Fecal shedding of thermophilic Campylobacter in a dairy herd producing raw milk for direct human consumption.

Factors affecting the thermophilic Campylobacter fecal shedding in Italian dairy farming conditions have been investigated in a 12-month longitudinal study performed in a dairy farm authorized to sell raw milk in Italy. Fifty animals were randomly selected from 140 adult and young animals, and fecal sampling was performed six times at two month intervals; additionally at each sampling, three trough water samples and two trough feed samples were collected for both adult and young animals. Samples were analyzed by real-time PCR and cultural examination. Overall 33 samples (9.7%) were positive for thermophilic Campylobacter by real-time PCR: 26 out of 280 (9.2%) fecal samples, six out of 36 water points (16.6%) and one of the 24 feed samples (4.2%). Campylobacter jejuni was isolated from 6 out of 280 samples; no other Campylobacter sp. was isolated. A higher but not significant positivity was observed in fecal samples of younger animals (11.33%% versus 6.92% of adult animals) and a higher and significant positivity was observed in water samples collected from the water troughs of young animals. A distinct temporal dynamic trend was observed during the study period for both cows and calves, with two prevalence peaks between November/December and between May/July. Several factors such as calving, housing practices, herd size, management practices forcing together a higher number of animals, variations in feeding or water source, which were previously reported as cause of temporal variation in different farming conditions, could be excluded as cause of the two seasonal peaks in this study. The factors affecting the seasonality of Campylobacter shedding in the dairy herds remain unclear and warrant further investigation. The results of the present study indicate that special attention should be paid to farm hygiene management in farms authorized to produce and sell raw milk with increased surveillance by the authorities in certain periods of the year

Chikungunya Virus in Aedes albopictus, Italy

Chikungunya Virus in Aedes albopictus, Ital

Antimicrobial resistance, biofilm synthesis and virulence genes in Salmonella isolated from pigs bred on intensive farms

Salmonella is the second cause of foodborne infection in humans in the USA and Europe. Pigs represent the second most important reservoir for the pathogen and the consumption of pork meat is a major risk factor for human salmonellosis. Here, we evaluated the virulence patterns of eleven Salmonella isolated from pigs (carcasses and faces) bred in intensive farms in the north of Italy. The two serotypes identified were S. Typhimurium and its monophasic variant 1,4,5,12:i:-. None of the isolates was an ESBL producer, as confirmed also by PCR. However, the presence of a multidrug resistant pattern was evident, with all the isolates being resistant to at least to five antimicrobial agents belonging to various classes. Moreover, six out of eleven isolates showed important resistance profiles, such as resistance against colistin and ciprofloxacin, with nine to twelve recorded resistances. The isolates were negative for the biofilm synthesis test, while four different virulotypes were characterized. All the isolates showed the presence of invA, hilA, stn, ssrA, sipC. One sample also harbored ssaR and spvC genes. One strain was positive for all the virulence genes tested and was resistant to 12 antimicrobial agents. The present study contributes new data to the surveillance program for antibiotic resistance. Furthermore, the presence of eleven highly virulent isolates poses concern for human health in relation to their diffusion in the environment

A human-neutral large carnivore? No patterns in the body mass of gray wolves across a gradient of anthropization

The gray wolf (Canis lupus) expanded its distribution in Europe over the last few decades. To better understand the extent to which wolves could re-occupy their historical range, it is important to test if anthropization can affect their fitness-related traits. After having accounted for ecologically relevant confounders, we assessed how anthropization influenced i) the growth of wolves during their first year of age (n = 53), ii) sexual dimorphism between male and female adult wolves (n = 121), in a sample of individuals that had been found dead in Italy between 1999 and 2021. Wolves in anthropized areas have a smaller overall variation in their body mass, during their first year of age. Because they already have slightly higher body weight at 3–5 months, possibly due to the availability of human-derived food sources. The difference in the body weight of adult females and males slightly increases with anthropization. However, this happens because of an increase in the body mass of males only, possibly due to sex-specific differences in dispersal and/or to “dispersal phenotypes”. Anthropization in Italy does not seem to have any clear, nor large, effect on the body mass of wolves. As body mass is in turn linked to important processes, like survival and reproduction, our findings indicates that wolves could potentially re-occupy most of their historical range in Europe, as anthropized landscapes do not seem to constrain such of an important life-history trait. Wolf management could therefore be needed across vast spatial scales and in anthropized areas prone to social conflicts

Quantitative risk assessment of human salmonellosis and listeriosis related to the consumption of raw milk in Italy.

