The gp135 of caprine arthritis encephalitis virus affords greater sensitivity than the p28 in immunodiffusion serology

The 135,000 mw glycoprotein (gp135) and the 28,000 mw internal protein (p28) of caprine arthritis encephalitis virus are major viral constituents in precipitin lines formed between crude antigen preparations and sera from infected goats. In testing 307 goat and sheep sera, 118 samples were positive in a gp135 assay and only 82 were positive in a p28 assay. However, some goat sera were found which reacted only with the p28 and therefore testing for antibody against both proteins may be necessary to identify a maximum number of virus infected goats by immunodiffusion.     

A retrospective study of spinal cord lesions in goats submitted to 3 veterinary diagnostic laboratories

A retrospective study of spinal cord lesions in goats was conducted to identify the range of lesions and diseases recognized and to make recommendations regarding the best tissues to examine and tests to conduct in order to maximize the likelihood of arriving at a definitive etiologic diagnosis in goats with clinical signs referable to the spinal cord. Twenty-seven goats with a spinal cord lesion were identified. The most common lesion recognized, in 13 of 27 goats, was degenerative myelopathy. Eight goats with degenerative myelopathy were diagnosed with copper deficiency. Non-suppurative inflammation due to caprine arthritis encephalitis virus, necrosis due to parasite larvae migration, and neoplasia were each diagnosed 3 times. Based on these findings, it is recommended that, in addition to careful handling and histologic examination of the spinal cord, samples of other tissues, including the brain, liver, and serum, be collected for ancillary testing if warranted.     

Evaluation of newly developed ‘six recombinant secretary proteins based ‘cocktail ELISA’ and ‘whole cell lysate’ based ‘indigenous ELISA’ and tissue microscopy’ with ‘Gold standard’ histo-pathology for the diagnosis of Johne’s disease in slaughtered goats and buffaloes

Three screening tests {(newly developed, six recombinant secretory proteins based 'cocktail ELISA', in-house robust 'indigenous ELISA' based on semi-purified protoplasmic antigens and tissue microscopy were evaluated with 'Gold standard', histo-pathology for the diagnosis of Johne's disease in goats and buffaloes. Serum and tissues {mesenteric lymph nodes and intestines) were driven from farmer's goats (n = 77) and buffaloes (n = 40) slaughtered for harvesting meat and farm goats (n = 77), died and necropsied. Twenty seven (35%) goats and 23 (57.5%) buffaloes were positive in all the four tests. Of 134 tissues screened by histo-pathology, 79.8% MLN and 76.8%, intestines, were positive for MAP infection. In tissue microscopy, 55.2 and 52.3%, goats and buffaloes were positive, respectively. Of 117 sera screened by i_ELISA, 58.4 and 70.0%, goats and buffaloes were positive, respectively. Whereas, c_ELISA detected 55.8 and 62.5%, goats and buffaloes, positives, respectively. Twelve tissues (70.5%) of goats necropsied were positive, both in tissue microscopy and histo-pathology. Most significant gross findings were serous atrophy of the fat and mild to moderate, diffuse thickening of terminal ileum, especially at ileo-caecal junction with or without transverse / longitudinal corrugations. In histo-pathology grade III and IV lesions were significantly low as compared to grade I and II. Of the four tests used for screening 268 samples, histo-pathology was most sensitive (78.3%), followed by i_ELISA (62.3%), c_ELISA (58.9%) and tissue microscopy (58.9%). Between two ELISA tests, c_ELISA using six recombinants secretory proteins, had higher specificity as compared to i_ELISA.     

Isolation of caprine arthritis encephalitis virus from goats in Mexico.

