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Simple Summary This paper describes an easy-to-build box for the noninvasive collection of feces from wild birds or small wild animals (up to 1 kg), including a plastic storage box, a plastic tray, and a vinyl-coated hardware cloth. This method could minimize potential contamination and allow for cross-study comparisons on gut microbiomes for wildlife medicine, conservation, ecology, and evolutionary biology. Abstract Gut microbial communities play important roles in host health, modulating development, nutrient acquisition, immune and metabolic regulation, behavior and diseases. Wildlife microbiome studies and host–microbe interaction and exploration might be an important goal for evolutionary biology, conservation, and ecology. Therefore, collection and sampling methods must be considered before choosing a microbiome-based research plan. Since the fecal microbial community reflects the true gut community better than that of cloacal swab samples and only few nondestructive methods have been described, we propose an easy-to-build box for a noninvasive fecal collection method. The main components of the collection box include a plastic storage box, a plastic tray, a vinyl-coated hardware cloth, and a 10% bleach solution. In the plastic box, the tray is positioned under the raised grate, where the bird is placed, to reduce the risk of contamination of the fecal samples. This procedure could simplify handling and processing phases in wild birds or other animals. It might represent a cheap and useful method for research studies, wildlife rescue center activities, veterinary practices, and conservation practitioners.

The scope of the present work is to illustrate the main idea and the realization process at the basis of Flying Outfit for Control of Unsafe Seagulls FOCUS project. The project has been realized by a consortium of companies in collaboration with the University of Naples "Federico II" and CeSMA (Centro Servizi Metrologici e Tecnologici Avanzati) center. The main objective of the research is the definition of a process able to provide a supply of services for the management of coexistence problems between human and unsafe seagulls by means of small remotely piloted aircraft. The main purpose is to overheat the seagull eggs in the nest by means of a microwave device so they are no longer fertile. This is considered as an acceptable way to reduce the risk of unsafe interaction among seagulls and humans. The paper describes the different steps needed for the realization of the mission and the small unmanned aircraft used for the monitoring, operating on the nest, and collecting of gull fecal samples. Finally, the software developed for image detection of seagulls nest is presented.

Antimicrobial resistance is a public health concern worldwide and it is largely attributed to the horizontal exchange of transferable genetic elements such as plasmids carrying integrons. Several studies have been conducted on livestock showing a correlation between the systemic use of antibiotics and the onset of resistant bacterial strains. In contrast, although companion birds are historically considered as an important reservoir for human health threats, little information on the antimicrobial resistance in these species is available in the literature. Therefore, this study was aimed at evaluating the antimicrobial resistance of Escherichia coli and Pseudomonasaeruginosa isolated from 755 companion birds. Cloacal samples were processed for E. coli and P. aeruginosa isolation and then all isolates were submitted to antimicrobial susceptibility testing. P. aeruginosa was isolated in 59/755 (7.8%) samples, whereas E. coli was isolated in 231/755 (30.7%) samples. Most strains showed multidrug resistance. This study highlights that companion birds may act as substantial reservoirs carrying antimicrobial resistance genes which could transfer directly or indirectly to humans and animals, and from a One Health perspective this risk should not be underestimated.

Sea turtles possess relevant characteristics to serve as sentinel species for monitoring the health of marine ecosystems, which is currently threatened. This study examined 35 loggerhead turtles from the western Mediterranean, focusing on the oral and cloacal prevalence of aerobic Gram-negative bacteria, their antibiotic resistance and the influence of several variables linked both to the animal and the environment (i.e. estimated life stage; area, season and cause of recovery; plastic ingestion). Conventional bacteriology methods led to the isolation of bacterial families commonly regarded as opportunistic pathogens (i.e. Aeromonadaceae; Enterobacteriaceae; Pseudomonadaceae; Shewanellaceae; Vibrionaceae), but pointing out sea turtles as carriers of potential zoonotic agents. The high rates of antibiotic resistance, here detected, raise important concerns on the dissemination of this phenomenon in marine environments. Moreover, several of the examined variables showed a significant influence on the prevalence of bacterial families, strengthening the role of sea turtles as mirrors of their ecosystems.

