Orf virus circulation in cattle in Turkey

Orf virus (ORFV) causes contagious skin disease that mainly affects sheep and goats with zoonotic potential. However, there is not enough information about the association between ORFV and occurrence of skin disease in cattle. The present study describes outbreaks of ORFV infection in cattle in different provinces that are located in the Aegean, Central Anatolian and Mediterranean regions of Turkey. During the months of June and August 2017, vesicular fluid and scab samples were collected from cattle which had proliferative skin lesions. First, presence of lumpy skin disease virus (LSDV) and bovine herpesvirus 2 (BoHV-2, known as the causative agent of pseudo-lumpy skin disease) were investigated by real time PCR and PCR, respectively. Then, samples tested for the presence of parapoxviruses by PCR using primers specific to major envelope protein gene (B2L). Parapoxvirus DNA was detected in investigated samples whereas LSDV and BoHV-2 DNA were not detected. The analysis of the B2L gene sequences revealed that cattle were infected with ORFV. The isolates in the present study shared 100% sequence identity at the nucleotide and amino acid level when compared with previously characterised Turkish field ORFV isolates from goats in 2016. Results of the study show unusual infection of cattle with ORFV, and suggest that ORFV jumps the host species barrier from goats to cattle.     

Infection of sheep and goats with bovid herpesvirus 2

Sheep and goats were shown to be susceptible to experimental infection with bovid herpesvirus 2 (BHV2), administered by either the intradermal or intravenous route. Lesions developing in sheep following intradermal inoculation of virus were similar to those in cattle inoculated intradermally, whereas the lesions in goats resolved without ulceration or scabbing. Disseminated circumscribed skin lesions developed in sheep and goats given BHV2 by the intravenous route. These lesions resolved in four to eight days without significant effect on the skin. BHV2 was isolated from skin lesions of sheep, goats and cattle that were infected intravenously, from sheep and cattle infected intradermally and from the leucocytes of the three species following intravenous inoculation of virus. Latent infection of sheep and goats was demonstrated following corticosteroid treatment 60 days after infection.     

Severe persistent orf in young goats.

Orf (contagious ecthyma) is a viral disease of small and wild ruminants, humans, and less frequently other species. In sheep and goats, the disease is characterized by the formation of vesiculo-proliferative lesions in the skin of lips and nostril. Here, a form of generalized orf in 16 goat kids from 2 different locations in west Texas is described. The disease was characterized by multifocal, severe, proliferative dermatitis that persisted from about 2 months of age until the goat kids were euthanized 3 months later. All affected goats were Boer or Boer crosses under 1 year of age. The mean immunoglobulin concentration in sera of affected goats was elevated compared with healthy control goats. Severe to moderate lymphadenomegaly of the nodes draining the areas of the skin affected with orf lesions was present in all 16 goat kids. Suppurative arthritis, chronic fibrinous pneumonia, and premature thymic involution were found in 3, 5, and 7 of the goat kids, respectively. The skin lesions of 3 goat kids were infested with larvae of the opportunistic black garbage fly (Ophira sp.). The orf virus was identified in skin lesions by isolation in Marbin-Darby ovine kidney cells, electron microscopy, and amplification of viral DNA by polymerase chain reaction. The orf virus was not detected in peripheral blood or lymph node mononuclear cells of any of the goats. Cross-neutralization experiments showed that an ovine orf virus antiserum raised in sheep was more effective in neutralizing a sheep orf virus isolate than a caprine orf virus isolate. The clinical and epidemiological characteristics of these orf cases may be the result of susceptibility factors within some individuals of the Boer breed of goats.     

The pathology of heartwater. III. A review

The pathological changes in cattle, sheep, goats, mice and various game species infected with Cowdria ruminantium are summarized. Macroscopical lesions in most animals include effusion of body cavities, oedema of the lungs and lymph nodes and splenomegaly. Apart from the presence of heartwater organisms in most organs the histopathological lesions are not striking. The ultrastructural lesions in the lungs of sheep and goats infected with the Ba11 3 strain of heartwater, and mice infected with the Welgevonden strain, are discussed. Damage to capillary endothelial cells of the alveoli is limited and the mild cytopathic changes in parasitized cells indicate that the damage caused by the organisms is most probably not responsible for the increased vascular permeability associated with the disease. Pathological changes in domestic ruminants and game animals are briefly compared.     

