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Inter-Laboratory Ring Trial to Evaluate Reverse Transcription Polymerase Chain Reaction Methods Used for Dolphin MorbillivirusDetection in Italy

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Advances in Microbiology 2019年第5期9卷 423-437页

作者： Zoccola Roberto Pautasso Alessandra Grattarola Carla Monnier Milena Miceli Ilaria Ingravalle Francesco Crescio Maria Ines Giorda Federica Masoero Loretta Zaccaria Guendalina Purpari Giuseppa Cersini Antonella Viscardi Maurizio Serracca Laura Padalino Iolanda Puggioni Giantonella Canonico Cristina Toffan Anna Centelleghe Cinzia Di Francesco Cristina Esmeralda Mazzariol Sandro Di Guardo Giovanni Mignone Walter Casalone Cristina Goria Maria istituto zooprofilattico sperimentale del Piemonte Liguria e Valle d’Aosta Torino Italy istituto zooprofilattico sperimentale dell’Abruzzo e del Molise Teramo Italy istituto zooprofilattico sperimentale della Sicilia Palermo Italy istituto zooprofilattico sperimentale del Lazio e della Toscana Roma Italy istituto zooprofilattico sperimentale del Mezzogiorno Portici (NA) Italy istituto zooprofilattico sperimentale del Piemonte Liguria e Valle d’Aosta La Spezia Italy istituto zooprofilattico sperimentale della Puglia e Basilicata Foggia Italy istituto zooprofilattico sperimentale della sardegna Sassari Italy istituto zooprofilattico sperimentale dell’ Umbria e delle Marche Ancona Italy istituto zooprofilattico sperimentale delle Venezie Legnaro (PD) Italy University of Padova Department of Comparative Biomedicine and Food Science (BCA) Legnaro (PD) Italy Faculty of Veterinary Medicine University of Teramo Teramo Italy istituto zooprofilattico sperimentale del Piemonte Liguria e Valle d’Aosta Imperia Liguria Italy

Dolphin Morbillivirus (DMV) is one of the most frequently detected pathogens in stranded cetacean specimens worldwide as well as in Italy. Due to the persistence of DMV in the Mediterranean Sea and to the lack of information about the efficiency of the available diagnostic techniques, the Italian National Reference Centre for diagnostic activities on dead stranded marine mammals (***) performed the first inter-laboratory ring trial with the aim to standardize a diagnostic biomolecular approach for DMV in Italy. Viral isolation is usually considered the “gold standard” for the definitive diagnosis of most pathogens, but it is not often feasible in DMV diagnosis, due to the poor preservation of virus-targeted tissues in stranded cetacean carcasses, as well as to the lack of appropriate sensitivity of cell lines towards DMV variability. Therefore direct viral detection on tissues by means of reverse transcription-PCR (RT-PCR) represents a valuable option for DMV infection’s diagnosis. For detecting DMV in cetacean die-offs occurred in the Mediterranean basin since 2013, *** developed an RT-PCR based method targeting to a 287 bp fragment of DMV nucleoprotein (N) gene. With the purpose to evaluate its performances in terms of accuracy (Se = sensitivity and Sp = specificity) and precision (reproducibility), it was submitted to a ring trial. So, 12 Public Laboratories belonging to the Italian dead stranded marine mammals diagnostic network were asked to analyze a panel of 40 samples (positive and negative for DMV, using different dilutions of a viral suspension obtained from a cell culture supernatant of a DMV strain) with the aforementioned technique. Furthermore, we also aimed at comparing the accuracy of other 7 molecular methods routinely applied for DMV detection in Italy. For this purpose, the second panel of identical 40 DMV +ve and -ve samples was provided to Laboratories that routinely used DMV detection methods other than those developed by ***.

关键词： Dolphin Morbillivirus RT-PCR Interlab Ring Test Method Comparison Stranded Cetaceans Mediterranean Sea

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Development and Validation of an Ion Chromatography Method for the Simultaneous Determination of Seven Food Additives in Cheeses

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Journal of Analytical Sciences, Methods and Instrumentation 2013年第3期3卷 30-37页

作者： Marco Iammarino Aurelia Di Taranto istituto zooprofilattico sperimentale della Puglia e della Basilicata Foggia Italy.

Cheeses are characterized by several chemical-physical properties that make it difficult for the microorganisms growth, consequently. The actual European legislation allows the addition of few food additives in this type of food products. In this work, the entire procedure of extraction, purification, chromatographic separations and quali/quantitative determination of seven food additives (sorbic acid, benzoic acid, lactic acid, acetic acid, nitrites, nitrates and phosphates) was developed and applied for the analysis of different types of cheese (mozzarella, cheese spread, semi-hard and hard cheeses). Through validation procedure it was possible to evaluate the most important validation parameters. Extended calibration curves (r > 0.990) were obtained for all the analyzed compounds. Recovery values ranged from 72.8% to 98.4% and a good repeatability was obtained, with precision levels in the range of 0.03% - 0.11% (n = 6). The potential and feasibility of the method were tested by analysing real samples, such as mozzarella, cheese spread, semi-hard and hard cheeses, confirming that the method is well suited to satisfy the demands for accurate confirmation analyses of seven food additives in cheeses, which is especially valuable in official check analyses and in monitoring schemes.

