Efficacy of ivermectin against Dirofilaria immitis larvae in dogs 30 and 45 days after induced infection

Forty-two Beagles, 14 to 15 weeks of age, were injected subcutaneously with 50 infective larvae of Dirofilaria immitis and were allotted by weight, within sex, to 6 treatment groups. Group 1 served as nonmedicated vehicle-treated controls; groups 2 through 5 were given an oral tablet form of ivermectin at dosages of 0.3 micrograms/kg, 1.0 micrograms/kg, 2.0 micrograms/kg, and 3.3 micrograms/kg at 30 days after inoculation; group 6 was given the 2.0 micrograms/kg dosage at 45 days after inoculation. Dogs were euthanatized and necropsied 154 days after treatment (day 139 for dogs in group 6) and examined for heartworms. On the numerical bases of helminths recovered in the groups, the efficacies for preventing heartworm maturation were 0% (group 2), 53.2% (group 3), 97.2% (group 4), 98.1% (group 5), and 63.8% (group 6). Drug-related adverse reactions were not detected.     

Effect of supplementing graded concentrations of non-phytate phosphorus on performance,egg quality and bone mineral variables in White Leghorn layers

1. An experiment was conducted to determine optimal non-phytate phosphorus (NPP) concentrations for White Leghorn (WL) layers (22–72 weeks) fed diet containing 38 g Ca/kg.

2. Eight diets with graded concentrations (1.5–3.25 g/kg in increments of 0.25 g) of NPP were prepared. Each diet was fed to eight pen replicates containing 88 birds in each. Performance data was evaluated in three different phases (phase I-22–37 weeks, phase II-38–53 weeks and phase III-54–72 weeks). Optimum levels of NPP were determined by fitting a quadratic polynomial (QP) regression model.

3. Egg production (EP) was not affected (P = 0.059) by the concentration of NPP and interaction between NPP and diet phase was non-significant, indicating that the lowest concentration (1.5 g/kg diet) of NPP used in the study was adequate across the three phases. However, EP was influenced by phase (P < 0.001).

4. Optimum concentration of NPP for feed intake (FI) was estimated to be 1.5, 1.71 and 2.40 g/kg diet during phases I, II and III, respectively. FI per egg mass (EM) or feed efficiency (FE) responded quadratically with NPP and also differed significantly between phases. Optimum concentration of NPP for FE during phases I, II and III was 1.5, 2.56 and 2.32 g/kg diet, respectively.

5. Egg weight (EW), EM, shell weight and thickness were not affected by NPP concentration although all of these variables (except shell weight) were influenced by phases.

6. Breaking strength of tibia and Ca contents in tibia ash were not affected by the concentration of NPP, but bone ash and P contents in tibia ash were influenced (P < 0.001) by NPP. Predicted optimal concentrations of NPP for responses for tibia ash at 44 or 72 weeks, tibia ash P at 44 weeks and tibia ash P at 72 weeks were 1.55, 2.63 and 1.5 g/kg diet, respectively.

7. Based on the results, it was concluded that WL layers required 1.5 g, 2.63 g and 2.4 g, respectively/kg diet during phase I, II and III with the calculated daily intake of 137.3, 278.3 and 262 mg NPP/b/d.     

Development of a polyclonal-antibody-based immunohistochemical method for the detection of type 2 porcine circovirus in formalin-fixed, paraffin-embedded tissue.

An immunohistochemical method for the detection of type 2 porcine circovirus (PCV2) in paraffin-embedded tissue was developed. Rabbits were inoculated with purified PCV2 to obtain a polyclonal antiserum. Antiserum was applied to sections of porcine tissue that contained lesions consistent with postweaning multisystemic wasting syndrome and in which PCV2 genome had been demonstrated by in situ hybridization. In all cases (18/18), the density and distribution of positive cells detected by in situ hybridization or immunohistochemistry were identical. The immunohistochemical method is more rapid and less expensive than in situ hybridization and is thus more suitable for routine diagnostic use.     

