Occurrence and spatial distribution of Toggenburg Orbivirus in Switzerland

Recently, a new member of the Bluetongue virus (BTV) serogroup named Toggenburg Orbivirus (TOV) in goats from Switzerland has been described. The epidemiology and host range of TOV are currently unknown. Since TOV causes cross-reactions in laboratory tests used for BTV diagnosis, this study was carried out in order to determine the spatial and temporal spread of TOV. Therefore, serum samples from a national survey in goats, collected during winter and spring 2008 in Switzerland, were serologically examined. Additionally, cattle and sheep from holdings with seropositive goats were tested for the presence of viral RNA and antibodies against BTV and TOV. All goat samples analysed within routine diagnostics at the Institute of Virology and Immunoprophylaxis from 2008 to 2009 were also tested for the presence of TOV. Finally, goat sera collected 1998 in the Canton of Ticino (TI) were analysed.Although the TOV index cases had been identified in flocks north of the Alps, no additional TOV-positive herds were found by serological testing in this region. In contrast, south of the Alps, i.e. in the Canton of Ticino (TI), an apparent seroprevalence of 49% in goats was found at animal and 60% at herd level. In the eastern and western part of the Swiss Alps 15.2% and 10% of tested goats were serologically positive, respectively. A within-herd prevalence of up to 100% was found in some of the positive flocks. The positive flocks in TI were mainly found in three of the five districts, but seropositive animals were identified in each district. Certain selected seropositive flocks were investigated virologically. By RT-qPCR and genome sequencing, the presence of TOV could be confirmed in all investigated seropositive flocks.By testing the goats within routine diagnostics, TOV genome was detected in one goat showing BT-like clinical symptoms from the central Alps and in three healthy animals imported from Germany.Although 3.8% of the sheep from flocks with TOV-positive goats or in contact with these animals showed a positive antibody reaction, TOV-specific RNA was not found in any of the tested sheep and also not in cattle from flocks with TOV-positive goats.Serological and virological test results from archived Swiss goat samples collected in 1998 indicated the presence of TOV already at that time, prior to any Bluetongue disease outbreak in this part of Europe. The results of this study demonstrate that TOV is widespread in certain parts of Switzerland and suggests that this virus has been present in the goat population for at least a decade, albeit without causing any disease signs.     

Lack of neutralizing antibodies to caprine arthritis-encephalitis lentivirus in persistently infected goats can be overcome by immunization with inactivated Mycobacterium tuberculosis

The pathogenesis of the persistent progressive diseases of sheep (visna-maedi) and goats (arthritis-encephalitis) is dependent on continuous replication of the causative lentiviruses. One subgroup of these viruses, Icelandic visna virus, accomplishes this form of replication by undergoing antigenic mutation. Mutant viruses arising late in the infection escape neutralization by antibodies directed to the parental virus. In contrast, we show here that viruses obtained from persistently infected sheep and goats with natural disease in this country do not induce virus-neutralizing antibodies, although antibodies to virus core proteins were produced. The lack of neutralizing antibodies was not overcome by hyperimmunization of animals with concentrated preparations of live or inactivated virus. Thus, failure to produce these specific antibodies was not due to lack of sufficient antigen or interference with the immune response because of the ability of these viruses to infect macrophages. The hyporesponsive state, however, was overcome by immunization of animals with virus and large amounts of inactivated Mycobacterium tuberculosis. Induction of agglutinating and neutralizing antibodies by this method was probably due to a unique form of antigen processing by macrophages activated by M. tuberculosis. Neutralizing antibodies were produced for the first time against the caprine arthritis-encephalitis virus by this method. These antibodies have similar biological properties to those induced by Icelandic visna virus. They belong to the immunoglobulin G1 subclass, they are effective against a narrow range of caprine arthritis-encephalitis viruses, and they identify (for the first time) antigenic variants among these caprine agents.     

