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.     

A blocking ELISA using a monoclonal antibody for the serological detection of Mycoplasma hyopneumoniae

A blocking ELISA was developed by using a monoclonal antibody (4082-05-344-18) which specifically detected an epitope on the Mycoplasma hyppneumoniae 40 kDa membrane protein without cross-reacting with M flocculare or M hyorhinis. The results obtained with sera from specific pathogen-free pigs inoculated with M flocculare or M hyorhinis confirmed the specificity of the assay. An immunoblotting procedure was used to characterise the antibody response of pigs experimentally infected with M hyopneumoniae. Antibodies to the 40 kDa antigen were detected two weeks after infection and remained as major markers for at least 20 weeks. Cross-reacting antibodies to this antigen were not detected in convalescent sera from piglets infected with M flocculare or M hyorhinis. Sera from experimentally infected pigs were compared by means of the blocking ELISA and an indirect ELISA. The kinetics of ELISA antibodies after experimental inoculation were also studied. The detection of antibody was rather more stable for a longer time with the blocking ELISA than with the indirect ELISA. In an evaluation of more than 1000 sera from the field there was excellent agreement between the two methods.     

Serological analysis of a small herd-sample to predict presence or absence of animals persistently infected with bovine viral diarrhoea virus (dvdv) in dairy herds

In 10 herds containing animals persistently infected (pi) with bovine viral diarrhoea virus (vvdv) and nine herds without such animals the probabilities of obtaining at least two antibody-positive animals in a test sample of three or five animals selected among animals six to 18 months old were calculated. Among herds with Pi animals these probabilities, with the exception of one herd, varied between 0·725 and 0·992 when samples of three animals were tested and between 0·977 and one when samples of five animals were tested. Among herds without Pi animals the probabilities varied between 0 and 0·015 when samples of three animals were tested and between 0 and 0·048 when samples of five animals were tested. Thus, based upon a few bloodsamples, herds with pi animals and herds without pi animals could be distinguished with a high degree of accuracy.     

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.     

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.     

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.     

Development of immune responsiveness to Ascarissuum antigens in pigs vaccinated with ultraviolet-attenuated eggs

Pigs fed $\text{Ascaris}$$\text{suum}$ eggs attenuated by short wave ultraviolet-radiation developed resistance to challenge infection. Per os inoculation of pigs on three successive weeks with 10,000 eggs irradiated to total exposures of 150,100 and 75 μW-min/cm², respectively, resulted in an 88% reduction in the number of larvae recovered from the lungs 7 days after challenge with 10,000 infective eggs. Peripheral blood lymphocytes (PBL) from vaccinated pigs were specifically stimulated in vitro to incorporate tritiated thymidine by egg hatching fluid (Ea) and by excretory-secretory products obtained from cultures in defined-media of second-stage larvae (L2) developing to third-stage (L2–3) and from cultures of third-stage larvae developing to fourth-stage (L3–4). PBL were also specifically stimulated by living L2. Ea and L2 stimulated pig PBL significantly at 7 days after the first inoculation; responses to L2–3 and L3–4 developed 7 days after a second inoculation. The antigen-responsive cells in the PBL population were non-immunoglobulin-bearing lymphocytes. Antibodies to Ea and L2–3 were not detected in the serum of vaccinated pigs, and only 3 of 7 pigs had low concentrations of serum antibodies to L3–4.     

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.     

Trials of an inactivated equid herpesvirus 1 vaccine: challenge with a subtype 1 virus

Vaccinated yearlings , two-year-old and in-foal pony mares with appropriate controls were exposed to aerosols of a subtype 1 virus one to two months after two or three vaccinations; all became infected. No obvious differences in the febrile responses, clinical signs and subsequent abortions were found between vaccinated and control mares. All vaccinated yearlings and two-year-old ponies developed a febrile respiratory disease but this was less severe than that suffered by the controls and the amounts and duration of virus shedding were reduced.     

Examination of milk samples from experimentally infected heifers by an N-acetyl-β-D-glucosaminidase test and an antigen-capture elisa

Milk samples collected from normal and experimentally infected quarters of five heifers throughout their first lactation were examined by bacterial culture, milk cell count (MCC), N-acetyl-beta-D-glucosaminidase (NAGase) test and a monoclonal antibody based antigen-capture ELISA. The results were analysed according to the presence (mastitis positive) or absence (mastitis negative) of bacteria on culture, and the numbers of false negative and false positive results of the other three tests defined in relation to this. Similar numbers of false negative results were observed with the MCC (20), NAGase (18) and ELISA (13). False positive results due to the physiological factors present in early lactation were evident in the MCC, prominent in the NAGase test and absent from the ELISA. The major difference in false positive results associated with experimental infection between the three tests was the more rapid return to negative values of the ELISA following resolution of infection, compared with MCC and NAGase.     

