PCR examination of bronchoalveolar lavage samples is a useful tool in pre-clinical diagnosis of ovine pulmonary adenocarcinoma (Jaagsiekte)

Ovine pulmonary adenocarcinoma (OPA) is a contagious lung tumour of sheep caused by Jaagsiekte sheep retrovirus (JSRV). The disease is a particular problem in flocks in many parts of the world. The aim of the study was to assess screening methods for individual animals as a prelude to future eradication trials. Results of histological examination were used as the standard to evaluate the relative sensitivity and specificity of an established heminested polymerase chain reaction (PCR) test for JSRV proviral DNA from blood and bronchoalveolar lavage (BAL) samples. PCR results from tissue samples are included as control data. PCR testing of blood samples was found to have an estimated sensitivity of only 10% (95% confidence interval (CI) 3-20) while the sensitivity of the PCR test on BAL samples was 89% (CI 79-96) in comparison to the results of histological examination. We conclude that PCR testing of BAL samples is an effective confirmatory test for sheep with suspected clinical OPA. It is also a useful tool for the pre-clinical identification of individual infected sheep within an infected flock and therefore may prove beneficial in future control or eradication programmes.     

Cryptosporidium parvum infection in orphan lambs on a farm open to the public

A longitudinal survey was undertaken on an open farm to investigate the occurrence of Cryptosporidium species infection in orphan lambs obtained from three local flocks. During an initial pilot study, Cryptosporidium oocysts were detected by a fluorescent antibody test (fat) in the faeces of two of 21 lambs aged between one and three weeks derived from one flock (flock A). Pooled pen samples of faeces were collected weekly from lambs derived from each flock; oocysts were detected by fat in 24 (49.0 per cent) of 49 samples from lambs from flock A, 18 (30.5 per cent) of 59 samples from lambs from flock B and 14 (29.8 per cent) of 47 samples from lambs from flock C. Oocyst counts of 1 x 10(3) to more than 2 x 10(6) per gram of faeces were detected in lambs up to 12 weeks old, with the peak counts occurring at six weeks of age in the lambs from flocks A and B and at four weeks of age in those from flock C. The oocysts were confirmed by molecular analysis as Cryptosporidium parvum. Virtually all the infections were subclinical.     

Use of serology and polymerase chain reaction for the rapid eradication of small ruminant lentivirus infections from a sheep flock: A case report

A SRLV-free sheep flock incurred infection which led to an SRLV infection rate of over 50% of the ewes (34/64) within a 30 months period, indicating that environmental conditions were favourable to transmission. An intensive regimen of sampling at short intervals and testing for SRLV antibodies and proviral DNA combined with strict management was implemented for the entire flock, lambs and yearlings included. This resulted in eradication of the infection within two testing and culling rounds with a 3 months interval. The additional value of the proviral DNA detection by PCR in identifying infected animals was clear in that nine infected animals were found that would have been missed if tested by serology alone. PCR also saved two lambs from being culled; they were sero-positive probably due to maternal antibodies, but not infected.     

Colostrum and milk can transmit jaagsiekte retrovirus to lambs

Ovine pulmonary adenocarcinoma (OPA) is a contagious disease caused by jaagsiekte sheep retrovirus (JSRV). In the three studies performed, we have obtained data of the importance of colostrum/milk (C/M) in the transmission of JSRV. In the first study, a group of sheep from a flock with a long history of OPA, samples from colostrum and peripheral blood leucocytes (PBLs) were collected. Two specific PCRs (U3-LTR and env of the JSRV) were carried out. Using U3PCR 8/34 sheep were positive in colostrum whereas with envPCR 7/34 were positive. From these animals only one was positive with U3PCR in the PBLs. Evidence of the transmission of JSRV infection by C/M was obtained in two more separate studies. In the second study, PBLs from five lambs from JSRV+ ewes and two from JSRV-ewes were tested by the U3PCR. They were fed C/M by their mothers during 3 months and slaughtered 7 months after birth. Three out of five lambs from the JSRV+ sheep become PBL positive at 3-4 months old and the other two were also positive at 4-6 months of age. One lamb of the JSRV-sheep became also PBL positive at an age of 3 months. In the third study, a group of lambs from JSRV negative mothers were fed with C/M from JSRV+ sheep and housed in separate unit. For comparison, another group of the same origin and maintained in another different unit, were fed with C/M containing a JSRV virus preparation. All lambs were blood sampled monthly and JSRV infection was detected as early as 15 days and several times onwards in both groups. Control groups fed with C/M from JSRV free flock and JSRV blood test negative sheep were always negative. Together these results indicate that suckling is an important natural transmission route for JSRV.     

