Short communication: phenotypic and genetic diversity of wild Lactococcus lactis isolated from traditional Pecorino cheeses of Tuscany.

Wild Lactococcus lactis isolates from traditional Pecorino cheeses in 4 regions of Tuscany were isolated and characterized to evaluate the diversity of autochthonous lactococci. Sixty strains of Lactococcus were clustered by the results of carbohydrate utilization and diagnostic enzyme activity. Twenty-one unique strains were then chosen for characterization of salt and temperature tolerance, as well as acidification and proteolytic activity in milk. Genetic analysis of these strains was performed via 16S ribosomal DNA sequencing and multilocus sequence typing (MLST) to elucidate diversity relative to their location of origin. Phylogenetic analysis showed distinct clustering by region within organism subspecies, and phenotypic properties demonstrated concomitant trends. Multilocus sequence typing thus allowed for the regional distinction of isolates separate from those of previous works, supporting the concept that distinctive regional qualities of cheeses are strongly influenced by microbial ecology.

Antibiotic resistance of Aeromonas ssp. strains isolated from Sparus aurata reared in Italian mariculture farms.

Selective pressure in the aquatic environment of intensive fish farms leads to acquired antibiotic resistance. This study used the broth microdilution method to measure minimum inhibitory concentrations (MICs) of 15 antibiotics against 104 Aeromonas spp. strains randomly selected among bacteria isolated from Sparus aurata reared in six Italian mariculture farms. The antimicrobial agents chosen were representative of those primarily used in aquaculture and human therapy and included oxolinic acid (OXA), ampicillin (AM), amoxicillin (AMX), cephalothin (CF), cloramphenicol (CL), erythromycin (E), florfenicol (FF), flumequine (FM), gentamicin (GM), kanamycin (K), oxytetracycline (OT), streptomycin (S), sulfadiazine (SZ), tetracycline (TE) and trimethoprim (TMP). The most prevalent species selected from positive samples was Aeromonas media (15 strains). The bacterial strains showed high resistance to SZ, AMX, AM, E, CF, S and TMP antibiotics. Conversely, TE and CL showed MIC90 values lower than breakpoints for susceptibility and many isolates were susceptible to OXA, GM, FF, FM, K and OT antibiotics. Almost all Aeromonas spp. strains showed multiple antibiotic resistance. Epidemiological cut-off values (ECVs) for Aeromonas spp. were based on the MIC distributions obtained. The results showed a high frequency of Aeromonas spp. contamination in Sparus aurata reared on the Italian coast and an elevated biodiversity in isolated bacterial strains. Aeromonas isolates comprise potentially pathogenic species for humans, often resistant to several antibiotics and able to transfer the genes responsible for antibiotic resistance to microorganisms pathogenic for humans throughout the food chain. The few ECV studies available on many antibiotics against Aeromonas spp. strains isolated from the aquaculture environment highlight the need for further research in this area, while regular monitoring programmes should be stepped up to check for antibiotic resistance.

Experimental infection of dogs with Borrelia burgdorferi.

Four beagle dogs were inoculated subcutaneously with the BITs1 Italian strain of Borrelia burgdorferi. Only one dog became infected and B burgdorferi was isolated from its blood and urine three and four weeks after infection. B burgdorferi antibodies were detected by immunofluorescence from four to 11 weeks after infection. An uninoculated dog kept in the same run as the infected dog, developed a positive serological response, but none of the five dogs showed clinical signs.

Leptospira interrogans in the genital tract of sheep. Research on ewes and rams experimentally infected with serovar hardjo (hardjobovis).

To verify if Leptospira hardjo can colonize the male and female genital organs of sheep, 9 animals (6 non pregnant ewes and 3 mature rams) were infected with a strain of L. hardjobovis recently recovered from the kidneys of a seropositive ewe. Postinfection controls (bacteriologic, serologic, immunohistochemistry and electron microscopy) failed to disclose the presence of leptospires in the uterus and oviducts, testicles, epididymis, prostate and bulbourethral glands of animals used for the experiment and slaughtered from 37 to 242 postinfection days. All animals showed a renal localization of L. hardjobovis lasting for the entire period of the study (over 8 months). These results emphasize the important role of sheep as maintenance hosts of the serovar.

Epididymitis by Brucella ovis: experimental infection in virgin ram lambs.

Ten sexually immature rams were experimentally infected with Brucella ovis, to verify the antibody kinetics and its localization in urinary and genital tracts. All animals became positive to the complement fixation test from the 2nd post infection (p.i.) week and reached the maximum titre (1:256) on the 4th p.i. week. Bacteriemia was demonstrated on 3rd, 4th and 5th p.i. weeks. Two animals, respectively slaughtered 11 and 13 weeks after the infection, showed macroscopic and microscopic genital lesions and the etiologic agent was cultured from their urine and genital organs.

Leptospira interrogans serovar hardjo in the kidneys and genital tracts of naturally infected sheep.

A bacteriological study was carried out to identify possible renal and/or genital carriers of Leptospira interrogans serovar hardjo. L. hardjo was found at slaughter in the kidneys of three seropositive ewes, but not in uterus or salpinges of these animals.

Raw milk for sale in Pisa province: biosecurity of dairy farms and hygienic evaluation of milk.

Selling raw milk by automatic dispenser on the farm is a good way to increase income. The aim of the present research is to evaluate both the biosecurity of dairy farms in the Pisa province and the hygienic quality of raw milk. Two farms, chosen because of previously analyzed results carried out on animals and milk, were monitored for 9 months according to the regional law DGR 381/2007. The results obtained showed that these farms presented good cattle health status. The raw milk tested was of a satisfactory hygienic quality, with great variability among milk samples in certain parameters, such as somatic cell count. This study confirmed the importance of consistent, ongoing control of safety conditions on dairy farms.