Use of pooled fecal culture for sensitive and economic detection of mycobacterium avium subsp. paratuberculosis infection in flocks of sheep.

Ovine Johne's disease, or paratuberculosis, occurs in many countries. In Australia, surveillance using serology is used as part of a control program, but the testing regime is costly relative to its sensitivity. For this reason, culturing of Mycobacterium avium subsp. paratuberculosis in fecal samples pooled from a number of sheep was evaluated. Initially, the effect of pooling on the sensitivity of fecal culture was evaluated using samples from 20 sheep with multibacillary paratuberculosis and 20 sheep with paucibacillary paratuberculosis, each confirmed histologically. All multibacillary cases and 50% of paucibacillary cases were detected by culturing of feces at a pooling rate of 1 infected plus 49 uninfected sheep. In a pilot-scale study in 1997, M. avium subsp. paratuberculosis was detected by pooled fecal culture on 93% of 27 infected farms which were identified originally based on history, clinical signs, and one or more rounds of testing using serologic and histopathologic examinations. Pooled fecal culture was compared with serologic examination for submissions from 335 farms where both tests had been conducted on the same sheep and was significantly more sensitive (P<0.001). Computer simulation of random sampling indicated that the testing of 6 pools of 50 sheep would provide 95% confidence in detecting > or =2% prevalence of infection. The estimated laboratory cost of pooled fecal culture when applied as a flock test is approximately 30% that of serologic examination, and sample collection costs are lower. It is recommended that pooled fecal culture replace serologic examination for detection of M. avium subsp. paratuberculosis infection at the flock level.

Assessment of gelatin gel and elastase tests for detection of protease activity of Dichelobacter nodosus isolates from ovine footrot.

Protease tests (the gelatin gel protease thermostability test, the elastase test or both) were performed on 4296 isolates of Dichelobacter nodosus derived from 452 outbreaks of ovine footrot occurring in New South Wales. Both tests showed a high level of repeatability. In the gelatin gel test, culture broths were heated for 16 min at 66.8 degrees C. Heated broths containing thermostable protease digested gelatin (positive gelatin gel test) while those broths containing thermolabile protease failed to digest gelatin (negative gelatin gel test). Gelatin gel positive isolates were unable to be graded into subcategories on the basis of the percentage stability of their protease. In the elastase test, the ability of isolates to digest (positive elastase test) or not digest elastin particles (negative test) was measured up to 28 days incubation. Individual elastase positive isolates yielded a graded result based on the number of days to reach a positive result. There was a very high level of agreement between the gelatin gel and the elastase tests consistent with their separating isolates into two groups based on protease activity (either gelatin gel positive and elastase positive or gelatin gel negative and elastase negative). Either test is suitable for use in footrol control and eradication schemes. The gelatin gel test provides clearcut separation of isolates into positive and negative categories and has the major advantage of yielding a more rapid result than the elastase test. The elastase test should be utilised where a graded assessment of protease activity is desired.

Meningoencephalitis and other conditions associated with Histophilus ovis infection in sheep.

Histophilus ovis was isolated from 29 sheep in 20 flocks and 2 artificial insemination (AI) centres in southern New South Wales from 1984 to 1990. The clinical and pathological findings were consistent with previous reports and included polyarthritis (7 flocks), epididymo-orchitis (5), meningoencephalitis (3), pneumonia (3), septicaemia (2), mastitis (1) and metritis (1). Six sheep had meningoencephalitis, a syndrome not previously associated with H ovis infection in sheep, which was similar pathologically to thromboembolic meningoencephalitis in cattle, caused by the related organism, Haemophilus somnus. H ovis was isolated from the semen of 12-month-old rams in a flock that had polyarthritis due to H ovis, in 4-month-old ram lambs and from the uterus of a ewe in a flock that had sporadic cases of H ovis septicaemia.

Yersinia species isolated from sheep with enterocolitis.