Molecular epidemiology of ovine herpesvirus type 2 infection in Kashmir, India

The aims of this investigation were to determine the prevalence of ovine herpesvirus type 2 (OvHV-2) (the causative agent of malignant catarrhal fever) infection in cattle, the carrier status of sheep and goats, and to define the pattern of acquisition of OvHV-2 in lambs under natural flock conditions in Kashmir, India. None of the buffy coat samples from 21 lambs contained OvHV-2 DNA sequences up to 28 days after birth, only one lamb had sequences of OvHV-2 DNA as early as 29 days after birth, and they were detected in the other 20 lambs when they were between 43 and 94 days of age. Sequences of OvHV-2 DNA were detected in buffy coat samples from 28 (85 per cent) of 33 adult sheep and in 16 (61 per cent) of 26 samples from adult goats by hemi-nested PCR. Seventeen (31 per cent) of 55 cattle with malignant catarrhal fever-like clinical signs had sequences of OvHV-2 DNA in their blood, and nine of the 17 died, all of them during the months of April to November, between November 2002 and March 2004. No clinical cases of sheep-associated malignant catarrhal fever was recorded during the months of December to March. The overall prevalence of OvHV-2 infection in the cattle in the region was estimated to be less than 1 per cent.     

Role of endogenous transplacental transmission in toxoplasmosis in sheep

To investigate the potential role of endogenous transplacental transmission of Toxoplasma gondii, 31 seropositive ewes presumed to be persistently infected with the parasite and 15 seronegative ewes were mated and monitored throughout pregnancy and lambing. Antibody titres were determined in precolostral sera from the liveborn lambs and in thoracic fluid from the dead lambs. A PCR for the B1 gene of T gondii was applied to the placentas from all the ewes and to the brains of the stillborn lambs. Samples of brain, lung, liver, spleen and heart from the dead lambs were examined by histopathology. No evidence of toxoplasmosis was detected by histopathology or PCR in any of the samples, but low titres of antibody to T gondii were detected in two liveborn, healthy offspring of a seropositive ewe by the immunofluorescent antibody test (3.2 per cent of pregnancies and 4.1 per cent of lambs in the seropositive group). Antibody to specific antigens of T gondii was demonstrated in sera from these two lambs by Western blotting.     

Parasitic otitis in sheep associated with psoroptes infestation: a clinical and epidemiological study.

Parasitic otitis associated with psoroptes infestation was diagnosed in a small pedigree flock of sheep with aural haematomas, abscessation and cauliflower ears. Thirteen of the 15 lambs were affected but the clinical signs were mild; small, discrete, crusty lesions on the inner aspect of the ear at the junction of its anterior and posterior borders were typical. Nine of the 20 adults were affected but the lesions were more severe. Eighteen of the 64 members of the breed society who responded to a telephone survey reported sheep with similar clinical signs in their flocks. The proportion of animals affected ranged from 1 to 60 per cent with a median value of 16 per cent.     

