Use of restriction fragment length polymorphism, immunoblotting, and polymerase chain reaction in the differentiation of avian poxviruses.

Restriction deoxyribonucleic acid (DNA) fragment profile analysis coupled with immunogenic protein profile analysis has provided useful information in determining the differences between vaccine strains and field isolates of fowlpox virus (FPV). The DNA of strains examined in this study clearly fell into 3 minor groups of restriction patterns similar but distinct from one another: restriction patterns exhibited by the vaccine strains except 1 vaccine strain, Vac-82; restriction profiles indicated by Vac-82 and field isolates FI-38 and FI-42; and restriction patterns indicated by field isolates FI-43, FI-51, FI-54, and FI-56. Furthermore, when the strains were analyzed and compared by immunoblotting analysis, they showed group differences similar to the differences in restriction profiles. Both techniques provided high sensitivity in verifying differences between vaccine strains and field isolates of FPV. The disparity found in restriction fragments or immunogenic protein profile between vaccine strains and field isolates does not exclude the appreciable high degree of DNA sequence conservation and homology. However, the minor disparity observed in these strains suggests a molecular basis for why vaccinated commercial flocks could have continually been infected by variant strains of FPV. A rapid and sensitive polymerase chain reaction method, which amplified a product from the 4b core protein gene of the FPV genome, was developed for identification and differentiation of members of the genus Avipoxvirus. Whereas total DNA from either vaccine strains or field isolates was used as template for amplifying a predicted product of 578 or 1409 bp, only cleavage of the amplified product (1409 bp) represented an additional detection technique for species differentiation. An attempt to distinguish between strains on the basis of amplification product was partially successful.     

Enzootic Bovine Leukosis

The author emphasizes the significance of enzootic bovine leukosis in Canada. He describes in detail diagnostic methods, various types of the disease and methods of transmission.

Various aspects of the disease in Canada are compared with those in other countries. Prevention and control are discussed in a Canadian context and include the current policies of the Government of Canada in relationship to this disease. The possibility of developing a certification program for herds free of the disease is also discussed. The paper includes incidence in various parts of Canada.

     

Clearance and shedding of infectious bovine rhinotracheitis virus from the nasal mucosa of immune and nonimmune calves

Infectious bovine rhinotracheitis virus was rapidly cleared from the nasal mucosa of calves after intranasal aerosol exposure. Nonimmune calves (experiment 1) cleared 10(9) plaque-forming units (PFU) of virus from the nasal mucosa in less than 4 hours and 10(6) PFU of virus in 1 hour. An eclipse phase followed the clearance of viral inoculum. Replicating virus was first detected at 9 hours. Viral titers increased stepwise until maximum was attained on postinoculation day 4. Virus persisted in the nasal mucus until day 12. Clinical signs of disease corresponded with the shedding of virus. In contrast to nonimmune calves, immune calves (experiment 2; same calves as in experiment 1, but 30 days after initial exposure) cleared 10(9) PFU of virus in 1 hour and 10(6) PFU of virus in less than 5 minutes. An abortive reinfection occurred after exposure of immune calves with 10(9) PFU of virus. Virus was first detected in these calves at 14 hours after exposure and was not detected beyond 24 hours after inoculation. Immune calves given 10(6) PFU of virus did not shed virus after clearance of inoculum. Clinical signs of infection were not observed in immune calves after viral challenge exposure. The date indicated that there was no detectable residual virus beyond 3 hours after the exposure.     

Comparison of inoculation regimes for the experimental production of swine erysipelas arthritis: II Serological findings in a gel diffusion precipitin test and enzyme-linked immunosorbent assay

The serological response of pigs to Erysipelothrix rhusiopathiae inoculation was monitored by a gel diffusion precipitin test (GDPT) using a crude, serotype-specific, autoclaved antigen and an enzyme-linked immunosorbent assay (ELISA) using a heat-extracted, alcohol precipitated and molecular seived antigen previously shown to react with serum from pigs infected with serotypes 1 or 2. All pigs receiving 3 or 5 weekly intravenous inoculations of either a highly virulent (VRS 229) or a lowly virulent isolate (VRS 252) produced GDPT-reactive antibody within 3 weeks, but only 44% were still reactive at 8 to 9.5 weeks. The ELISA response was significantly higher in pigs inoculated with the highly virulent strain, and was similar in pigs receiving 3 or 5 doses of either strain. In a dose-response trial, after 3 doses of VRS 229, GDPT reactivity occurred earlier and was stronger in pigs given higher doses of E. rhusiopathiae, but the response peaked 3 to 5 weeks after the start of challenge and was short lived. GDPT reactivity correlated with dose, but not with the severity of arthritis. The ELISA demonstrated specific IgG antibody was present by 2 weeks, and persisted to at least 11 weeks. The ELISA reactivity was significantly higher in pigs with arthritis than in pigs that received low doses and were not arthritic. Within groups of pigs with arthritis a significant, dose dependent, linear ELISA response developed but did not correlate with the presence or degree of arthritis at slaughter. Non-arthritic pigs had similar low ELISA responses to uninoculated controls.     

Effect of field pea level on intake, digestion, microbial efficiency, ruminal fermentation, and in situ disappearance in beef steers fed growing diets

Effects of increasing level of field pea (variety: Profi) on intake, digestion, microbial efficiency, and ruminal fermentation were evaluated in beef steers fed growing diets. Four ruminally and duodenally cannulated crossbred beef steers (367+/-48 kg initial BW) were used in a 4 x 4 Latin square. The control diet consisted of 50% corn, 23% corn silage, 23% alfalfa hay, and 4% supplement (DM basis). Treatments were field pea replacing corn at 0, 33, 67, or 100%. Diets were formulated to contain a minimum of 12% CP, 0.62% Ca, 0.3% P, and 0.8% K (DM basis). Each period was 14 d long. Steers were adapted to the diets for 9 d. On d 10 to 14, intakes were measured. Field pea was incubated in situ, beginning on d 10, for 0, 2, 4, 8, 12, 16, 24, 36, 48, 72, and 96 h. Bags were inserted in reverse order, and all bags were removed at 0 h. Ruminal fluid was collected and pH recorded at -2, 0, 2, 4, 6, 8, 10, and 12 h after feeding on d 13. Duodenal samples were taken for three consecutive days beginning on d 10 in a manner that allowed for a collection to take place every other hour over a 24-h period. Linear, quadratic, and cubic contrasts were used to compare treatments. There were no differences in DMI (12.46 kg/d, 3.16% BW; P > 0.46). Ruminal dry matter fill (P = 0.02) and mean ruminal pH (P = 0.009) decreased linearly with increasing field pea level. Ruminal ammonia-N (P < 0.001) and total VFA concentrations (P = 0.01) increased linearly with increasing field pea level. Total-tract disappearance of OM (P = 0.03), N (P = 0.01), NDF (P = 0.02), and ADF (P = 0.05) increased linearly with an increasing field pea level. There were no differences in total-tract disappearance of starch (P = 0.35). True ruminal N disappearance increased linearly (P < 0.001) with increasing field pea level. There were no differences in ruminal disappearance of OM (P = 0.79), starch (P = 0.77), NDF (P = 0.21), or ADF (P = 0.77). Treatment did not affect microbial efficiency (P = 0.27). Field pea is a highly digestible, nutrient-dense legume grain that ferments rapidly in the rumen. Because of their relatively high level of protein, including field peas in growing diets will decrease the need for protein supplementation. Based on these data, it seems that field pea is a suitable substitute for corn in growing diets.