A lentivirus was isolated from 2 goats in Mexico that were seropositive to caprine arthritis encephalitis virus (CAEV) by the agar gel immunodiffusion (AGID) test. The lentivirus was identified as CAEV by the observation of giant multinucleated cells (syncytia) in goat synovial membrane (GSM) monolayers co-cultivated with blood mononuclear (BMN) cells from the seropositive goats, and by amplifying a DNA segment of the CAEV gag gene using the polymerase chain reaction (PCR) technique. Subsequently, cell supernatants from the GSM cells co-cultivated with BMN cells were used to infect 2 CAEV-seronegative goats. These goats seroconverted to CAEV as determined by the AGID test, and CAEV was re-isolated from these goats. One of the goats developed polyarthritis 8 mo after inoculation. Previous serological surveys indicate that infection with CAEV is prevalent among goats in Mexico. To our knowledge this is the first report of CAEV isolation in Mexico. Because of globalization of markets and increased trading among nations, the rapid identification and reporting of diseases such as CAEV are important to prevent the dissemination of these diseases.     

A latex agglutination test for field diagnosis of contagious caprine pleuropneumonia

Latex beads were sensitised with a polysaccharide isolated from a F38 culture supernatant and used in a slide agglutination test to detect serum antibodies in goats with contagious caprine pleuropneumonia. The latex agglutination test detected antibodies in the sera of goats by 22 +/- 2 (mean +/- 1 sd) days after contact exposure to contagious caprine pleuropneumonia, whereas the complement-fixation test detected antibodies by 24 +/- 4 days after contact exposure. Both tests were negative with 181 sera from a farm which was free of the disease. When the same tests were done on 763 sera from two different farms with outbreaks of classical contagious caprine pleuropneumonia, 63 per cent were positive by the latex agglutination test and 23 per cent were positive by the complement-fixation test. Besides being more sensitive than complement fixation, the latex agglutination test can be performed in the field using undiluted serum or whole blood and a result obtained within two minutes.     

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.     

Evidence of mixed infection of peste des petits ruminants virus and bluetongue virus in a flock of goats as confirmed by detection of antigen, antibody and nucleic acid of both the viruses

A mixed infection with peste des petits ruminants virus (PPRV) and bluetongue virus (BTV) occurred in goats which exhibited symptoms characteristic of PPR. A number of samples were collected from ailing or dead goats for labrotory diagnosis. Antibody to BTV and PPRV was detected in sera samples by competitive ELISA. No PPRV antigen was detected in tissue samples like lung and spleen, however, presence of PPRV antigen in some sera samples was confirmed by sandwich ELISA. All the blood samples collected from the ailing animals were found positive for BTV antigen by a sandwich ELISA. BTV- and PPRV nucleic acids were amplified from the pooled blood and tissue samples respectively by RT-PCR assays. The identity of the amplicons was confirmed by cloning and sequencing. All these tests confirm that the goats were infected with PPRV and BTV simultaneously. Isolation of viruses from the clinical samples is underway.     

Acute arthritis in caprine arthritis-encephalitis virus challenge exposure of vaccinated or persistently infected goats

Goats vaccinated with inactivated caprine arthritis-encephalitis virus (CAEV) developed more severe arthritis after infectious CAEV challenge exposure than did goats vaccinated with tissue culture medium. Arthritis also developed more rapidly in the group vaccinated with inactivated virus. In another experiment, goats with persistent CAEV infection developed acute arthritis after at least 2 injections of infectious CAEV at monthly intervals. In this experiment, the control group consisted of goats with persistent CAEV that were given tissue culture medium. Seemingly, the immune response to CAEV is an important cause of the CAEV-induced arthritis.     

Evaluation of diagnostic tests for the detection of classical swine fever in the field without a gold standard.