The sugar glider (Petaurus breviceps) is a small, arboreal, nocturnal, gliding mammalian possum belonging to the marsupial infraclass. Exotic marsupials, including sugar gliders, are becoming popular companion pets and, consequently, the risk of potential infections that can be transmitted to humans should be investigated. Data on the role of the sugar glider as a possible carrier of pathogenic and zoonotic bacteria are scarce and fragmentary. Therefore, this study is aimed at evaluating the prevalence of potentially zoonotic bacteria (Salmonella spp., Escherichia coli, Campylobacter spp., Pseudomonas spp., Klebsiella spp., Listeria monocytogenes and Yersinia enterocolitica) in 64 sugar gliders kept as pets in Italy. The highest prevalence of infection pertained to members of the family Enterobacteriaceae, in particular Citrobacter spp. (50%), Enterobacter spp. (28·1%) and Klebsiella pneumoniae (15·6%); Pseudomonas aeruginosa was isolated from 10 out of 64 samples (15·6%). All strains of Klebsiella pneumoniae exhibited some level of resistance to multiple antimicrobials (ampicillin, amoxicillin-clavulanic acid and doxycycline). SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study show that sugar gliders may act as carriers of potentially pathogenic agents for humans and other animal species, therefore caution should be exercised in the handling and contact with these animals.

PubMedID: 30953839 A total of 476 European isolates (310 Cryptococcus neoformans var. grubii, 150 C. neoformans var. neoformans, and 16 C. gattii species complex) from both clinical and environmental sources were analyzed by multi-locus sequence typing. Phylogenetic and population genetic analyses were performed. Sequence analysis identified 74 sequence types among C. neoformans var. neoformans (VNIV), 65 among C. neoformans var. grubii (56 VNI, 8 VNII, 1 VNB), and 5 among the C. gattii species complex (4 VGI and 1 VGIV) isolates. ST23 was the most frequent genotype (22%) among VNI isolates which were mostly grouped in a large clonal cluster including 50% of isolates. Among VNIV isolates, a predominant genotype was not identified. A high percentage of autochthonous STs were identified in both VNI (71%) and VNIV (96%) group of isolates. The 16 European C. gattii species complex isolates analyzed in the present study originated all from the environment and all belonged to a large cluster endemic in the Mediterranean area. Population genetic analysis confirmed that VNI group of isolates were characterized by low variability and clonal expansion while VNIV by a higher variability and a number of recombination events. However, when VNI and VNIV environmental isolates were compared, they showed a similar population structure with a high percentage of shared mutations and the absence of fixed mutations. Also linkage disequilibrium analysis reveals differences between clinical and environmental isolates showing a key role of PLB1 allele combinations in host infection as well as the key role of LAC1 allele combinations for survival of the fungus in the environment. The present study shows that genetic comparison of clinical and environmental isolates represents a first step to understand the genetic characteristics that cause the shift of some genotypes from a saprophytic to a parasitic life style. © 2019 Elsevier Inc. Fundação para a Ciência e a Tecnologia National and Kapodistrian University of Athens APP1031943 70/3/6915, 70/4/5905 POCI-01-0145-FEDER-006958, UID/AGR/04033/2019 We thank Dr. Kwon Chung and Prof. Dromer for contribution in the critical reading of the manuscript. The findings and conclusions in this manuscript are those of the authors and do not necessarily represent the official position of the Centers for Disease Control and Prevention. W Meyer and S Lockhart were supported by an NH&MRC grant # APP1031943 . AC Sampaio was supported by Fundação para a Ciência e Tecnologia and FEDER/COMPETE/POCI provided to CITAB under the projects POCI-01-0145-FEDER-006958 and UID/AGR/04033/2019. A Velegraki was supported by SARG K.A.70/4/5905 and 70/3/6915, National and Kapodistrian University of Athens and the Bodosakis Foundation , Athens, Greece.

Background New epidemiological data on bacterial and parasitic infections in 24 Italian wall lizards, namely Podarcis sicula (mainland population) and P. sicula klemmerii (insular population) in southern Italy were provided. To achieve this goal, samples were collected from individuals belonging to the two populations and analysed by microbiological and parasitological methods. Results A wide range of bacteria (e.g. Pantoea spp., Citrobacter spp., Morganella spp., Pseudomonas, Enterobacter spp., Staphylococcus spp. and Escherichia coli) and parasites (e.g. Ophionyssus natricis, coccidia, Dicrocoelidae) were detected in both P. sicula and P. sicula klemmerii individuals. Insular population presented similar bacterial and parasitic diversity to its mainland counterpart. Ampicillin was the antimicrobial with the highest resistance rate. Conclusion This study highlighted various bacteria and parasites, some of them potentially zoonotic. Further studies are needed to better understand the epidemiology and transmission routes of these pathogens along with their impact on the welfare and behaviour of Italian wall lizards.