Two quantitative risk assessment (RA) models were developed to describe the risk of salmonellosis and listeriosis linked to consumption of raw milk sold in vending machines in Italy. Exposure assessment considered the official microbiological records monitoring raw milk samples from vending machines performed by the regional veterinary authorities from 2008 to 2011, microbial growth during storage, destruction experiments, consumption frequency of raw milk, serving size, and consumption preference. Two separate RA models were developed: one for the consumption of boiled milk and the other for the consumption of raw milk. The RA models predicted no human listeriosis cases per year either in the best or worst storage conditions and with or without boiling raw milk, whereas the annual estimated cases of salmonellosis depend on the dose-response relationships used in the model, the milk storage conditions, and consumer behavior in relation to boiling raw milk or not. For example, the estimated salmonellosis cases ranged from no expected cases, assuming that the entire population boiled milk before consumption, to a maximum of 980,128 cases, assuming that the entire population drank raw milk without boiling, in the worst milk storage conditions, and with the lowest dose-response model. The findings of this study clearly show how consumer behavior could affect the probability and number of salmonellosis cases and in general, the risk of illness. Hence, the proposed RA models emphasize yet again that boiling milk before drinking is a simple yet effective tool to protect consumers against the risk of illness inherent in the consumption of raw milk. The models may also offer risk managers a useful tool to identify or implement appropriate measures to control the risk of acquiring foodborne pathogens. Quantification of the risks associated with raw milk consumption is necessary from a public health perspective

Men and wolves: Anthropogenic causes are an important driver of wolf mortality in human-dominated landscapes in Italy

Over the last 40 years the gray wolf (Canis lupus) re-colonized its historical range in Italy increasing human-predator interactions. However, temporal and spatial trends in wolf mortality, including direct and indirect persecution, were never summarized. This study aims to fill this gap by focusing on the situation of Tuscany and Emilia-Romagna regions, hosting a significant proportion of the Italian wolf population, by: (i) identifying the prevalent causes of wolf mortality, (ii) summarizing their temporal and spatial patterns and (iii) applying spatially-explicit Generalized Linear Models to predict wolf persecution. Between October 2005 and February 2021, 212 wolf carcasses were collected and subjected to necropsy, being involved in collisions with vehicles (n = 104), poisoned (n = 45), wounded with gunshot (n = 24) or blunt objects (n = 4) and being hanged (n = 2). The proportion of illegally killed wolves did not increase through time. Most persecution events occurred between October and February. None of our candidate models outperformed a null model and covariates such as the density of sheep farms, number of predations on livestock, or human density were never associated to the probability of having illegally killed wolves, at the municipal scale. Our findings show that conventional correlates of wolf persecution, combined with a supposedly high proportion of non-retrieved carcasses, fail to predict illegal wolf killings in areas where the species have become ubiquitous. The widespread spatial distribution of illegal killings indicates that persecution probably arises from multiple kinds of conflicts with humans, beyond those with husbandry. Wolf conservation in Italy should thus address cryptic wolf killings with multi-disciplinary approaches, such as shared national protocols, socioecological studies, the support of experts’ experience and effective sampling schemes for the detection of carcasses

A Probable Fatal Case of Oleander (Nerium oleander) Poisoning on a Cattle Farm: A New Method of Detection and Quantification of the Oleandrin Toxin in Rumen

Oleander (Nerium oleander) is an ornamental plant common in tropical and sub-tropical regions that is becoming increasingly widespread, even in temperate regions. Oleander poisoning may occur in animals and humans. The main active components contained in the plant are cardiac glycosides belonging to the class of cardenolides that are toxic to many species, from human to insects. This work describes a case of oleander poisoning that occurred on a small cattle farm and resulted in the fatality of all six resident animals. Furthermore, the investigation of the poisonous agent is described, with particular focus on the characterization of the oleandrin toxin that was recovered from the forage and rumen contents. The innovation of this study is the first description of the detection and quantification of the oleandrin toxin by liquid chromatography-high resolution mass spectrometry (LC-HRMS) in rumen

