An ELISA for porcine reproductive and respiratory syndrome: production of antigen of high quality.

A reliable method was developed to produce a viral antigen preparation from porcine reproductive and respiratory syndrome virus (PRRSV) infected MARC-145 cells by solubilizing the virus with Triton X-100. This method eliminated problems previously encountered with high background reactions associated with PRRSV antigen or cell control antigen. With this new antigen, an indirect enzyme-linked immunosorbent assay (ELISA) was adapted to detect swine serum anti-body against PRRSV. In the ELISA, non-specific reactions associated with test serum samples have been eliminated by utilizing an effective blocking serum diluent. The ELISA is more sensitive than an indirect immunofluorescent assay (IFA), particularly with late-infection sera, while maintaining a high diagnostic specificity. In a comparison of IFA and ELISA using sera collected from 250 pigs of various ages from 5 herds that had PRRS histories, IFA revealed 178 positive samples and 72 negative samples. All of the IFA-positive sera were proven to be ELISA reactors. However, nearly one-half (34/72) of the IFA-negative samples were also ELISA reactors. The diagnostic specificity and sensitivity of the ELISA were 100% and 96.6% with 257 serum samples collected from known healthy PRRS-negative swine herds and 57 sera collected from infected swine at 6 to 56 days after infection, respectively. The ELISA is technically superior to IFA, time-efficient and cost-effective, and suitable for testing of a large number of samples over a short period of time.     

Outcome of surgical correction of meconium impactions in 8 foals

Meconium impactions are only rarely refractory to medical therapy. The purpose of this paper is to examine the outcome of 8 foals that required an exploratory celiotomy to correct a meconium impaction. Between 1984 and 1992, 24 foals were referred with a primary diagnosis of meconium impaction. All foals were treated medically prior to and following referral. Of the 24 foals, 8 had impactions requiring surgical intervention. Exploratory celiotomies were performed, and the impaction was reduced manually or by enterotomy. Follow up information was available on 7 foals. All survived surgery and were discharged. Four of the 8 foals matured and raced. Two foals were euthanatized due to extensive serosal adhesions and one foal was euthanatized due to an unrelated orthopedic condition. Our results support the decision for an exploratory surgery only after aggressive medical therapy has failed. Several treatment options have been developed in recent years that have reduced the number of foals that may require surgery.     

Specific serum antibody responses in channel catfish (Ictalurus punctatus) provide limited protection against Streptococcus ictaluri challenge

Passive immunization studies were conducted to determine the role of specific antibodies in immunity to Streptococcus ictaluri. Adult channel catfish (Ictalurus punctatus) were injected i.p. with tryptic soy broth as control or with 1.5 × 10(7)colony-forming units (cfu) S. ictaluri/fish at 0, 30, and 60 d, and serum was collected 90 d after the original challenge. Fish were passively immunized by i.p. injection with serum from the tryptic soy broth (TSB) control group, anti-S. ictaluri serum from fish immunized three times and sampled at 90 d (SSI), or heat-inactivated anti-S. ictaluri serum from fish immunized three times and sampled at 90 d (HISSI). These passively immunized fish were then challenged 72 h later with 1.5 × 10(8)cfu S. ictaluri/fish. Over 21 d, the mean cumulative percent survival was 43.3 (TSB), 63.3 (SSI), and 50.0 (HISSI). A significant difference in cumulative percent survival was noted between the TSB and the HISSI groups, and significant differences were noted between these groups and the SSI group. Serum obtained from immunized fish 72 h after passive immunization exhibited increased anti-S. ictaluri antibody levels. Twenty-one days after the challenge, the HISSI and SSI group antibody levels significantly increased above their corresponding pre-challenge levels. No significant (r(2)=0.0806; P<0.5985) correlation between increased pre-challenge specific serum antibody levels and survival after challenge was demonstrated when analyzing the control and passive immunization groups. The results indicate that both specific anti-S. ictaluri antibodies and non-specific immune responses are important for protection against S. ictaluri.     