Genotypic characterisation and cluster analysis of Campylobacter jejuni isolates from domestic pets, human clinical cases and retail food

The genetic similarity of Campylobacter jejuni isolates from pets, compared to human clinical cases and retail food isolates collected in Ireland over 2001-2006 was investigated by cluster analysis of pulsed-field gel electrophoresis (PFGE) fingerprinting profiles. Comparison of the PFGE profiles of 60 pet isolates and 109 human isolates revealed that seven (4.1%) profiles were grouped in clusters including at least one human and one pet C. jejuni isolate. In total six (1.6%) of 60 pet and 310 food profiles were in clusters with at least one food and one pet C. jejuni isolate. The detection of only a small number of genetically indistinguishable isolates by PFGE profile cluster analysis from pets and from humans with enteritis in this study suggests that pets are unlikely to be an important reservoir for human campylobacteriosis in Ireland. However, genetically indistinguishable isolates were detected and C. jejuni from pets may circulate and may contribute to clinical infections in humans. In addition, contaminated food fed to pets may be a potential source of Campylobacter infection in pets, which may subsequently pose a risk to humans.     

Analysis of differential protein expression in Actinobacillus pleuropneumoniae by Surface Enhanced Laser Desorption Ionisation--ProteinChip (SELDI) technology

Actinobacillus pleuropneumoniae (APP) is the aetiological agent of porcine pleuropneumonia. An increased understanding of its molecular basis of pathogenicity and vaccine development will be facilitated by the availability of sequence data from a complete genome which, by analogy to other bacteria, is predicted to encode many proteins in the molecular mass range 3-20kDa. However, conventional techniques to study bacterial protein expression, such as SDS-PAGE and 2-dimensional electrophoresis, typically focus on the 15-200kDa range. In this study we have evaluated Surface Enhanced Laser Desorption Ionisation-ProteinChip (SELDI) technology for the analysis of protein expression, in particular those of <20kDa, of APP grown under different environmental conditions. Cytoplasmic/periplasmic and outer membrane protein fractions were obtained from the APP wildtype serotype 1 strain 4074 grown in Brain Heart Infusion (BHI) broth (+different concentrations of NAD), BHI containing pig serum or defined medium. Optimum conditions for SELDI profiles included a sample size of 1 microg and the use of sinapinic acid as the energy absorbing matrix. In the <20kDa range, the SELDI profiles obtained from wild-type bacteria grown in rich medium plus 33-66% pig serum were most similar to those grown in defined medium. The SELDI profiles of extracts of the wild-type and of an rpoE mutant were similar although there were clear differences. The results suggest that SELDI is a useful complementary approach to conventional proteomic analytical methods with APP, and presumably other bacterial pathogens, being particularly suited for analysis of proteins in the <20kDa mass range.     

Immune dynamics following infection of avian macrophages and epithelial cells with typhoidal and non-typhoidal Salmonella enterica serovars; bacterial invasion and persistence, nitric oxide and oxygen production, differential host gene expression, NF-κB signalling and cell cytotoxicity

Poultry-derived food is a common source of infection of human with the non-host-adapted salmonellae while fowl typhoid and pullorum disease are serious diseases in poultry. Development of novel immune-based control strategies against Salmonella infection necessitates a better understanding of the host-pathogen interactions at the cellular level. Intestinal epithelial cells are the first line of defence against enteric infections and the role of macrophages is crucial in Salmonella infection and pathogenesis. While gene expression following Salmonella infection has been investigated, a comparison between different serovars has not been, as yet, extensively studied in poultry. In this study, chicken macrophage-like cells (HD11) and chick kidney epithelial cells (CKC) were used to study and compare the immune responses and mechanisms that develop after infection with different Salmonella serotypes. Salmonella serovars Typhimurium, Enteritidis, Hadar and Infantis showed a greater level of invasion and/or uptake characters when compared with S. Pullorum or S. Gallinarum. Nitrate and reactive oxygen species were greater in Salmonella-infected HD11 cells with the expression of iNOS and nuclear factor-κB by chicken macrophages infected with both systemic and broad host range serovars. HD11 cells revealed higher mRNA gene expression for CXCLi2, IL-6 and iNOS genes in response to S. Enteritidis infection when compared to S. Pullorum-infected cells. S. Typhimurium- and S. Hadar-infected HD11 showed higher gene expression for CXCLi2 versus S. Pullorum-infected cells. Higher mRNA gene expression levels of pro-inflammatory cytokine IL-6, chemokines CXCLi1 and CXCLi2 and iNOS genes were detected in S. Typhimurium- and S. Enteritidis-infected CKC followed by S. Hadar and S. Infantis while no significant changes were observed in S. Pullorum or S. Gallinarum-infected CKC.