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The assessment of red fox population density is considered relevant to the surveillance of zoonotic agents vectored by this species. However, density is difficult to estimate reliably, since the ecological plasticity and elusive behavior of this carnivore hinder classic methods of inference. In this study, red fox population density was estimated using a non-invasive molecular spatial capture-recapture (SCR) approach in two study areas: one in a known hotspot of the zoonotic cestode Echinococcus multilocularis, and another naïve to the parasite. Parasitological investigations on collected samples confirmed the presence of the parasite exclusively in the former area; the SCR results indicated a higher fox population density in the control area than in the hotspot, suggesting either that the relationship between fox density and parasite prevalence is not linear and/or the existence of other latent factors supporting the parasitic cycle in the known focus. In addition, fox spotlight count data for the two study areas were used to estimate the index of kilometric abundance (IKA). Although this method is cheaper and less time-consuming than SCR, IKA values were the highest in the areas with the lower molecular SCR density estimates, confirming that IKA should be regarded as a relative index only. [ABSTRACT FROM AUTHOR]

Methicillin-resistant Staphylococcus aureus (MRSA) is seldom reported in dairy cattle yet when it enters a herd it spreads among animals, causing sub-clinical intramammary infection and providing a long-term reservoir of exposure to farmworkers. Clinical samples from dairy herds can be used for scanning surveillance based on at-risk sampling. To estimate MRSA prevalence in dairy herds reporting mastitis (all mastitis) in north-east Italy we set up an observational study from March 2016 to October 2017 (19 months) by selecting 977 dairy herds among those submitting milk samples to the IZSVe laboratories (north-east Italy) for screening of mastitis agents. The size of herd sample was intended to provide a prevalence estimate supposing the highest uncertainty on true prevalence, 95% confidence and 7% precision; to avoid over-clustering the sample size was 3 times multiplied and herds were sampled only once over the study period. The herd sample enrolled in the study was distributed over the reference population of the study area according to single provinces and representing the average of the proportion of herds and dairy cows of the province. Milk from clinical samples was pooled and inoculated in cefoxitin and aztreonam selective medium. S. aureus colonies tested mecA-positive and resistant to oxacillin and/or cefoxitin were classified as MRSA, mecA-negative were further screened for mecC. MRSA was detected in pooled milk from clinical samples of three herds out of 977 selected. In two herds MRSA was characterized by spa typing and multilocus sequence typing (MLST), and in both MRSA ST398 spa type t034 was detected. Two dairy herds were further investigated by thorough within herd sampling and MRSA detected in single cows, environment and fomites. Given the low (0.3%) prevalence of MRSA contaminated dairy herds the screening of MRSA among mastitis agents should be performed by using high sensitive laboratory methods. [ABSTRACT FROM AUTHOR]

