Genetic Diversity of Australian Bacillus anthracis Isolates Revealed by Multiple-Locus Variable-Number Tandem Repeat Analysis.

Outbreaks of anthrax occur sporadically in Australia and most commonly in the "anthrax belt", a region which extends from southern Queensland through the centre of New South Wales and into northern Victoria. Little is known about the epidemiological links between Bacillus anthracis isolates taken from different outbreaks and the diversity of strains within Australia. We used multiple-locus variable-number tandem repeat analysis employing 25 markers (MLVA25) to genotype 99 B. anthracis isolates from an archival collection of Australian isolates. MLVA25 genotyping revealed eight unique genotypes which clustered within the previously defined A3 genotype of B. anthracis. Genotyping of B. anthracis strains from outbreaks of disease in Victoria identified the presence of multiple genotypes associated with these outbreaks. The geographical distribution of genotypes within Australia suggests that a single genotype was introduced into the eastern states of Australia, followed by the spread and localised differentiation of the pathogen (MLVA25 genotypes MG1-MG6) throughout the anthrax belt. In contrast, unexplained occurrences of disease in areas outside of this anthrax belt which are associated with different genotypes, (MLVA25 genotypes MG7 and MG8) indicate separate introductions of B. anthracis into Australia.

Abalone viral ganglioneuritis: establishment and use of an experimental immersion challenge system for the study of abalone herpes virus infections in Australian abalone.

In late 2005, acute mortalities occurred in abalone on farms located in Victoria, Australia. Disease was associated with infection by an abalone herpes virus (AbHV). Subsequently, starting in 2006, the disease (abalone viral ganglioneuritis; AVG) was discovered in wild abalone in Victorian open waters. Currently, it continues to spread, albeit at a slow rate, along the Victorian coast-line. Here, we report on experimental transmission trials that were carried out by immersion using water into which diseased abalone had shed infectious viral particles. At various time points following exposure, naïve abalone were assessed by an AbHV-specific real-time PCR and histological analyses including in situ hybridization (ISH). Results demonstrated that while exposed abalone began displaying clinical signs of the disease from 60 hours post exposure (hpe), they tested positive for the presence of viral DNA at 36 hpe. Of further interest, the AbHV DNA probe used in the ISH assay detected the virus as early as 48 hpe.

Development and validation of a TaqMan PCR assay for the Australian abalone herpes-like virus.

The recent emergence of a herpes-like virus in both farmed and wild populations of abalone in Victoria, Australia, has been associated with high mortality rates in animals of all ages. Based on viral genome sequence information, a virus-specific real-time TaqMan assay was developed for detection and identification of the abalone herpes-like virus (AbHV). The assay was shown to be specific as it did not detect other viruses from either the Herpesvirales or the Iridovirales orders which have genome sequence similarities. However, the TaqMan assay was able to detect DNA from the Taiwanese abalone herpes-like virus, suggesting a relationship between the Taiwanese and Australian viruses. In addition, the assay detected < 300 copies of recombinant plasmid DNA per reaction. Performance characteristics for the AbHV TaqMan assay were established using 1673 samples from different abalone populations in Victoria and Tasmania. The highest diagnostic sensitivity and specificity were 96.7 (95% CI: 82.7 to 99.4) and 99.7 (95% CI: 99.3 to 99.9), respectively, at a threshold cycle (C(T)) value of 35.8. The results from 2 separate laboratories indicated good repeatability and reproducibility. This molecular assay has already proven useful in confirming presumptive diagnosis (based on the presence of ganglioneuritis) of diseased abalone in Victorian waters as well as being a tool for surveillance of wild abalone stocks in other parts of Australia.