Identification of toxigenic Pasteurella multocida in atrophic rhinitis of pigs by in vitro characterisation

Toxigenic strains of Pasteurella multocida were readily differentiated from non-toxigenic strains by an agarose overlay method using bovine turbinate cells or bovine lung cells. Cells which were young and densely confluent were best suited to this assay. The incubation period required to distinguish toxigenic strains was dependent on the confluence of the monolayers, which was affected by the seeding rate, cell passage level and growth time prior to overlay. The agarose overlay method correctly identified 11 of 11 reference strains of Pasteurella multocida, and visible cytotoxic changes were present in the monolayers after 48 to 65 h. Outbreaks of the enzootic form of atrophic rhinitis in 2 New South Wales piggeries were associated with the isolation of toxigenic type D strains of P. multocida.     

The effects of antigenic competition on the efficacy of multivalent footrot vaccines

SUMMARY A multivalent footrot vaccine has been developed, containing pilus antigens produced in recombinant Pseudomonas aeruginosa and representing all nine serogroups of Dichelobacter (Bacteroides) nodosus commonly recognised in the field. The responses of sheep to the multivalent vaccine have been compared with those to monovalent vaccines representing only a single serogroup. Antigenie competition between serogroups occurred in sheep immunised with the multivalent formulation, but high levels of protection were still achieved. The study showed that in multivalent footrot vaccines, antigenic competition is predominantly due to the presence of a family of immunologically-related pilus antigens rather than to interference by extraneous proteins.     

Cryptosporidium parvum infection in orphan lambs on a farm open to the public

A longitudinal survey was undertaken on an open farm to investigate the occurrence of Cryptosporidium species infection in orphan lambs obtained from three local flocks. During an initial pilot study, Cryptosporidium oocysts were detected by a fluorescent antibody test (fat) in the faeces of two of 21 lambs aged between one and three weeks derived from one flock (flock A). Pooled pen samples of faeces were collected weekly from lambs derived from each flock; oocysts were detected by fat in 24 (49.0 per cent) of 49 samples from lambs from flock A, 18 (30.5 per cent) of 59 samples from lambs from flock B and 14 (29.8 per cent) of 47 samples from lambs from flock C. Oocyst counts of 1 x 10(3) to more than 2 x 10(6) per gram of faeces were detected in lambs up to 12 weeks old, with the peak counts occurring at six weeks of age in the lambs from flocks A and B and at four weeks of age in those from flock C. The oocysts were confirmed by molecular analysis as Cryptosporidium parvum. Virtually all the infections were subclinical.     

Assessing the convergence of Markov Chain Monte Carlo methods: an example from evaluation of diagnostic tests in absence of a gold standard

The accessibility of Markov Chain Monte Carlo (MCMC) methods for statistical inference have improved with the advent of general purpose software. This enables researchers with limited statistical skills to perform Bayesian analysis. Using MCMC sampling to do statistical inference requires convergence of the MCMC chain to its stationary distribution. There is no certain way to prove convergence; it is only possible to ascertain when convergence definitely has not been achieved. These methods are rather subjective and not implemented as automatic safeguards in general MCMC software. This paper considers a pragmatic approach towards assessing the convergence of MCMC methods illustrated by a Bayesian analysis of the Hui–Walter model for evaluating diagnostic tests in the absence of a gold standard. The Hui–Walter model has two optimal solutions, a property which causes problems with convergence when the solutions are sufficiently close in the parameter space. Using simulated data we demonstrate tools to assess the convergence and mixing of MCMC chains using examples with and without convergence. Suggestions to remedy the situation when the MCMC sampler fails to converge are given. The epidemiological implications of the two solutions of the Hui–Walter model are discussed.     

Calcium chelate is as effective as phosphite in controlling Phytophthora root rot in glasshouse trials

Species in the genus Phytophthora cause significant economic losses in crops and damage to forests and natural ecosystems worldwide. Currently, phosphite is the most effective chemical for disease management, but excessive phosphite concentrations can result in phytotoxicity in plants and the development of tolerance by the pathogen. Two newly developed metal chelates and phosphite (alone and in combination) were tested for their in vitro and in planta efficacy against Phytophthora cinnamomi. In glasshouse trials, 0.25% and 0.5% of each chemical treatment (phosphite, Ca chelate, Zn chelate) and Ca chelate + phosphite were used as a foliar application on 3-month-old seedlings of Banksia grandis (experiment not repeated) and Eucalyptus marginata, prior to inoculation with P. cinnamomi. All noninoculated control plants remained healthy, while significant root damage and reduction of dry root weights were observed for inoculated untreated plants. Individually, phosphite and Ca chelate significantly reduced root lesion development of P. cinnamomi compared to the control, with Ca chelate attaining superior results to phosphite at the same concentration. In combination, Ca chelate + phosphite had the largest reduction in root lesion development in both plant species; however, this result has not yet been replicated but did reflect previous in vitro results. The Zn chelate applications were not effective. Ca chelate has the potential to be developed as a fungicide to control Phytophthora species.     

Exophiala angulospora Causes Systemic Mycosis in Atlantic Halibut: a Case Report

Filamentous black yeasts from the genus Exophiala are ubiquitous, opportunistic pathogens causing both superficial and systemic mycoses in warm- and cold-blooded animals. Infections by black yeasts have been reported relatively frequently in a variety of captive and farmed freshwater and marine fishes. In November 2012, moribund and recently dead, farm-raised Atlantic Halibut Hippoglossus hippoglossus were necropsied to determine the cause of death. Histopathology revealed that three of seven fish were affected by a combination of an ascending trans-ductual granulomatous mycotic nephritis, necrotizing histiocytic encephalitis, and in one fish the addition of a fibrogranulomatous submucosal branchitis. Microbial cultures of kidney using selective mycotic media revealed pure growth of a black-pigmenting septated agent. Application of molecular and phenotypic taxonomy methodologies determined that all three isolates were genetically consistent with Exophiala angulospora. This is the first report of E. angulospora as the causal agent of systemic mycosis in Atlantic Halibut.

Received January 27, 2014; accepted July 23, 2014