Fundamental niche prediction of Cryptococcus neoformans and Cryptococcus gattii in Europe is an important tool to understand where these pathogenic yeasts have a high probability to survive in the environment and therefore to identify the areas with high risk of infection. In this study, occurrence data for C. neoformans and C. gattii were compared by MaxEnt software with several bioclimatic conditions as well as with soil characteristics and land use. The results showed that C. gattii distribution can be predicted with high probability along the Mediterranean coast. The analysis of variables showed that its distribution is limited by low temperatures during the coldest season, and by heavy precipitations in the driest season. C. neoformans var. grubii is able to colonize the same areas of C. gattii but is more tolerant to cold winter temperatures and summer precipitations. In contrast, the C. neoformans var. neoformans map was completely different. The best conditions for its survival were displayed in sub-continental areas and not along the Mediterranean coasts. In conclusion, we produced for the first time detailed prediction maps of the species and varieties of the C. neoformans and C. gattii species complex in Europe and Mediterranean area.

The aim of this study was to propose the use of a new rapid and user-friendly diagnostic tool for the detection of Macrorhabdus ornithogaster infection in birds. The current report focuses on the diagnostic feasibility of different methods, with particular emphasis on the application of the mini-Flotac technique for the diagnosis of M . ornithogaster infection. The mini-Flotac method is particularly tailored for epidemiological monitoring and surveillance, where large numbers of fecal samples must be rapidly, yet reliably, examined. Gram staining, as the standard method, was used to validate the reliability of the mini-Flotac method. This tool has not yet been used in avian species or in the diagnosis of yeast infections. In our study, M. ornithogaster showed excellent performance in a flotation assay, which had not been demonstrated previously. Our results suggest that the mini-Flotac method is a valid, sensitive, and potentially low-cost alternative technique for use in the diagnosis of this yeast infection in birds.

Avian species are considered as the main reservoir of Campylobacter spp. However, few data are available on the presence of this microorganism in pet birds. This study was therefore performed to determine the prevalence of Campylobacter spp. in pet birds bred in southern Italy. Faecal samples were collected from 88 cages housing different species of pet birds and examined by bacteriological culture and polymerase chain reaction. A total of 13.6% of the cage samples were positive for Campylobacter coli. Other Campylobacter spp. were not found. The study shows that C. coli can be isolated from the cages of apparently healthy pet birds, which should therefore be considered as potential carriers of C. coli and a possible source of infection for humans and companion animals.

A recent review on the prevalence of zoonotic bacteria in organic and conventional farms of cattle, pigs and poultry in the United States and Europe showed no differences between the presence of zoonotic agents and the type of the farm management [1, 2]. This study was undertaken both to evaluate the presence of zoonotic infectious agents not included in the National Control Planes for the animals presents in Farms open to public (FoP) and, consequently, to emphasize the importance of the role of the veterinary doctor with training in interspecies relationship in this context. From September 2014 to July 2016 thirty-two FoP of Campania Region have been chosen basing on the size farm population (at least 25 subjects from three different species) and then sampled. In each farm, in respect of animal welfare, rectal and cloacal swabs were collected in order to isolate Salmonella spp., Shigatoxins producing Escherichia coli (STEC) and thermotolerant Campylobacter by cultural and molecular methods. A total of 800 animal divided in dogs (34), cats (45), horses (82), donkeys (91), cattle (127), sheep (62), goats (73), pigs (41), leporidae (173), henhouse (32) and "other samples" such as eggs and other animal species (40) were sampled. In our study, all farms were positive to STEC and/or thermotolerant Campylobacter, conversely they were all time negative for Salmonella spp. In particular, a total of 124/800 (15.5%) strains of STEC were isolated primarily from poultry and donkeys followed by other animal species. Finally, 127/800 (15.87%) strains of thermotolerant Campylobacter were isolated primarily from poultry followed by other animal species [48/127 (37.79 %) C. jejuni (mainly from donkeys) and 79/127 (62.20%) C. coli (mainly from poultry)]. All positive animals to STEC and thermotolerant Campylobacter were clinically healthy. The healthcare monitoring carried out in FoP of Campania region, where the animals are raised primarily for zooanthropological purposes, has provided interesting data increasing the international scientific literature in the field of zoonoses. In this study some of the major gastrointestinal zoonoses linked to animals in the FoP were considered as well as the role of the veterinary doctor in the prevention and control of zoonotic diseases and in health education of the operator in the FoP. In fact, both the operators and visitors (families, school groups, children, elderly etc.) often underestimates the potential health risks caused by contact with FoP animals.

