Closed-circuit system for the depuration of mussels experimentally contaminated with hepatitis A virus.

In Italy, the consumption of raw or slightly cooked mussels represents the most important risk factor for the transmission of hepatitis A virus (HAV). Although there exist effective methods for the bacterial depuration of contaminated mussels, these methods are poorly effective on enteric viruses. The objective of the present study was to evaluate the effectiveness of a closed-circuit depuration system that uses both ozone and UV light for disinfecting water and that allows salinity and temperature, important parameters for the metabolism of mussels (Mytilus galloprovincialis), to be maintained at constant levels. The results showed that this depuration method decreased the viral load (from 1.72 log 50% tissue culture infective dose [TClD 50 ] ml -1 to <1 log TCID 50 ml -1 within 24 h and from 3.82 log TCID 50 ml -1 to <1 log TCID 50 ml -1 within 48 h). However, in both cases, after 120 h of depuration, a residual amount of virus capable of replicating in cells was detected. These results show that depuration, even if performed with advanced systems, may not guarantee the absence of virus

Development of rapid methods for determination of Salmonella in meat products

The aim of the present study was the evaluation of two different techniques to detect Salmonella in meat: an ELISA assay using electrochemical detection coupled with a FIA system and a PCR method. A sandwich format was chosen for ELISA assay, using monoclonal and polyclonal antibodies against salmonella. A 35-cycle PCR was carried out for the research of genomic fragment. The assays were used to analyse samples of pork, chicken and bovine meat experimentally contaminated with different concentrations of S. Enteritidis. Results show that both methods were efficient, sensitive and rapid. After only 4 hours of incubation in pre-enrichment broth (buffered peptone water) it was possible to detect salmonella in meat experimentally contaminated with low concentrations (1-10 cells in 25 g) both with the ELISA assay and the PCR method

Hepatitis E Virus Occurrence in Pigs Slaughtered in Italy

In Europe, foodborne transmission has been clearly associated to sporadic cases and small clusters of hepatitis E in humans linked to the consumption of contaminated pig liver sausages, raw venison, or undercooked wild boar meat. In Europe, zoonotic HEV-genotype 3 strains are widespread in pig farms but little information is available on the prevalence of HEV positive pigs at slaughterhouse. In the present study, the prevalence of HEV-RNA positive pigs was assessed on 585 animals from 4 abattoirs located across Italy. Twenty-one pigs (3.6%) tested positive for HEV in either feces or liver by real-time RT-PCR. In these 21 pigs, eight diaphragm muscles resulted positive for HEV-RNA. Among animals collected in one abattoir, 4 out of 91 plasma tested positive for HEV-RNA. ELISA tests for the detection of total antibodies against HEV showed a high seroprevalence (76.8%), confirming the frequent exposure of pigs to the virus. The phylogenetic analyses conducted on sequences of both ORF1 and ORF2 fragments, shows the circulation of HEV-3c and of a novel unclassified subtype. This study provides information on HEV occurrence in pigs at the slaughterhouse, confirming that muscles are rarely contaminated by HEV-RNA compared to liver, which is the most frequently positive for HEV

salmonella enterica control in stick carrots through incorporation of coriander seeds essential oil in sustainable washing treatments

Chemical disinfectants represent one of the commonly used practice in minimal processed vegetables food-chain. However, the scarce safety and sustainability of these agents force food industry to move toward more sustainable "green washing solutions". Among the latter, while the application of plant derivates for the control of several pathogens is already well-known, specific information on the potential anti-Salmonella activity of Coriandrum sativum seeds derivates are still limited and were therefore investigated in this study. In detail, Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) of different coriander seed derivates (i.e. essential oil, hydrosol and ethanolic extract) were determined by broth dilution against six Salmonella enterica strains isolated from fresh and minimally processed fruits and vegetables. Only the essential oil (EO) was effective in vitro with strain-dependent results. In addition, when mixed in co-culture, the strains were more sensitive to the essential oil treatment. Chemical investigations allowed to define (s)-(+)-linalool as major compound, and to underline interesting phenolic content with correlated antioxidant capacity. A cocktail of three strains of different serovars was selected and employed for a preliminary in situ trial on stick carrots. The obtained results allowed to establish that the application of coriander seed EO at concentrations of 5 µL mL-1 was able to reduce and contain the growth of the Salmonella cocktail up to 24 hours at 10°C. Good sensory evaluation results were obtained by applying this EO concentration as washing treatment, especially in terms of color parameter. Further studies should be undertaken to emphasize the upstream activity, improving the formulation or exploiting a combined effect with other sanitizers or treatments (e.g. physical treatments). The present study contributes to the knowledge on coriander derivates activity against Salmonella spp. and on the potential application as sustainable washing treatment in removing this pathogen from fresh cut carrots

Foodborne botulism associated with home-preserved turnip tops in Italy

In Italy, foodborne botulism is a rare disease mainly due to home-preserved food. Inthe case reported here, clinical diagnosis was performed on the basis of clinical signsand referred consumption of home-preserved turnip tops in oil. Definitive diagnosis wasperformed by detection of botulinum toxin in sera and neuro-toxigenic organisms instools and leftover food. This case report highlights the need of a high medical awareness,prompt clinical diagnosis, and synergic collaboration among the health authorities for acorrect management of botulism as well as disease containment

First isolation of Salmonella enterica serovar Napoli from wild birds in Italy

AbstractSalmonella enterica serovar Napoli (S. Napoli) is an emerging serovar in Italy. It accounts for 2-4% of all serovars isolated from human infections. The zoonotic origin of this serovar is still unknown and this makes difficult to apply any control intervention. We report here the isolation of S. Napoli from a river nightingale (Cettia cetti, Temminck 1820) which represents the first description of this serovar from wild birds. This finding adds knowledge to the ecology of S. Napoli and addresses further studies aimed to assess the epidemiologic link between S. Napoli isolated from wild birds, food, environmental sources and human infections.

