Experimental infection of neonatal pigs with CNF toxin-producing strains of Escherichia coli

To examine and compare the pathogenicity of cytotoxic necrotising factor (CNF)-producing Escherichia coli, two litters of piglets were infected orally with 10¹⁰E coli O88 or 10¹⁰E coli O32 strains. Of the six piglets infected with E coli O88, two died within 24 hours and three developed blood-stained diarrhoea. The other piglets were killed one, five, six and eight days after infection, when bacterial cultures indicated an overwhelming bacteraemic infection with E coli O88 in the early stages followed by clearance through the large intestine. The pathological changes consisted of an early enteritis, progressing to enterocolitis and a bacteraemic spread to the lungs. The histopathological changes were characteristic of toxaemic effects in brain, heart, liver and kidney, and characterised by congestion, oedema and exudation. Infection with E coli O32 produced a milder but similar enterocolitis, also with bacterial colonisation in the lungs. The histopathological findings again reflected a toxaemia. The enteritis was more persistent after E coli O32 infection and the strain persisted in large numbers in the intestine. No evidence of bacterial adherence to the intestinal mucosa was found with either strain. Enteroinvasion was only evident in one E coli O884-nfected piglet, but the consistent occurrence of interstitial pneumonia showed the predilection of these organisms for the lung. The results confirm the toxigenic properties of CNF⁺E coli and suggest an important role for this organism in enteric infection of young pigs.     

Serum and mucosal antibody responses against Mycoplasma hyopneumoniae following intraperitoneal vaccination and challenge of pigs with M hyopneumoniae

Pigs were immunised intraperitoneally when six weeks old and again at about 10 weeks old with killed Mycoplasma hyopneumoniae antigen prepared in an oil adjuvant. The pigs were challenged with live M hyopneumoniae (Beaufort strain) at between 11 and 15 weeks old. Antigen specific antibody levels for both IgG and IgA classes in serum and respiratory tract secretion were monitored over time. In serum anti-M hyopneumoniae antibody was detected shortly after the second intraperitoneal vaccination and was largely IgG. In respiratory tract secretion the response was observed after challenge, and was primarily IgA. Anti-M hyopneumoniae antibody-containing cells and their immunoglobulin class specificity were monitored in lung and tracheal lamina propria. In lung the majority of anti-M hyopneumoniae-containing cells were IgG, whereas in the tracheal lamina propria the majority were IgA. These results are discussed in terms of the use of intraperitoneal vaccination for the control of M hyopneumoniae infection.     

Seroepidemiologic study of natural transmission of Mycoplasma hyopneumoniae in a swine herd

A cohort of 57 pigs in a farrow-to-finish swine herd with mild clinical mycoplasmal disease was followed to determine patterns of seroconversion to Mycoplasma hyopneumoniae (MH), detected with an enzyme-linked immunosorbent assay (ELISA). Survival analysis was used to evaluate the relationship between time to seroconversion and possible risk factors for MH infection (or enzootic pneumonia).

Pigs were housed in outdoor pens at approximately 9 weeks of age, when passively acquired MH antibodies had decayed. From 9 to 11 weeks of age and during a 5 week period, pigs were exposed by direct (nose-to-nose) or indirect contact to older seropositive gilts. Blood samples were collected from each pig at 3 week intervals until market age, when they were either slaughtered or selected for breeding. Antibody concentration was measured as the ratio of optical densities of the serum sample to the positive control (S/P). Based on the sample distribution of S/P ratios from pigs in an MH-free herd, pigs were considered positive when S/P ratios were greater than 0.34. At the beginning of the study, all pigs were seronegative to MH. Seroconversion was first detected after 21 days, and was most frequent about 11 weeks after exposure to older seropositive gilts. By the end of the study, 11 pigs (19%) had seroconverted, with S/P ratios ranging from 0.40 to 1.11. The presence of gross lung lesions showed a moderate to good agreement with ELISA results (K = 0.62). Histologic lesions were evident in virtually all slaughtered pigs, ranging from mild, non MH-specific lesions to severe lesions typical of MH infection. No secondary respiratory pathogens were isolated. Clinical signs were mild and there was no significant difference (P > 0.4) in weight gain between seropositive and seronegative pigs, or between pigs with and without lung lesions. A Cox regression model was fitted to the seroconversion data, and opportunity of contact (direct or indirect) was the only significant variable. After adjustment for breed and antibody S/P ratio prior to exposure, pigs in direct contact with seropositive gilts were seven times more likely to seroconvert than those in only indirect contact.     

