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Grattarola C.,University of Bologna | Giorda F.,University of Bologna | Iulini B.,University of Bologna | Pintore M.D.,University of Bologna | And 15 more authors.
Diseases of Aquatic Organisms | Year: 2016

Listeria monocytogenes, Toxoplasma gondii and Brucella spp. can infect a wide range of species, including humans. In cetaceans, meningoencephalitis has been associated with T. gondii and Brucella spp. infection, whereas to our knowledge, L. monocytogenes infection has not previously been reported. Meningoencephalitis and L. monocytogenes, T. gondii and Brucella spp. were identified by means of both direct and indirect laboratory techniques in an adult female striped dolphin Stenella coeruleoalba found stranded in January 2015 on the Ligurian Sea coast, northwestern Italy. The animal was emaciated, and histopathology disclosed severe meningoencephalitis. The nature of the inflammatory response and intra-lesional protozoa were consistent with a mixed infection by L. monocytogenes, T. gondii and Brucella spp. We believe this is an unprecedented case of infection by 3 zoonotic pathogens and also the first bacteriologically confirmed case report of neurolisteriosis in cetaceans. Cerebral toxoplasmosis and neurobrucellosis may have led to the animal's disorientation and stranding, with L. monocytogenes having likely exacerbated the coinfection leading to the demise of this dolphin. © Inter-Research 2016. Source

The multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA) is a genetic typing method based on the evaluation of the number of repeated sequences in multiple selected loci of microbial DNA. Although several MLVA typing panels have been proposed for brucellae, the 16-loci panel is recognized as the standard genotyping method, also used for the Brucella international online database. This chapter describes a high-throughput MLVA-16 protocol using multiplex PCRs and multicolor capillary electrophoresis. © Springer Science+Business Media New York 2015. Source

Garofolo G.,National and OIE Reference Laboratory for Brucellosis | Fasanella A.,Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata | Di Giannatale E.,National and OIE Reference Laboratory for Brucellosis | Platone I.,National and OIE Reference Laboratory for Brucellosis | And 5 more authors.
BMC Research Notes | Year: 2016

Background: Human brucellosis cases are still reported each year in Sweden despite eradication of the disease in animals. Epidemiological investigation has never been conducted to trace back the source of human infection in the country. The purpose of the study was to identify the source of infection for 16 human brucellosis cases that occurred in Sweden, during the period 2008-2012. Results: The isolates were identified as Brucella melitensis and MLVA-16 genotyping revealed 14 different genotypes of East Mediterranean and Africa lineages. We also reported one case of laboratory-acquired brucellosis (LAB) that was shown to be epidemiological linked to one of the cases in the current study. Conclusions: Brucella melitensis was the only species diagnosed, confirming its highest zoonotic potential in the genus Brucella, and MLVA-16 results demonstrated that the cases of brucellosis in Sweden herein investigated, are imported and linked to travel in the Middle East and Africa. Due to its zoonotic concerns, any acute febrile illness linked to recent travel within those regions should be investigated for brucellosis and samples should be processed according to biosafety level 3 regulations. © 2016 The Author(s). Source

Garofolo G.,National and OIE Reference Laboratory for Brucellosis | Di Giannatale E.,National and OIE Reference Laboratory for Brucellosis | De Massis F.,National and OIE Reference Laboratory for Brucellosis | Zilli K.,National and OIE Reference Laboratory for Brucellosis | And 4 more authors.
Infection, Genetics and Evolution | Year: 2013

Despite the eradication of brucellosis from most of Europe, the disease remains relatively common in a variety of livestock in southern European countries. It is therefore surprising that with such high prevalence rates, there have been few genetic characterizations of brucellosis outbreaks in this region. We conducted a genetic assessment of 206 isolates of Brucella abortus and B. melitensis from Italy using Variable Number Tandem Repeats (VNTRs). We determined genetic diversity and geographic distribution of these Brucella VNTR genotypes from 160 farms in eight regions of Southern Italy in a fine-scale analysis using 16 VNTR loci in a MLVA-16 methodology. In a broad scale analysis, we then used a reduced dataset of 11 VNTR loci (MLVA-11) to compare genotypes from Italy to a global database. In the 84 isolates of B. melitensis, there were 56 genotypes using MLVA-16; 43 of these genotypes were found only once. At a broad scale, 81 of these isolates were part of an Italian sub-group within the West Mediterranean group. One of the two B. melitensis isolates from a human patient shared the same genotype as a livestock isolate, suggesting a possible epidemiological connection. In 122 B. abortus isolates, there were 34 genotypes by MLVA-16; 16 of these genotypes were found only once. At a broad scale with MLVA-11, one genotype was predominant, comprising 77.8% of the isolates and was distributed throughout Southern Italy. These data on the current lineages of Brucella present in Italy should form the basis for epidemiological studies of Brucella throughout the country, while placing these strains in a global context. © 2013 Elsevier B.V. Source

Del Chierico F.,Unit of Parasitology and Unit of Metagenomics | Ancora M.,National and OIE Reference Laboratory for Brucellosis | Marcacci M.,National and OIE Reference Laboratory for Brucellosis | Camma C.,National and OIE Reference Laboratory for Brucellosis | And 2 more authors.
Methods in Molecular Biology | Year: 2015

The next-generation sequencing (NGS) technologies are revolutionary tools which have made possible achieving remarkable advances in genetics since the beginning of the twenty-first century. Thanks to the possibility to produce large amount of sequence data, these tools are going to completely substitute other high-throughput technologies. Moreover, the large applications of NGS protocols are increasing the genetic decoding of biological systems through studies of genome anatomy and gene mapping, coupled to the transcriptome pictures. The application of NGS pipelines such as (1) de-novo genomic sequencing by mate-paired and whole-genome shotgun strategies; (2) specific gene sequencing on large bacterial communities; and (3) RNA-seq methods including whole transcriptome sequencing and Serial Analysis of Gene Expression (Sage-analysis) are fundamental in the genome-wide fields like metagenomics. Recently, the availability of these advanced protocols has allowed to overcome the usual sequencing technical issues related to the mapping specificity over standard shotgun library sequencing, the detection of large structural genomes variations and bridging sequencing gaps, as well as more precise gene annotation. In this chapter we will discuss how to manage a successful NGS pipeline from the planning of sequencing projects through the choice of the platforms up to the data analysis management. © Springer Science+Business Media New York 2015. Source

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