Differentiation of porcine Eimeria spp. by morphologic algorithms

Oocysts of Eimeria spp. were isolated from feces of naturally infected sows by conventional flotation. Saturated sodium chloride solution was superior to zinc chloride, zinc chloride/sodium chloride or sugar solution to isolate oocysts. Seven species, namely Eimeria scabra, E. polita, E. perminuta, E. debliecki, E. suis, E. porci and E. spinosa, were identified. The dimensions of oocysts (n = 4088) and sporocysts (n = 3594) were measured with an image analysis system; colour and shape of oocysts were estimated and transformed to numerical values. Of the 4088 oocysts approximately 99% were allocated to the correct species by algorithms calculated on the basis of these values. Rough-walled oocysts (E. scabra, E. polita, E. perminuta, E. spinosa) could be distinguished without previous sporulation in most cases (>97%). Smooth-walled oocysts require sporulation for further classification and were accurately allocated to species in at least 93% of cases.     

Cytotoxicity for porcine islet cells by complement of six animal species

Complement-mediated cytotoxicity for porcine islet cells (PICs) was evaluated using sera of six animal species. Then soluble complement receptor type-1 (sCR1) as an anti-complement agent was added to those sera, and the changes in 50% hemolytic unit of complement serum (CH50) and cytotoxic effect of those sera on PICs were examined. All the sera except for that of pig showed cytotoxicity. However, the extent of toxicity was considerably different between species. In the rat and human serum, sCR1 significantly reduced CH50 and cytotoxicity, however in the dog serum, sCR1 had no suppressive effects. These results may suggest that complement contribute to humoral cytotoxicity for PICs as a main factor, and the compatibility of complement with PICs differs between animal species.     

Pseudorabies virus, porcine parvovirus, and porcine enterovirus interactions with the zona pellucida of the porcine embryo

Porcine embryos (n = 93) were incubated on cell monolayers that had been previously inoculated with pseudorabies virus, porcine parvovirus (PPV), or each of 2 porcine enteroviruses. After 2, 24, or 48 hours of incubation, the embryos were fixed in glutaraldehyde and examined by electron microscopic procedures. It was found that pseudorabies virus adsorbed to the zona pellucida (ZP) and entered sperm tracks in the ZP. The PPV and both enteroviruses entered pores in the ZP and were associated with sperm that were at or near the outer surface of the ZP. In addition, PPV was seen enmeshed in cellular debris on the outer surface of the ZP. Evidence of a productive viral infection of the blastomeres of the embryos was not found.     

The protective effect of two porcine enterovirus vaccines in swine.

The virus neutralizing substance in the gastrointestinal tract of swine vaccinated in different ways with porcine enterovirus strain T80 was characterized by tests for enhancement and absorption of virus neutralizing activity by class specific antiporcine Ig antisera. The gastrointestinal virus neutralizing activity of piglets which were vaccinated with live virus orally resided predominantly in the IgA class, although some activity was present also in the IgM and IgG classes. The serum virus neutralizing activity of this group was present in all three classes but primarily in the IgG class. The IgA in the serum of this group was presumably of gut origin. However, in piglets vaccinated with live virus intramuscularly, with formaldehyde-inactivated virus orally or intramuscularly or with ethylenimine-inactivated virus by both oral and subcutaneous routes, both serum and gastrointestinal virus neutralizing activity were attributable predominantly to antibodies of the IgG and IgM classes. None possessed serum IgA. There was evidence also for the presence of Ig fragments in some gastrointestinal extracts.     

Differentiation of twelve Actinobacillus pleuropneumoniae serotypes by outer membrane lipoprotein gene-based restriction fragment length polymorphism

Twelve Actinobacillus pleuropneumoniae serotypes were differentiated by restriction fragment length polymorphism (RFLP) of polymerase chain reaction (PCR)-amplified fragments from the outer membrane lipoprotein (omlA) gene. All 12 reference serotypes and 80 field isolates produced the expected 950-base pair (bp) fragment of the omlA gene by PCR. Combining the RFLP patterns obtained with SfaNI, Bst71I, AluI, NciI, nine distinct patterns were observed in the 12 serotype reference strains. The PCR-based RFLP analysis of omlA genes allows differentiation among the 12 serotypes, with the exception of group 1 (serotypes 1, 9 and 11), and group 2 (serotypes 2 and 8). When the PCR products from the 70 field isolates were subjected to RFLP analysis, 68 showed the same RFLP patterns as their respective serotype reference strain. Two isolates that could not be typed had the same RFLP patterns as those of serotype 5. These results suggest that PCR-based RFLP analysis of the omlA genes may be of value in differentiating among 12 A. pleuropneumoniae serotypes.