H5 antibody detection by blocking enzyme-linked immunosorbent assay using a monoclonal antibody

Many commercial enzyme-linked immunosorbent assays (ELISAs) are unable to differentiate antibody responses to different avian influenza virus (AIV) subtypes. Developing an ELISA for specifically detecting the H5 antibody is the purpose of this study. Four monoclonal antibodies (Mabs) were raised using A/duck/Yunlin/04 (H5N2). They were confirmed as being specific to H5. Two of these antibodies showed hemagglutination inhibition (HI) activity using the HI test. Using immunodot blot assays, three Mabs recognized both Eurasian and American H5, whereas the other Mab recognized only the tested Eurasian H5 virus. When testing denatured H5 antigen, one of the Mabs lost its antigen binding activity using Western blotting. For detecting the H5 humoral response in serum, one monoclonal antibody was purified and labeled with horseradish peroxidase to set up a blocking ELISA. Chicken sera that blocked H5 Mab binding by > 29% were considered H5 antibody positive. Inhibition percentages for sera from chickens infected with other AIV subtypes, H1 to H15, were < 29%. This blocking ELISA was used for 478 field chicken serum samples. The results showed that the sensitivity and specificity of this ELISA were 98.3% (232/236) and 95.9% (232/242), respectively. This blocking ELISA could be used specifically for detecting the H5 humoral responses in chickens.     

An ELISA test to detect antibody to Bordetella bronchiseptica

An enzyme-linked immunosorbant assay (ELISA) was developed to measure antibody to $\text{Bordetella bronchiseptica}$ in dogs. The ELISA test was more rapid and sensitive and required 50 to 150 times less antigen than the amount of antigen required for the conventional tube agglutination test. A survey of 50 canine serum samples using ELISA suggested that 8% of all sera had titers greater than 1:64, 56% had titers of 1:8 to 1:64, and 36% had titers of less than 1:8. The mean titer of survey sera was 1:46 and the median titer was 1:16. Serum antibody responses in dogs inoculated with a commercially available bacterin were compared with responses in dogs inoculated with experimental endotoxin depleted bacterin.     

Responses of ponies to equid herpesvirus-1 Iscom vaccination and challenge with virus of the homologous strain

An experimental (Iscom) vaccine previously shown to protect hamsters from lethal challenge with equid herpesvirus-1 (Ehv-1), was tested in horses. Vaccination with Ehv-1 Iscoms induced serum antibodies to the major virus glycoproteins gp10, 13, 14, 17, 18 and 21/22a, whereas antibody responses to gp2 were weak or absent. High levels of virus neutralising antibody of long duration were induced, but did not prevent challenge infection with virus of the homologous strain. However, in the vaccinated ponies there was a significant reduction in clinical signs, nasal virus excretion and cell associated viraemia compared with age-matched unvaccinated controls. There was a strong correlation between pre-challenge levels of serum virus neutralising antibody and the duration and total amount of virus excreted from the nasopharynx.     

Lack of immunostimulatory effect of N-acetyl muramyl-l-alanyl-d-isoglutamine on selected chicken immune functions

Synthetic N-acetyl muramyl-l-alanyl-d-isoglutamine, syn. muramyl-dipeptide (MDP) was found to be immunostimulatory in several experimental animal species. In order to determine the influence of MDP on the chicken immune response, different doses (0.05–0.2 mg) of this compound were administered to 6-week old chickens, and cellular as well as humoral immune functions were tested. Neither the immune response against sheep red blood cells or Newcastle disease virus (strain Hitchner B 1), nor the ability to reject skin grafts, or to react in the delayed hypersensitivity (tuberculin) test, were affected significantly under the experimental conditions employed. This study reveals little evidence for parallels between the ability of the chicken immune system and the immune system of other animal species examined so far, to develop enhanced immune reactions under the influence of MDP.     

