The in vitro cytotoxicity of native and modified trichloroacetic acid-extracted antigens from Fusobacterium necrophorum for mouse peritoneal macrophages

A Boivin lipopolysaccharide-protein complex (LPS-P), Westphal LPS (W-LPS), Boivin LPS (B-LPS), and a lipid A-associated protein (LAP) were extracted from Fusobacterium necrophorum bov 5 strain and chemically analyzed for protein, total nitrogen, hydrolyzed hexosamine, and nucleic acid content. Dose and time effects on the in vitro cytotoxicity of these preparations for mouse peritoneal macrophages were determined. Macrophage monolayers were exposed to 2.0 ml of 10, 100, 250, 500, or 1000 μg/ml solutions of the individual preparations for 1, 2, 4, or 6 hr at 37°C, and relative toxicities were determined; B-LPS> LPS-P> LAP, W-LPS. The effects of heat, NaOH, and trypsin and pronase on the native antigens were determined. Heat treatment had no effect on the cytotoxicity of B-LPS, but increased the toxicity of LPS-P and LAP for macrophages. Alkaline treatment decreased the cytotoxicity of B-LPS and increased the toxicity of LPS-P and LAP for macrophages. Enzyme treatment had no effect on LPS-P toxicity. The relationship of lipid A and protein content to toxicity for macrophages is discussed. Lipid A appears to be important in the cytotoxicity of B-LPS, and the protein components of LPS-P and LAP may contribute to their toxic activity. The ability of these bacterial cellular antigens to destroy phagocytic cells may facilitate the establishment of necrotic abscesses in susceptible hosts.     

Milk antibodies against Ostertagia ostertagi: relationships with milk IgG and production parameters in lactating dairy cattle

The present study was carried out to evaluate the relationship between milk optical density ratios (ODRs) from an indirect Ostertagia ostertagi ELISA, total milk IgG levels and milk production and then establish a correction factor to adjust ODR. Five hundred and sixty composite milk samples collected from 358 cows on four dairy herds in June and August 2002 were used in this analysis. The average ODR was 0.34. A positive correlation was found between ODR and IgG values in milk, days in milk, age and log transformed somatic cell counts (SCC). However, ODR was negatively correlated with milk production. The IgG levels and ODR values were constant from 30 to 200 days in milk. However, ODRs increased from 200 days until the end of the lactation. After controlling for age, season, herd and SCC, an increase in milk production of 13 kg/day was associated with a reduction in ODR values of 0.052. The results of the present study suggest that ODR values are not greatly influenced by production factors. ODR follow the same pattern as the IgG variation across lactation and could be adjusted in order to compare ODR values obtained from high producing cows with those obtained from low producing animals.     

Poxviruses as vaccine vectors

The discovery of Jenner in 1798 founded the science of immunology and eventually led to smallpox eradication from the earth in 1980 after a world-wide vaccination campaign with vaccinia virus (another poxvirus) and paradoxically, despite the eradication of smallpox, there has been an explosion of interest in vaccinia virus in the eighties. This interest has stemmed in part from the application of molecular genetics to clone and express foreign genes from recombinant vaccinia viruses. Vaccinia is also gaining renewed interest due to bioterrorism.

These recombinant viruses have multiple applications in research and vaccinology and led to the development of vectored vaccines, such as the recombinant vaccinia rabies vaccine used to eliminate rabies in Western Europe and, more recently, in the United States. Secondly, alternative poxvirus vectors, such as avipox viruses, were proved to be even safer and efficacious non-replicating vectors (suiciole vectors) when used in non-avian species.     

New gene-based approaches for an AIDS vaccine

Vaccine approaches against AIDS have focused on inducing cellular immune responses, since many studies revealed the role of T cell responses in the control of human immunodeficiency virus or simian immunodeficiency virus (SIV) infections. The experimental infection of rhesus macaques with SIV or chimeric SHIV is routinely used as a model for AIDS. In such models, DNA immunization is a tool to elicit specific T cell responses and to study their protective efficacy. DNA immunogenicity in primates depends on parameters such as level of antigen expression, choice of the antigen among SIV proteins, use of fusion proteins, route of immunization, and addition of adjuvants. Recent results suggest that priming with DNA and boosting with attenuated recombinant viral vectors, each expressing corresponding SIV antigens, leads to improved specific immunity and, in some cases, affords protection against pathogenic challenge. After preclinical evaluations, DNA has entered clinical trials for a therapeutic or prophylactic gene-based AIDS vaccine.     

Corynebacterium equi: An interhost review with emphasis on the foal

A review of the recent and/or significant literature concerning Corynebacterium equi, including its morphologic, biochemical, immunological, and pathological characteristics in the foal, humans and other animals is presented. The similarity in the tissue responses of mammalian hosts to C. equi and Mycobacterium spp. is discussed.

The antigenic structure and virulence factors associated with C. equi. other corynebacteria and mycobacteria are compared.

The immunological aspects of resistance to C. equi are considered. The evidence suggests that the major immune response elicited in the foal by C. equi is cell-mediated. However, the immunopathogenic mechanism needs clarification. Areas of future research are suggested.     