关键词： Dairy Products Cheese Food Additives Food Preservatives Nitrites Nitrates Ion Chromatography Validation

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Recent Epidemic-Like Anthrax Outbreaks in Italy: What Are the Probable Causes?

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Open Journal of Veterinary Medicine 2012年第2期2卷 74-76页

作者： Lucia Palazzo Esterina De Carlo Gianfranco Santagada Luigina Serrecchia Angela Aceti Achille Guarino Rosanna Adone Antonio Fasanella ISS-istituto Superiore di Sanità Roma Italy istituto zooprofilattico sperimentale del Mezzogiorno Napoli Italy istituto zooprofilattico sperimentale della Puglia e della Basilicata Foggia Italy

In Italy, in an area of about 50 km2 between Basilicata and Campania Southern regions, 28 outbreaks of anthrax occurred from August 28th to September 27th 2011. Different species were affected: laboratory tests confirmed anthrax in cattle, horses and sheep. The genetic analysis of strains isolated from infected animals indicated that outbreaks occurring in the two regions were not correlated. Intriguingly, the incidence was highly significant in horses compared to that of cattle and sheep or goats, which were prevalent species in the animal population. In addition, allinfected horses and many cattle developed a fatal, sub-acute form of anthrax, characterized by the presence of massive edema, usually absent in hyperacute forms. The characteristics of these outbreaks suggested a possible role of tabanids as vectors in the transmission of B. anthracis like-epidemic infection.

关键词： Anthrax Epidemic Edema Horses Tabanids

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Diagnosis and treatment of unusual multiorgan Echinococcus hydatid cysts

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Current Urology 2024年第1期18卷 75-77页

作者： Giovanni Drocchi Cinzia Santucciu Scilla Mastrandrea Francesco Sanguedolce Massimo Madonia Department of Medical Surgical and Experimental SciencesUniversita degli Studi di SassariSassariItaly OIE and National Reference Laboratory for Echinococcosis(CeNRE) Istituto Zooprofilattico Sperimentale della SardegnaSassariItaly Department of Infectious disease Sassari University HospitalSassariItaly

Background:Echinococcosis is a zoonotic disease caused by Echinococcus ***,the liver is the most affected organ,accounting for approximately 70%to 85%of *** lungs represent 20%of the *** than 10%are found in other sites,including the bone,brain,spleen,and *** presentation:A young man was referred to a local hospital for dull pain in the right flank,fever,and mild *** tomography showed 2 large cysts:the first involved the left lung and measured 130×90.5×120 mm,whereas the second cyst was located in the right kidney and measured 130×100×120 *** was performed to remove both lesions in 2 separate surgical ***:Echinococcosis is a compulsorily notifiable *** between medical doctors from different specializations is necessary.A multidisciplinary approach is important for the correct therapeutic management of the ***,the high possibility of recurrence makes the long-term follow-up mandatory.

关键词： Cyst Echinococcosis Infection Kidney Nephrectomy

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The cold chain and the COVID-19 pandemic:an unusual increase in histamine content in fish samples collected in Southern Italy during lockdown

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Food Quality and Safety 2021年第4期5卷 329-335页

作者： Francesco Giuseppe Gailuzzo Gaetano Cammilleri Antonello Cicero Licia Pantano Andrea Pulvirenti Andrea Macaluso Nicola Cicero Vittorio Calabrese Vincenzo Ferrantelli istituto zooprofilattico sperimentale della Sicilia PalermoItaly Dipartimento di Scienze della Vita Universita degli studi di Modena e Reggio EmiliaModenaItaly Dipartimento SASTAS Universita degli studi di MessinaMessinaItaly Dipartiment'o di Scienze biomediche e biotecnologiche Universita degli studi di CataniaCataniaItaly

Objectives:We analysed 900 samples of fresh(250)and processed(650)fish products collected in Sicily(Southern Italy)in 2020 during the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)pandemic(hereafter:COVID-19).Materials and methods:The samples were divided temporally based on four phases relating to the various restrictions imposed by the Italian government in this *** validated method of ultrahigh-performance liquid chromatography combined with a diode array detector was then employed for the ***:The samples collected during the Phase I lockdown period and after it had ended(Phase II)revealed significant increases in the mean histamine levels:(41.89±87.58)mg/kg and(24.91±76.76)mg/kg,*** 11(1.3%of the total)fresh fish samples that were identified as being non-compliant with Regulation(EC)No.2073/2005 were only found during these two *** the processed samples were always *** histamine values decreased as the restrictions eased,achieving a mean value of(11.16±9.3)mg/kg(Phase III).Conclusions:There was an increase in the incidence offish samples that were non-compliant with Regulation(EC)No.2073/2005 compared to previous surveillance *** results provide a first report on the effect of lockdown measures on food safety and the cold *** findings must cause food safety operators to intensify their controls over fresh fish products in such periods to safeguard consumer *** studies are required to evaluate whether the same trend would be observed with other food contaminants.

