Serovar profiling of Haemophilus parasuis on Australian farms by sampling live pigs

Objective Investigate the diversity of serovars of Haemophilus parasuis (Hps) present in Australian pig herds. Design Nasal swabs were used to obtain multiple isolates of Hps, which were grouped first by genotyping and then by serotyping representative isolates. Procedure Swabs were taken from the nasal cavity of just-weaned healthy pigs from multiparous sows on 12 farms and from post-weaned pigs of multiparous sows on 1 farm. On 5 of the 13 farms, nasal swabs were also obtained from pigs showing clinical signs suggestive of Glässer's disease. On a further 7 farms, nasal swabs were obtained only from pigs with clinical signs suggestive of Glässer's disease. Results A total of 556 Hps isolates were genotyped, and 150 isolates were serotyped. Hps was detected on 19 of the 20 farms, including 2 farms with a long history of freedom from Glässer's disease. Isolates of Hps belonging to serovars regarded as potentially pathogenic were obtained from healthy pigs at weaning on 8 of the 10 farms with a history of Glässer's disease outbreaks. Sampling 213 sick pigs yielded 115 isolates of Hps, 99 of which belonged to serovars that were either potentially pathogenic or of unknown pathogenicity. Only 16 isolates from these 213 sick pigs were of a serovar known to be non-pathogenic. Conclusion Healthy pigs contain a range of Hps serovars, even on farms free of Glässer's disease. Nasal swabbing of both healthy and sick pigs seems a useful method of serovar profiling of farms.     

Effects of increasing days of exposure to prepartum diets on the concentration of certain blood metabolites in dairy cows

Objective To evaluate the effect of the number of days exposure to a prepartum transition diet on mineral, energy and protein metabolism in dairy cows. Design A prospective cohort study. Procedures The prepartum transition diet consisted of ryegrass pasture, cereal hay, grain, grain by-product, protein meals, BioChlor®, rumen modifiers, minerals and vitamins and contained 13.9% crude protein, 10.0 MJ metabolisable energy/kg and a dietary cation anion difference of −35 meq/kg dry matter. Forty cows were bled bi-weekly from their introduction to the prepartum transition diet until day 35 of lactation. Blood samples were submitted for estimation of a range of metabolites. Cubic smoothed splines were fitted to scatterplots of metabolite concentration as a function of day relative to calving and the area under the curve (AUC) was calculated. Linear regression modelling determined the effect of days exposed, age, body condition score (BSC) and calving day on AUC. Results The prepartum AUC of blood phosphorus and beta-hydroxybutyrate increased and the AUC of blood calcium and cholesterol decreased with increasing days exposed to the diet. The postpartum AUC of beta-hydroxybutyrate and non-esterified fatty acids decreased with increasing days exposed but increased with days exposed × age and days exposed × BCS, respectively. The AUC of other metabolites did not vary significantly with number of days exposed. Conclusions Increasing exposure to the prepartum diet significantly altered the AUC of blood metabolites associated with mineral, energy and protein metabolism in ways consistent with improvements in production and reproduction previously reported and the result suggests links between energy, protein and skeletal metabolism.     

Hsp70 enhances presentation of FMDV antigen to bovine CD4+ T cells in vitro

Foot-and-mouth disease virus (FMDV) is the causative agent of a highly contagious acute vesicular disease affecting cloven-hoofed animals, including cattle, sheep and pigs. The current vaccine induces a rapid humoral response, but the duration of the protective antibody response is variable, possibly associated with a variable specific CD4⁺ T cell response. We investigated the use of heat shock protein 70 (Hsp70) as a molecular chaperone to target viral antigen to the Major Histocompatibility Complex (MHC) class II pathway of antigen presenting cells and generate enhanced MHC II-restricted CD4⁺ T cell responses in cattle. Monocytes and CD4⁺ T cells from FMDV vaccinated cattle were stimulated in vitro with complexes of Hsp70 and FMDV peptide, or peptide alone. Hsp70 was found to consistently improve the presentation of a 25-mer FMDV peptide to CD4⁺ T cells, as measured by T cell proliferation. Complex formation was required for the enhanced effects and Hsp70 alone did not stimulate proliferation. This study provides further evidence that Hsp70:peptide complexes can enhance antigen-specific CD4⁺ T cell responses in vitro for an important pathogen of livestock.     

Estimated frequency of the canine hyperuricosuria mutation in different dog breeds

Background: Hyperuricosuria is a condition that predisposes dogs to urate urolithiasis. A mutation that causes canine hyperuricosuria was previously identified in 3 unrelated dog breeds. The occurrence of the mutation in additional breeds was not determined. Hypothesis/Objectives: Identify additional breeds that have the hyperuricosuria mutation and estimate the mutant allele frequency in those breeds. Animals: Three thousand five hundred and thirty dogs from 127 different breeds were screened for the hyperuricosuria mutation. Methods: DNA samples were genotyped by pyrosequencing and allele‐specific polymerase chain reaction methods. Results: Mutant allele frequencies that range from 0.001 to 0.15 were identified in the American Staffordshire Terrier, Australian Shepherd, German Shepherd Dog, Giant Schnauzer, Parson (Jack) Russell Terrier, Labrador Retriever, Large Munsterlander, Pomeranian, South African Boerboel, and Weimaraner breeds. Conclusions and Clinical Importance: The hyperuricosuria mutation has been identified in several unrelated dog breeds. The mutant allele frequencies vary among breeds and can be used to determine an appropriate breeding plan for each breed. A DNA test is available and may be used by breeders to decrease the mutant allele frequency in breeds that carry the mutation. In addition, veterinarians may use the test as a diagnostic tool to identify the cause of urate urolithiasis.     