Evidence of Common Isolates of Streptococcus agalactiae in Bovines and Humans in Emilia Romagna Region (Northern Italy)

Streptococcus agalactiae (group B Streptococcus, GBS) is one of the most important agents of bovine mastitis and causes remarkable direct and indirect economic losses to the livestock sector. Moreover, this species can cause severe human diseases in susceptible individuals. To investigate the zoonotic potential of S. agalactiae, 203 sympatric isolates from both humans and cattle, isolated in the same time frame (2018) and in the same geographic area (Emilia Romagna region, Northern Italy), were characterized by molecular capsular typing (MCT), pilus island typing (PI), and multi-locus sequence typing (MLST). In addition, antibiotic-resistant phenotypes were investigated. The distribution of the allelic profiles obtained by combining the three genotyping methods (MCT-PI-MLST) resulted in 64 possible genotypes, with greater genetic variability among the human compared to the bovine isolates. Although the combined methods had a high discriminatory power (>96,2%), five genotypes were observed in both species (20,9% of the total isolates). Furthermore, some of these strains shared the same antibiotic resistance profiles. The finding of human and bovine isolates with common genotypes and antibiotic resistance profiles supports the hypothesis of interspecies transmission of S. agalactiae between bovines and humans

Environmental contamination and colonization in pig by methicillin-resistant Staphylococcus aureus (MRSA)

Le attività di ricerca condotte e riportate negli studi sperimentali costituiscono a parere del Dottorando un utile approfondimento sulle conoscenze dell’epidemiologia di Staphylococcus aureus meticillino resistente MRSA nelle fase di produzione primaria del suino. Dallo studio sulla contaminazione ambientale è emerso che le operazioni di lavaggio e disinfezione (C&D) applicati negli allevamenti suinicoli sono in grado di ridurre la contaminazione ambientale da MRSA ma non di eliminarla. D’altronde, il risultato atteso di tali interventi è la riduzione della carica infettante ambientale piuttosto che la sterilizzazione degli ambienti. Assai rilevante appare il fatto che l’efficacia delle pratiche di C&D non sia risultata omogenea nelle diverse fasi ma si sia dimostrata significativamente superiore nelle sale parto rispetto alle altre strutture di allevamento. Nell’allevamento suinicolo convenzionale italiano il rigore e l’intensità con cui sono realizzate e operazioni di C&D è sicuramente superiore nelle sale parto, pertanto, si può concludere che una rigorosa applicazione di tali operazioni potrebbe ridurre il challenge ambientale a cui sono esposti gli animali nei nostri allevamenti. Un altro risultato importante di questo primo studio è stato quello di evidenziare come il livello di contaminazione degli ambienti di allevamento non sia omogeneo in quanto, nelle fasi di ingrasso, il livello del challenge ambientale diminuisce rispetto alle prime fasi di accrescimento. Il motivo di questo andamento non appare chiaro anche se è possibile ipotizzare una relazione con la pressione selettiva esercitata dai trattamenti antibiotici che tende a diminuire in modo ragguardevole con l’avanzare dell’età degli animali. Lo studio longitudinale, nel quale la contaminazione degli ambienti è stata studiata parallelamente alla colonizzazione nasale degli animali, ha fornito ulteriori spunti di riflessione. In particolare, si è registrato un andamento sovrapponibile fra il tasso di portatori nasali ed il tasso di campioni ambientali positivi. Inoltre, è stato rilevato un drastico aumento del numero dei portatori nasali fra la fase sottoscrofa e la fine del periodo di svezzamento. Tale rapida diffusione della colonizzazione nasale è risultata coincidente con due fattori: 1) lo spostamento degli animali da un ambiente negativo per MRSA ad un reparto di svezzamento contaminato pur se sottoposto a C&D, 2) la somministrazione di medicazioni antibiotiche per via orale. Il numero di soggetti portatori a livello nasale si è mantenuto a livelli elevatissimi (fino al 100% dei testati) per tutto il periodo coincidente con questa pratica terapeutica e si è ridotto drasticamente (attorno al 20%) dopo l’interruzione delle medicazioni orali, nei reparti di ingrasso. Questi risultati permettono di formulare le conclusioni che l’ambiente contaminato possa fungere da punto di partenza per la diffusione della colonizzazione di un gruppo e che l’effetto della pressione selettiva dei trattamenti di massa sia tutt’altro che trascurabile. Oltre quanto già esposto circa dell’importanza dell’applicazione rigorosa delle operazioni di C&D, appare quindi evidente che il tasso dei soggetti portatori potrebbe essere contenuto se i trattamenti di massa fossero ridotti e limitati allo stretto necessario. Lo studio che ha coinvolto alcuni allevamenti e gli animali da essi provenienti in fase di macellazione ha permesso di confermare quanto evidenziato da Battisti et al (2010) circa una elevata positività in termini di colonizzazione nasale delle partite di suini macellati. Infatti, tutti i 5 allevamenti (di cui era già noto lo stato di positività per MRSA) hanno dato origine a partite colonizzate. Un risultato meno atteso è stato quello riguardante la percentuale di carcasse che sono risultate contaminate alla fine della catena di macellazione, da un minimo del 20% ad un massimo del 60%, a seconda delle partite. Da ciò appare evidente che la macellazione di partite colonizzate dà origine a carcasse contaminate. Le tipizzazioni biomolecolari sui ceppi isolati in quest’ultimo studio hanno confermato la notevole eterogeneità dei ceppi di MRSA diffusi nell’allevamento suinicolo italiano. Infatti, accanto al predominante ST 398, tipicamente associato al suino e ad altre specie di animali da reddito, è stata conferma la presenza di altri ST fra cui ST1, ritenuto fino a poco tempo fa strettamente adattato all’uomo. È inoltre emerso come taluni ceppi riscontrati nei suini macellati non siano fra quelli rinvenuti negli allevamenti di origine. L’importanza dell’igiene in tutte le fasi post allevamento (trasporto, permanenza nelle stalle di sosta, macellazione) è nota ed ampiamente documentata per altri importanti agenti batterici trasmissibili dal suino all’uomo, in particolare per Salmonella. I risultati di questo lavoro permettono di ipotizzare, parimenti, un ruolo di tali fasi sulla contaminazione finale da MRSA delle carcasse. Seppure ad oggi la trasmissione dal suino all’uomo tramite prodotti carnei non sia stata dimostrata, ulteriori approfondimenti sul destino di questa contaminazione nella filiera meriterebbe ulteriori approfondimenti