关键词： Histamine cold-chain food safety COVID-19

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Species Identification by Means of Mitochondrial Cytochrome bDNA Sequencing in Processed Anchovy, Sardine and Tuna Products

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Food and Nutrition Sciences 2018年第4期9卷 369-375页

作者： Anna Cutarelli Giorgio Galiero Federico Capuano Federica Corrado istituto zooprofilattico sperimentale del Mezzogiorno Napoli Italy

Identifying the contents of processed food products is essential to correct labelling. In processed foodstuffs, species identification through morphological analysis is difficult. Several factors hinder the identification of fish species in processed foods: proteins or other materials subjected to analysis may be denatured during heat treatments;the presence of other ingredients (e.g., olive and other vegetable oils) may interfere with the analysis. Consequently, possible frauds perpetrated by replacing valuable species with less precious ones may go undetected. In most processed samples (e.g. canned products), DNA is degraded into small fragments, which considerably reduces the sensitivity of molecular analysis. The main goal of our research was to develop an analytical method able to identify fish species in highly processed products, such as canned fish. The assay was developed by combining an effective method of DNA recovery from samples with the detection of small-sized sequences of the mitochondrial Cytb gene. This method appears particularly suitable when morphological characterization is difficult, to carry out such as in canned products where DNA is degraded or present in small quantities. We have analyzed 60 samples of seafood commercial products identifying 3 different genera and five different species. All analyzed samples revealed a correct species declaration, for one sample we highlighted important commercial fraud. We also used bio-informatic identification systems for the Sequence Alignment and the construction of phylogenetic tree to better confirm the revealed fraud.

关键词： Species Identification Processed Seafood Commercial Fraud Dna Sequencing Cytochrome b (Cytb)

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Effectiveness of Thymus vulgarisEssential Oil in the Treatment of Skin Infections in Dogs

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Open Journal of Veterinary Medicine 2017年第6期7卷 63-75页

作者： Vincenzo Naccari Bianca Maria Orlandella Vittorio Fisichella Francesco Naccari Santo Caracappa istituto zooprofilattico sperimentale della Sicilia “A. Mirri” Palermo Italy Department of Veterinary Sciences University of Messina Messina Italy

Currently, there are no data in the literature on the topical treatment with Thymus vulgaris essential oil (EO) in bacterial skin infections of dogs. This study evaluates the effectiveness of Thymus vulgaris EO for topical use in dogs with skin 18 half-breed dogs, affected by skin infections housed in a rescue shelter were studied. The bacteria isolated from these dogs were identified by MALDI-TOF-MS. The susceptibility of the isolated microorganisms to Thymus vulgaris L. EO was estimated in vitro by bacteriological test (CLSI 2015), in comparison to some antimicrobials drugs (amoxicillin-clavulanic acid, doxycy-cline, thiamphenicol and marbofloxacin) and to Citrus bergamia EO. The dogs, divided in two groups at random, were treated topically for 7 days with Thymus vulgaris L. EO (Group 1: n. 10 animals) and Citrus bergamia Risso e Poiteau EO (Group 2: n. 8 animals) respectively. The bacteria isolated were: Staphylococcus pseudintermedius (7 samples), Staphylococcus sciuri (4), ESBL Escherichia coli (3) and Proteus mirabilis (4). In all animals treated with Thymus vulgaris EO, the clinical signs decreased rapidly within 5 days from the administration, with complete remission 7 days after the treatment. No bacterial growth was observed from skin swabs after 7 days of treatment. None of the treated animals showed local or general side effects. The use of Thymus vulgaris EO could be a possible alternative or additional treatment to antibiotics in dermatological infections, particularly in cases refractory to conventional therapy.