Susceptibility of various ceil culture systems to pseudorabies virus

A comparative study was carried out to determine the susceptibility of five different cell lines to pseudorabies virus (PRV), a herpes virus of pigs. The cell systems tested were swine testicle (ST), mink lung (ML), equine dermal (ED), porcine kidney (PK₁₅), and bovine turbinate (BT) cells. Virus titers obtained were 10^4.88, 10^4.38, 10^3.75, 10^2.63, and 10^0.25 for ML, ST, PK₁₅, BT and ED cells, respectively indicating that ML, ST, and PK₁₅ are optimal cell lines for the growth of PRV whereas BT and ED are not very sensitive.     

Host immune responses after administration of inactivated Venezuelan equine encephalomyelitis virus vaccines—III. Kinetics for neutralizing antibody immunoglobulin class responses in donors and adoptively-immunized recipients

C57B16 mice were immunized with either live, attenuated TC-83 strain VEE virus vaccine or formalin-inactivated VEE vaccine combined with Bordetella pertussis. The kinetics of specific donor and adoptively-immunized recipient anti-VEE neutralizing antibody responses were studied. Donor mice immunized with either live or inactivated VEE virus vaccine combined with potent adjuvants develop specific anti-VEE IgM and IgG responses as early as 7 days post-immunization. Anti-VEE IgM antibody responses comprise the majority of anti-VEE neutralizing antibody at this early time period. By 14 to 21 days post-immunization, anti-VEE IgG responses predominated. When adoptively-immunized recipients were studied, the anti-VEE IgM to IgG predominance seen in donors early after administration was reversed, and for each time-period studied, recipients' serum anti-VEE antibody class responses consisted principally of IgG rather than IgM antibody. Since T-cells cooperation with B-cells is critical in the IgM-IgG antibody shift, these studies support the critical role T-cells exert in adoptive transfer in a murine model of experimental VEE infection. Furthermore, immunization with either live or inactivated VEE vaccine coupled to a potent adjuvant induce comparable donor and adoptively-immunized recipient anti-VEE antibody class responses.     

Serotypes of Escherichia coli strains isolated from piglets with diarrhea in swine industrial farms

Serotypes of E. coli strains isolated from piglets, which died with symptoms of diarrhea in 9 swine industrial farms, were determined. Large numbers of serotypes (from 16 to 27) in individual farms were detected. The sets of serotypes from 9 investigated farms differed among each other significantly, depending on the farm and time of examination. It was found that more than one serotype of E. coli may exist in the pig body and contribute to the development of disease. The predominant serotypes, i.e. those comprising more than 10% of serologically determined strains, were found to exist in 6 of the investigated farms and not in the remaining ones. Among the predominant serotypes, particularly important seem to be strains with K88 antigen. For prophylaxis of piglet colibacteriosis in industrial farms in Poland two vaccines for sows are recommended: one containing the K88 antigen only and the other the following serotypes: 0149:K91,K88; 020:K57; 020:K83; 0157:K88; 01:K1; 0136:K78; 024:K?; 078:K80 and 0118:K? Strains belonging to these serotypes were the most prevalent in our strain collection.     

Specific identification of Pasteurella multocida serogroup-A isolates by PCR assay

A polymerase chain reaction (PCR) assay targeting the hyaC-hyaD gene was developed and used to identify strains of Pasteurella multocida belonging to serogroup-A. A set of serogroup-specific-PCR primers amplified a 564 bp product from genomic DNA prepared from bacterial cells or directly from bacterial colonies. This method detected as low as 10 ng of bacterial DNA and had a specificity of 100% for P. multocida serogroup-A. A nested PCR method yielded a single 374 bp product. All fifty isolates were also shown to be identical by restriction fragment length polymorphism (RFLP) analysis of the PCR products after digestion with BglII.     

Evaluation of the chemoprophylactic efficacy of 10% long acting injectable moxidectin against gastrointestinal nematode infections in calves in Belgium

The chemoprophylactic efficacy of a single dose of the 10% long acting (LA) injectable formulation of moxidectin on nematode infections in calves, was evaluated. Two similar groups of 11 female, first grazing season Holstein calves were turned out in early May on separate plots of a single, naturally infected pasture. Until 56 days post-treatment (pt), the percentage reduction in faecal egg output was 100%, remaining above 90% during the entire trial, except for day 126 pt. More than 90% of the larvae in the treated group were identified as Cooperia until 140 days after treatment and more than 70% during the rest of the trial, whereas in the control group Cooperia was the most abundant species until day 84 pt and Ostertagia from 126 days pt onwards. The reduction in faecal egg output in the treated group was reflected in the mean pepsinogen levels being below the pathogenic threshold at the end of the grazing season (1.8 units of tysrosine (U tyr)) and the absence of diarrhoea during the second half of the grazing season. In the control group pepsinogen levels remained high (mean: 5.5 U tyr) and prolonged diarrhoea occurred in the second half of the grazing season. Furthermore, the weight gain for the treated group at the end of the grazing season was 41.9 kg higher than for the control group. At necropsy, the reduction in O. ostertagi worm burden in the treated group was 97.5% compared to the control group, while the reduction in C. oncophora worm burden was 57%. An additional benefit of the long acting parasitological control, was reduced pasture contamination.