Shiga toxin-producing Escherichia coli in meat and vegetable products in Emilia Romagna Region, years 2012-2013

In 2012-2013 Emilia-Romagna Region introduced a monitoring plan for Shiga toxin-producing <em>Escherichia coli</em> (STEC) in foodstuff. Six hundred eighty-nine meat samples and 273 fruit and vegetable products were analyzed according to ISOTS13136. Pre-enriched samples were tested by multiplex real time PCR targeting the virulence genes <em>eae</em>, <em>stx1</em> and <em>stx2</em>. <em>Stx2</em> positive samples were investigated for the presence of serogroup O104 associated gene. O103, O111, O145, O157, O26 associated genes were tested on samples positive for <em>stx</em> in association with <em>eae</em> gene. Isolation of E. coli strains was attempted from samples positive for serogroup-associated genes. Thirtyfour meat products (4.9%) resulted positive for <em>stx1</em> and/or <em>stx2</em> genes and 46 (6.7%) for <em>stx1</em> and/or <em>stx2</em> genes in association with <em>eae</em> gene. Forty-five (6.5%) samples resulted positive at least at one serogroup. Serogroup O103, O104, O111, O145, O157 and O26 genes were detected respectively in 1.3, 0.3, 0.1, 3.9, 2.9 and 2.5% samples; 0.6% samples resulted positive for STEC isolation (2 <em>E. coli</em> O103 and 2 <em>E. coli</em> O157). It is worth noting that STEC virulence genes were detected at high frequency (19%) in fresh pork meat sausages. Four (1.5%) vegetable samples were positive for <em>stx1</em> and/or <em>stx2</em> genes and 1 (0.4%) for <em>stx1</em> and/or <em>stx2</em> genes in association with <em>eae</em> gene; none resulted positive for the tested serogroups. Only a low number of samples positive by molecular methods were confirmed by cultural isolation. It is therefore of the uttermost importance for appropriate risk management, to be fully aware of the meaning of the analytical result