Risk factors for Toxoplasma gondii infection in dogs: a serological survey.

BACKGROUND: Dogs, as well as a wide variety of other warm-blooded animals, act as intermediate host of Toxoplasma gondii. In dogs, most cases of toxoplasmosis are subclinical, although clinical disease has been sporadically reported. Beyond its role in diagnostic pathways, seropositivity also functions as a reflection of the parasite's spread within the dog's living environment. The aim of the present study was to evaluate the possible risk factor associated with seropositivity to T. gondii in dogs in Central-Northern Italy, analysing 120 dogs sera for the presence of IgG antibodies by indirect fluorescence antibody test (IFAT). RESULTS: The population examined was composed of 54.2% hunting dogs, 24.2% companion dogs, 14.2% truffle dogs and 7.5% watchdogs. Thirty-four (29.2%) dogs tested positive for T. gondii IgG, with titres ranging from 1:40 to 1:1280. Seroprevalence and antibodies titres were not related to dog gender, age or function. The logistic regression and ordered logistic regression results indicated that seroprevalence, and antibody titres were significantly higher in dogs cohabiting with cats, exhibiting coprophagy habits, and living constantly outdoors. Notably, the lifestyle factor showed the highest odds-ratios in the study: dogs living constantly outdoors were found to be at approximately 5 times greater risk of testing positive and having higher antibody titres compared to dogs living both indoors and outdoors. CONCLUSION: Both logistic and ordered logistic regression results support the key role of living with cats, engaging in coprophagy behaviours, and maintaining an outdoor lifestyle in increasing the risk of T. gondii infection in dogs. These identified risk factors collectively suggest that both ingesting oocysts, as observed through cat cohabitation and coprophagy, and engaging in predatory behaviours, as possible for outdoor living dogs, are indicating likely sources of T. gondii infection in this host species.

"Till death do us apart": The common destiny of brown hare and its parasite community.

Although parasites may threaten individual hosts' survival and reproduction, their role as an essential part of ecosystem functioning and biodiversity has been recognized. In Northern Italy, the hare population has evidently declined since 2008. This paper aims to assess the relationship between host demographic trends and helminth parasite community diversity in a two-year survey in Northern Italy to evaluate the impact of parasites on hosts and confirm the hypothesis that endangered host populations are poor in parasites. In 2013 and 2015 the viscera of 54 and 61 hares legally hunted in agro-ecosystems of the Po Plain were collected. This area is characterized by heavy anthropic pressure: more than 60% of the landscape is represented by agricultural or urbanized territories. No intestinal cestodes were detected. Trichuris sp. and Micipsella numidica were collected in 2015 only; Trichostrongylus retortaeformis, Taenia pisiformis cysticercosis and bronchopulmonary lesions caused by small strongyles were observed in both years. The richness and evenness appeared increased in the second year of the survey, although lower than those obtained by literature data from similar populations examined in the last two decades of 1900. The dominant helminth, T. retortaeformis, was more abundant in individuals with higher weight, while pathological findings were mostly unrelated to this nematode infection; this is consistent with a reduced action, even no harm, of the parasites on the individual host or population level. Our results suggest that the conservation of hosts, environment, and parasites cannot be achieved separately and that parasites and wildlife hosts' destinies are intimately linked, confirming the complexity of ecosystems and the need to contemplate parasite biodiversity in conservation strategies.

Helminth biocoenosis of Lepus europaeus meridiei (Hilzheimer, 1906) from Pianosa island, Italy.

Pianosa is a 10 km2 Italian island in the Tyrrhenian Sea which is part of the Tuscan Archipelago National Park. In this island lives a brown hare population which, according to the literature, belongs to the ancestral taxon Lepus europaeus meridei that offers a unique opportunity to observe how the parasite biocoenosis shapes in condition of isolation, limited space availability and high population density. The aim of this work is to describe the helminth component community of a non-managed, isolated, and dense hare population, evaluating host-parasite relationship and parasite community structure. All 26 analyzed hares (13 males and 13 females) were in good physical conditions, and all of them harboured exclusively the nematode Protostrongylus oryctolagi only. This is the first report of this lungworm species in Italy. The estimated overall abundance was 48.15 worms per examined hare (range 3-258, median 50) and the parasites were unevenly distributed across host population, with few hosts having most parasites (aggregated or overdispersed distribution). No significant relationship was detected between the number of isolated parasites and hare sex and weigh. The effect of the isolation of Pianosa's hare population seems to have acted reducing parasite richness, while the high host density is probably the cause of the high prevalence and abundance of the single helminth species collected. In conclusion, despite the low impact of parasites confirmed also by the overdispersed parasite distribution, the low diversity of the studied parasite community sounds a warning for the management of the hare population and the whole Pianosa's ecosystem.