关键词： Bacteriological Skin Infections Dog Thymus vulgaris Essential Oil

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Monitoring Plan on PCDD/Fs and DL-PCBs Milk Contamination in Campania Region (Italy)

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International Journal of Analytical Mass Spectrometry and Chromatography 2015年第2期3卷 32-36页

作者： Francesco Paolo Serpe Alfredo Scaramuzzo Pasquale Maglio Sara Lambiase Mauro Esposito istituto zooprofilattico sperimentale del Mezzogiorno Portici Italy

During the carrying out of the monitoring plan as part of the Regional Law n. 3/2005 emerged, in 2008, the well known “buffalo milk crisis” that led to the enactment of other monitoring plans targeted to the research of dioxins in food. From the last monitoring plan on PCDD/Fs and DL-PCBs in Campania Region, named as “Surveillance plan 2011-2014”, all dairy products from N. 50 farms intended to the production of bovine, buffalo and ovine milk, resulted compliant to the maximum levels fixed by EU in Regulation 1259/2011, suggesting the effectiveness of the restrictive measures on farming practices applied after the sanitary emergency of 2008. On the other hand, the action limits reported in EU Recommendations 516/2011 and 711/2013 were exceeded in two sheep farms and two buffalo farms, to represent the critical issues that nowadays exist.

关键词： Dioxin Campania Region Italy Milk

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Some pathogenic characters of paratyphoid Salmonella enterica strains isolated from poultry

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Asian Pacific Journal of Tropical Medicine 2017年第12期10卷 1161-1166页

作者： Fabrizio Bertelloni Giovanni Tosi Paola Massi Laura Fiorentini Maria Parigi Domenico Cerri Valentina Virginia Ebani Department of Veterinary Science University of PisaViale Delle Piagge 256124 Pisa Italy istituto zooprofilattico sperimentale della Lombardia e Dell'Emilia Romagna Sezione Diagnostica di Forl(i)Via Don E. Servadei 3E/3F47122 Forl(i)(FC) Italy

Objective: To investigate some pathogenic characters of Salmonella enterica strains isolated from ***: Twenty-three genetically distinct Salmonella enterica strains, of different serovars and pulsotype, were examined for virulence traits. Resistance to gastric acid environment was estimated by measuring the percentage of survived bacterial cells after exposure for 2 h to a synthetic gastric juice. Strains were analyzed with PCR for the presence of the following virulence genes: mgtC and rhu M located on SPI-3, sopB and pipB located on SPI-5, Salmonella virulence plasmid(spv)R(spvR), spvB and spvC located on Salmonella plasmid virulence and sodCI, sopE, and gipA located on ***, resistance to 21 antibiotics was tested with Kirby–Bauer ***: A percentage of 82.60% of strains were resistant to gastric environment after induction and 60.87% of the strains exhibited constitutive resistance too. Nineteen different virulence profiles were detected. The phage related genes sodCI and sopE and the plasmid mediated operon spvR, spvB and spvC(spvRBC) were detected in 82.60%,47.82% and 52.17% of strains, respectively. Typhimurium and Enteritidis strains showed the highest number of virulence genes. Twenty-one different antibiotic resistance profiles were obtained and two isolates(Typhimurium and Enteritidis) resulted sensible to all the tested molecules. The ampicillin, streptomycin, sulfonamide and tetracycline resistance profile was detected in seven isolates(30.43%).Conclusion: Our results show that paratyphoid Salmonella strains with several characters of pathogenicity, that may be cause of severe pathology in animals and humans, are circulating among poultry.

关键词： Paratyphoid salmonellae Poultry Gastric acid resistance Virulence genes Antimicrobial resistance

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Isolation of Bacillus cereusin a Facility Preparing School Meals

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Food and Nutrition Sciences 2020年第3期11卷 186-193页

作者： Francesca Garofalo Anna Cutarelli Rita Nappi Assunta De Lella Marcella Palomba Salvatore Capo Angela Michela Immacolata Montone Loredana Biondi Federica Corrado Food Inspection Department Istituto Zooprofilattico Sperimentale del Mezzogiorno Portici Italy

Food safety is a fundamental requirement in mass catering, as large numbers of meals are served each day to potentially vulnerable consumers, such as children. Food Business Operators implement plans for the microbiological monitoring of the meals prepared and served in the catering sector, and for the swab-sampling of surfaces. From January 2018 to June 2019, our laboratory analyzed both food and swab samples from four catering facilities. Considering the EFSA 2018 data, we specifically focused on samples analyzed for Bacillus cereus. Our data substantially showed episodic contamination due to a piece of equipment that is not usually subjected to microbiological control, thus suggesting that every aspect should be scrutinized in order to identify critical points. While Bacillus cereus is widespread in nature and common in soil, it is adapted for growth in the intestinal tracts of insects and mammals. It is often present in a variety of foods, and may cause an emetic or a diarrheal type of food-associated illness. B. cereus produces several toxins. Multiplex PCR enables seven toxin genes to be detected (hblC, hblD, hblA, nheA, nheB, nheC and cytK).

关键词： Bacillus cereus Food-Borne Disease School Meals Enterotoxin Genes

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