Intradermal DNA vaccination in ear pinnae is an efficient route to protect cats against rabies virus

A DNA vaccine against rabies (pGQH) was administrated to cats in order to examine different administration routes. Four groups of three cats each were inoculated with pGQH as follows: group A, intramuscularly (IM), 100 microg; group B, intranasally (IN), 100 microg; group C, intradermally into ear pinnae (ID-EP), 100 microg, and group D, IM, 200 microL of phosphate buffer solution (PBS) alone (control group). Blood was drawn on days 0, 30, 60, 90, 120, 150, and 180. Groups A, B, and C received a booster on day 30. At day 200 all animals were challenged. A passive transfer of cat sera, as well as a viral challenge, was performed in mice. The results displayed that neutralizing antibody titers were higher in cats of group C (ID-EP) showing high early titers (> 2 IU) and the highest titer was on day 120 (> 14 IU). In group B (IN), two out of three cats seroconverted on day 30 (> 0.5 IU), the third cat seroconverted until day 60 (> 0.5 IU). In contrast, the lowest levels of neutralizing antibodies were detected in group A (IM). The control group showed no anti-rabies antibodies. Groups A (IM) and D (control) succumbed after lethal challenge. All animals from the ID-EP group (C) survived, only one individual from the IN (B) group died. Mice that received cat sera from ID-EP, IM, and IN groups survived and were protected (30/30 survivors). Mice groups that received pre-immunization sera from cats were not protected (0/30 survivors). This study demonstrates that pGQH immunization was successful when it was administrated ID-EP, and acceptable through the IN route. The IM route, however, was not effective in cats. For vaccination, the IN route seems attractive due to its accessibility for application, but it seems to activate seroconversion slowly. The best route to promote anti-rabies antibody titers was the ID-EP route. This practical and efficient route should be further studied.     

The effect of repeated doses of toltrazuril on coccidial oocyst output and weight gain in suckling lambs

Coccidial oocyst output and liveweight were monitored in 43 pairs of lamb twins over a 10-week period during September-November 1992. One of each set of twins was treated orally with toltrazuril (20 mg/kg) weekly while the other was an untreated control. The oocyst output was significantly lower (P < 0.01) and the weight gain over the trial period was significantly greater (P < 0.05) in the treated group than in the control group.     

Protection of sheep by vaccination against experimental challenge with Leptospira borgpetersenii serovar Hardjo and L. interrogans serovar Pomona

AIMS: To evaluate a multivalent leptospiral and clostridial vaccine for prevention of renal colonisation and urinary shedding in sheep, following experimental challenge with New Zealand strains of Leptospira borgpetersenii serovar Hardjo type Hardjobovis and L. interrogans serovar Pomona.

METHODS: Two separate but similarly designed studies were conducted. In both studies, Romney-cross lambs, aged 9–11 weeks, were randomly allocated to a vaccinated group and a control group. Vaccinated lambs each received two 1.5-mL S/C doses of a multivalent leptospiral and clostridial vaccine, 4 weeks apart, and animals in the control groups received the same dose of saline. Groups of 12 vaccinated and 12 control lambs were randomly selected in each study for challenge with serovars Hardjo or Pomona. Challenge was initiated 16 weeks following the second vaccination with three daily doses of live leptospires by intranasal and conjunctival routes. Following challenge, urine samples were collected weekly for 6 weeks, for dark field microscopy and leptospiral culture; 6 weeks after challenge the lambs were slaughtered and kidneys collected for leptospiral culture.

RESULTS: In lambs challenged with serovar Hardjo, 8/12 unvaccinated lambs had ≥1 urine or kidney sample that was positive for leptospires following culture, compared with 0/12 lambs in the vaccinated group (p=0.001). In lambs challenged with serovar Pomona, 9/12 unvaccinated lambs had ≥1 urine or kidney sample that was positive following culture, compared with 0/12 lambs in the vaccinated group (p<0.001). Prevention of renal colonisation and urinary shedding, expressed as the prevented fraction, was 100 (95% CI=61.7–100)% and 100 (95% CI=68.3–100)% against challenge with serovars Hardjo and Pomona, respectively, at 4 months after vaccination.

