phylogenetic trees, Mongolian LSDVs (2021) were clustered together with the Chinese (2020) and Vietnamese isolates (2020). This is the first report alarming the LSD outbreak in Mongolia that was confirmed by our study. The newly isolated viruses would be a useful base for developing diagnostic tools and inactivated vaccine technology. A large-scale study of LSD is next priority for establishing successful control strategy of further disease outbreak