Deep sequencing targeted for strain identification of trypanosomatids

Trypanosomatidae is a group of kinetoplastid excavates distinguished by having only a single flagellum. All members are exclusively parasitic, found primarily in insects. The custom reads form Ilumina sequencing platform are aligned to reference genomes of different Leishmania species taken from TriTryp dabatase to be used for bioinformatics analysis.peer-reviewe

Genetic database development for the characterisation of Sicilian sheep population

The most representative sheep in Sicily are Belicina, Comisana, Pinzirita, Barbarescaand the crossbred derived sheep from all this species. In this study, the allelic frequencies of the Sicilian sheep population were investigated. It currently represents the best way to determine the genetic identity and/or family even with limited amounts of sample or when the DNA is degraded. The aim of the study was to provide a reference data bank and to evaluate a microsatellite panel for pedigree analysis as suggested by the International Society for Animal Genetics (ISAG). There are various studies on European sheep, but few datasets were developed on the population of Sicilian sheep. The reference database will include allele frequencies at each locus and will determine genetic parameters for Sicilian ovine species selection. Our results indicated that Hardy Weinberg equilibrium was not always maintained. These results could be explained by a non-random mating. The database is useful to investigate the relationship, the parentage the meat traceability and in disease control programs. The standardized panels of allele frequencies represent a molecular fingerprinting characterizing the subjects with very high definition level and can be useful to control all the livestock. The parentage identification could be important for the veterinary police to investigate the theft or the animal substitutions in the Sicilian farms

Taxidermal Preparations of an Extinct Sicilian Wolf Population (Canis lupus Linnaeus, 1758): Biological Studies and Restoration

In last decades, biotechnologies have enabled a deep development of knowledge in all fields of life sciences, allowing the collection of a wide range of data. Furthermore, the relationship between scientific investigations and conservation actions of cultural heritage is increasingly consolidated and now it is one of the peculiar points both for sustainable restoration and use of the cultural asset, also evaluating the cultural assets as source of biological information. In this study, Sicilian wolf (Canis lupus) specimens conserved in Sicilian Natural Museums, become a source of biological information on Sicilian biodiversity, establishing specific conservative protocol developed cooperatively with Sicilian Natural Museums. Using Next-Generation Sequencing technique, ancient DNA (aDNA) molecules extracted from nail were utilized to perform the complete mitochondrial DNA genome sequence. Phylogenetic analyses showed that this genome, which was aligned with a number of historical and extant complete wolf and dog mtDNAs sampled worldwide, was closely related to an Italian wolf mtDNA genome. These mtDNA findings suggest that wolves probably colonized Sicily from southern Italy toward the end of the last Pleistocene glacial maximum, when the Strait of Messina was almost totally dry. Furthermore, a conservative restoration of this peculiar natural specimen (taxidermal preparation) has been performed, basing on: recovery of unglued or unstitched parts; revitalization of the colors of the mucous membranes; restoration of deformed or broken parts of the body, washing of the coat and polishing of the fur; specific treatments against woodworms, moths, and mites; brushing and repositioning of the hair; cleaning, restoration, or creation of adequate supports or display case. The conservative intervention that should be periodically performed on each taxidermized specimen allows the protection against biodeterioration or re-infestation by parasites over time

Genetic tools discriminate strains of Leishmania infantum isolated from humans and dogs in Sicily, Italy.

BackgroundLeishmaniasis is one of the most important vector-borne diseases and it represents a serious world health problem affecting millions of people. High levels of Leishmania infections, affecting both humans and animals, are recognized among Italian regions. Among these, Sicily has one of the highest prevalence of Leishmania infection.Methodology/principal findingsSeventy-eight Leishmania strains isolated from human and animal samples across Sicily, were analyzed for the polymorphic k26-gene and genotypes were assigned according to the size of the PCR products. A multilocus microsatellite typing (MLMT) approach based on the analysis of 11 independent loci was used to investigate populations structure and genetic diversity of the isolated strains. Six L. infantum reference strains were included in the analysis for comparison. Bayesian clustering analysis of microsatellite data showed that all the isolated strains clustered in two genetically distinct populations, corresponding to human and canine isolates respectively. A further subdivision was observed between the two main groups, giving a good correlation between human strains and their geographic origin, conversely canine population showed a great genetic variability diffused in the territory.Conclusions/significanceAmong the 78 Leishmania isolates, K26 analysis detected 71 samples (91%) as MON-1 zymodeme, confirming it as the predominant strain in Mediterranean area and 7 human samples (9%) as non-MON-1. MLMT gives important insights into the epidemiology of leishmaniases and allows characterization of different strains to a higher resolution than possible with zymodeme typing. Two main populations presented a strong correlation respect to the different hosts, exhibiting a co-circulation of two distinct populations of L. infantum. The population found in infected humans exhibited a correlation with geographic origin. These clusters could represent a geographically restricted population of strains with the same or related genotypes. This study can contribute to an understanding of Leishmania epidemiology, including the spread of reservoirs and sand fly vectors in the different foci of infection, characterizing parasites within the different hosts