Comparative molecular characterization of Corynebacterium pseudotuberculosis of different origin

Ribotyping and susceptibility to 17 antimicrobial agents were used to compare 37 isolates of Corynebacterium pseudotuberculosis (28 from horses, 1 from cattle, 3 from sheep and 5 from goats) derived from various types of lesions, and different geographic locations. According to the presence of nitrate reductase, all but one isolate from horses reduced nitrate (nitrate-positive), whereas all isolates from sheep and goats were unable to reduce nitrate (nitrate-negative). The ribotype of the nitrate-negative isolate from a horse with ulcerative lymphangitis was identical to all the other isolates from horses, and different than the ribotype of nitrate-negative isolates from sheep and goats. Ribotyping with one of the restriction endonucleases, Apa I, revealed differences between, but not within, the two biotypes. However, ribotyping with Pst I endonuclease revealed one variant within the equine biotype and one variant within the ovine biotype. The minimum inhibitory concentration (MIC; μg/ml) of antimicrobial agents against isolates from nitrate-negative and nitrate-positive groups was very similar, with the exception of isolates from sheep and goats which had a higher MIC for amikacin than isolates from horses and cattle.     

Seroprevalence of ovine progressive pneumonia virus in various domestic and wild animal species, and species susceptibility to the virus

Ovine progressive pneumonia is caused by a lentivirus of known infectivity only for sheep and goats. Virus susceptibility of 11 other species of animals was examined. Species included cattle, chickens, deer, dogs, goats, hamsters, horses, mice, pigs, rabbits, and rats. Of these species, only goats and rabbits could be experimentally infected with the virus. The infection in rabbits was acute, and virus did not persist or induce antibody production as it does in sheep and goats. Sera obtained from several people working in close contact with the virus and from several wild species, with unknown exposure history, were tested for antibodies to viral antigens. All results were negative. Knowledge of the host range of this virus is important for scientific studies and for virus eradication programs.     

Characterization of bovine herpesviruses isolated from six sheep and four goats by restriction endonuclease analysis and radioimmunoprecipitation

Viral DNA from 10 herpesviruses isolated from 6 sheep and 4 goats were examined by restriction endonuclease analysis with respect to their relatedness to one another; to bovine herpesvirus type 6 (BHV-6), also known as caprine herpesvirus; and to 2 strains of bovine herpesvirus type 1 (BHV-1), known as infectious bovine rhinotracheitis virus (IBRV) and infectious pustular vulvovaginitis virus (IPVV). Viral proteins from the isolates were examined by radioimmunoprecipitation with anti-BHV-1/IBRV gnotobiotic calf (bovine) serum, anti-BHV-1/IBRV bovine hyperimmune serum, and anti-BHV-6 rabbit serum to evaluate their antigenic relatedness to each other. The goat isolates were obtained from animals with various disease conditions including respiratory tract disorders, vulvovaginitis, and wart-like lesions on the eyelid. The other isolates were from domestic sheep and came from aborted fetuses or from sheep with fatal pneumonia or proliferative lesions around lips and nose. All of the goats and 4 of the sheep from which the viral isolates were obtained had comingled with cattle. Purified DNA from each of the 10 field isolates and from BHV-1/IBRV, BHV-1/IPVV, and BHV-6 caprine herpesvirus was cleaved with restriction endonuclease Pst I. Five of 6 sheep isolates and 3 of 4 goat isolates yielded unique restriction patterns, ie, patterns that differed from each other by one or more bands. Sheep isolate DNA patterns were different from goat isolate patterns, and all restriction endonuclease analysis patterns were similar to the pattern for BHV-1/IBRV, but different from that for BHV-1/IPVV or for BHV-6.(ABSTRACT TRUNCATED AT 250 WORDS)     

Experimental infection of sheep by caprine arthritis-encephalitis virus and goats by progressive pneumonia virus

The lentiviruses, caprine arthritis-encephalitis virus (CAEV) and progressive pneumonia virus (PPV) of sheep, cause major diseases in their respective hosts; however, the infectivity of these viruses for closely related species has not been determined. Experiments were conducted to determine whether CAEV would infect sheep and whether PPV would infect goats. Upon inoculation with CAEV, lambs developed a nonsuppurative arthritis and antibody to CAEV, and the virus was isolated up to 4 months later. Exposure of 3 lambs to CAEV-infected adult goats did not lead to demonstrable infection after 18 months. Young goats inoculated with PPV replicated the virus and developed arthritis and antiviral antibody. These results demonstrate that these distinctly different lentiviruses may infect and cause diseases in species other than their accustomed host. Presently used techniques may not be effective in differentiating which lentivirus is responsible for infection of sheep and goats. Our results also indicate that mixing sheep and goats may adversely influence attempts to eradicate lentiviruses from these species.     