The incidence of campylobacteriosis have increased in both developed and developing countries over the last 10 years [1]. Several avian species are considered the main reservoirs of Campylobacter spp. [2,3]. Nevertheless, current scientific knowledge on the presence of Campylobacter spp. in pet birds is scarce. To address this lack of information, the present study was undertaken with the aim to evaluate the presence of these microorganisms in pet birds bred in southern Italy. To achieve this goal, 14 bird farms located in the Campania region (southern Italy) were visited. In each farm, bird farm population ranged between 20 to 100 animals belonging to the families of Estrildidae, Fringillidae and Psittacidae, from which 33, 28, and 27 pooled faecal samples were collected respectively. Specifically, eighty-eight cages housing a total of 225 captive birds were examined. The cage was used as an epidemiological unit, and each cage housed from 1 to 5 birds. Each sample was stored in Amies Charcoal Transport Medium at 4°C, transported to the laboratory, and analyzed within 2 h of collection by cultural and molecular methods. A total of 12/88 (13.6%) cages were positive for Campylobacter spp. which was identified as C. coli. In particular, 7/33 (21.2%) cages came from Estrildidae and 5/27 (18.5%) cages came from Psittacidae family. The twentyeight cages coming from Fringillidae family were consistently negative. our results demonstrate that C. coli may be found in the intestines of apparently healthy pet birds which could be considered as a further potential carrier of C. coli for humans and other companion animals. The adoption of good hygiene practices when handling pet birds should be, therefore, promoted.

To evaluate the presence of Campylobacter spp. and related cdt genes, cloacal swabs were collected from 70 common teals (Anas crecca) and analyzed by culture methods and polymerase chain reaction. In addition, C. jejuni were examined also for the presence of wlaN gene. This is believed to be the first report of Campylobacter spp. in common teal and our results confirm the very common occurrence of C. jejuni (n=40) and C. coli (n=13) in waterfowls. Furthermore, the cdt genes were frequently present in both C. jejuni and C. coli isolated. Moreover, seven C. jejuni isolates carried also the wlaN gene which is presumably involved in the expression of ganglioside mimics in Guillain-Barré syndrome.

The study was aimed at evaluating the prevalence of enteropathogenic bacteria (i.e. Campylobacter spp., shigatoxin-producing Escherichia coli, Salmonella spp.) in common quail (Coturnix coturnix). To achieve this goal, 70 common quails were collected during the hunting season in the Campania region (southern Italy). From each bird, cloacal swab samples were collected and subjected to culture methods, polymerase chain reaction and serotyping. The results of the present study showed a prevalence of 21.4% and 5.7% for Campylobacter spp. and shigatoxin-producing E. coli, respectively. In contrast, no Salmonella spp. was isolated. These findings show that common quail, as migratory birds, may constitute an environmental carrier of these pathogens representing a source of infection for other birds, livestock and humans.

Recently, several studies have demonstrated that pigeon is an important reservoir of Shiga toxin-producing E. coli O157:H7. The aim of this study was to evaluate the presence of this pathogen in urban pigeons in the city of Napoli. The sampling was carried out during the period November 2005/July 2006. The city was subdivided in 56 quadrants by Geographical Information System. Each quadrant was analysed three times. From each quadrant, 3 pigeons were analysed by cloacal swabs. A total of 504 cloacal swabs was obtained. We isolated four E. coli O157:H7 strains. By multiplex PCR, all strains carried eae and stx2 genes, whereas only one strain carried the stx1 gene. 2/4 isolated strains carried hly gene which is considered a hallmark of human pathogenic strains. Our results indicate that pigeon faces are a source of E. coli O157:H7 for birds, mammals and humans.