Distribution of Deer Keds (Diptera: Hippoboscidae) in Free-Living Cervids of the Tuscan-Emilian Apennines, Central Italy, and Establishment of the Allochthonous Ectoparasite Lipoptena fortisetosa.

Lipoptena fortisetosa and L. cervi are hematophagous ectoparasites belonging to the Hippoboscidae family and preferentially living on cervids. In recent years, they have received specific attention due to the great increase in the abundance of their host species, and to their medical and veterinary importance as possible vectors of pathogens harmful to humans and animals. The aim of this study was to investigate the parasitism level of both of these flies on their main hosts in Italy, which are red deer, fallow deer, and roe deer, and to highlight a possible preference for a species, sex, or age class among the hosts. Deer keds were collected by examining 326 cervids hunted in the Tuscan-Emilian Apennines. Outcomes showed that L. fortisetosa has greatly spread throughout the study area, where it competes with the autochthonous L. cervi. Moreover, red deer was the favored host species of both ectoparasites, while different preferences for host sex and age classes were observed in the two hippoboscids. The regular monitoring of deer ked populations, especially the allochthonous L. fortisetosa, which is continuously spreading in Europe, is recommended to expand the knowledge on these parasitic species that are potentially dangerous to public health.

Use of Flubendazole and Fenbendazole for Treatment of Lung Severe Infection by the Gapeworm Cyathostoma bronchialis (Nematoda: Syngamidae) in Branta hutchinsii, Anser indicus and B. leucopsis Exotic Geese: An Interesting Case.

A 6-year-old female goose (Branta hutchinsii) from a group of ornamental exotic geese was found dead due to severe respiratory failure, followed by emission of haemorrhagic sputum and blood clots from the beak and nostrils, and then collapse. At necropsy, the cause of death was attributed to a total of 76 helminth parasites found in the trachea and lungs, then identified as Cyathostoma bronchialis. The flock was initially treated by feed with flubendazole (1200 g/1000 kg/feed) for 7 consecutive days but, at the reappearance of the respiratory symptoms 10 days after, the animals were given fenbendazole by drinking water (300 mg/L) for 7 consecutive days, but at the reappearance of the respiratory symptoms 10 days after, the animals were given fenbendazole via drinking water (300 mg/L) for 7 consecutive days. Despite these treatments, the respiratory symptoms continued to relapse 10-15 days after the end of drug administration. In the literature, there are no data regarding drugs for the treatment of C. bronchialis infestations in geese, and the use of these drugs in exotic birds occurs as "off-label" use. This case study provides information on C. bronchialis life cycle, which is still poorly studied and poorly documented today. In particular, the case provides useful suggestions for evaluating an appropriate protocol for the treatment of C. bronchialis in geese.

Survey on Endoparasites of Dairy Goats in North-Eastern Italy Using a Farm-Tailored Monitoring Approach.

With the spread of anthelmintic resistance (AR), endoparasite monitoring consolidates its role for a more sustainable targeting of treatments. A survey on endoparasites in dairy goat farms of north-eastern Italy was conducted to test a monitoring approach based on a farm-tailored sample size. Farm management and parasites control practices were investigated in 20 farms through a questionnaire survey. Further, fecal samples were collected (November 2018-September 2019) from 264 animals from 13 farms and were analyzed individually with a modified McMaster method and subsequently pooled to perform a coproculture. Coccidia (78.4%), gastrointestinal strongyles (37.9%), Strongyloides (28.4%), Skrjabinema (18.9%), Trichuris (8.0%) and Nematodirus/Marshallagia (0.4%) were identified. Abundances were higher for coccidia and gastrointestinal strongyles. Haemonchus (71%) was the dominant gastrointestinal nematode. Pasture and age class resulted in the main risk factors at the multivariable analysis through a negative binomial regression model. Results from farm monitoring indicate that our approach can be a cost-effective decision tool to target treatments more effectively, but farmers need to be educated about the importance of parasitological testing, which is currently scarcely implemented, against the risk of AR.