Detection of Leptospira spp. in semen and vaginal fluids of goats and sheep by polymerase chain reaction

Thirteen goat herds and seven sheep flocks in the state of Rio de Janeiro, Brazil were screened for leptospirosis. From the three herds and three flocks with greatest seroreactivity, 19 goats (16 females and three bucks) and 40 sheep (26 ewes and 14 rams) that were seropositive (specific anti-Leptospira titres > or =400, based on a microscopic agglutination test), were selected for more detailed studies. From those animals, samples of vaginal fluids or semen were collected for bacteriological and molecular assays. For both species of animals, the most prevalent reactions were to serovars Hardjo, Shermani, and Grippotyphosa. Although leptospires were detected by darkfield microscopy in three vaginal fluid samples (from two goats and one ewe), pure isolates were not obtained by bacteriological culture of vaginal fluids or semen. However, seven vaginal fluid samples (from four goats and three ewes) and six semen samples (all from rams) were positive on polymerase chain reaction (PCR). Based on these findings, in addition to analogous findings in cattle, we inferred that there is potential for venereal transmission of leptospirosis in small ruminants.     

Peste des Petits Ruminants Virus Tissue Tropism and Pathogenesis in Sheep and Goats following Experimental Infection

Peste des petits ruminants (PPR) is a viral disease which primarily affects small ruminants, causing significant economic losses for the livestock industry in developing countries. It is endemic in Saharan and sub-Saharan Africa, the Middle East and the Indian sub-continent. The primary hosts for peste des petits ruminants virus (PPRV) are goats and sheep; however recent models studying the pathology, disease progression and viremia of PPRV have focused primarily on goat models. This study evaluates the tissue tropism and pathogenesis of PPR following experimental infection of sheep and goats using a quantitative time-course study. Upon infection with a virulent strain of PPRV, both sheep and goats developed clinical signs and lesions typical of PPR, although sheep displayed milder clinical disease compared to goats. Tissue tropism of PPRV was evaluated by real-time RT-PCR and immunohistochemistry. Lymph nodes, lymphoid tissue and digestive tract organs were the predominant sites of virus replication. The results presented in this study provide models for the comparative evaluation of PPRV pathogenesis and tissue tropism in both sheep and goats. These models are suitable for the establishment of experimental parameters necessary for the evaluation of vaccines, as well as further studies into PPRV-host interactions.     

Sequence homology between cloned caprine arthritis encephalitis virus and visna virus, two neurotropic lentiviruses.

Caprine arthritis encephalitis virus (CAEV) is an exogenous, nononcogenic retrovirus which causes neurological disease and crippling arthritis in goats. A complete CAEV genome was cloned from unintegrated viral DNA in two fragments of 9.4 and 0.4 kilobases in length, respectively. The biological activity of these clones was tested by ligation of the fragments followed by transfection onto goat synovial membrane cells; infectious virus was recovered. Cloned CAEV and visna virus, a related neurotropic virus of sheep, were compared by heteroduplex and molecular hybridization analyses. These data demonstrated that the greatest overall conservation of nucleotide sequences occurred in the gag and pol gene regions and two smaller regions, sor and the putative tat gene. The region of greatest divergence occurred in the env gene and, in particular, was localized primarily in the region coding for the glycosylated outer membrane protein. These findings and the recently demonstrated genetic relationship of visna virus, CAEV, and human T-cell lymphotropic virus type III, the etiologic agent of the acquired immune deficiency syndrome, may have important implications concerning the biological properties of these related viruses for human and veterinary medicine.     

Corynebacterium Pseudotuberculosis Infection in Goats IV.

Adult animals from 2 herds were examined clinically and serologically, 5 (Herd A) and 4 (Herd B) times during the same period of 3% years. Serum samples were examined for antibodies against Gorynebac-terium pseudotuberculosis using the bacterial agglutination test (BAT) and the hemolysis inhibition test (HIT). The results of the first examination showed that no animal in Herd A was seropositive, while in Herd B 1 animal showed a high positive titre in BAT. The prevalence of animals with superficial swellings was then 2 % in Herd A and 4 % in Herd B. In both herds, the prevalence of animals with superficial swellings and seropositive reactions increased during the following 1–2 yeairs. About 30 % of animals had superficial lesions and close to 100 % were seropositive. The proportion, of animals with superficial swellings and seropositive reactions was almost constant on subsequent examinations. In some of the animals, superficial swellings were found during 2 or more of the examinations, a few animals having such lesions at the same site on both or all occasions. Animals in Herd A became infected through grazing together with goats from infected herds. Caseous lymphadenitis was introduced into Herd B by animals obtained from infected herds.     

