Evaluation of the humoral and cellular immune response to different antigens of Corynebacterium pseudotuberculosis in Canindé goats and their potential protection against caseous lymphadenitis

Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis, a disease that affects goats and sheep, and can cause severe economic losses. In this study, four different antigenic extracts were obtained from the attenuated strain T1, which was isolated in the state of Bahia (Brazil). Forty-four Canindé breed goats were divided in five groups, each receiving a different antigen solution and saline buffer as a control. The humoral response was monitored through the identification of specific IgG by indirect ELISA and Western Blotting, and the production of IFN-gamma was followed in order to observe the activation of cellular response. After twelve weeks of antigen inoculation, the animals were challenged with 2 x 10(5)CFU of a wild strain, also isolated in Bahia, and necropsy was performed on all animals twelve weeks afterwards. It was observed that the attenuated bacteria gave a protection of 33.3%, in addition to the weak humoral response elicited. Animals inoculated with secreted antigen associated with Freund's incomplete adjuvant and oligodeoxynucleotide containing unmethylated CpG dinucleotides (CpG ODN) showed a strong humoral response, but this inoculation could not prevent the spread of challenge bacteria in the majority of animals. These results demonstrate the immunogenic potential of the attenuated T1 strain in the development of a vaccine against caseous lymphadenitis in goats.     

Comparison of Closely Related Orthopoxvirus Isolates by Random Amplified Polymorphic DNA and Restriction Fragment Length Polymorphism Analysis

The reliability and reproducibility of random amplified polymorphic DNA analysis (RAPD) was compared with restriction fragment length polymorphism (RFLP) by analysing three virus strains isolated from zoo animals in Berlin and three isolates which were cultivated from pets from Northern Germany. The RAPD technique was evaluated as a reliable tool with good reproducibility of the patterns for each virus strain investigated. Problems of interpretation due to inconsistent intensity of bands in different polymerase chain reaction runs may arise for less experienced personnel. The RAPD analysis can be performed within one working day and needs less DNA compared with RFLP so costs will be reduced. The obvious advantage of RFLP is that the pattern can be traced to the recognition site of the restriction enzyme whereas the RAPD primer sequence is not present in the orthopoxvirus genome at all. To the authors knowledge, the RAPD technique has never been applied in DNA viruses before and they conclude that this technique is a useful tool for the discrimination of closely related cowpoxviruses.     

Evaluation of five serological tests for the diagnosis of porcine brucellosis in French Polynesia

Porcine brucellosis due to Brucella suis biovar 1 raises important issues for pig breeders in French Polynesia. In this region, the disease is enzootic, spreads silently and engenders economic losses in infected farms as well as sporadic human cases. While serological tests are essential in surveillance and control programmes of animal diseases, to date none of the available tests have been shown to be reliable enough to be used as a gold standard in routine individual diagnosis of porcine brucellosis. Few studies about the estimation of the sensitivity and the specificity of porcine brucellosis screening tests have been published, none of them dealing with French Polynesia. The studied population included 1,595 pigs from French Polynesia. Five tests were evaluated: Rose Bengal test, fluorescence polarisation assay, indirect ELISA, and two competitive ELISAs (C-ELISA). The sensitivity and the specificity of each test were estimated. C-ELISA₂ was the most sensitive test (Se C-ELISA₂?=?0.954 [0.889; 0.992] 95 % credibility interval (CrI)) while both C-ELISA and Rose Bengal test (RBT) were the most specific ones (Sp C-ELISA₁?=?0.856 [0.806; 0.915] 95 % CrI; Sp C-ELISA₂?=?0.849 [0.817; 0.879] 95 % CrI; Sp RBT?=?0.853 [0.812; 0.898] 95 % CrI).     

Usefulness of serological examinations in the analysis of Salmonella infections in pig herds

The development of the antibody concentration against lipopolysaccharide (LPS) of S. Typhimurium und S. Choleraesuis in rearing pigs during the fattening period and in breeding sows of the corresponding age was recorded. The studies revealed the following results. Antibodies of isotypes IgG1 and IgG2 revealed a more pronounced specificity against the according Salmonella serovar than IgM antibodies. The calculated "antibody percent value" based on the total amount of Salmonella antibodies is mainly determined by the IgM antibodies in sera and meat juice, respectively. In fattening pigs a significant increase of antibodies against IgM and total Ig was observed between week 3 and 10 after beginning of the rearing period. In breeding pigs this increase was detectable already earlier. In only 3 out of 10 groups an increase of IgG1 and IgG2 was also seen. The detected significant increase of total Ig and IgM in the other groups might be the result of a less intensive exposure to salmonellas or it might be due to an increase of unspecific antibodies induced by other antigens. Serological investigations represent a valuable tool to record the intensity and development in time of the Salmonella exposure in pigs farms. Examination of total Ig is an appropriate method to detect pig herds with a high level of Salmonella exposure, for detailed epidemiological studies in pig farms the examination of antibody isotypes will give more comprehensive information.     