Ultrasonography of the rumen in 30 Saanen goats

This study describes the results of ultrasonographic examination of the rumen in 30 healthy Saanen goats. A linear or convex transducer with a variable frequency of 5 to 13 MHz was used to scan standing, non-sedated goats. The location and size of the rumen, the distance between the wall of the rumen and abdominal wall and the appearance and size of the gas, fibre mat and fluid layers of the ruminal contents were assessed. The rumen was seen as a large organ medial to the left abdominal wall. The wall of the rumen appeared as a thick echogenic line. The longitudinal groove was seen as an echogenic notch, which divided the rumen into the dorsal and ventral sacs. The rumen could be visualized from the 9th to 12th intercostal space (ICS) and flank on the left side in all the goats. The rumen was largest in the 12th ICS at 41.6 ± 5.13 cm and smallest in the 8th ICS at 11.3 ± 4.29 cm. The dorsal sac of the rumen was largest in the left cranial flank (17.4 ± 4.43 cm) and the ventral sac was largest in the 12th ICS on the left (29.1 ± 6.03 cm). In the cranial left flank, the rumen was situated immediately adjacent to the abdominal wall in all the goats. The spleen was located between the rumen and abdominal wall in the 8th to 12 th ICS in many of the goats. The gas, fibre mat and fluid layers of the ruminal contents could be visualized in all the goats. The gas layer was 9.9 ± 3.05 cm, the fibre mat layer 16.0 ± 4.55 cm and the fluid layer 12.2 ± 5.57 cm.     

Decreased serum apolipoprotein A-I concentrations in cows infected with Salmonella typhimurium.

Serum apolipoprotein A-I concentrations in cows infected with Salmonella Typhimurium were evaluated to assess its relevance in salmonellosis. Apolipoprotein A-I has been shown in rats to be secreted by the intestine as well as the liver. Clinical symptoms such as diarrhea revealed an outbreak of salmonellosis in 22 cows on a farm, and sera were obtained at 6 (acute phase), 16, 28 (convalescent period) and 42 d (postconvalescent period) after the outbreak. Apolipoprotein A-I concentrations (mean +/- SD, mg/mL), determined by ELISA, were 0.598 +/- 0.497 (day 6), 0.111 +/- 0.060 (day 16), 0.432 +/- 0.311 (day 28) and 0.727 +/- 0.516 (day 42). Compared with the concentration at day 42, those at 16 and 28 d were significantly (P < 0.01, P < 0.05) lower, but that at day 6 was not. The serum concentration of apolipoprotein B-100 (of liver origin in cattle) was unaltered during the course of salmonellosis. The concentration of apolipoprotein A-I was positively correlated with those of serum total cholesterol (r = 0.589, P < 0.01) and phospholipids (r = 0.590, P < 0.01). These results suggest that apolipoprotein A-I in cattle is in part of intestinal origin, and also that its decreased serum concentration in salmonellosis can be attributed to the reduced intestinal synthesis or secretion of this apolipoprotein. Moreover, as a potential carrier for dietary lipids such as cholesterol, determination of serum apolipoprotein A-I concentration is suggested to be useful when assessing the nutritional status of the affected cows.     

Disseminated Mycobacterium paratuberculosis infection in a cow

A cow with chronic diarrhea and weight loss caused by localization of Mycobacterium paratuberculosis in the intestinal tract (Johne's disease) had gross and microscopic changes indicative of a disseminated infection. A direct association between the remote lesions and the intestinal infection was shown by isolation of M paratuberculosis from renal tissue, detection of intracellular M paratuberculosis antigen(s), using an indirect immunoperoxidase method, and by the characteristic granulomatous nature of the lesions. This case illustrates the potential for extra-intestinal lesions in M paratuberculosis infection of cattle and should cause veterinarians to consider mycobacterial disease when confronted with multinodular lesions of the bovine kidney. The immunoperoxidase method was useful in determining the cause of the inflammatory lesion in which intact organisms were not evident.