CONCLUSIONS AND CLINICAL RELEVANCE: Use of a multivalent leptospiral and clostridial vaccine demonstrated protection against challenge from New Zealand strains of serovars of Hardjo and Pomona 4 months after vaccination in lambs first vaccinated at 9–11 weeks of age. Further studies are required to assess the duration of immunity against challenge in sheep.     

Effect of time of progesterone supplementation on serum progesterone and the conception rate of cooled Holstein heifers during the summer

To investigate the effects of progesterone supplementation at two different times on serum progesterone (P₄) concentration, conception rate and resynchronization of cooled Holstein heifers in summer, 90 heifers were randomly assigned to two groups: (i) heifers subjected to TAI (timed artificial insemination) and progesterone supplementation from days 4 to 14 after TAI (S_1; n = 45); and (ii) heifers under the same TAI protocol as S₁ and progesterone supplementation from days 17 to 22 after TAI (S₂; n = 45). The groups S₁ and S₂ were cooled 10 days before and 21 days after TAI. Respiratory rate, body surface temperature, vaginal temperature and rectal temperature recorded during the experiment were not different (P > 0.05) between S₁ and S₂ groups. Progesterone concentration was not different (P > 0.05) in S₁ compared to S₂. The conception rates on days 30 and 55 were similar between groups (P > 0.05). Progesterone supplementation did not increase either conception rate or concentrations of P₄ in heifers during the summer. Heifers not pregnant to first service in the group S₂ were resynchronized (77.7%) for a second breeding.     

Hepatitis B Surface Antigen S Gene is an Effective Carrier Molecule for Developing GnRH DNA Immunocastration Vaccine in Mice

Relatively molecular mass of GnRH antigens is small and hence needs to couple to a large carrier molecule to enhance its immunogenicity. This study investigated whether hepatitis B surface antigen S (HBsAg‐S) gene can be used as an effective carrier molecule for developing GnRH DNA immunocastration vaccine. Two copies of human GnRH gene were fused with HBsAg‐S gene for constructing a recombinant plasmid pVAX‐HBsAg‐S‐2GnRH that coded for 27 kDa target fusion protein. Ten male mice were divided into two equal groups, treatment and control. The vaccine (50 μg/mice) prepared in saline solution was injected into male mice at weeks 0, 1, 2, 4 and 7 of the experiment. Vaccine's efficacy was evaluated in terms of GnRH‐specific IgG antibody response, plasma testosterone levels, testicular weight and extent of the testicular tissue damage. The specific anti‐GnRH antibody titre in vaccinated animals was significantly higher than in controls in only 4th week of immunization (p < 0.05). In addition, vaccinated animals showed lower testicular weight than those of the controls (p < 0.05). Spermatogenesis in seminiferous tubules in vaccinated animals was suppressed. In conclusion, in this study, the engineered plasmid to be used as a GnRH DNA vaccine induced antibody response and suppressed spermatogenesis in mice. This suggests that HBsAg‐S gene can be an effective carrier molecule for developing GnRH DNA immunocastration vaccine when relatively molecular mass of the aimed antigens is small.     

Subclinical infection with the nematode Trichostrongylus colubriformis increases gastrointestinal tract leucine metabolism and reduces availability of leucine for other tissues

Gastrointestinal (GI) tract leucine metabolism was measured in 6- to 9-mo-old lambs subjected to trickle infection with Trichostrongylus colubriformis larvae and in separate animals that were not infected. Animals prepared with a jejunal catheter and with indwelling catheters into the aorta and the portal- (PDV) and mesenteric- (MDV) drained viscera were infused simultaneously with [1-13C] and [5,5,5-2H3] leucine to determine GI tract sequestration of leucine from arterial and luminal amino acid pools by tracer and tracee arteriovenous concentration differences. Leucine oxidative losses and net fluxes were also determined across the GI tract. Infection had no detectable effect on whole-body leucine flux, but it increased total GI tract leucine sequestration by 24% (P<.05) and GI tract oxidative losses of leucine by 22 to 41% (P<.01). Net PDV fluxes of leucine were decreased by 20 to 32% during the infection. The infection did not alter either the proportion of precursor leucine used by GI tract metabolism that was derived from the arterial leucine pool (.84 to .88) or the proportional sequestration of digesta-derived leucine during "first pass" absorptive metabolism (.12 to .18). These findings help to elucidate the metabolic basis for the reduced growth rates and nitrogen retention observed when animals are subjected to subclinical nematode infection.     