The pathology of heartwater. I. A study of mice infected with the Welgevonden strain of Cowdria ruminantium

Gross and microscopical lesions in mice intravenously infected with the Welgevonden strain of Cowdria ruminantium closely resembled the lesions described in cattle, sheep and goats. A high concentration of organisms was present in alveolar endothelial cells. Cytopathic changes in parasitized and non-parasitized endothelial cells and the morphology of the organisms are described and compared with the Ball3 strain of C. ruminantium. Possible mechanisms in the development of the lung oedema are considered and the role of mice as animal model is discussed.     

Bovine viral diarrhoea virus infection in cattle,sheep and goats in northern Tanzania

Virological and serological investigations were carried out in cattle, sheep and goats raised in the northern part of Tanzania in order to explore the possibility of bovine viral diarrhoea (BVD) virus cycling within these animal species. Two noncytopathogenic BVD virus isolates (A4/5/Tan86 and A4/10/Tan86) were obtained from sera sampled from inapparently infected indigenous (zebu) cattle originating from Kiteto district in Arusha region. No BVD virus was isolated from any of the sheep or goat sera. Seroepidemiological investigations revealed widespread prevalence of neutralising antibodies to BVD virus not only in cattle but also in sheep and goats. The seropositive rates are discussed in relation to previous observations in Tanzania and other parts of the world and to the livestock husbandry practices in northern Tanzania.     

牛结节性皮肤病病毒感染MDBK细胞后转录组差异表达分析

旨在探索牛结节性皮肤病病毒（lumpy skin disease virus,LSDV）与宿主细胞的相互作用,采集了具有典型临床症状的牛皮肤病变组织,接种MDBK细胞进行病毒分离,采用透射电镜观察病变组织及病毒感染后的MDBK细胞,同时对感染后24 h的MDBK细胞进行转录组学分析,筛选差异表达基因并将其按功能分类,分析它们参与的细胞信号通路。结果显示,LSDV在电镜下呈椭圆形,病毒粒子大小约320 nm×260 nm。转录组分析发现,与未处理组相比,显著差异表达的基因共499个,其中,112个基因上调表达,387个基因下调表达。通过GO和KEGG分析得知,差异表达基因主要富集在从细胞表面传导信号至细胞核内部的MAPK信号通路,其中,参与天然免疫且涉及细胞凋亡的MAP3K1基因表达呈现显著下调,提示病毒感染会影响宿主细胞的迁移与凋亡等过程。显著变化基因包括HIF1A、RORA、LRRK2、AP3B1和ANO6等,分别涉及到调节血管内皮生长因子、炎症反应、细胞增殖分化以及信号转导等功能,除此之外,不饱和脂肪酸及胆汁分泌两个信号通路也发生了显著变化。本研究为进一步深入探究LSDV的致病机制奠定了基础。     

The role of goats as reservoir hosts for bovine herpes virus 1 under field conditions

Bovine herpesvirus 1 (BoHV1) is the cause of economically significant viral infections in cattle. Respiratory symptoms associated with the infection are known as Infectious Bovine Rhinotracheitis (IBR). Sheep and goats are less sensitive to the infection although their role in inter-species viral transmission under field conditions is subject to controversy. The objective of this study was to investigate seroprevalence of BoHV1 infections in cattle, sheep, and goats raised together for at least a year. Blood serum samples were taken from 226 cattle, 1.053 sheep, and 277 goats from 17 small- to medium-scale farms. BoHV1-specific antibody presence and titers were determined using virus neutralization test. In total, 73 of the 226 cattle (32.3%) were seropositive. The infection was detected in 13 of the 17 farms. Infection rates ranged from 5.8 to 88.8%. Only one of the 1053 sheep (0.09%) was seropositive. However, 58 of the 277 (20.9%) goats were seropositive. Goat samples taken from 8 of the 17 farms were seropositive with infection rates ranging from 17 to 38.9%. Statistical analysis showed a significant correlation in infection rates between cattle and goats but not sheep. These results suggest that goats may be more sensitive to the BHV1 infection than sheep and the role of goats as possible reservoirs for BoHV1 in the control and eradication of BHV1 in cattle should be considered in future studies.