Cysticercosis by Taenia pisiformis in Brown Hare (Lepus europaeus) in Northern Italy: Epidemiologic and pathologic features.

In Northern Italy, a hastening of hare population decline was noticeable from 2008. In the same year hunters reported a sudden increase of hares infected by Taenia sp. larvae, whose morphology was consistent with T. pisiformis cysticerci. The aim of the survey was: i)to identify the parasites through morphological features and molecular techniques; ii)to quantify the prevalence and abundance of cysticerci in hunted hares; iii)to describe pathological aspects of parasite-induced lesions; iv)to evaluate the short-term trend of the infection comparing two different hunting seasons; v)to highlight possible relationship between T. pisiformis infection and hare-related variables. In 2013, 2015 the viscera of 54 and 61 hares legally hunted in agro-ecosystems of the Po Plain were collected. Peritoneum, liver and lungs were examined for cysticercosis; abundance was estimated counting superficial parasites in liver; parasites were microscopically identified by shape and measure of both large and small hooks. One cysticercus from each hare was analized by a PCR targeting Taeniid species and then sequenced. Frozen liver, lungs and gastrointestinal peritoneum were macroscopically observed and, after thawing, representative samples from the available organs were collected for histologic examination to verify parasitic cysts and the subsequent damage of the involved organs. Sex, weight and age class of the animals were recorded. Generalized linear models were used for statistical analysis. T. pisiformis was isolated in 8 hares in 2013 (prevalence 14.8%; abundance range: 0-400; mean abundance 17.8) and in 2 hares in 2015 (prevalence 3.28%; abundance range: 0-180; mean abundance 3.22). Identification was confirmed morphologically and by PCR. The DNA sequencing confirmed T. pisiformis in all samples. The sequences were all identical each-other. Infection was significantly related with adult age class, sampling year and low full-weight. Epidemiological and pathological pattern suggest both a possible role on host population health and a tendency toward host-parasite equilibrium.

Are small strongyles (Cyathostominae) involved in horse colic occurrence?

Strongyle infections have been traditionally regarded as a possible cause of colic in horses. Aim of the study was the comparison of parasitological status between subjects with or without colic syndrome, with particular attention to small strongyle infections. Coprological analyses were performed on 86 horses: 43 with colic and 43 controls. Strongyle eggs were found in 34/86 horses (prevalence 39.5%), the mean number of strongyles eggs per gram of faeces (EPG)2 was 145.34 (standard deviation 398.28). All those 34 positive animals had small strongyles infections. Negative binomial multiple regression highlighted no influence of horse sex on strongyle EPG, while there was a negative relationship between age and EPG (p <0.05); the same analysis revealed a significant difference of EPG (p <0.05) between control horses (mean EPG=178.1; standard deviation: 411.4) and horses with surgical colic (mean EPG=68.6 standard deviation: 259.8) when controlling for S. vulgaris presence including it in the model. On the contrary, the intensity of infection in horses with non-surgical colic (mean EPG=154.5; standard deviation: 480.4) did not significantly differed from controls. Similar results were obtained having estimated cyathostomine EPGs as dependent variable. Multinomial logistic regression confirmed the negative relationship between cyathostonine presence and surgical colic occurrence. It is possible that (1) the presence of adult luminal parasites, would have a protective effect against the pathogenic action exerted by the development and emergence of small strongyles larvae from intestinal mucosa; (2) the management practices able to reduce the risk of colics are the same that cause higher exposure to strongyle infective larvae.

Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii Circulation in a Dairy Farm and Sources of Milk Contamination.

Even though dairy cows are known carriers of Arcobacter species and raw or minimally processed foods are recognized as the main sources of human Arcobacter infections in industrialized countries, data on Arcobacter excretion patterns in cows and in milk are scant. This study aimed to identify potentially pathogenic Arcobacter species in a dairy herd and to investigate the routes of Arcobacter transmission among animals and the potential sources of cattle infection and milk contamination. A strategy of sampling the same 50 dairy animals, feed, water, and milk every month for a 10-month period, as well as the sampling of quarter milk, animal teats, the milking environment, and animals living on the farm (pigeons and cats), was used to evaluate, by pulsed-field gel electrophoresis (PFGE), the characteristic patterns in animals, their living environment, and the raw milk they produced. Of the 463 samples collected, 105 (22.6%) were positive for Arcobacter spp. by culture examination. All the matrices except quarter milk and pigeon gut samples were positive, with prevalences ranging from 15 to 83% depending on the sample. Only three Arcobacter species, Arcobacter cryaerophilus (54.2%), A. butzleri (34.2%), and A. skirrowii (32.3%), were detected. PFGE analysis of 370 isolates from positive samples provided strong evidence of Arcobacter circulation in the herd: cattle likely acquire the microorganisms by orofecal transmission, either by direct contact or from the environment, or both. Water appears to be a major source of animal infection. Raw milk produced by the farm and collected from a bulk tank was frequently contaminated (80%) by A. butzleri; our PFGE findings excluded primary contamination of milk, whereas teats and milking machine surfaces could be sources of Arcobacter milk contamination.