Border disease without nervous signs or fleece changes

A natural infection with border disease virus occurred in a flock on low ground in Argyll in the spring of 1984. The outbreak was unusual in that the typical clinical signs of border disease, ie, tremor and, or, fleece changes were not present; manifestations of disease were restricted to abortion and the birth of small weak lambs. The disease was shown to have been introduced to the flock by four healthy ewes persistently infected with border disease virus among a group of 39 purchased in October 1983. Further investigations in late August 1984 detected viraemia in six of seven ill-thriven lambs and four of 24 apparently healthy lambs. Attempted 'natural vaccination' of susceptible sheep by mixing them at grass for three months with groups of ewes and lambs known to contain virus excretors was largely unsuccessful as only four of 22 'sentinel' sheep seroconverted. In October 1984 the persistently infected purchased animals and all that year's lamb crop were removed from the farm. No disease occurred in 1985 when the lambing percentage was 129 per cent compared with 100 per cent in 1984. Two of the four persistently infected purchased ewes were mated at Moredun Research Institute in December 1984 and both produced healthy but persistently infected lambs.     

Uterine tube abnormalities as a cause of bovine infertility

A total of 2000 bovine reproductive tracts (1000 parous and 1000 nulliparous) from freshly slaughtered animals were examined for uterine tube abnormalities and lesions. Tubal lesions were recorded in 180 tracts (9.0 per cent). More parous tracts (6.4 per cent) had tubal lesions than nulliparous tracts (2.6 per cent). The most frequently identified lesion was ovarobursal adhesions which comprised 80 per cent of all abnormalities and were identified in 6.85 per cent of all the genital tracts examined. The uterine tubes were insufflated with carbon dioxide gas to a maximum pressure of 300 mm Hg; 22 were found to be occluded even though they were macroscopically normal.     

Measurements of footpad dermatitis in broiler chickens at processing plants

A four-point photographic scale was developed to score the severity of lesions of footpad dermatitis (FPD) in broiler chickens. There was a linear relationship between the square root of the relative area of the lesions and their scores, and good agreement between different assessors using the scale. The scale was used to assess samples of 100 birds from each of 190 flocks slaughtered at two uk processing plants in 2002 and 2003; 12 of the flocks (6.3 per cent) had no lesions, but the others had lesions of different prevalences and severity. The maximum proportion of affected birds in a flock was 92 per cent. In the 178 affected flocks, 84.0 per cent of the birds had no lesions and 16.0 per cent had some evidence of FPD. The overall unweighted prevalence of birds with FPD in all 190 flocks sampled was 18.1 per cent; 10.2 per cent had only small lesions, on average equivalent in area to 2.1 per cent of the total area of the foot, 6.2 per cent had lesions on average equivalent to 6.6 per cent of the area of the foot, and 1.7 per cent had lesions on average equivalent to 21.5 per cent of the area of the foot. There were differences between the two plants in the overall prevalence and severity of FPD, but this may have been due to the fact that the plants were sampled at different times of the year.     

Use of sentinel lambs for early monitoring of the South Powys Hydatidosis Control Scheme: prevalence of Echinococcus granulosus and some other helminths.

Sentinel lambs were used to identify young Echinococcus granulosus infections in sheep, to provide an early indication of the progress of the South Powys Hydatidosis Control Scheme. Four sentinel lambs were purchased on each of 60 farms, from inside and outside the control area; they were examined when approximately six, 10 and 15 months of age. Gross examination, thin slicing of organs and histological examination of the lesions in the viscera revealed no E granulosus hydatid cysts in lambs born within the control area, whereas 25 per cent of the 15-month-old lambs from outside the area harboured E granulosus cysts (less than 1 to 2 mm in diameter). Lambs from E granulosus infected farms had significantly higher anti-E granulosus ELISA antibody titres than lambs from uninfected farms. It was concluded that within one year of beginning to treat dogs with praziquantel every six weeks the transmission of E granulosus to sheep had ceased. In contrast, this treatment did not prevent infections with Taenia hydatigena or T ovis; an examination of the 240 lambs revealed T hydatigena in 33.3 per cent of them, Tovis in 4.2 per cent, Dictyocaulus filaria in 12.1 per cent and Meullerius capillaris in 49.2 per cent.     