Presence of antigen sensitized leukocytes in carp (Cyprinus carpio L) following bath immunization against Flexibacter columnaris

Bath immunization of carp (Cyprinus carpio L) resulted in protection of fish at natural challenge. Stimulation of leukocytes derived from thymus, spleen, anterior kidney and mid-kidney of fish immunized with Flexibacter columnaris bacterin revealed the presence of antigen sensitized cells in all lymphoid tissues except the anterior kidney. After 28 days a response was obtained in thymus and spleen leukocyte cultures.     

Bovine babesiosis: The immunization of cattle with fractions of erythrocytes infected with Babesia bovis (syn B. argentina)

Soluble antigen which protected susceptible cattle against challenge with $\text{Babesia bovis}$ was extracted from $\text{B. bovis}$-infected erythrocytes by sonic disintegration and separation of the soluble from the insoluble matter by ultracentrifugation. The material was then fractionated by the precipitation of fibrinogen-like proteins. The precipitate contained the babesial antigens that were located on the stroma of the infected erythrocytes. Antigen originally located on the parasite remained in solution. Both fractions conferred protection on splenectomized calves against challenge with $\text{B. bovis}$. However, the fraction containing the parasite antigens appeared to have more potential for development as a killed vaccine because it was not heavily contaminated with antigenic material from bovine erythrocytes.     

Brucella abortus antigen-induced lymphocyte reactivity in guinea pigs

A whole blood lymphyocyte transformation (WBLT) assay was used to detect anti-brucella lymphocyte reactivity in guinea pigs. Brucella antigens stimulated an antigen-specific lymphoproliferative response in WBLT assays from $\text{Brucella abortus}$ infected guinea pigs. The response was best detected from 6 to 16 weeks after challenge inoculation with viable $\text{B. abortus}$ 2308. Lymphocytes were not stimulated by unrelated bacterial antigens and control animals did not respond to the Brucella antigens. The responding cell population was characterized as mostly T lymphocytes. The WBLT assay was found to be specific for the detection of anti-brucella lymphocyte reactivity. However, a negative response was not definitive, which indicated a need for repeated testing to establish that a guinea pig did not have anti-brucella lymphocyte sensitization.     

Effects of dietary supplementation of l-carnitine on the reproductive performance of sows in production stocks

Investigations by Robinson [Robinson, D.L., 2007. Days to calving in artificially inseminated cattle: comparison of potential traits. Livestock Science 110, 174–180] concluded that the most useful trait for assessing fertility of artificially inseminated (AI) beef cows is AI days to calving (AIDC), a trait that mimics days to calving for naturally mated cows. Various fixed and random effects were fitted to AIDC to determine the best way of modelling lactation status of the cow, the effect of service sire, using smaller contemporary groups and lowering the penalty value for non-calvers. Fitting the time interval between calving and the start of mating either as a 10-level factor or a cubic spline function explained considerably more variation than fitting the standard 2-level factor (wet or dry). Estimated permanent environmental effects of the cow were considerably reduced. This suggests that, if a cow calves late in the season (less than 60 days before she is inseminated), her fertility is reduced. Models should therefore account for this effect. If fitted, service sire explained 1.6% of phenotypic variation, compared to a much larger sire × contemporary group interaction (3% of phenotypic variation). It is therefore important to account for sire × contemporary group interactions. When the fertility of service sires is not being evaluated, service sires could be incorporated into the definition of contemporary groups. Ideally, breeders should be encouraged to formally record contemporary (or mating) groups. Reducing the size of contemporary groups (inferred from the data) by limiting the time interval between first and last inseminations from 120 to 60 days had only a marginal effect as did reducing the penalty for non-calvers from 21 to 10 days.