Paratuberculosis (Johne's disease) in bighorn sheep and a Rocky Mountain goat in Colorado.

Between May, 1972 and February, 1978, six cases of paratuberculosis (Johne's Disease) caused by Mycobacterium paratuberculosis were diagnosed in free-ranging Rocky Mountain bighorn sheep (Ovis canadensis) and one Rocky Mountain goat (Oreamnos americanus) on or near Mt. Evans in Colorado. Diagnosis of paratuberculosis was based on gross and histopathologic examination of the animals and by isolation of M. paratuberculosis from three sheep and the goat. The clinical signs and pathologic changes seen in the bighorn sheep resembled those described in cattle, while the lesions in the goat were similar to those described for domestic sheep and goats.     

Pestivirus Infections in Norway. Serological Investigations in Cattle,Sheep and Pigs

Serum samples from 1133 dairy cows (187 herds), 3712 ewes (103 flocks) and 1317 adult pigs (877 herds), were tested for neutralizing antibodies against the NADL strain of bovine virus diarrhoea virus. The prevalence rate of seropositive animals was 18.5% in cattle, 4.5% in sheep and 2.2% in pigs, such seroreactors being found in 28 % of the cattle herds and 18 % of the sheep flocks. In all three species the rate showed considerable herd and geographical variation. In cattle the seroreactor rate was similar in herds with normal reproduction and in 62 herds with problems of repeat breeding. Of 31 pig sera containing antibodies against the NADL strain, 27 were also positive in a neutralization test for antibodies against swine fever virus (Baker strain). However, all sera showed a higher titre of antibodies against the bovine strain than against the swine fever virus. It was concluded that the immune response of the pigs had been induced by ruminant pestivirus, and not by swine fever virus.     

Characterization of parapoxviruses circulating among wild Japanese serows (Capricornis crispus)

We antigenically and molecularly compared 5 parapoxvirus isolates and 7 viral DNA samples from clinical lesions of Japanese serows with 3 viruses from sheep and goats. All isolates from Japanese serows except one, Ishikawa-S, reacted with six monoclonal antibodies to orf virus (ORFV). Restriction endonuclease analysis using amplified viral DNA showed the ORFV-specific pattern in all samples except Ishikawa-S, which showed a bovine papular stomatitis virus (BPSV)-specific pattern. Partial nucleotide sequences of the envelope genes were determined and those of all samples from Japanese serows and sheep except Ishikawa-S were completely identical and also had high identities with the goat virus. These findings suggest that parapoxvirus infection in Japanese serows might be mainly caused by ORFV and accidentally by BPSV. The envelope gene sequenced here seems to be conserved in Japanese ORFVs.     

Functional Murine Leukemia Virus Vectors Pseudotyped with the Visna Virus Envelope Show Expanded Visna Virus Cell Tropism

Pseudotype virus vectors serve as a powerful tool for the study of virus receptor usage and entry. We describe the development of murine leukemia virus (MuLV) particles pseudotyped with the visna virus envelope glycoprotein and encoding a green fluorescent protein reporter as a tool to study the expression of the visna virus receptor. Functional MuLV/visna virus pseudotypes were obtained when the cytoplasmic tail of the visna virus envelope TM protein was truncated to 3, 7, or 11 amino acids in length. MuLV/visna virus particles were used to transduce a panel of cell types from various organisms, including sheep, goat, human, hamster, mouse, monkey, and quail. The majority of the cells examined were susceptible to MuLV/visna pseudotype viruses, supporting the notion that the visna virus cellular receptor is a widely expressed protein found in many species. Of 16 different cell types tested, only mouse embryo fibroblast NIH 3T3 cells, hamster ovary CHO cells, and the human promonocyte cell line U937 cells were not susceptible to transduction by the pseudotyped virus. The production of functional MuLV/visna virus pseudotypes has provided a sensitive, biologically relevant system to study visna virus cell entry and envelope-receptor interactions.     