The Workshop on Animal Botulism in Europe

A workshop on animal botulism was held in Uppsala, Sweden, in June 2012. Its purpose was to explore the current status of the disease in Europe by gathering the European experts in animal botulism and to raise awareness of the disease among veterinarians and others involved in biopreparedness. Animal botulism is underreported and underdiagnosed, but an increasing number of reports, as well as the information gathered from this workshop, show that it is an emerging problem in Europe. The workshop was divided into 4 sessions: animal botulism in Europe, the bacteria behind the disease, detection and diagnostics, and European collaboration and surveillance. An electronic survey was conducted before the workshop to identify the 3 most needed discussion points, which were: prevention, preparedness and outbreak response; detection and diagnostics; and European collaboration and surveillance. The main conclusions drawn from these discussions were that there is an urgent need to replace the mouse bioassay for botulinum toxin detection with an in vitro test and that there is a need for a European network to function as a reference laboratory, which could also organize a European supply of botulinum antitoxin and vaccines. The foundation of such a network was discussed, and the proposals are presented here along with the outcome of discussions and a summary of the workshop itself

Animal Botulism Outcomes in the AniBioThreat Project

Botulism disease in both humans and animals is a worldwide concern. Botulinum neurotoxins produced by Clostridium botulinum and other Clostridium species are the most potent biological substances known and are responsible for flaccid paralysis leading to a high mortality rate. Clostridium botulinum and botulinum neurotoxins are considered potential weapons for bioterrorism and have been included in the Australia Group List of Biological Agents. In 2010 the European Commission (DG Justice, Freedom and Security) funded a 3-year project named AniBioThreat to improve the EU's capacity to counter animal bioterrorism threats. A detection portfolio with screening methods for botulism agents and incidents was needed to improve tracking and tracing of accidental and deliberate contamination of the feed and food chain with botulinum neurotoxins and other Clostridia. The complexity of this threat required acquiring new genetic information to better understand the diversity of these Clostridia and develop detection methods targeting both highly specific genetic markers of these Clostridia and the neurotoxins they are able to produce. Several European institutes participating in the AniBioThreat project collaborated on this program to achieve these objectives. Their scientific developments are discussed here

Separated by a Common Language: Awareness of Term Usage Differences Between Languages and Disciplines in Biopreparedness

Preparedness for bioterrorism is based on communication between people in organizations who are educated and trained in several disciplines, including law enforcement, health, and science. Various backgrounds, cultures, and vocabularies generate difficulties in understanding and interpretating terms and concepts, which may impair communication. This is especially true in emergency situations, in which the need for clarity and consistency is vital. The EU project AniBio- Threat initiated methods and made a rough estimate of the terms and concepts that are crucial for an incident, and a pilot database with key terms and definitions has been constructed. Analysis of collected terms and sources has shown that many of the participating organizations use various international standards in their area of expertise. The same term often represents different concepts in the standards from different sectors, or, alternatively, different terms were used to represent the same or similar concepts. The use of conflicting terminology can be problematic for decision makers and communicators in planning and prevention or when handling an incident. Since the CBRN area has roots in multiple disciplines, each with its own evolving terminology, it may not be realistic to achieve unequivocal communication through a standardized vocabulary and joint definitions for words from common language. We suggest that a communication strategy should include awareness of alternative definitions and ontologies and the ability to talk and write without relying on the implicit knowledge underlying specialized jargon. Consequently, cross-disciplinary communication skills should be part of training of personnel in the CBRN field. In addition, a searchable repository of terms and definitions from relevant organizations and authorities would be a valuable addition to existing glossaries for improving awareness concerning bioterrorism prevention planning

Management of Animal Botulism Outbreaks: From Clinical Suspicion to Practical Countermeasures to Prevent or Minimize Outbreaks

Botulism is a severe neuroparalytic disease that affects humans, all warm-blooded animals, and some fishes. The disease is caused by exposure to toxins produced by Clostridium botulinum and other botulinum toxin–producing clostridia. Botulism in animals represents a severe environmental and economic concern because of its high mortality rate. Moreover, meat or other products from affected animals entering the food chain may result in a public health problem. To this end, early diagnosis is crucial to define and apply appropriate veterinary public health measures. Clinical diagnosis is based on clinical findings eliminating other causes of neuromuscular disorders and on the absence of internal lesions observed during postmortem examination. Since clinical signs alone are often insufficient to make a definitive diagnosis, laboratory confirmation is required. Botulinum antitoxin administration and supportive therapies are used to treat sick animals. Once the diagnosis has been made, euthanasia is frequently advisable. Vaccine administration is subject to health authorities' permission, and it is restricted to a small number of animal species. Several measures can be adopted to prevent or minimize outbreaks. In this article we outline all phases of management of animal botulism outbreaks occurring in wet wild birds, poultry, cattle, horses, and fur farm animals