We report a case of polyostotic chondroblastic osteosarcoma in a kestrel ( Falco tinnunculus ) admitted to the Wildlife Rehabilitation and Rescue Center (Naples, Italy). A consolidated fracture of the left tibiotarsus bone and a deviation of the limb were evident. After radiographic, cytologic, and histopathologic examinations, a diagnosis of polyostotic chondroblastic osteosarcoma was made. To our knowledge, this is the first report on polyostotic chondroblastic osteosarcoma in a kestrel.

The gut microbiota plays a crucial role in the bi-directional gut–brain axis, a communication that integrates the gut and central nervous system (CNS) activities. Animal studies reveal that gut bacteria influence behaviour, Brain-Derived Neurotrophic Factor (BDNF) levels and serotonin metabolism. In the present study, we report for the first time an analysis of the microbiota–gut–brain axis in zebrafish (Danio rerio). After 28 days of dietary administration with the probiotic Lactobacillus rhamnosus IMC 501, we found differences in shoaling behaviour, brain expression levels of bdnf and of genes involved in serotonin signalling/metabolism between control and treated zebrafish group. In addition, in microbiota we found a significant increase of Firmicutes and a trending reduction of Proteobacteria. This study demonstrates that selected microbes can be used to modulate endogenous neuroactive molecules in zebrafish.

In order to elucidate the distribution of Cryptococcus neoformans and C. gattii in the Mediterranean basin, an extensive environmental survey was carried out during 2012-2015. A total of 302 sites located in 12 countries were sampled, 6436 samples from 3765 trees were collected and 5% of trees were found to be colonized by cryptococcal yeasts. Cryptococcus neoformans was isolated from 177 trees and C. gattii from 13. Cryptococcus neoformans colonized 27% of Ceratonia, 10% of Olea, Platanus and Prunus trees and a lower percentage of other tree genera. The 13 C. gattii isolates were collected from five Eucalyptus, four Ceratonia, two Pinus and two Olea trees. Cryptococcus neoformans was distributed all around the Mediterranean basin, whereas C. gattii was isolated in Greece, Southern Italy and Spain, in agreement with previous findings from both clinical and environmental sources. Among C. neoformans isolates, VNI was the prevalent molecular type but VNII, VNIV and VNIII hybrid strains were also isolated. With the exception of a single VGIV isolate, all C. gattii isolates were VGI. The results confirmed the presence of both Cryptococcus species in the Mediterranean environment, and showed that both carob and olive trees represent an important niche for these yeasts. United States Department of Health & Human Services National Institutes of Health (NIH) - USA - ZIAAI000744 NIH National Institute of Allergy & Infectious Diseases (NIAID) CITAB - UID/AGR/04033/2013 National Health and Medical Research Council (NHMRC) of Australia - APP1031943 Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES) - 9313133

A total of 169 faecal samples were collected from migrating birds, belonging to the Order of Passeriformes, in Campania region in order to isolate Campylobacter spp. Campylobacter spp. were isolated from 39 of the 169 birds examined (23.1%). Among these 36 were identified as C. jejuni and the remaining strains were identified as Campylobacter coli. Given the high isolation rates wild birds could be considered natural reservoir of infection.

A total of 1,000 rectal samples were collected from rabbits coming from 25 rabbit farms in southern Italy. All samples were processed for isolation of Salmonella spp. by standard culture method based on the ISO 6579:2002 method. Salmonella spp. was isolated from 1/25 rabbit farms analyzed. In particular, four out of 1,000 rectal swab samples, taken from young rabbits, were serotyped as S. Typhimurium and phage typed as S. Typhimurium DT104. All the isolates were resistant to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline (ACSSuT pentaresistance type). The findings of the present study suggest the rabbit as potential carrier of S. Typhimurium DT104.

Aim: To estimate the prevalence of thermotolerant Campylobacter spp. in commercially reared partridges (Perdix perdix) in southern Italy. Methods and Results: Cloacal swabs of partridges (n = 240), equally distributed between male and female birds, from a game bird farm located in the Southern Italy were examined for the prevalence of thermotolerant Campylobacter spp. The samples were processed in order to detect thermotolerant Campylobacter spp. by culture methods. The positive samples were then confirmed by multiplex polymerase chain reaction. Thermotolerant Campylobacter spp. were isolated from 118 (49·2%) of the 240 cloacal swabs examined. As proved by PCR, 100% of the strains were identified as Campylobacter coli (118/118), and 15 (12·7%) out of the 118 positive samples were also positive for Campylobacter jejuni. In contrast, Campylobacter lari was not identified. Adult partridges showed a significantly higher prevalence (P