A serologic investigation of epizootic hemorrhagic disease virus in China between 2014 and 2019

Epizootic hemorrhagic disease virus (EHDV) is a member of the genus Orbivirus, family Sedoreoviridae. It was firstly recognized in 1955 to cause a highly fatal disease of wild white-tailed deer in America. So far, EHDV was detected and isolated in many wild or domestic ruminants, and widely distributed all over the world. Although the domestic cattle and sheep infected by EHDV were usually asymptomatic or subclinical, several outbreaks of epizootic hemorrhagic disease (EHD) in deer and cattle had been reported. Many EHDV strains were isolated and sequenced in last two decades in China, which promoted a general serologic investigation of EHDV in China. In this study, 18,122 sera were collected from asymptomatic or subclinical domestic ruminants (cattle, cow, yaks, sheep, goats, and deer) in 116 regions belonging to 15 provinces in China. All the sera were tested by EHDV C-ELISA, and the results were obtained by big data analysis. EHDV infections were detected in the 14 of 15 provinces, and only Tibet (average altitude ≥ 4000 m) which was the highest province in China was free of EHDV. The numbers of seropositive collections in both bovine and goat/sheep were in an inverse proportion to the latitude. However, the seropositive rates in bovine were ranged from 0% to 100%, while the seropositive rates in goat/sheep were no more than 50%. The results suggested that bovine was obviously more susceptive for EHDV infection than goat and sheep, therefore might be a major reservoir of EHDV in China. The prevalence of EHDV was consistent with the distribution of Culicoides which were known as the sole insect vectors of EHDV. In particular, the seropositive rates of EHDV were very high in the southern provinces, which required the enhanced surveillance in the future.     

Evaluation of an indigenous ELISA for diagnosis of Johne’s disease and its comparison with commercial kits

Country lacks sensitive and indigenous diagnostic kits for the screening of goats and sheep against Johne’s disease. Therefore an indigenous ELISA kit was developed using protoplasmic antigen from native Mycobacterium avium subspecies paratuberculosis ‘Bison Type’ strain of goat origin (Kit 1). In the present study, kit 1 and two commercial kits (kit 2 and 3) were evaluated with respect to ‘Gold Standard’ fecal culture in 71 animals (55 goats and 16 sheep). Kit 1 using indigenous antigen (protoplasmic antigen) was sensitive at very low concentration (0.1 μgm / well) as compared to purified commercial protoplasmic antigen (4 μgm / well) used in kit 2, in the Type 1 reactors (strong positive as positive). Screening of 71 animals by fecal culture detected 38.0% animals (goats-40.0%, sheep-31.2%) as positive (clinical shedders of bacilli) from these farm animals. Of the farm animals located at Central Institute for Research on Goats, herds of goat were endemic whereas, sheep flocks were comparatively resistant to Johne’s disease. The 29.5 and 61.9, 15.4 and 57.7 and 4.2 and 14.0% animals (goats and sheep) were in the category of sero-reactors type 1 and 2 of the ELISA kits 1, 2 and 3, respectively. In the type 1 sero-reactors, sensitivity and specificity of kit 1, 2 and 3 was 53.7 and 86.0, 17.8 and 86.0 and 3.5 and 94.7%, respectively. Indigenous ELISA test (kit 1) was significantly superior for the screening of goatherds and sheep flocks against JD as compared to commercial ELISA kits (Kit 2 and 3). In comparison to kit 2 and 3, kit 1 had highest sensitivity, comparable specificity and substantial to nearly perfect proportional agreement (Kappa Scores) with respect to ‘Gold standard’ fecal culture in goats and sheep. Disease being endemic in herds and flocks screened using ELISA kits, Type I sero-rectors had better correlation with fecal culture in comparison to Type II sero-reactors therefore, used for estimation of sero-prevalence. Newly developed Indigenous ELISA kit was simple, inexpensive, sensitive and reliable for screening of goats and sheep population against Johne’s disease. The study reports high prevalence of Johne’s disease in farm goatherds and sheep flocks, using sensitive tests (fecal culture and ELISA kit). Results of Type 1 reaction in kit 1 were optimally correlated with culture and were good for estimating the sero-prevalence. For controlling Johne’s disease in endemic herds initial removal of the animals in strong positive category (Tyep 1 reactors), may help to remove heavy shedders.     