Preliminary assessment of serum clusterin as a potential biomarker for canine lymphoma

Clusterin (CLU), also known as apolipoprotein J, is a widely expressed, heterodimeric, glycoprotein, important in tumourigenesis, apoptosis and immunoregulation. In humans, CLU expression has been associated with anaplastic large cell and Hodgkin's lymphoma. In this study, serum CLU levels in dogs with multicentric lymphoma (MLSA) were compared with healthy control dogs, using both western blot and enzyme‐linked immunosorbent assay (ELISA). Western blot confirmed the presence of CLU in dog sera at the predicted molecular weight and the relative levels detected correlated with the levels detected by ELISA. CLU level analysis by ELISA found treatment naïve dogs with MLSA had a significantly (P < .001) lower serum CLU level compared with healthy controls. However, there was no significant difference between MLSA dogs prior to treatment and in complete remission. The wide variation in serum CLU levels may limit its potential as a single candidate biomarker for MLSA, although any prognostic predictive value of serum CLU concentrations has yet to be assessed.     

重组牛生长激素（rbST）和季节对泌乳牛血清和乳中胰岛素样生长因子Ⅰ（IGF-Ⅰ）和Ⅱ（IGF-Ⅱ）的影响

进行两个试验,研究热应激（S）和冷应激（W）条件TrbST对母牛血清和乳中IGF的影响,每个试验各有试牛12头。每个试验分成两个阶段：农场阶段（30d）和气候室阶段（CC,30d）。每个试验设对照组和试验组,每组各有试牛6头,对照组母牛每天注射盐水：rbST处理组每天注射rbST、牛痘苗和USAN（注射量为25mg·头^-1）。在农场阶段,每周收集1次血样和乳样（上午和下午收集后混合）;CC阶段,每两天收集1次血样,每天收集1次乳样（上午和下午收集后混合）。结果显示,注射rbST的牛,血浆IGF—Ⅰ和IGF-Ⅱ浓度升高。研究发现,对照组和rbST处理组血浆IGF-Ⅰ浓度呈现明显的季节性变化规律,但是血浆IGF-Ⅱ浓度无季节性变化规律。在夏季,尽管日粮采食量和能量平衡急剧降低,但是对照组和rbST处理组血浆IGF-Ⅰ浓度升高。rbST处理或季节因素对乳中IGF—Ⅰ和IGF-Ⅱ浓度无影响。虽然rbST处理对乳中IGF-Ⅰ和IGF—Ⅱ浓度无影响,但是总IGF量随奶产量的增加而增加。血浆IGF—Ⅰ的季节性变化规律可能是由于生长激素-IGF轴偶联的季节性差异造成的。尽管热应激下诱导能量负平衡,但是没有发现生长激素-IGF-Ⅰ轴解偶联。     

Serological characterisation of Haemophilus parasuis isolates from Australian pigs

A total of 31 isolates of Haemophilus parasuis obtained from Australian pigs were serotyped by the Kielstein-Rapp-Gabrielson scheme. The isolates were assigned to serovar 1 (1 isolate), serovar 2 (1 isolate), serovar 4 (4 isolates), serovar 5 (7 isolates), serovar 9 (2 isolates), serovar 10/7 (4 isolates), serovar 12 (1 isolate) and serovar 13 (6 isolates). The remaining 5 isolates could not be assigned to a serovar. Two different serovars (5 and 13) were detected in one herd. The only 2 isolates obtained from clinically normal pigs (from the same herd) were serovar 9. The common serovars were isolated from pigs with pneumonia as well as from pigs with conditions of the Glässer's disease type. The serological heterogeneity amongst Australian isolates of H parasuis has important implications for the use of vaccines to control Glässer's disease.     

Heterozygote detection for maple syrup urine disease in cattle

SUMMARY The clinical, pathological and biochemical manifestations of maple syrup urine disease (MSUD) are similar in Poll Hereford and Poll Shorthorn X Poll Hereford calves. No significant differences were observed in branched-chain amino acid concentrations in plasma, or of branched-chain keto acid dehydrogenase activity in fibroblasts, between Poll Herefords homozygous normal and heterozygous for the mutation responsible for MSUD. Haemopoietic chimerism resulted in incorrect diagnosis of the MSUD genotype in 30% of non-identical twins when blood DNA was analysed using allele-specific amplification. Hair roots are shown to be a suitable source of target DNA for genotyping Poll Hereford cattle for the MSUD mutation. Twelve of 203 (5.8%) aged Poll Hereford bulls, sampled at saleyards during the last 4 months of 1993, were found to be heterozygous for the mutation. In contrast, the mutant sequence was detected in only 1 of 150 (0.7%) 2- and 3-year-old Poll Hereford bulls offered for sale at 2 stud sales held during 1993, suggesting that the prevalence of the disease may decline over the next few years.