This study was conducted with the aim to evaluate the prevalence of thermotolerant Campylobacter spp. in pheasant (Phasianus colchicus) in Southern Italy. To achieve this goal, 60 cloacal swabs were collected. Campylobacter spp. were isolated from 52 out of 60 cloacal swabs tested. As proved by PCR, 100% of the strains were identified as C. coli (52/52), and 10 out of the 52 (19.2%) positive samples were also positive to C. jejuni. The pheasant, can be considered, at least theoretically, as potential Campylobacter carriers.

This study was aimed at evaluating the prevalence of Campylobacter spp., Escherichia coli O157, Salmonella spp., and related virulence factors (the cdt, stx, and eae genes) in urban pigeons of the coastal area of the Campania region (southern Italy). To achieve this goal, cloacal swab samples from a total of 1800 urban pigeons were collected and subjected to culture methods, PCR, and serotyping. The results of the present study showed a prevalence of 48.3% (870/1800), 7.8% (141/1800), and 0.9% (16/1800), for C. jejuni, E. coli O157, and S. Typhimurium, respectively. All C. jejuni isolates (870/870) carried cdt genes, whereas all E. coli O157 isolates carried stx genes, and 14.9% (21/141) carried the eae gene. These findings clearly show that urban pigeons in the coastal area of the Campania region may constitute an environmental reservoir of these pathogens, thus representing a source of infection for other birds, livestock, and humans.

SUMMARY A total of 170 birds of prey admitted to two Wildlife Rescue and Rehabilitation Centers of Italy were examined. Birds were divided by diurnal (n = 15) and nocturnal (n = 7) species, sampled by cloacal swabs, and examined for Campylobacter spp. by cultural and molecular methods. Campylobacter spp. were isolated in 43 out of the 170 (25.3%) birds of prey examined. Among these, 43/43 (100%) were identified as Campylobacter jejuni and 10/43 (23.3%) were identified as Campylobacter coli recovered from mixed infections. Diurnal birds of prey showed a significantly higher prevalence value (P = 0.0006) for Campylobacter spp. than did nocturnal birds of prey.

Forty common swifts (Apus apus), synanthropic birds living in an urban environment closely with humans and other animals, were hospitalized in the public veterinary hospital of the Regional Reference Center of Urban Veterinary Hygiene located in Naples, Campania Region, Italy. Each bird was sampled for bacteriological analyses. Out of 40 common swifts examined, eight were found positive for Salmonella enterica serovar Infantis although no sign of salmonellosis (e.g. diarrhoea) was shown. This is believed to be the first report of Salmonella spp. infection in common swifts. Our results suggest this avian species as a novel potential reservoir for one of most important Salmonella serovars.

This research investigated the prevalence of gastrointestinal (GI) strongyles and Escherichia coli O157 in naturally infected sheep, as well as the possible correlation between the pathogens examined. A total of 314 sheep, randomly selected from 21 farms located in southern Italy, were examined. GI strongyles and E. coli O157 were detected by using the FLOTAC double technique and culture media, respectively. GI strongyles were detected on 19/21 farms (90.5%) and E. coli O157 on 12/21 (57.4%). At the animal level, GI strongyles were detected from 193/314 (61.5%) sheep analysed, whereas E. coli O157 was isolated from 20/314 (6.4%) sheep. Statistical analysis performed at animal-level showed a negative correlation between E. coli O157 and GI strongyle EPGs (Spearman’s ρ = -0.128; P = 0.03). Caution should be exercised in interpreting the research findings because a number of different confounding factors possibly influenced the trend of negative correlation between the two pathogenic agents investigated. Further studies, including molecular diagnostics, production data and multivariable analytical approaches, are needed to assess the actual impact of multiple pathogen infections in grazing sheep and other livestock species.