Serological Evidence of Rift Valley Fever Virus Circulation in Sheep and Goats in Zambézia Province,Mozambique

Rift Valley fever (RVF) is endemic in most parts of Africa and has also been reported to occur in the Arabian Peninsula. It is responsible for significant morbidity and mortality, particularly in livestock, but also in humans. During the last two decades several outbreaks of RVF have been reported in countries in Southern Africa. In contrast to other countries, no clinical disease has been reported in Mozambique during this period. In a serological study conducted in 2007 in five districts of Zambézia Province, Mozambique, of a total of 654 small ruminants sampled (277 sheep and 377 goats), 35.8% of sheep sera and 21.2% of goat sera were positive for RVF virus (RVFV) antibodies in a virus neutralization test (VN) and in an IgG enzyme-linked immunosorbent assay (ELISA). In 2010, a cross-sectional survey was conducted in 313 sheep and 449 goats in two districts of the same province. This study revealed an overall seropositivity rate of 9.2% in sheep and 11.6% in goat and an increased likelihood of being seropositive in older animals (OR = 7.3; p<0.001) using an IgG ELISA. 29 out of 240 animals assessed for RVF specific IgM by ELISA were positive, suggesting recent exposure to RVFV. However, a longitudinal study carried out between September 2010 and April 2011 in a cohort of 125 of these animals (74 sheep and 51 goats) failed to demonstrate seroconversion. The results of the study indicate that RVFV circulates sub-clinically in domestic small ruminants in Zambézia Province.     

Corynebacterium Pseudotuberculosis Infection in Goats V.

In 2 goat herds, one infected with Gorynebacterium pseudo-tuberculosis and one free from the infection,, goats were examined for superficial swellings on the shoulder and chest. All animals in this study had been vaccinated against paratuberculosis before the age of 4 weeks. The vaccine had been applied subcutaneously behind the shoulder. Twenty-two of 40 (55 %) and 31 of 45 (69 %) goats had such lesions in the infected and non-infected herds, respectively. The difference between the herds was not significant, P > 0.05. Swellings found behind the shoulder in 19 goat carcasses derived from 4 herds in which G. pseudotuberculosis infection occurred were examined bacteriologically. No bacteria could be isolated from such lesions in 15 animals, while G. pseudotuberculosis in pure culture was isolated from 3 carcasses, and a mixed bacterial flora from the re-maining carcass. Bacteria could not be isolated from lesions situated behind the shoulder in 7 carcasses from 3 herds free from G. pseudo-tuberculosis infection. It is concluded that most swellings on the shoulder and chest in goats were granulomas resulting from vaccination against paratuber-culosis.     

Attempted cross-transmission of coccidia between sheep and goats and description of Eimeria ovinoidalis sp. n.

Attempted infection of 2 young lambs with oocysts of Eimeria christenseni from a goat was unsuccessful. Negative results were obtained also when young kids were fed oocysts of Eimeria ninakohlyakimovae from sheep. There was no difficulty in infecting lambs with the sheep coccidium resembling E. ninakohlyakimovae nor goats with the goat coccidium E. christenseni. Oocysts from the goat measured 38.4 X 26.7 microns, but were easily distinguished from Eimeria ahsata from the sheep by sporocyst size and shape, and from Eimeria ovina by oocyst size. Eimeria ninakohlyakimovae-like oocysts from sheep averaged 23.0 X 18.2 microns and were morphologically indistinguishable from previously reported goat coccidia. Since no cross infections of sheep and goats could be accomplished with oocysts of Eimeria sp. characteristic of one or the other host, I concluded that sheep coccidia previously known as E. ninakohlykimovae are distinct from morphologically similar goat coccidia and therefore constitute a separate species. Since the name E. ninakohlyakimovae was first used for coccidia from the goat, the sheep coccidium is renamed Eimeria ovinoidalis with oocyst structure and endogenous stages similar to those previously described from the sheep.     

Attempted Cross-Transmission of Coccidia Between Sheep and Goats and Description of Eimeria ovinoidalis sp. n.