Close cohabitation between pet and their owners allows transmission of Escherichia coli from dogs to humans and vice versa. E. coli carrying extended-spectrum β-lactamases (ESBL-E.coli) pose a serious risk of therapy failure in humans and strict surveillance of any ESBL-E.coli reservoir is of importance for risk mitigation. Aim of this preliminary work was to describe the β-lactamases involved in ESBL-E.coli isolates from dogs and to verify the occurrence of mobile colistin resistance genes mcr-1 and mcr-2. In 2016-2017, for 16 months, 303 dogs were selected by systematic sampling among dog carcasses or dog faecal samples received for necropsy or parasitology screening at the IZSVE laboratories (north-east Italy). Presumptive ESBL-E.coli were isolated from faecal samples or intestinal content by selective media added with cefotaxime. One presumptive ESBL-E.coli isolate per dog was further investigated by antimicrobial susceptibility testing and molecular methods to confirm β-lactamase resistance and detect carbapenemases. Among selected dogs 15% (45/303) harbored ESBL-E.coli, while no carbapenem-resistant isolates were found. Isolates were screened for AmpC and ESBL by conventional PCR, and further in-depth identification of the bla genes was performed by sequencing. Overall 16/45 isolates harbored blaCTX-M1 group genes, 15/45 of which blaCTX-M15, 6/45 isolates harbored blaCMY-2, 4/45 blaTEM-1 and 2/45 blaSHV-12. In the remaining 17/45 isolates various associations of β-lactamases were found: the blaCTX-M15 and blaTEM-1 association was detected in 9/45 isolates, the blaTEM-1 and blaCMY-2 association in 5/45 isolates, the blaTEM-1 and blaSHV-12 association in 2/45 and the blaSHV-12 and blaCTX-M1 group was detected in 1/45 isolate. Four/45 ESBL-E.coli, 2 blaCTX-M15, two blaTEM-1 and blaCMY-2 and one blaCMY-2, belonged to B2 phylogenetic group, the most represented among human ESBL-E.coli isolates. Three isolates harbored mcr-1, two were associated to blaCTX-M15, belonging to phylogroups B1 and D, and one to blaTEM-1 belonging to phylogroup E. Our results show moderate variability of bla involved in ESBL-E.coli from dogs and confirm that in this pet animal, as in humans, TEM-type and SHV-type are outnumbered by the CTX-M ESBL class. The occurrence of mcr-1 in three ESBL-E.coli isolates is of concern, yet the phylogenetic group points to the animal reservoir origin. [ABSTRACT FROM AUTHOR]

Paratuberculosis (or Johne’s disease) is an infectious disease which affects mainly ruminants and it is caused by Mycobacterium avium subsp. paratuberculosis (MAP). During a culling program (years 2011–2015) aimed at controlling the red deer ( Cervus elaphus ) population in Stelvio National Park (Italian Alps), where paratuberculosis was already described in this species, 382 tissue samples from the Lombardy Region and 102 fecal specimens from the Autonomous Province of Bolzano were analyzed by PCR. Of these, 77 samples (20.16%) from the Lombardy area and 19 specimens (18.63%) from the Bolzano area resulted PCR positive. The cultural test was carried out on PCR positive samples (n = 96), enabling the isolation of 19 MAP field strains which were genotyped using MIRU-VNTR typing and Short Sequence repeats (SSRs). Our results suggest that all isolates share an identical VNTR profile corresponding to the INMV1 genotype. The only variation was on the locus SSR2, but the utility of this last locus has already been questioned because of its instability. Overall, these data suggest a common clonal origin and host adaptation during the diffusion of paratuberculosis in this population. Finally, this profile is the same as that which has already been described in the cattle population in Northern Italy, suggesting a possible inter-species disease transmission pattern from wildlife to domestic ruminants and vice versa . [ABSTRACT FROM AUTHOR]

Background: Paenibacillus larvae is the etiological agent of American foulbrood (AFB), a widespread and severe bacterial brood disease of honey bees. The genomic characterization of P. larvae strains by enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) is able to differentiate four genotypes (ERIC I, ERIC II, ERIC III, ERIC IV). The information on the presence of P. larvae ERIC genotypes worldwide is few., Objectives: We have characterized P. larvae strains isolated in Italy from AFB outbreaks to obtain information on ERIC genotypes and phenotypes of the strains circulating in the country., Methods: A total of 117 P. larvae isolates from 115 AFB outbreaks occurring in 2008-2012 were subjected to phenotypic and genetic characterization., Results: The genomic characterization allowed the identification of ERIC I and ERIC II genotypes. Examining the data of Northern and Central Italy separately it was noted that in Northern Italy most outbreaks were caused by the ERIC I genotype (78.6%), followed by the ERIC II genotype (18.6%) and by co-infections (ERIC I + ERIC II) (2.6%). In Central Italy, only outbreaks caused by the ERIC I genotype were observed. With regard to phenotypic characteristics all examined strains of ERIC II genotype fermented fructose while no strains of ERIC I genotype possessed this ability., Conclusion: Both P. larvae ERIC I and ERIC II genotypes were isolated from the AFB outbreaks, but ERIC II genotype was isolated only in Northern Italy. The fermentation of fructose seems to be a genotype-specific biochemical marker.