This paper describes the parasitologic findings for 60 synanthropic bird carcasses recovered in the Campania region of southern Italy. Birds consisted of 20 yellow-legged gulls (Larus michahellis), 15 rock pigeons (Columba livia), 15 common kestrels (Falco tinnunculus), and 10 carrion crows (Corvus corone). Each carcass was examined to detect the presence of ectoparasites and then necropsied to detect helminths. Ectoparasites occurred in 100% of the birds examined. In particular, chewing lice were recovered with a prevalence of 100%, whereas Pseudolynchia canariensis (Hippoboscidae) were found only in pigeons with a prevalence of 80%. Regarding endoparasites, a total of seven helminth species were identified: three nematodes (Ascaridia columbae, Capillaria columbae, Physaloptera alata), one cestoda (Raillietina tetragona), one trematoda (Cardiocephalus longicollis), and two acanthocephalans (Centrorhynchus globocaudatus and Centrorhynchus buteonis). The findings of the present study add data to the parasitologic scenario of synanthropic birds. This is important because parasitic infection can lead to serious health problems when combined with other factors and may affect flying performance and predatory effectiveness.

As no data are available on the prevalence of cytolethal distending toxin (cdt) genes carried by Campylobacter spp. in laying hens, this study was conducted with the aim to evaluate the prevalence of both Campylobacter spp. and cdt genes in 1680 laying hens from four different farms. The samples were analyzed by culture methods and by polymerase chain reaction. Campylobacter spp. were isolated from 1097/1680 cloacal swabs. Among the isolates, 913 were identified as Campylobacter jejuni whereas 345 were identified as Campylobacter coli. All isolates carried cdt genes. The results presented here confirm the very common occurrence of C. jejuni and C. coli in laying hens and underline that the cdt genes may also be frequently present in both C. jejuni and C. coli isolates from laying hens.

Thermotolerant Campylobacter spp. were isolated from 118/240 (49.2%) rectal swabs from commercially farmed hares (Lepus europaeus) in southern Italy. Using multiplex PCR, Campylobacter coli was identified in 118/118 (100%) positive samples, while 17/118 (14.4%) positive samples were also positive for Campylobacter jejuni. Adult hares had a higher prevalence of infection with Campylobacter spp. than juvenile hares.

To evaluate the presence of Escherichia coli O157, cloacal swabs were collected from 60 adult frogs kept as pets and analyzed by culture methods and multiplex polymerase chain reaction for stx1, stx2, and E. coli attaching and effacing (eae) and hly genes. The pathogen was isolated from two samples and both carried stx2 gene. This is believed to be the first report of natural E. coli O157 infection in living frogs.

Connexin (Cx) channels are sites of cytoplasmic communication between contacting cells. Evidence indicates that the opening of hemichannels occurs under both physiological and pathological conditions. In this paper, the involvement of Cx-43 hemichannels is demonstrated in the pathogenesis of Yersinia . Parental HeLa cells and transfected HeLa cells stably expressing Cx-43 (HCx43) were infected with Yersinia enterocolitica , and bacterial uptake was measured by the colony-forming unit method. Bacterial uptake was higher in HCx43 cells than in parental cells and was inhibited by the Cx channel blocker, 18-α-glycyrrhetinic acid (AGA). The inhibitory effect of AGA was more pronounced on the Y. enterocolitica uptake by HCx43 cells than by parental cells. The ability of HCx43 cells to incorporate the permeable fluorescent tracer Lucifer Yellow (LY) was assessed. Dye incorporation was inhibited by AGA, whereas Y. enterocolitica infection of HCx43 cells increased LY incorporation. Western blotting analysis demonstrated that Y. enterocolitica infection of HCx43 cells induced tyrosine phosphorylation of Cx-43, thus supporting a critical role for Cx-43 in the strategies exploited by bacterial pathogens to invade non-phagocytic cells.

Rabbit sera (n=1600) from 40 commercial farms were submitted to a serological screening for Encephalitozoon cuniculi by an enzyme-linked immunosorbent assay (ELISA) and a carbon immunoassay (CIA test). Antibodies anti-Encephalitozoon cuniculi were found in 505/1600 (31.6%) sera analysed, and all the farms (100%) resulted positive. Rabbits older than 4 months showed a significantly higher seropositivity for E. cuniculi (chi-squared test: p

Pet rabbits (n = 125) from Southern Italy were submitted to a serological screening for Encephalitozoon cuniculi, using an enzyme-linked immunosorbent assay (ELISA) and a carbon immunoassay (CIA). Seventy-eight examined rabbits showed clinical signs suggestive of encephalitozoonosis (head tilt, ataxia, paralysis, cataracts, uveitis, polyuria and polydipsia), whereas 47 were healthy rabbits. Antibodies anti-E. cuniculi were found in 84/125 (67.2%) sera analysed. The results of the chi-squared test showed that sex and health status had no significant effect (P > 0.05) on E. cuniculi seropositivity; however, rabbits older than 4 months had a seropositivity for E. cuniculi significantly (P < 0.05) higher than that of rabbits aged up to 4 months. The results of the present survey reinforce the assumption that rabbit may be indicated as the main reservoir of E. cuniculi; therefore, routine screening examinations in pet rabbits are strongly advised considering the zoonotic potential of this parasite.