Knowledge of the sensitivity of diagnostic tests for infectious diseases under field conditions can be used to design a surveillance program that increases the effectiveness of the control policy. In this study, the sensitivity of tests for the detection of classical swine fever (CSF) virus (CSFV) under field conditions was estimated without knowledge of the true disease status of the animals tested. During the CSF epidemic of 1997-1998 in The Netherlands, tonsil samples from pigs of CSF suspect farms were collected for laboratory diagnosis of CSE These specimens were tested in a fluorescence antibody test (FAT1) for the presence of CSFV antigen. When at least 1 specimen in a particular sample series from a farm was positive, this farm was declared CSFV infected. Specimens of that series, either FAT1 negative (98) or FAT1 positive (127), were subsequently tested again (FAT2). After that, a suspension was made of the remaining tissue, and this suspension was evaluated with a virus isolation test. In total, 225 tonsil specimens were examined. A statistical model was formulated, and the sensitivity of the 3 tests and the prevalence of positive specimens in the sample were estimated by the method of maximum likelihood. The sensitivity of the FAT1, the test that was used for confirmation of CSFV infection in a pig herd, was approximately 78% (95% confidence interval [CI] = 62-92%). The effectiveness of the selection process of animals on the farm by the veterinarian was estimated to be 77% (64-87%). The sensitivity of the combination of FAT1 and FAT2 (60%) indicates that at least 5 animals should be selected on a CSF-suspect farm to gain a detection probability for CSFV of 99%.     

Caprine arthritis-encephalitis virus infection of goats in South Australia

Caprine arthritis-encephalitis virus (CAEV) infection of dairy goats was shown by virus isolation and serology to be widespread in South Australia. CAEV was isolated at necropsy from 24 of 27 dairy goats with swollen joints from 13 herds, and from 9 of 30 liver dairy goats in 7 herds. Virus was isolated most frequently from synovial membranes, and occasionally from mammary glands, mammary lymph nodes, choroid plexus, lungs, spleen, bone marrow, salivary glands, leucocytes, synovial fluid and milk. Antibody to CAEV was detected in the serum of 13 of 17 of the necropsied goats tested in a single-line gel diffusion test, and in another 3 retested with a modified double-line technique. Serum antibody was also demonstrated in 61 of 77 dairy goat herds, many with histories of arthritis. In 1984 to 1986 the annual number of serologically positive serums and proportions of the numbers tested were 134 (40%), 121 (45%) and 42 (18%), respectively. CAEV was isolated from leucocytes of 8 live goats in 6 of these herds. In fibre goats antibody was detected in the serum of 25 Angora and 19 crossbreds (0.1%) from the 33,279 Angora, 1,705 Cashmere, 8,715 crossbred and 904 feral goats tested.     

Phylogenetic analysis of SRLV sequences from an arthritic sheep outbreak demonstrates the introduction of CAEV-like viruses among Spanish sheep

Small ruminant lentiviruses (SRLVs) cause different clinical forms of disease in sheep and goats. So far in Spain, Maedi visna virus-like (MVV-like) sequences have been found in both species, and the arthritic SRLV disease has never been found in sheep until a recent outbreak. Knowing that arthritis is common in goats, it was of interest to determine if the genetic type of the virus involved in the sheep arthritis outbreak was caprine arthritis encephalitis virus-like (CAEV-like) rather than MVV-like. Alignment and phylogenetic analyses on nucleotide and deduced amino acid sequences from SRLV of this outbreak, allowed a B2 genetic subgroup assignment of these SRLV, compatible with a correspondence between the virus genetic type and the disease form. Furthermore, an isolate was obtained from the arthritic outbreak, its full genome was CAEV-like but the pol integrase region was MVV-like. Although its LTR lacked a U3 repeat sequence and had a deletion in the R region, which has been proposed to reduce viral replication rate, its phenotype in sheep skin fibroblast cultures was rapid/high, thus it appeared to have adapted to sheep cells. This outbreak study represents the first report on CAEV-like genetic findings and complete genome analysis among Spanish small ruminants.     

某规模化猪场伪狂犬病的诊断

河南平顶山某猪场母猪出现较严重的流产和产死胎现象,且50日龄~70日龄仔猪出现神经症状,根据临床表现初步诊断为伪狂犬病。为排除猪繁殖与呼吸综合征和猪瘟,进行了实验室诊断。应用ELISA方法检测发病保育猪及母猪血清的伪狂犬病病毒野毒株gE抗体,并对发病仔猪病料进行了伪狂犬病病毒(PRV)、猪繁殖与呼吸综合征病毒(PRRSV)和猪瘟病毒(CSFV)的实时荧光定量PCR检测。结果显示,伪狂犬病病毒野毒抗体阳性,实时荧光定量PCR检测确定仔猪病料中PRV核酸阳性,PRRSV和CSFV核酸阴性。结合临床症状及实验室检测,确诊该猪场发生的是猪伪狂犬病。     