Effect of passive transfer of maternal immune components on infection with ovine herpesvirus 2 in lambs

OBJECTIVE: To define the role of passively tranferred immunity in protection against early infection with ovine herpesvirus 2 (OvHV-2) in lambs. ANIMALS: 15 adult sheep and 34 lambs. PROCEDURES: 2 groups of animals were used, including 15 lambs born to OvHV-2-free ewes and 19 lambs born to OvHV-2-positive ewes. After nursing colostrum, all lambs and their dams were introduced into a flock positive for OvHV-2. Blood was obtained from the lambs every 2 weeks and examined by PCR assay and competitive inhibition ELISA. RESULTS: None of the animals had positive results by PCR analysis for samples obtained approximately 2 weeks after introduction into the flock. In the group of lambs from OvHV-2-infected ewes, 5 of 19 had positive results at 1 month of age and 17 of 19 by 5 months of age. In the group of offspring from OvHV-2-negative ewes, only 1 of 15 had positive results at 1 month of age, and the number reached 12 of 15 by 5 months of age. All lambs in both groups had positive results by 6 months. An active antibody response to the virus was detected in animals within 3 weeks after viral DNA became detectable in the blood. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis suggests that passively transferred immunity does not play an important role in the delay of infection with OvHV-2 in lambs. Age also does not seem to influence susceptibility. The rate of infection in young lambs may simply be a reflection of the intensity of viral exposure in their environment.     

Farm and slaughter survey of bovine tuberculosis in captive deer in Switzerland

In 1998, a survey was conducted by postal questionnaire to gather basic knowledge about the management, health and productivity of captive deer in Switzerland. In addition, lymph nodes were collected from slaughtered deer from 124 of the 262 holdings surveyed, and tested for Mycobacterium bovis and Mycobacterium tuberculosis. The total farmed deer population was 8389 animals kept on 485 holdings; 87 per cent were fallow deer, 8 per cent red deer, 4 per cent sika deer, and there were small numbers of other species. The median herd sizes were 12 for fallow deer and eight for red deer. Few owners had handling facilities or crushes. In none of the lymph nodes examined were lesions typical of bovine tuberculosis observed, and neither M bovis nor M tuberculosis was cultivated from any of the samples.     

A survey of more than 11 years of neurologic diseases of ruminants with special reference to transmissible spongiform encephalopathies (TSEs) in Greece

The first cases of scrapie were detected in Greece in a flock of sheep in October 1986. All the animals of the affected flock and all sheep in two flocks that were in contact were killed and buried. A systematic investigation of all available cases with signs indicating a neurological disease started in sheep and goats in late 1986, as well as in cattle in 1989. The investigation was based on clinical examination, necropsy or macroscopical examination of the brain and viscera, and histological examination of the brain in all animals except those with coenurosis. Histological examinations of specimens from the spinal cord and other tissues, and if considered necessary bacteriological, toxicological and serological examinations were also carried out. In October 1997, scrapie was diagnosed in sheep of a second flock (a mixed flock of sheep and goats), grazing in a pasture close to the place where scrapie was initially detected. All animals of the second flock were also killed and buried. Diagnosis in the first flock was based on clinical signs and histological lesions, and in the second immunoblotting was also used. Distinctive lesions of scrapie were found in the brain and/or the spinal cord of eight sheep with clinical signs from the two flocks. The lesions were revealed in the brain stem and/or in the cervical spinal cord, and tended to be symmetrical. In one sheep, severe lesions in the cortex of cerebral hemispheres and of the cerebellum were also found. In the brain of two sheep from the second flock the pathological isoform of PrP protein was detected. Despite the eradication scheme applied, scrapie in sheep reappeared after 11 years in a place close to where it occurred initially. This may indicate that the effectiveness of the eradication scheme implemented was not adequate and additional approaches may be needed.     