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

Factors affecting the fecal shedding of thermophilic Campylobacter in Italian dairy farms were investigated in a 12-month longitudinal study performed on a dairy farm authorized to sell raw milk in Italy. Fifty animals were randomly selected from 140 adult and young animals, and fecal samples were collected six times at 2-month intervals. At each sampling time, three trough water samples and two trough feed samples also were collected for both adult and young animals. Samples were analyzed with real-time PCR assay and culture examination. Overall, 33 samples (9.7%) were positive for thermophilic Campylobacter by real-time PCR: 26 (9.2%) of 280 fecal samples, 6 (16.6%) of 36 water samples, and 1 (4.2%) of 24 feed samples. Campylobacter jejuni was isolated from 6 of 280 samples; no other Campylobacter species was isolated. A higher (but not significantly) number of positive fecal samples were found in younger animals (11.33 versus 6.92% of adult animals), and a significantly higher number of positive water samples were collected from the water troughs of young animals. A distinct temporal trend was observed during the study period for both cows and calves, with two prevalence peaks between November and December and between May and July. Several factors such as calving, housing practices, herd size, management practices forcing together a higher number of animals, and variations in feed or water sources (previously reported as a cause of temporal variation in different farming conditions) were excluded as the 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 on farms authorized to produce and sell raw milk, with increased surveillance by the authorities at certain times of the year.

First detection of Arcobacter sp. in Eurasian collared doves (Streptopelia decaocto).

Data about the presence of Arcobacter in wild birds are currently lacking. In this study cloacal swabs from 95 collared doves (Streptopelia decaocto), submitted to the Department of Veterinary Medical Sciences (Bologna, Italy) between 2011 and 2013 from various urban and suburban areas of the Emilia-Romagna region (Northern Italy), were tested for the presence of Arcobacter sp. by a rRNA 23S nested Polymerase Chain Reaction (PCR). Eighteen out of 95 (19%) samples showed the expected PCR product. Further cultural and molecular studies are needed to assess the Arcobacter prevalence in wild birds and elucidate their potential epidemiological role as source of animal and human infections.

Temporal Variation of Faecal Shedding of Escherichia coli 0157:H7 in A Dairy Herd Producing Raw Milk for Direct Human Consumption.

The objective of this study was to analyse over time the evolution of E. coli O157:H7 faecal shedding in a dairy herd producing raw milk for direct human consumption. The study was performed between October 2012 and September 2013 in an average size Italian dairy farm where animals are housed inside the barn all over the year. The farm housed about 140 animals during the study - 70 cows and 70 calves and heifers. Twenty-six animals were randomly selected from both the cows and young animals group, and faecal sampling was performed rectally six times two months apart in each animal. Eleven animals were culled during the study and a total of 285 faecal samples were collected. At each faecal sampling, three trough water samples and two trough feed samples were also collected for a total of 36 water samples and 24 feed samples. Samples were analysed by real time polymerase chain reaction (RT-PCR) and culture. Overall, 16 (5.6%) faecal samples were positive for E. coli O157 by RT-PCR. Cultural examination found 9 (3.1%) samples positive for E. coli O157; all the isolates were positive for stx1, stx 2 and eae genes. One (4.1%) feed sample was positive for E. coli O157 by RT-PCR; none of the water samples was positive for E. coli O157. The model highlighted a general significant reduction of the number of positive samples observed during the study from the first to the sixth sampling (P=0.000) and a positive relation between the presence of positive samples and average environmental temperature (P=0.003). The results of the study showed that in an Italian dairy farm housing animals all year, faecal shedding of E. coli O157 followed the same temporal trend reported for other types of farming. The enhanced faecal shedding during warmer months may have a significant impact on environmental contamination and the safety of raw milk and its byproducts.