Accuracy of an anaemia scoring chart applied on goats in sub-humid Kenya and its potential for control of Haemonchus contortus infections

We tested the practical application of an anaemia scoring chart (the FAMACHA chart) as a method for controlling Haemonchus contortus in goats kept under smallholder conditions in a sub-humid area of Central Kenya. The objectives were: (1) to test the accuracy of the FAMACHA chart in identifying anaemic goats (PCV< or =18); (2) to quantify the proportion of goats left untreated at farm level when using the chart. On each of two farms, Small East African goats of various ages were allocated to two treatment groups; a FAMACHA group (F1 (n=34) and F2 (n=31) on farms 1 and 2, respectively) and a control group (C1 (n=34) and C2 (n=30)). In F1 and F2 goats with a FAMACHA score of 3, 4 or 5 were treated with anthelmintic after scoring. In C1 and C2 goats were treated every 4 weeks from 15 February to 20 July. Every 2 weeks all goats were scored with the FAMACHA chart and weighed. Furthermore, faecal samples were collected for faecal egg counts (FEC) and blood samples were collected for packed cell volume (PCV) determination. H. contortus was found to be the predominant nematode on both farms. The mean FECs were higher on farm 1 compared to farm 2, while in contrast the mean PCV levels were lowest on farm 2. The latter was most likely due to the presence of Fasciola spp., flea and tick infections on farm 2. The accuracy of the chart was evaluated by using PCV as the gold standard for anaemia (PCV< or =18%). The mean percentage of false-negative scorings per sampling was 0.7% on farm 1 and 1.6% on farm 2, while the mean percentage of false-positive scorings was 9.7% and 21.4%, respectively. It is most likely that the accuracy of the chart was negatively affected by the concurrent parasite infections on farm 2. The mean proportion of untreated goats per sampling was 89% and 77% on farm 1 and farm 2. It was concluded that the FAMACHA chart can be a valuable tool for decision-making in control of H. contortus in goats kept under smallholder conditions, without morbidity or mortality unacceptable to the farmer. The application may further reduce the risk of development of anthelmintic resistance by increasing refugia. However, caution should be taken under conditions where other anaemia-causing parasites are present (e.g. Fasciola spp. and ecto-parasites), since this possibly decreases the accuracy of the FAMACHA chart.     

Sheep-associated malignant catarrhal fever in a petting zoo.

In a privately owned petting zoo in Arizona, 17 deer from five different species, white-tailed deer (Odocoileus virginianus), Reeve's muntjac (Muntiacus reevesi), mule deer (Odocoileus hemionus), reindeer (Rangifer tarandus), and axis deer (Axis axis), died of suspected malignant catarrhal fever (MCF) over a period from late 1992 to early 1995. A PCR assay specific for ovine herpesvirus 2, the putative causative agent of sheep-associated MCF, and a competitive-inhibition enzyme-linked immunosorbent assay based on a monoclonal antibody specific to an epitope conserved among all known MCF viral isolates were used to investigate the outbreak. Ovine herpesvirus 2 DNA sequences were detected by PCR from fresh-frozen and/or formalin-fixed, paraffin-embedded tissue samples in seven deer out of eight available animals previously suspected as cases by histopathology. A high seroprevalence to the virus was found among mouflon (Ovis musimon, 80%) and pygmy goats (Capra hircus, 61%), both of which were present on the farm during the outbreak. Sixteen percent of fallow deer (Dama dama) were also seropositive to the virus. After removal of the mouflon and positive pygmy goats, no further MCF cases occurred on the farm, confirming the importance of careful management to avoid mixing clinically susceptible species with carrier species. Until better control measures are available, adherence to this practice is necessary if MCF is to be prevented in intense exposure environments such as zoos and densely populated animal parks.     

