Epidemiological surveillance for <i>Trichinella britovi</i> infection in free-ranging pigs of Sardinia

The aim of the present work was to investigate on Trichinella sp. infection in free-ranging pigs of the Orgosolo municipality

Caratteri fenotipici/genotipici per la produzione di Alginato negli isolati clinici di Pseudomonas aeruginosa presenti negli allevamenti ovini

Introduzione - In ambito sanitario il genere Pseudomonas è temuto per la sua resistenza agli antibiotici e agli antimicrobici, resistenza dovuta alla capacità di formare biofilm protettivo. P. aeruginosa è il patogeno più comune isolato nelle infezioni nosocomiali e in campo veterinario è di notevole importanza in bovini, ovini e caprini, dove può causare mastiti ambientali; è un piccolo bacillo Gram negativo caratterizzato da una capsula spessa ricoperta di alginato. L’alginato è il componente principale della matrice extracellulare del biofilm; ha una funzione protettiva in un ambiente relativamente ostile, in cui i batteri sono continuamente sottoposti a stress ossidativo e attaccati dal sistema immunitario. P. aeruginosa presenta un fenotipo non mucoide sensibile agli antibiotici e un fenotipo mucoide resistente agli antimicrobici. In alcuni casi, la conversione del fenotipo non mucoide nel fenotipo mucoide è causata da mutazioni che si verificano in due distinti loci cromosomici denominati MUC. L’inattivazione in vitro di mucA in P. aeruginosa POA1 (non mucoso) produce i ceppi Alg+; questo sembra indicare, quindi, che mucA agisce come un regolatore negativo della produzione di alginato perché può legare e sequestrare il fattore 22 attraverso il dominio citoplasmatico N-terminale. L’obiettivo del nostro lavoro è un’analisi dell’influenza delle mutazioni del gene mucA nei ceppi mucoidi di P. aeruginosa

Pseudomonas infections in surgery practice needs for innovative antimicrobial procedures?

Pseudomonas genus is known and studied from many years and it’s currently widely to the attention in microbiological and in medical clinical area for severe infections occur in patients who are already illness or who have a weak immune system, such as long term hospitalized subject. This bacteria group, normally is isolated in the environment, such as in soil, water, plants and in nosocomial area as well as in veterinary field such as intensive no controlled farming [1]. Pseudomonas spp. represents a Gram negative opportunistic pathogens bacteria group with a great, outstanding adaptability in the different environmental conditions. In fact, recentstudies by using the comparative genomics on different strains of P. aeruginosa isolated in human and veterinary field have identified some key genes involved in the shift environment-status vs. pathogen-status of these bacteria

High prevalence of co-infection with multiple Torque teno sus virus species in Italian pig herds.

Torque teno viruses (TTVs) are a large group of vertebrate-infecting small viruses with circular single-stranded DNA, classified in the Anelloviridae family. In swine, two genetically distinct species, Torque teno sus virus 1a (TTSuV1a) and 1b (TTSuV1b) are currently grouped into the genus Iotatorquevirus. More recently, a novel Torque teno sus virus species, named Torque teno sus virus k2b (TTSuVk2b), has been included with Torque teno sus virus k2a (TTSuVk2a) into the genus Kappatorquevirus. In the present study, TTSuV1 (TTSuV1a and TTSuV1b), TTSuVk2a and TTSuVk2b prevalence was evaluated in 721 serum samples of healthy pigs from Sardinian farms, insular Italy. This is the largest study to date on the presence of TTSuV in healthy pigs in Italy. The global prevalence of infection was 83.2% (600/721), being 62.3% (449/721), 60.6% (437/721), and 11.5% (83/721) the prevalence of TTSuV1, TTSuVk2a and TTSuVk2b, respectively. The rate of co-infection with two and/or three species was also calculated, and data show that co-infections were significantly more frequent than infections with single species, and that TTSuV1+TTSuVk2a double infection was the prevalent combination (35.4%). Quantitative results obtained using species-specific real time-qPCR evidenced the highest mean levels of viremia in the TTSuV1 subgroup, and the lowest in the TTSuVk2b subgroup. Interestingly, multiple infections with distinct TTSuV species seemed to significantly affect the DNA load and specifically, data highlighted that double infection with TTSuVk2a increased the viral titers of TTSuV1, likewise the co-infection with TTSuVk2b increased the titers of TTSuVk2a

Use of PCR-restriction fragment length polymorphism analysis for identification of yeast species isolated from bovine intramammary infection

