Search

Tick-borne diseases are among the major widespread emerging zoonotic diseases, and their circulation in the environment is influenced by a broad range of abiotic and biotic factors, including the abundance of vectors and vertebrate hosts. In this study, we estimated the prevalence of tick-borne pathogens and the impact of wildlife head count on their circulation in a lowland natural area in northwestern Italy. We collected ticks and camera trap pictures from 14 sampling points every 2 weeks for 1 year and identified pathogens through molecular analyses: Babesia capreoli, B. microti-like, Borrelia burgdorferi sensu lato (s.l.), Rickettsia of the spotted fever group (SFG), Theileria capreoli, and Anaplasma phagocytophilum. We modeled the presence of B. capreoli, B. microti-like, B. burgdorferi s.l., and SFG Rickettsia on head counts of wild ungulates and mesocarnivores. We tested a global model including all collected ticks, as well as a model focusing solely on Ixodes ricinus nymphs, the species, and the developmental stage most associated with zoonotic infection risk. The highest prevalence was obtained for B. microti-like (13%) and SFG Rickettsia (11%), and, for most pathogens, no differences were detected among tick species and their developmental stages. Mesocarnivores showed an additive effect on B. microti-like and B. burgdorferi s.l., while wild ungulates, non-competent for transmission of our target pathogens, showed a dilutive effect. These findings confirm the circulation of relevant tick-borne pathogens in the study area and show the use of camera trap data in predicting tick-borne pathogens’ risk by targeting host species which may have an indirect impact and are more easily addressed by monitoring and control strategies.

Bartonella is a genus of bacteria known to cause various rare but potentially dangerous diseases in humans and wildlife. The objective of this study was to investigate the presence of Bartonella spp. in red foxes (Vulpes vulpes) from Piedmont and Aosta Valley (NW Italy) and to explore the potential association between environmental humidity and Bartonella infection using remote sensing data. A total of 114 spleen samples were collected from hunted red foxes and screened for Bartonella spp. DNA using a qPCR assay targeting the ssrA locus. Samples that tested positive were further analyzed using end-point PCR to detect the ssrA, gltA, and rpoB loci. The overall prevalence of Bartonella spp. infection was found to be 7.9% (9/114), with a prevalence of 6.9% (7/101) in foxes from Piedmont and 15.4% (2/13) in foxes from Aosta Valley. Sequencing results identified Bartonella schoenbuchensis R1 as the most commonly isolated species (5/9, 62.5%), followed by Candidatus “Bartonella gerbillinarum” in two samples (2/9, 28.6%). To investigate the relationship between environmental factors and Bartonella infection, data from NASA USGS Landsat missions (TOA collection) from 2011 to 2022 were processed using Google Earth Engine. The Tasseled Cap Wetness Index (TCW), an indicator of landscape moisture, was calculated for each meteorological season. The study found that Bartonella spp. infections in foxes were positively associated with higher TCW values (>0.7). Canonical Correspondence Analysis further demonstrated a strong link between pathogen prevalence and municipal-level TCW, suggesting that TCW could be used as a parameter to facilitate disease management and control. This study provides a starting point for a more comprehensive epidemiological assessment of Bartonella spp. on a national scale and highlights the potential role of environmental humidity in influencing pathogen distribution.

Abstract Background This study aimed to evaluate the effects of Hermetia illucens (Black soldier fly-BSF) and Tenebrio molitor (Yellow mealworm-YMW) live larvae as a new nutritional model on duck’s gut health, considering gut histomorphometry, mucin composition, cytokines transcription levels, and microbiota. A total of 126, 3-days-old, females Muscovy ducks were randomly allotted to three dietary treatments (6 replicates/treatment, 7 birds/pen): (i) C: basal diet; (ii) BSF: C + BSF live larvae; (iii) YMW: C + YMW live larvae. BSF and YMW live larvae were administered on top of the basal diet, based on the 5% of the expected daily feed intake. The live weight, average daily gain, average daily feed intake and feed conversion ratio were evaluated for the whole experimental period. On day 52, 12 ducks/treatment (2 birds/replicate) were slaughtered and samples of duodenum, jejunum, ileum, spleen, liver, thymus and bursa of Fabricius were collected for histomorphometry. Mucin composition was evaluated in the small intestine through histochemical staining while jejunal MUC-2 and cytokines transcription levels were evaluated by rt-qPCR. Cecal microbiota was also analyzed by means of 16 S rRNA gene sequencing. Results Birds’ growth performance and histomorphometry were not influenced by diet, with a proximo-distal decreasing gradient from duodenum to ileum (p

Hiking trails may act as hotspots at the wildlife–human interface, posing an acarological risk for people and their pets. Ticks that are maintained in the environment by wild animals may quest on people walking along the trails. Assessing the risk of tick bites for people involved in outdoor activities is a further step in mitigating the risk of tick-borne diseases. This work describes the variation of tick abundance along a gradient of distances from hiking trails, where wildlife passage is favored by higher accessibility. Hiking trails with dense vegetation on the sides were sampled for ticks along a 100 m dragging transect, located in a natural park in North-Western Italy. Additional transects were replicated at 1, 2 and 4 m away from the trail on both sides. After morphological identification, descriptive statistics and modeling were applied to determine the abundance patterns across distances. Larvae were most abundant near the trail, peaking at 1 m and dropping sharply at further distances. Nymphs showed a more gradual and consistent decrease at progressing distance from the trail. Few adults were collected, preventing the identification of a clear trend. With higher tick abundance, the immediate vicinity of hiking trails may represent a source of acarological risk for humans and pets.