     

藏系绵羊VEGF基因的克隆、同源性分析及其表达水平的实时荧光定量PCR检测

采用PCR技术从藏系绵羊皮肤组织细胞中扩增出了血管内皮细胞生长因子(VEGF)基因,长度为573 bp,共编码190个氨基酸;序列比对发现,VEGF基因在所选择的多个物种之间具有高度同源性,相似性在100%～74.5%之间,其中藏系绵羊与普通绵羊的VEGF基因序列完全相同,其次为牛和猪,相似性分别为99.1%和96.2%,而与鸡的同源性最低,相似性为74.5%。采用实时荧光定量PCR SYBR Green I荧光染料法,对藏系绵羊皮肤组织细胞中VEGF基因的表达水平进行了相对定量分析,建立了快速检测VEGF基因表达量的方法。检测结果表明:给妊娠后期藏系母羊补饲自制的营养舔砖和颗粒饲料后,所产羔羊体内VEGF基因的表达水平分别为对照组的11.5倍和0.85倍。为提高羔羊皮肤组织中VEGF基因mRNA的表达水平,对妊娠后期藏系母羊补饲自制的营养舔砖,效果优于补饲颗粒饲料。     

Use of the Capripoxvirus homologue of Vaccinia virus 30 kDa RNA polymerase subunit (RPO30) gene as a novel diagnostic and genotyping target: development of a classical PCR method to differentiate Goat poxvirus from Sheep poxvirus

Sheep poxvirus (SPPV), Goat poxvirus (GTPV) and Lumpy skin disease virus (LSDV) are Capripoxviruses (CaPVs) responsible for causing severe poxvirus disease in sheep, goats and cattle, respectively. Serological differentiation of CaPVs is not possible and strain identification has relied on the implicitly accepted hypothesis that the viruses show well defined host specificity. However, it is now known that cross infections can occur and authentication of identity based on the host animal species from which the strain was first isolated, is not valid and should be replaced with molecular techniques to allow unequivocal strain differentiation. To identify a diagnostic target for strain genotyping, the CaPV homologue of the Vaccinia virus E4L gene which encodes the 30 kDa DNA-dependent RNA polymerase subunit, RPO30 was analyzed. Forty-six isolates from different hosts and geographical origins were included. Most CaPVs fit into one of the three different groups according to their host origins: the SPPV, the GTPV and the LSDV group. A unique 21-nucleotide deletion was found in all SPPV isolates which was exploited to develop a RPO30-based classical PCR test to differentiate SPPV from GTPV that will allow rapid differential diagnosis of disease during CaPV outbreaks in small ruminants.     

Molecular epidemiology of Echinococcosis from food producing animals in north India

Echinococcosis is an important medical, veterinary and economic concern in India. Ten cysts were randomly selected from each intermediate host species (cattle, buffalo, sheep, goat and pigs). Either the germinal layer (sterile cysts) or protoscoleces (fertile cysts) were collected for molecular characterization. A 434 base pair fragment of the mitochondrial cytochrome oxidase-1 gene was amplified using PCR from each isolate. Ten representative samples (2 from each intermediate host species) were sequenced in both the directions from which readable sequences were obtained from nine for phylogenetic analysis (NCBI, Blast). Phylogenetic analysis of cytochrome oxidase I gene revealed that seven (77.7%) isolates, from cattle (2), pigs (2), buffaloes (1) and goat (2) were clustered with the Indian Buffalo (G3) strain of Echinococcus granulosus, while two (22.2%) isolates from sheep were clustered with the sheep strain (G1) of E. granulosus. Phylogenetic analysis of the cytochrome oxidase-1 gene revealed that the buffalo strain (G3) and common sheep strain (G1) are cycling among livestock in north India and that these strains are highly adapted to cattle, buffalo, sheep, goats and pigs.     

IS900 restriction fragment length polymorphism (RFLP) analysis of Mycobacterium avium subsp. paratuberculosis isolates from goats and cattle in Norway

In Norway, paratuberculosis has been frequently diagnosed in goats, while cattle have been almost free of the infection. This difference in prevalence between goats and cattle has led to speculations about the existence of a Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis) isolate that is non-pathogenic for cattle. There is little information available on genotypic variation of M. a. paratuberculosis isolated from animals in Norway. In the present study, genotypic information on 51 isolates from goats and four isolates from cattle in Norway was obtained by use of IS900 restriction fragment length polymorphism (RFLP) analysis. All isolates from cattle and 84% of the isolates from goats had the same RFLP pattern (B-C1). Five RFLP patterns not previously detected were found. No genotypic variation that could explain a difference in host origin was found between the isolates from cattle and the majority of the Norwegian goat isolates. This lack of difference indicates that the most common M. a. paratuberculosis isolates in Norway may infect both cattle and goats.     