In 40 submissions to the Regional Veterinary Laboratory (RVL) Wagga Wagga from sheep in southern New South Wales from 1981 to 1989, 53 isolates of Yersinia sp were recovered from 45 sheep in 37 flocks. Of 53 isolates, 26 were identified as Y. pseudotuberculosis, 20 as Y. enterocolitica, 5 as Y. intermedia and 2 as Y. frederiksenii. Twelve isolates of Y. pseudotuberculosis tested in the slide agglutination test all belonged to serotype III. The 20 Y. enterocolitica isolates were categorised biochemically as biotype 5 strains and, of 6 isolates serotyped, all belonged to serogroup 2,3. Outbreaks of yersiniosis were most common in late winter and early spring and affected flocks often had experienced a change in husbandry. Infection with Yersinia sp was associated with diarrhoea, illthrift and mortality. At necropsy, congestion and occasionally thickening of the intestinal mucosa were observed in affected sheep. Gastrointestinal nematodiasis and coccidiosis often were concurrent findings. The characteristic histological lesion in sheep infected with Y. pseudotuberculosis was acute segmental suppurative erosive enterocolitis. There were no lesions consistently associated with Y. enterocolitica, Y. intermedia or Y. frederiksenii.

Clostridial myocarditis in lambs.

Five outbreaks of myocarditis were investigated in young sheep. They occurred during late winter and spring when there was lush growth of pasture following a prolonged period of drought. Clinically the disease was characterised by sudden death and pathological findings were dominated by acute multifocal locally extensive necrotising and haemorrhagic myocarditis. A fluorescent antibody technique was used to demonstrate the presence of Clostridium chauvoei in paraffin embedded sections of myocardium from 4 of the outbreaks.

Examination of rams culled during an ovine brucellosis accredited free flock scheme.

During the early stages of an Ovine Brucellosis Accredited Free Flock Scheme in New South Wales 62 rams were examined to determine the status of their flocks of origin with regard to infection with Brucella ovis. Forty rams were selected because they were either single reactors to low titre or one of a small number of reactors in a B. ovis complement fixation test of the whole ram flock. Twenty two rams were selected because they had palpable abnormalities within the scrotum but were negative serologically. After serological, pathological, bacteriological and histological examinations they were classified in the ensuing categories: 7 positive, 7 inconclusive, 26 false positive and 22 with other lesions. The usefulness of this classification, particularly within the accreditation scheme is discussed.

An association between encephalomyocarditis virus infection and reproductive failure in pigs.

An outbreak of reproductive failure, characterised by mummified foetuses and stillbirths, was investigated in an intensive piggery. Six foetuses that died towards the end of gestation had multifocal myocardial necrosis and encephalomyocarditis virus was recovered from 4 of these foetuses but not from 6 mummified foetuses. There was also a significant increase in failure of conception or early embryonic deaths in sows mated at the same time as sows which produced affected litters.

Cryptosporidial infection of piglets.

Cryptosporidia were detected in the brush border of villous epithelium in the small intestine of 4 piglets 2 to 9 weeks old. Although the 4 piglets had been suffering from diarrhoea no specific role could be attributed to the cryptosporidia. Other than damage to the brush border there were no significant histological abnormalities in the small intestine associated with the presence of cryptosporidia.

The New South Wales mastitis control program. 2. Effect upon bacterial infections.

Bacteriological examinations were made on quarter samples from cows in 35 herds over a 3 year period to monitor the response in a mastitis control program. Initially, Staphylococcus aureus predominated in 32 of the herds and the mean herd prevalence was 26%. The control measures halved this rate but there was considerable variation in response between herds. The decline occurred rapidly and there was a significant reduction (P less than 0.01) by 3 months. Streptococcus agalactiae predominated in 3 herds and the overall infection rate was 4.9%. Control measures eliminated the infection completely from most herds but reinfection occurred in 2 herds. The greatest decline occurred in the first 6 months and was significant (P less than 0.05). The measures had little effect upon Str. uberis and Str. dysgalactiae which remained fairly consistently at low levels. Initially, strains of Staph. aureus resistant to penicillin were dominant in most herds. In a minority of herds strains resistant to streptomycin predominated and in these herds there was a concurrent resistance to penicillin. These patterns did not change greatly over the control period. Resistance by Str. agalactiae to streptomycin occurred in most herds at the start of the program.