Rectal swab samples were collected from 60 red squirrels (Sciurus vulgaris) from July 2006 to April 2007 in Southern Italy. Samples were tested for Campylobacter jejuni and C. coli by culture methods and suspected colonies were then confirmed by polymerase chain reaction. C. jejuni was detected in 5/60 (8.3%) samples examined but infection status was not related to age or sex and C. coli was not isolated. This is believed to be the first report of C. jejuni infection in the red squirrel.

Skin, lymph node (popliteal), and bone marrow samples were collected from 50 red foxes (Vulpes vulpes) from May 2004 to May 2005 in southern Italy. Samples were tested for Leishmania infantum by polymerase chain reaction (PCR). The parasite was detected by PCR from 20 of 50 (40%) fox carcasses. All 20 positive cases were PCR-positive from lymph node and bone marrow samples, whereas 17 of 20 positive cases were PCR-positive from skin samples. Infection status was not related to age or sex. This is the first report of leishmaniasis in red foxes in Italy based on PCR results, and these results reinforce the assumption that this wild canid can serve as a reservoir for Leishmania.

During the period January 2004 to December 2005 four different ostrich farms were investigated to evaluate the presence of thermotolerant Campylobacter. A total of 150 ostriches were examined and all were found to be clinically healthy. Campylobacter spp. were isolated from 60/150 cloacal swabs. Among the isolates, 48 were identified as Campylobacter jejuni whereas the remainder were identified as Campylobacter coli. Campylobacter lari was not isolated. The ostriches, although apparently in a healthy body condition, can be considered, theoretically, as potential Campylobacter carriers.

Aims: To evaluate the presence of Shiga toxin-producing strains of Escherichia coli (STEC) of the O157:H7 serotype in living layer hens so as to analyse the role of this avian species as potential reservoir. Methods and Results: Cloacal swabs were collected between November 2004 and November 2005 from four intensive management layer hen farms and analysed for STEC O157:H7 by immunomagnetic separation methods and multiplex polymerase chain reaction for stx1 and/or stx2, the E. coli attaching and effacing (eae) and hly genes. STEC was detected in 26 of the 720 samples. Conclusions: The layer hens analysed were shown to carry STEC O157:H7. The presence of this bacterium in living layer hen farms investigated did not result in any detectable increase in gastrointestinal disease in this species. Significance and Impact of the Study: Living layer hens are a novel potential reservoir of E. coli O157:H7.

In this study, three structurally distinct disintegrins (flavoridin, echistatin, kistrin) were used as molecular probes to further characterize the molecular mechanisms underlying Yersinia enterocolitica infection of host cells. The activity of the three disintegrins on Y. enterocolitica uptake into fibronectin-adherent HeLa cells was evaluated at disintegrin doses which were non-cytotoxic and unable to induce cell detachment. Flavoridin resulted to be the most effective in inhibiting bacterial entry into host cells; echistatin was almost 50% less effective than flavoridin, whereas kistrin was definitely inactive. Our results suggest that alpha(5)beta(1) integrin receptor, which binds flavoridin with higher affinity than the other two disintegrins, plays a major role in Y. enterocolitica uptake into HeLa cells. Furthermore, flavoridin binding to this integrin prevented the disruption of the functional complex FAK-Cas, which occurs in the Y. enterocolitica uptake process.

The present study was undertaken with the aim to evaluate the prevalence of thermotolerant Campylobacter spp. in living pheasants in Italy. To achieve this goal, a total of 240 living pheasants, equally shared between female and male birds, were examined. Thermotolerant Campylobacter spp. was isolated in 104 out of 204 (43.3%) living pheasants analysed. Campylobacter coli (100%) and Campylobacter jejuni (13.5%) were identified by polymerase chain reaction. Adult pheasants showed a significantly higher prevalence value (P < 0.05) than younger pheasants.