Effects of maternal selenium supply and plane of nutrition during gestation on passive transfer of immunity and health in neonatal lambs

To investigate the influence of maternal Se supply and plane of nutrition on lamb morbidity, mortality, and passive transfer of IgG, pregnant ewe lambs were used in 2 experiments with 2 × 3 factorial treatment arrangements. Supplementation of Se began at breeding and was either adequate Se (ASe, 9.5 μg/kg of BW) or high Se (HSe, 81.8 μg/kg of BW) in Exp. 1 or ASe (11.5 μg/kg of BW) or HSe (77.0 μg/kg of BW) in Exp. 2. On d 50 or 40 of gestation for Exp. 1 or 2, respectively, ewes were assigned randomly to 1 of 3 nutritional planes: 60% (RES), 100% (control, CON), or 140% (HI) of NRC requirements. This resulted in the following treatments: ASe-RES, ASe-CON, ASe-HI, HSe-RES, HSe-CON, and HSe-HI. Upon parturition, lambs were separated from their dams and serum samples obtained. Lambs were fed artificial colostrum for the first 20 h and then placed on milk replacer and grain pellets until completion of the study (Exp. 1, 57 d; Exp. 2, 21 d). Twenty-four hours after parturition, lamb serum samples were collected for IgG analysis. All lambs were reared similarly and morbidity and mortality assessed. Main effects were considered significant when P ≤ 0.05. In Exp. 1, there was a Se × plane of nutrition interaction (P ≤ 0.01) for lamb morbidity from birth to weaning and for 24-h IgG concentration. Lambs from ASe-RES and HSe-HI ewes were treated more frequently (P < 0.01) for respiratory and gastrointestinal disease, and lambs from HSe-HI ewes had the smallest (P < 0.01) 24-h serum IgG concentration. In Exp. 1, lambs from HI ewes also had the greatest (P < 0.01) mortality rates from birth to weaning compared with lambs from CON and RES ewes. In Exp. 2, there was an effect (P < 0.01) of maternal plane of nutrition with lambs from RES ewes having increased 24-h IgG compared with lambs from CON and HI ewes. There was no effect of maternal Se supplementation on lamb 24-h IgG in Exp. 2; however, there was a Se × plane of nutrition interaction (P < 0.01) for morbidity. From birth to 21 d of age, lambs from ASe-CON ewes had fewer (P < 0.01) treatment days compared with lambs from any of the other treatment groups. There also tended (P = 0.08) to be an effect of maternal Se supplementation on lamb mortality with increased mortality observed in lambs from HSe ewes. Results from the studies show a restricted maternal plane of nutrition can increase lamb serum IgG concentration. Selenium results were not consistent between the 2 experiments and may be due to differences in maternal Se.     

The pharmacokinetics and residues of clenbuterol in veal calves.

Seven female Brown Swiss calves were used to study the pharmacokinetics of clenbuterol after an effective anabolic dosage of 5 micrograms/kg of BW was given twice daily for 3 wk. Analyses of clenbuterol concentrations in different tissues was done by enzyme immunoassay (EIA). Tissue samples were taken from three calves on the last day of administration and from two more after 3.5 or 14 d of clenbuterol withdrawal. The rate of clenbuterol elimination was dependent on time and tissue. Clenbuterol concentrations in the lung dropped from a mean of 76 ng/g to a level of less than .08 ng/g after 14 d, whereas in the liver the clenbuterol concentrations decreased from 46 ng/g to .6 ng/g within 14 d of withdrawal. Highest levels were always found in the eye: 118 ng/g, 57.5 ng/g, and 15.1 ng/g after 0, 3.5, and 14 d of withdrawal, respectively. These data reveal that different compartments contribute to the elimination of clenbuterol; therefore, concentrations in urine do not follow first order kinetics. An initial rapid decline in the concentration of clenbuterol in urine with a half-life of 10 h is followed by a slower elimination with a half-life of about 2.5 d. Treatments using the anabolic dose of 5 micrograms/kg of BW require longer withdrawal times than the therapeutic dose (.8 micrograms/kg BW).