A field study of watery mouth: clinical, epidemiological, biochemical, haematological and bacteriological observations

More than 500 cases of watery mouth in newborn lambs were recorded in 11 flocks and a detailed examination was performed in more than 200 cases. The condition occurred predominantly in lambs aged 24 to 48 hours. The incidence in a flock varied from 1 per cent to 24 per cent and the mortality in affected lambs varied from 7 per cent to 83 per cent. A bacteraemia was found in 38 per cent of the affected lambs. There were no consistent abnormalities of clinical biochemistry or haematology. Watery mouth was more common in twins and especially in triplets than in single lambs. In twins the highest incidence of watery mouth was recorded in lambs born to ewes in poor body condition. Antibiotic given orally within 15 minutes of birth greatly reduced the incidence of watery mouth.     

Evaluation of indirect biomarkers for detecting corticosteroids used as illegal growth promoters in beef cattle

The histological status of the thymus, blood cortisol concentration and circulating neutrophil:lymphocyte ratio were evaluated in 349 slaughtered beef cattle, to assess the potential of these parameters as indirect biomarkers of the illegal use of corticosteroids in meat production. The livers of 20 of the animals were analysed chemically for residues of corticosteroids. The morphology of the thymus was examined for adipose tissue infiltration, cortical atrophy and 'starry sky' appearance, and on the basis of these characteristics, the animals were considered to be negative, suspected or positive for illegal corticosteroid treatment. The animals considered to be negative had a mean cortisol concentration that was significantly higher (29 ng/ml) than that of the animals suspected for corticosteroid treatment (22 ng/ml). Using the chemical analysis as the gold standard for identifying illegally treated animals, the histological examination of the thymus had a sensitivity of 100 per cent and a specificity of 85 per cent. The samples that were positive by chemical analysis had cortisol concentrations of less than 2.0 ng/ml, whereas the mean cortisol concentration of the negative samples was 10.3 ng/ml.     

Evaluation of the prevalence and onset of lung lesions and their impact on growth of lambs

OBJECTIVE: To determine the prevalence and temporal onset of lung lesions in lambs and the impact of lung lesions on growth of affected lambs. ANIMALS: 259 crossbred wether lambs from a single flock in the upper Midwestern United States. PROCEDURE: An observational study was conducted. Lambs born in the spring and fall were slaughtered at finished weight or at a predetermined time point. Lungs of each lamb were examined and classified as normal, moderate lesions (consolidation > 5% but < or = 50% of any lobe), or severe lesions (consolidation > 50% of any lobe). Data were examined to detect effects of prevalence or severity of lung lesions on growth and carcass traits. RESULTS: 57 of 89 (64%) spring-born lambs had lung lesions characterized by consolidation of lung tissue. A small number of lambs had pulmonary adhesions or active abscesses. In contrast, only 31 of 108 (29%) fall-born lambs had lung lesions. Severe lung lesions were associated with a significant reduction in average daily gain. Severe lung lesions were not detected until the middle of the finishing period and were associated with culture of Mannheimia haemolytica or Pasteurella multocida. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of results indicates that the prevalence of severe lung lesions can be quite high in lambs. Severe lung lesions can lead to greatly decreased growth performance of lambs.     

