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Fasanella A.,Anthrax Reference Institute of Italy | Garofolo G.,Anthrax Reference Institute of Italy | Hossain M.J.,International Center for Diarrheal Disease Research | Shamsuddin M.,Bangladesh Agricultural University | And 2 more authors.
Epidemiology and Infection | Year: 2013

In Bangladesh from 1 July to 30 September 2010 there were 104 animal cases of anthrax and 607 associated human cases. This investigation was conducted in Sirajganj district in December 2010, on eight farms where animal cases had occurred. Bacillus anthracis was recovered from soil samples and turbinate bones on six farms. Canonical single nucleotide polymorphism (SNP) analysis showed that all the isolates belonged to the major lineage A, sublineage A.Br.001/002 of China and South East Asia while a multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) with 15 VNTRs demonstrated three unique genotypes. The single nucleotide repeat (SNR) analyses showed two SNR types in 97 out of 99 isolates; nevertheless, due to its higher discriminatory power the presence of two isolates with different SNR-type polymorphisms were detected within two MLVA genotypes. The epidemic occurred during the monsoon season, a time of extensive flooding, suggesting that the source was contaminated feed, not grazing, which is supported by the genetic variance. © 2012 Cambridge University Press.

Mullins J.C.,University of Florida | Garofolo G.,Istituto Zooprofilattico Sperimentale dellAbruzzo e Del Molise G. Caporale | Garofolo G.,Anthrax Reference Institute of Italy | Van Ert M.,University of Florida | And 4 more authors.
PLoS ONE | Year: 2013

We modeled the ecological niche of a globally successful Bacillus anthracis sublineage in the United States, Italy and Kazakhstan to better understand the geographic distribution of anthrax and potential associations between regional populations and ecology. Country-specific ecological-niche models were developed and reciprocally transferred to the other countries to determine if pathogen presence could be accurately predicted on novel landscapes. Native models accurately predicted endemic areas within each country, but transferred models failed to predict known occurrences in the outside countries. While the effects of variable selection and limitations of the genetic data should be considered, results suggest differing ecological associations for the B. anthracis populations within each country and may reflect niche specialization within the sublineage. Our findings provide guidance for developing accurate ecological niche models for this pathogen; models should be developed regionally, on the native landscape, and with consideration to population genetics Further genomic analysis will improve our understanding of the genetic-ecological dynamics of B. anthracis across these countries and may lead to more refined predictive models for surveillance and proactive vaccination programs. Further studies should evaluate the impact of variable selection of native and transferred models. © 2013 Mullins et al.

Garofolo G.,Anthrax Reference Institute of Italy | Galante D.,Anthrax Reference Institute of Italy | Serrecchia L.,Anthrax Reference Institute of Italy | Buonavoglia D.,University of Bari | Fasanella A.,Anthrax Reference Institute of Italy
Journal of Microbiological Methods | Year: 2011

In the present study, a Taqman allelic discrimination assay based on three SNPs of the TPI gene is described. It was used as a differential diagnostic tool to detect blackleg and malignant edema. Sudden deaths of grazing ruminants, such as cattle, sheep and goats, which show clinical signs related to hyperacute infective processes, encouraged the development of a rapid and precise diagnostic molecular method. Specific primers and probes for Clostridium septicum and Clostridium chauvoei were designed on the basis of the TPI gene sequence. The multiplex PCR was tested on the DNA of a total of 57 strains, including 24 Clostridium chauvoei, 20 Clostridium septicum, 1 Bacillus anthracis and 12 other Clostridium spp. The DNA samples from Clostridium chauvoei and Clostridium septicum strains were amplified. Amplification of other DNA samples was not observed, with the exception of Clostridium tertium, which showed a weak positive signal. To avoid misdiagnosis, a confirmatory assay based on a Sybr green real time PCR was proposed. The authors confirmed the efficacy and the specificity of the test used in this study, which proved to be a useful tool for the diagnosis of clostridiosis that are often diagnosed using only traditional tools. © 2010 Elsevier B.V.

Fasanella A.,Anthrax Reference Institute of Italy | Di Taranto P.,Anthrax Reference Institute of Italy | Battisti A.,Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana | Longobardi C.,Azienda Sanitaria Firenze | And 3 more authors.
Giornale Italiano di Medicina Tropicale | Year: 2011

Anthrax represents a danger to humans especially when it occurs in areas considered to be free and in atypical seasons and climatic conditions. The underestimation of disease may lead health workers to misdiagnosis and, consequently, to an inappropriate management of affected carcasses with a consequent and inevitable increase in the risk of human infection. The analysis of soil and water samples collected in the Mugello Natural Park (Tuscany - Italy), considered an anthrax free area, showed that Bacillus anthracis was isolated from 4 of 6 investigated sites where in the past were registered several deaths of animals. This result was obtained thank to a new investigation approach including the analysis of same environmental factors that tend to favour the isolation of B. anthracis from soil.

Fasanella A.,Anthrax Reference Institute of Italy | Garofolo G.,Anthrax Reference Institute of Italy | Galante D.,Anthrax Reference Institute of Italy | Quaranta V.,Anthrax Reference Institute of Italy | And 4 more authors.
New Microbiologica | Year: 2010

Anthrax is a disease of humans and animals caused by the encapsulated, spore-forming Bacillus anthracis. In Italy, anthrax is normally a sporadic disease. During the summer 2004, anthrax broke out in the Basilicata, in southern Italy, a region with a low prevalence of anthrax in which vaccination had been suspended since 1998. The disease involved several animals in few weeks and in a large area. Over 41 days, 81 cattle died, as well as 15 sheep, 9 goats, 11 horses and 8 deer. The Multiple-locus Variable-Number Tandem Repeats Analysis (MLVA) showed that all the 53 isolates belonged to the Cluster Ala, genotype 1. The results of the Single Nucleotide Repeats (SNRs) Analysis showed that 48/53 B.anthacis strains belonged to a single clonal lineage, the sub-genotype sgt - eB. Two sporadic mutants, sgt - eB,ml and sgt- eB, m2, were isolated, only one managing to infect other herds. Factors that could have contributed to the spread of infection, such as the transmission of spores by insect vectors and the favourable weather conditions were evaluated.

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