The immune response of goats vaccinated with low and high doses of Brucella melitensis Rev 1

The serological response, lymphocyte reactivity, and dermal hypersensitivity reactions of goats vaccinated with high and low doses of $\text{Brucella melitensis}$ Rev 1 organisms to Brucella antigens were studied. Antibodies to soluble antigen A2 were detectable by immunoelectrophoresis (IE), appeared later than agglutinating antibodies but disappeared faster in most vaccinated animals. Anti-A2 antibodies appeared earlier in goats which received the high dose. Antibodies to polysaccharide B antigen were not detected. Lymphocytes reactive to Brucella antigens in lymphocyte transformation assays appeared at variable times after vaccination. In contrast to the humoral response to antigen A2, the appearance of circulating, reactive lymphocytes was not dependent on vaccine doses. All vaccinated animals demonstrated dermal hypersensitivity reactions to one of the antigenic extracts. Skin reactions peaked at 24 hrs post inoculation. The reactions were elicited when all but one goat had undetectable anti-A2 antibodies and no circulating, reactive lymphocytes as measured in whole blood lymphocyte transformation assay.     

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.     

Coccidiosis: T-lymphocyte-dependent effects of infection with Eimeria nieschulzi in rats

The intestinal pathology caused by infection with $\text{Eimeria nieschulzi}$ was investigated and comparisons were made between the effects in athymic nude (rnu/rnu) rats and their heterozygous (rnu/+) litter-mates. Most of the changes noted, i.e. increase in gut weight, partial villous atrophy and increased numbers of mast, goblet and pyroninophilic cells were shown to be largely or wholely thymus dependent. The numbers of intraepithelial lymphocytes were decreased in both groups during the period of study. The peripheral blood leucocyte response was similar in both groups of rats during a primary infection but differed after a challenge inoculum, indicating that the secondary type of response which occurred in the rnu/+ rats was thymus dependent, as is resistance to reinfection.     

The mucosal humoral immune response of the horse to infective challenge and vaccination with equine herpesvirus-1 antigens.

Equine herpesvirus-1 (EHV-1) remains a frequent cause of upper respiratory tract infection and abortion in horses worldwide. However, little is known about the local antibody response elicited in the upper airways of horses following exposure to EHV-1. This study analysed the mucosal humoral immune response of weanling foals following experimental infection with virulent EHV-1, or vaccination with either of 2 commercial vaccines. Twenty weanlings were assigned to 5 groups and were inoculated with, or vaccinated against, EHV-1 following different regimens. Finally, all weanlings were simultaneously challenged intranasally with virulent EHV-1 Army 183 (A183). Nasal wash and serum samples were collected at regular intervals until 13 weeks after final challenge. Nasal washes were assayed for EHV-1-specific equine IgGa, IgGb, IgG(T), IgA, IgM and total virus-specific antibody using an indirect, quantitative ELISA. Total serum antibody responses were also monitored, and clinical signs of EHV-disease were recorded for each individual. Virus-specific IgA dominated the mucosal antibody response elicited in weanlings inoculated with A183, being detectable at up to 3.1 microg/mg total IgA 13 weeks after challenge. Neither inactivated EHV-1 administered i.m., nor attenuated EHV-1 administered intranasally induced detectable mucosal antibodies. EHV-1-specific mucosal antibodies impeded EHV-1 plaque formation in vitro. Such virus-neutralising antibody probably contributes to a reduction of shedding of EHV-1 from the respiratory tract of virus-infected horses.     

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.     

Lack of reactivity of sera from Theileriaparva-infected and recovered cattle against cell membrane antigens of Theileria parva transformed cell lines

Sera from $\text{Theileria}$$\text{parva}$ infected, recovered and rechallenged cattle were tested in complement-dependent cytotoxicity, membrane immunofluorescence and antibody-dependent cellular cytotoxicity assays for the presence of antibodies against cell membrane antigens of $\text{T}$. $\text{parva}$ transformed cell lines.In the complement-dependent antibody-mediated cytotoxicity assay, sera from lethally infected animals were negative. Some recovered cattle showed a positive reaction, but such reactions were also observed when an eland cell line infected with $\text{T}$. $\text{taurotragi}$, and bovine lymphoblastoid cells were used as targets. Reaction was less against Ig-negative peripheral blood lymphocytes.Evidence is presented that these reactions could be evoked by attachment of immune complexes to Fc-receptors. It is concluded that cattle exposed to $\text{T}$. $\text{parva}$ infection do not develop antibodies against specific $\text{T}$. $\text{parva}$ (or $\text{T}$. $\text{parva}$-induced) cell surface antigens.