Serologic and virologic investigations into pestivirus infection in wild and domestic ruminants in the Pyrenees (NE Spain)

An outbreak of disease associated to a border disease virus was described in the Southern chamois (Rupicapra pyrenaica) in Spain in 2002. Sera and/or spleen samples from 57 mouflon, 15 red deer, 21 roe deer, 3 fallow deer, 55 sheep, 32 cattle, and 68 goats sharing the chamois habitat were studied. An antibody ELISA test yielded an inconclusive result in 2 mouflon and positive results in 5 goat sera. Comparative virus neutralization tests were performed on the 2 inconclusive mouflons, 3 of the 5 seropositive goats, 55 sheep and 32 cattle, using 6 pestivirus strains. Positive results were obtained in 1 mouflon, 2 goats, 69% of sheep and 78% of cattle. Virological investigations performed with an antigen ELISA test yielded negative results in 21 goats and 39 mouflons, the result in 1 mouflon being inconclusive. PCR performed on 12 goats and the inconclusive mouflon gave negative results. These results suggested that it is unlikely that chamois BDV is infecting wild and domestic ruminants.     

Duration of bluetongue viraemia and serological responses in experimentally infected European breeds of sheep and goats

The duration of viraemia and the serological responses were studied in two breeds of sheep and two breeds of goats, experimentally infected with bluetongue (BT) virus serotype 4. Viraemia, detectable by cell culture and embryonated chicken egg inoculation, lasted from the third to sixth day until the 27th-54th day post infection (p.i.). Significant differences between sheep and goats were not recorded. Lesbos sheep and goats together appeared to have significantly longer viraemias (n = 9, mean 41.3 days) than east-Friesian sheep and Saanen goats (n = 10, mean 30.4 days, p = 0.0039). Serological response was studied by competitive ELISA (c-ELISA) and agar gel immunodiffusion (AGID) tests. The c-ELISA was more sensitive in detecting BT virus antibodies in all animals than the AGID tests. No significant differences were observed between sheep and goats or between breeds. The epidemiological significance of subclinical infection and the extended BT virus viraemias in Lesbos sheep and goats, in relation to the maintenance of the virus and to overwintering is discussed.     

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 and aetiology of lung lesions in goats infected with caprine arthritis-encephalitis virus

Fifty dairy goats, of various ages, sexes and breeds were selected for examination on the basis of positive serological reactions to caprine arthritis-encephalitis virus (CAEV). Thirty-one had lung lesions including chronic interstitial pneumonia of caudal or cranioventral lobes, bronchopneumonia, verminous pneumonia, pulmonary cryptococcosis or combinations of these. The only infective agent recovered from all the chronic interstitial pneumonia cases examined was CAEV, which was also recovered from lung tissue of 3 goats with arthritis but no lung lesions. The presence of CAEV in lavaged alveolar macrophages from normal lung tissue and from lungs affected with chronic interstitial pneumonia and verminous pneumonia, and the demonstration of a marked increase in nonspecific esterase staining macrophages in areas of chronic interstitial pneumonia, are discussed in relation to the aetiology of the pneumonia.     

Vulvovaginitis of goats due to a herpesvirus

Two concurrent outbreaks of genital disease in goats were associated with infection by a herpesvirus that was isolated from vulval and vaginal lesions of affected does. Serum neutralising antibody to the virus was present both in goats with the clinical disease and some unaffected goats. Of 19 goat herds examined only 4 had serum neutralising antibody positive goats with low (5%) to high (60%) incidence of infection. The virus isolate was characterised as a herpesvirus on its physico-chemical and morphological features. It contained DNA and was inactivated at low pH and by treatment with lipid solvents and trypsin. The virus particles were icosahedral, consisting of a nucleocapsid surrounded by an envelope membrane and measured approximately 150 nm in diameter. The virus was serologically related to a New Zealand isolate of caprine herpesvirus (NZ-CpHV), associated with similar genital disease, and was distinct from bovine herpes virus-1 (BHV-1) showing a one way neutralisation pattern.