[Occupational brucellosis in the veterinary service of the Local Health Service in the Abruzzo Region (Italy)]. / Brucellosi professionale nel Servizio Veterinario di una ASL della Regione Abruzzo.

During the health surveillance of the employees in the veterinary service of a Local Sanitary Service, we came across a medical case of occupational brucellosis in a veterinarian, identified by seriological data (Wright' seriological diagnosis + brucellosis specific test with identification of IgM and IgG), in a lack of significant anamnesis, anamnestic negativity of dangerous behaviours. Two medical cases of the sole seriological positivity have been identified in a veterinarian and in a health vigililant. In our Local Health Service the medical cases stated across the population have been 6 (in the period from 2000 to August 2007). The epidemiological flow of the animal brucellosis in the regional area, and in particular in our territory, shows one low incidence of positive stock farms, low incidence of oxes, sheep and goats infected. In Italy the medical cases of brucellosis denounced and approved from the National Institute for work-related Injury Insurance have been 38 out of 41, in the period from 1996 to 2006. This zoonosis represents a problem of relief in the sector the breeding of the animals and the butchery of the meats in all countries; the occupational health physician, within the area of prevention, has to exercise a specific health surveillance of biological risk and take part in specific programs of information and formation, in particular on the use of individual protection devices and correct behavioural rules.

Multiplex mtDNA coding region SNP assays for molecular dissection of haplogroups U/K and J/T.

Mitochondrial DNA (mtDNA) U/K and J/T are sister haplogroups within the superhaplogroup R. They are both common in Europe, with a combined overall frequency similar to the one reported for H, the most common European haplogroup (40-50%). In this study, we selected 159 Italian subjects, already ascribed to U/K and J/T by RFLP typing, and assigned each mtDNA to specific clades/subclades by investigating at least one diagnostic coding region SNP. For each sister haplogroup, one multiplex PCR and one SNaPshot minisequencing reaction were set up targeting 16 U/K and 7 J/T coding region SNPs. Each mtDNA sample was clearly assigned to a specific subclade, which could be further subdivided into several minor sub-branches according to peculiar HVS I/II motifs. Such a molecular dissection of haplogroups U/K and J/T could be extremely useful to reduce the overall analysis time and labor intensive sequencing procedures in high volume forensic casework, for example when it is important to rapidly exclude samples in order to restrict the number of suspects.

Subtyping mtDNA haplogroup H by SNaPshot minisequencing and its application in forensic individual identification.

Sequence variation of the hypervariable segments (HVS) I/II of mitochondrial DNA (mtDNA) and the haplogroup affiliation were determined in a sample of 271 Italian subjects. This analysis showed that 42% of the individuals could be ascribed to H, the most frequent haplogroup in European Caucasian populations. This fraction was then screened for specific single nucleotide polymorphisms located in the coding region to identify H subclades H1-H15. We set up two multiplex polymerase chain reactions and specific SNaPshot assays to investigate the frequency distribution of these subgroups in our population sample and to examine their usefulness in discriminating among commonly shared HVS I/II sequences. This allowed the assignment of a large portion of the mtDNAs ( approximately 70%) to specific subhaplogroups, with H1 and H5 being the most represented. About two-thirds of the individuals sharing common HVS I/II sequences were subdivided and ascribed to specific H subhaplogroups with a significant reduction of the frequencies of the most common mtDNA haplotypes. Haplogroup H subtyping could thus be extremely useful in forensic identification when many samples have to be analysed and compared, avoiding excessive time-consuming and labor-intensive sequencing analysis.

Polymorphism of the mitochondrial DNA control region in Italians.

Mitochondrial DNA sequences of the hypervariable regions HVI and HVII were analysed in 83 Caucasians living in central Italy to expand the database for forensic identification purposes, and 75 different haplotypes resulting from 62 polymorphic positions in HVI and 44 in HVII were observed. The most frequent haplotype (263G, 309.1C, 315.1C) was shared by 7 individuals, 2 haplotypes were shared by 2 individuals, and 72 were unique. The genetic diversity was found to be 0.99 and the random match probability 1.9%. A condition of sequence heteroplasmy was found in only one case at nt 16311, whereas a length heteroplasmy was found in the homopolymeric stretch of cytosines 303-315. Our results indicate that in direct sequencing beyond the poly-cytosine stretch, the overlap is due to length heteroplasmy, whereas the blurred signal occurs when the stretch is composed of more than 10 cytosines.