Effects of elicitors on trichothecene accumulation and <Emphasis Type="Italic">Tri</Emphasis> genes expression in potato tubers inoculated with <Emphasis Type="Italic">Fusarium sulphureum</Emphasis>

Postharvest losses due to pathogens are a major concern in agriculture and therefore new strategies to reduce these losses while making sure that the treated products are safe for the consumer, are of paramount importance. Chemical fungicides treatment is not only unsafe but also leads to pathogen developing resistance. Therefore, products that can reduce the development of dry rot of potato and mycotoxin accumulation, and be safe are needed. A better understanding of the induction of trichothecene biosynthesis is essential to reduce trichothecene production. The effects of three elicitors such as β-amino butyric acid (BABA), sodium silicate and chitosan, on the suppression of lesion development, trichothecene accumulation, and expression of Tri genes were assessed in potato tubers inoculated with Fusarium sulphureum. The results showed that lesion diameters were significantly reduced after treating with BABA at 100 mM for 3 d, sodium silicate at 100 mM for 2 d and chitosan at 0.50% for 3 d. Tri gene expressions were significantly down-regulated in inoculated tubers after elicitor treatments, and trichothecene accumulation were also suppressed. Meanwhile, the levels of trichothecene accumulation and Tri genes expression showed cumulative changes with the incubation time, extending after elicitor treatments. In addition, elicitor applications reduced more for type A trichothecene (T-2, DAS) than type B trichothecene (3ADON, Fus-X). It is possible that elicitors triggered downstream resistance genes to produce resistance related metabolites that suppressed the biomass of F. sulphureum, resulting in reduced Tri gene expressions and trichothecene accumulation.     

Transglutaminase treatment of wheat and maize prolamins of bread increases the serum IgA reactivity of celiac disease patients

Celiac disease (CD) is mediated by IgA antibodies to wheat gliadins and tissue transglutaminase (tTG). As tTG is homologous to microbial transglutaminase (mTG) used to improve foodstuff quality, it could elicit the immune response of celiac patients. This study evaluated the reactivity of IgA of celiac patients to prolamins of wheat and gluten-free (maize and rice flours) breads mTG-treated or not. Prolamins extracted from wheat and gluten-free breads were analyzed by ELISA and immunodetected on membranes with individual or pooled sera from nine celiac patients recently diagnosed. Sera pool IgA titers were higher against prolamins of mTG-treated wheat or gluten-free breads than against mTG-untreated, mainly due to two individual patients' sera. The electrophoretic pattern of gluten-free bread prolamins was changed by the mTG treatment, and a new 31000 band originated in maize was recognized by three CD patients' IgA.     

The Role of Uterine and Ovarian Hormones in Luteolysis: A Comparison among Species

Luteolytic mechanisms have evolved in mammals to improve reproductive efficiency. The hormonal interactions that control the onset and progress of luteolysis are complex. They involve endocrine and paracrine signals that link the corpus luteum, uterus and posterior pituitary gland. Current concepts concerning these interactions will be examined in the five major domestic ungulate species commonly raised in Europe and North America (cattle, sheep, goats, pigs and horses). Some of these interactions are similar across species. All five depend on prostaglandin F_2α secreted from the uterus, to induce luteolysis. Three hormones, progesterone, estradiol and oxytocin interact to regulate uterine secretion of PGF_2α. Oxytocin is an acute stimulus for uterine PGF_2α secretion. Progesterone and estradiol interact to regulate uterine secretary responsiveness to oxytocin. Precisely how these hormones interact varies across species.