This study reports a rapid PCR-based technique using a one-enzyme RFLP for discrimination of yeasts isolated from bovine clinical and subclinical mastitis milk samples. We analyzed a total of 1,486 milk samples collected over 1 yr in south Sardinia and northern Italy, and 142 yeast strains were preliminarily grouped based on their cultural morphology and physiological characteristics. Assimilation tests were conducted using the identification kit API ID 32C and APILAB Plus software (bioMerieux, Marcy l'Etoile, France). For PCR-RFLP analysis, the 18S-ITS1-5.8S ribosomal(r) DNA region was amplified and then digested with HaeIII, and dendrogram analysis of RFLP fragments was carried out. Furthermore, within each of the groups identified by the API or PCR-RFLP methods, the identification of isolates was confirmed by sequencing of the D1/D2 region using an ABI,Prism 310 automatic sequencer (Applied Biosystems, Foster City, CA). The combined phenotypic and molecular approach enabled the identification of 17 yeast species belonging to the genera Candida (47.9%), Cryptococcus (21.1%), Trichosporon (19.7%), Geotrichum (7.1%), and Rhodotorula (4.2%). All Candida species were correctly identified by the API test and their identification confirmed by sequencing. All strains identified with the API system as Geotrichum candidum, Cryptococcus uniguttulatus, and Rhodotorula glutinis also produced characteristic restriction patterns and were confirmed as Galactomyces geotrichum (a teleomorph of G. candidum), Filobasidium uniguttulatum (teleomorph of Crypt. uniguttulatus), and R. glutinis, respectively, by D1/D2 rDNA sequencing. With regard to the genus Trichosporon, preliminary identification by API was problematic, whereas the RFLP technique used in this study gave characteristic restriction profiles for each species. Moreover, sequencing of the D1/D2 region allowed not only successful identification of Trichosporon gracile where API could not, but also correct identification of misidentified isolates. In conclusion, the 18S-ITS1-5.8S region appears to be useful in detecting genetic variability among yeast species, which is valuable for taxonomic purposes and for species identification. We have established an RFLP database for yeast species identified in milk samples using the software GelCompar II and the RFLP database constitutes an initial method for veterinary yeast identification

LABORATORY DIAGNOSIS OF LEISHMANIA INFANTUM INFECTION

INTRODUZIONE La leishmaniosi è una zoonosi parassitaria causata da protozoi flagellati del genere Leishmania. In Italia la sola specie presente è L. infantum appartenente al complex donovani, i cui ceppi viscerotropi sono responsabili della leishmaniosi viscerale umana e della leishmaniosi canina, mentre i ceppi dermotropi sono responsabili della leishmaniosi cutanea nell'uomo. Nel nostro paese i vettori accertati appartengono al genere Phlebotomus e il cane rappresenta l'unico serbatoio di rilevanza epidemiologica. Le forme flagellate del parassita si moltiplicano nell’intestino dell’insetto vettore e la femmina del flebotomo può inocularle nella sede intradermica dell’ospite durante il pasto di sangue. Nell’ uomo la sintomatologia e i segni clinici risultano multiformi e talvolta difficili da inquadrare, per cui la diagnosi di laboratorio tramite sistemi cito-istologici, sierologici o molecolari, può rappresentare un valido aiuto per il clinico. Lo scopo del lavoro è stato verificare un sistema molecolare per il rilevamento di L. infantum, basato sulla tecnologia della PCR real time

Antimicrobial susceptibility pattern to disinfectants in Pseudomonas aeruginosa strains isolated from dairy sheep breeds in Sardinia

SUMMARY Introduction - Pseudomonas aeruginosa is still one of the leading causes of morbidity in dairy animals and particularly associated with severe clinical forms of mastitis. Its outstanding capacity to evade the activity of antimicrobial prophylaxis makes this species one of the most feared pathogens. In this context the World Health Organization (WHO) has reported this pathogen in a catalogue of bacteria that pose the greatest threat to human health. Thus, an accurate disinfection procedure is needed to avoid the incidence of Pseudomonas- related diseases in veterinary as well as in human medicine. Aim - The aim of this study was to evaluate the antimicrobial resistance patterns to common disinfectants in P. aeruginosa strains isolated from ovine breeding farms in Italy. Materials and methods - In this study a set of 44 clinical P. aeruginosa strains isolated from 11 different sheep breeding farms with clinical cases of severe mastitis were analyzed and evaluated for susceptibility to 4 different disinfectants: Benzalkonium Chloride (BZC), Chlorhexidine (CHX), Sodium Hypochlorite (NaClO) and Hydrogen Peroxide (H2O2). The capacity of these disinfectants to inhibit P. aeruginosa was evaluated by using standard broth microdilution methods. Results and discussion - The results showed that CHX and NaClO were the most active against this bacterium, but a great number of multi-disinfectant resistant strains were observed altogether, especially if we consider BZC and H2O2, with 55% of drug resistant strains. Moreover, a correlation was evaluated between H2O2 MIC values and: (i) percentage of mucoid strains, (ii) increase in resistance patterns to other disinfectants (Pearson’s 2 was 0.029 for CHX). Conclusion - The work suggests the crucial role of disinfection in veterinary medicine and raises concern about the possibility of an inadequate prophylaxis through the use of Hydrogen Peroxide on breeding farms contaminated with Pseudomonas spp