Ovine progressive pneumonia: pathologic and virologic studies on the naturally occurring disease

Pathologic and virologic studies were conducted on 13 mature ewes with serum precipitin antibodies to progressive pneumonia virus (PPV). Pulmonary lesions of ovine progressive pneumonia were found in 4 sheep, a meningoencephalitis resembling visna in 1 sheep, chronic proliferative carpal arthritis in 2, and massive lymphoid proliferation in the mammary gland in 3. Virus producing cytopathic effect typical of PPV was isolated from the lungs, mediastinal lymph node, spleen, and choroid plexus of 4 sheep and from the carpal synovium of 2 sheep with chronic carpal arthritis. Three viral isolates selected for further study were antigenically related to visna virus by immunofluorescence and immunodiffusion, but these 3 isolates were not neutralized by antisera to reference strains of visna virus. Seemingly, infection of sheep by ovine retroviruses is common in the United States, and these viruses are capable of causing disease in more than 1 organ system.     

Cultural characteristics and virulence of strains of Fusobacterium necrophorum isolated from the feet of cattle and sheep

Sixty-one isolates of Fusobacterium necrophorum were recovered for study. Thirty-one were obtained from lesions of foot abscess in cattle (25) and sheep (6), 28 were from interdigital lesions in cattle and 2 were from the normal interdigital skin of cattle. The majority of isolates from lesions of foot abscess were virulent, belonged to biotype AB (Fievez 1963), produced flat, irregular shaped, greyish colonies and haemolysis on blood agar, and grew as turbid filamentous suspensions in liquid media. They produced a soluble exotoxin, a leucocidin, and were pathogenic for cattle and mice. Virulent isolates also produced a haemolysin which most readily lysed bovine, equine and chicken erythrocytes; those from sheep were less susceptible while those of rabbit and pig were the most resistant. Isolates recovered from lesions of the feet not classified as foot abscess and from clinically normal feet were predominantly of the B biotype and caused few experimental lesions, produced convex, round, yellow colonies, flocculated and sedimented while growing in liquid medium and produced little or no haemolysin or leucocidin. Routine differentiation between virulent and non-virulent bovine isolates of F. necrophorum could be achieved by assessing the colour, morphology, and degree of haemolytic activity of colonies grown on blood agar.     

血管内皮生长因子在绵羊肺脏中的表达分布研究

目的研究血管内皮生长因子（vascular endothelial growth factor,VEGF）在绵羊肺脏中的表达分布特征。方法取成年绵羊肺脏组织,制备石蜡切片,利用HE染色法观察绵羊肺脏组织的形态结构,采用免疫组织化学方法检测VEGF在绵羊肺脏组织中的分布。结果肺脏的各类型细胞均可见VEGF表达,在绵羊肺脏导气部的细支气管和终末细支气管的上皮细胞,呼吸部的肺泡管和呼吸性细支气管的上皮细胞,以及肺的血管内皮细胞均可检测到VEGF的强阳性表达信号。结论VEGF广泛分布于绵羊肺脏组织中,对其形态结构和功能的维持具有重要作用。     

Peste des petits ruminants infection in domestic ruminants in Sudan

The existence of peste des petits ruminants (PPR) in domestic ruminants and camels in Sudan during 2008–2012 was investigated. Lung tissues and serum samples were randomly collected from sheep, goats, cattle, and camels at different areas of Sudan. A total of 12,384 serum samples were collected from clinically healthy 7413 sheep, 1988 camels, 1501 cattle, 1459 goats, and 23 gazelles at different areas in the Sudan. They were examined for PPR antibodies using competitive ELISA (cELISA). The overall detected seroprevalence of PPR in tested sera was 49.4%; seroprevalence values within species were 67.1, 48.2, 25.8, 2.1, and 21.7% in sheep, goat, cattle, camels, and gazelles, respectively. The highest seroprevalence (68.1%) was observed in sera collected from Darfur states, then the central states (54.3%). A total of 1276 lung tissue samples (623 sheep, 324 cattle, 220 camels, and 109 goats) were collected. The majority of lung samples were collected from clinically healthy animals that showed lesions on PM in slaughterhouses (95%) and during PPR outbreaks; samples were tested for PPR antigen using immunocapture ELISA (IcELISA). PPR antigen was detected in 233 out of the 1276 tested samples (18.3%). Positive results were observed in samples collected from clinically healthy and diseased animals. The observed prevalence values in each species were 33.6, 21.1, 15.4, and 12.3% in camel, goat, sheep, and cattle, respectively. PPR antigen was detected in samples from different areas; however, the highest prevalence (63.9%) was found in samples collected from the eastern states, then Khartoum state (28%). Trials for virus isolation were done in different cell cultures. Out of 30 IcELISA-positive samples inoculated in primary bovine and ovine kidney cells, Vero cells, the PPR virus was successfully isolated from 15 (eight sheep, five camels, and two goats) samples in the three cell culture types. Using RT-PCR, PPRV nucleic acid was detected in all 25 IcELISA-positive tested samples.