Quantification of the relative efficiency of factory surveillance in the disclosure of tuberculosis lesions in attested Irish cattle

In Ireland, factory surveillance of cattle for gross lesions is an important supplementary method for detecting herds infected with bovine tuberculosis (tb), and in recent years between 27 and 46 per cent of all new herd breakdowns in any year have been detected by this method. The aim of this study was to determine the relative efficiency of factories in detecting lesions among attested cattle slaughtered during 2003 and 2004. National databases were available on animal slaughter, programmes of tuberculin testing for bovine tb and laboratory confirmation of suspected lesions. Factories were ranked according to their submission risk (number of animals submitted with lesions/number of attested animals killed) and confirmation risk (number of animals with laboratory-confirmed lesions/number of animals submitted with lesions), adjusting for the risk profile of the animals slaughtered, including potential confounding factors such as their age and sex, whether they were purchased or homebred, the test history of their herd, the prevalence of bovine tb in the area and the season of slaughter. Approximately 3.7 million cattle were slaughtered in 42 Irish export-licensed factories during the two years. Complete data were available for 2,374,987 animals from 84,510 attested herds in 2845 District Electoral Divisions. Samples from 7398 animals with suspected tb lesions were submitted for laboratory examination; 4767 (64.4 per cent) were positive, 2011 were negative and 620 were inconclusive. The average unadjusted submission risk for all the factories was 22 per 10,000, ranging from 0 to 58 per 10,000. The unadjusted factory confirmation risk (excluding factories that had sent in fewer than 10 lesions) varied between 34.3 per cent and 86.3 per cent. The unadjusted and adjusted submission and confirmation risks were highly correlated, and animal-related factors (including their characteristics and origin) therefore did not contribute to the variations in factory-level submission and confirmation risks.     

Pathological,serological, and virological findings in sheep infected simultaneously with <Emphasis Type="Italic">Bluetongue</Emphasis>, <Emphasis Type="Italic">Peste-des-petits-ruminants</Emphasis>, and <Emphasis Type="Italic">Sheeppox viruses</Emphasis>

In this study, pathological, serological and virological examinations were performed on 15 sheep from a flock of 250 sheep and lambs that suffer from simultaneous naturally occurring BTV, PPRV and SPV outbreaks. SPV was diagnosed macroscopically and histopathologically, BTV was diagnosed by ELISA, and PPRV was diagnosed pathologically and by ELISA. Clinically fever, diarrhea, depression, polypnea, conjunctivitis, lacrimation, rhinitis, erosive stomatitis, edema of eyelids, photophobia, cutaneous eruption with erythematous areas especially noticeable in wool-free parts of the body and axilla lesions evolving into papules were observed. At necropsy, the most effected organs were lungs and gut. Subepicardial hemorrhages were also commonly seen. While typical pox lesions were observed in some lambs, usually fibrinous pleuropneumonia was more prominent lung lesion. SPV and PPRV lesions were seen at the histopathological examination of the lesioned tissues, BT lesions were mild than SPV and PPRV microscopically. Serum and leukocyte samples of 15 animals were examined for PPRV and BTV by ELISA; 5 samples were positive for PPRV and 6 BTV, 4 were positive for both PPRV and BTV simultaneously. One hundred animals died, most were lambs. Mortality rates were 100% in lambs and 80% in the herd.     

Production of malignant catarrhal fever virus-free sheep

The study was designed to better define the variables affecting the success of the establishment of ovine herpesvirus 2 (OHV-2)-free sheep flocks. A total of 38 lambs born to OHV-2-positive ewes was selected and divided into four groups. Three groups of 10 lambs each were separated from the positive ewes at 2, 2.5 and 3 months of age, respectively, and maintained in isolation facilities. One group of eight remained in the positive flock as controls. Peripheral blood samples from each lamb were examined regularly by PCR for OHV-2 DNA. All lambs (100%) that were weaned and maintained in isolation from the ages of 2, 2.5 and 3 months remained negative until the termination of the experiment at 1 year of age. One lamb was discovered to be PCR-positive on the day of isolation at 2.5 months of age, and was promptly removed from the isolation group. In contrast, all lambs (100%) that remained with the flock became PCR-positive by 6 months of age. The data confirmed that, with rare exceptions, separation of lambs from OHV-2 infected animals at around 2 months of age reliably yields OHV-2-free sheep. Appropriate PCR monitoring will enable the rare exceptions to be removed from the group, and is recommended as a safety measure.