SYNOPSIS.
Attempted infection of 2 young lambs with oocysts of Eimeria christenseni from a goat was unsuccessful. Negative results were obtained also when young kids were fed oocysts of Eimeria ninakohlyakimovae from sheep. There was no difficulty in infecting lambs with the sheep coccidium resembling E. ninakohlyakimovae nor goats with the goat coccidium E. christenseni. Oocysts from the goat measured 38.4 × 26.7 m, but were easily distinguished from Eimeria ahsata from the sheep by sporocyst size and shape, and from Eimeria ovina by oocyst size. Eimeria ninakohlyakimovae -like oocysts from sheep averaged 23.0 ×18.2 m and were morphologically indistinguishable from previously reported goat coccidia.
Since no cross infections of sheep and goats could be accomplished with oocysts of Eimeria sp. characteristic of one or the other host, I concluded that sheep coccidia previously known as E. ninakohlyakimovae are distinct from morphologically similar goat coccidia and therefore constitute a separate species. Since the name E. ninakohlyakimovae was first used for coccidia from the goat, the sheep coccidium is renamed Eimeria ovinoidalis with oocyst structure and endogenous stages similar to those previously described from the sheep.     

Corynebacterium Pseudotuberculosis Infection in Goats II.

The precalen-ce of caseous lymphadenitis was surveyed in 36 goat herds in Northern Norway. In each herd, information concerning the occurrence of the disease was obtained from the farmer. Adult animals (1 year of age or older) in 35 herds were examined for superficial swellings, and serum samples were collected from most animals in the herds. The sera were examined for antibodies to Corynebacterium pseudotuber-culosis using the bacterial agglutination test (BAT) and the hemolysis inhibition test (HIT). Gaseous lymphadenitis was diagnosed with certainty in 19 herds. Information from the farmers indicated that the disease indeed oc-curred in these herds, and that the majority had been infected with the disease for many years. The herds had apparently become infected through contact with animals from infected herds. Clinical examina-tions were carried out in 18 of these herds and superficial swellings were found in 26 % of the examined animals. The prevalence of ani-mals with lesions varied from 11 to 40 % among the herds. Of the animals in these herds, 81 % were positive in BAT and 84 % in HIT. The prevalence of positive animals varied from 26 to 99 % in BAT and 28 to 99 % in HIT. The prevalence of seropositive animals was lowest in a herd in which animals were kept separately in stalls. Caseous lymphadenitis could not be diagnosed in 16 herds. In-formation from the farmers indicated that the disease indeed seemed to be absent in 14 of these herds. These 14 herds had no history of contact with animals from herds considered to be infected. However, in the remaining 2 herds, the farmers were somewhat uncertain about the occurrence of the disease. One of these 2 herds had a history of contact with infected herds through participation in a goat “breeding circle”. Only a few of the animals were, however, seropositive and all these had low antibody titres. In 1 newly established herd, a single animal showed a high posi-tive titre in BAT only. All the other animals were negative in both tests. This particular herd consisted of animals obtained both from herds with caseous lymphadenitis and from herds in which the disease was not considered to occur.     

Corynebacterium Pseudotuberculosis Infection in Goats III.

Serological examinations for Gorynebacterium pseudotuberculosis in-fection were carried out in 14 known positive herds and in 1 herd that had beeni recently established through purchase of animals from different herds. Serum samples were collected sequentially on up to 4 occasions from animals during their first year of life. In most herds, these animals were examined clinically for superficial swellings once or twice during the same period. The adult goats in 8 infected herds were examined both clinically and serologically. Age distribution was similar in each of these herds. All serum samples were examined for antibodies against Gorynebac-terium pseudotuberculosis in both the bacterial agglutination test (BAT) and the hemolysis inhibition test (HIT). The proportion of kids which were seropositive in both tests de-creased to zero at 4 months of age. At the age of 11–12 months, the proportion of seropositive animals was about 50 % in the BAT and 30 % in the HIT. When examined when housed for the winter at the age of about 8 months (mean age), the prevalence of animals with superficial swellings was 7 %. At the age of about 1 year (mean age), 29 % of the examined animals had such lesions. In the recently established herd, only a few of the yearlings were seropositive. Titre values in BAT and HIT increased until 6 years of age. Anti-body titre values were significantly lower in yearlings than in older goats in both tests, P < 0.005. No significant difference in the propor-tion of goats with superficial swellings was seen at the different ages.