Abstract Background The ticks Ixodes ricinus and Dermacentor reticulatus are two of the most important vectors in Europe. Climate niche modelling has been used in many studies to attempt to explain their distribution and to predict changes under a range of climate change scenarios. The aim of this study was to assess the ability of different climate niche modelling approaches to explain the known distribution of I. ricinus and D. reticulatus in Europe. Methods A series of climate niche models, using different combinations of input data, were constructed and assessed. Species occurrence records obtained from systematic literature searches and Global Biodiversity Information Facility data were thinned to different degrees to remove sampling spatial bias. Four sources of climate data were used: bioclimatic variables, WorldClim, TerraClimate and MODIS satellite-derived data. Eight different model training extents were examined and three modelling frameworks were used: maximum entropy, generalised additive models and random forest models. The results were validated through internal cross-validation, comparison with an external independent dataset and expert opinion. Results The performance metrics and predictive ability of the different modelling approaches varied significantly within and between each species. Different combinations were better able to define the distribution of each of the two species. However, no single approach was considered fully able to capture the known distribution of the species. When considering the mean of the performance metrics of internal and external validation, 24 models for I. ricinus and 11 models for D. reticulatus of the 96 constructed were considered adequate according to the following criteria: area under the receiver-operating characteristic curve > 0.7; true skill statistic > 0.4; Miller’s calibration slope 0.25 above or below 1; Boyce index > 0.9; omission rate

The increase in acarological risk of tick bites is significantly driven by profound changes in landscape, which alter the density and distribution of wildlife that support tick populations. As a result of habitat shifts and land abandonment, which create environments conducive to tick proliferation, the risk of disease transmission to humans and animals is increasing. In this context, it is important to explore tick ecology by applying a comprehensive methodology. In this study, we examined the relationship between wildlife temporal occupancy and tick abundance in two distinct regions: an alpine hunting district and a natural park in the Apennines. For each sampling point, we calculated wildlife temporal occupancy from camera trap pictures and estimated ticks’ abundance from dragging transects in the area immediately surrounding camera traps. In modelling the relationship between those two variables, we included abiotic factors such as saturation deficit, normalized difference vegetation index, and altitude. Results show the importance of altitude and wildlife temporal occupancy (itself related to different habitat and land management characteristics) on the ecology of questing ticks. If employed in management decisions for natural environments, such information is useful to modulate the acarological risk and thus the risk of tick-borne pathogens’ transmission.

In Italy, there are hundreds of research, monitoring, and surveillance activities targeting emerging and re-emerging pathogens. These activities heavily rely on hunters for sample collection and early identification of morbidity/mortality events. The objective of this review is to describe and quantify the contribution of hunters in the context of disease research, monitoring, and surveillance in wild populations. A literature review and descriptive summary statistics were performed following PRISMA-2020 guidelines; articles were obtained from major scientific databases, abstracts from national and international conferences, proceedings, graduate-level theses from online library repositories, and direct contact with academic experts. The contribution of hunters in terms of sample collection for health-related activities on wildlife amounts to 400,000 sampled animals. Wild boars were involved in 158 surveillance systems/research studies, followed by red deer (71), foxes (63), and roe deer (59). The pathogens under surveillance were mainly zoonotic (Salmonella spp.), emerging (Hepatitis E virus), and/or vector-borne (West Nile virus). The temporal distribution of scientific papers followed a positive trend that reflects the growing interest in wildlife from different sectors. These results highlight how wildlife health-related efforts are a paradigm of the concept of One Health, in which the role of hunters is crucial to ensure sample availability, and it constitutes the base of much current wildlife health research, monitoring, and surveillance.

European hedgehogs (Erinaceus europaeus) are nocturnal insectivores frequently found in urban areas. In the last decades, their population has declined in various European countries and human activities have emerged as significant contributors to this trend. While the literature has mainly focused on trauma as the major cause of mortality, few authors have considered pathological findings. The present study is based on the results of full post-mortem examinations performed on 162 European hedgehogs in Italy and 109 in Switzerland. Unlike in previous studies, the main cause of mortality was infectious diseases (60.5%), followed by traumatic insults (27.7%). The lungs were the main organ affected, showing mostly lymphoplasmacytic (45.9%), granulomatous (18.1%) or suppurative (8.2%) pneumonia. Nematodes were detected in 57.2% of all lungs and were significantly associated with pneumonia (p-value < 0.001). To our knowledge, this is the first study to report infectious diseases as the main cause of hedgehog death, emphasizing the need for wildlife rescue centers to adopt appropriate diagnostic and therapeutic measures. Further research is necessary to determine the broad range of infectious agents that affect this species and elucidate their interplay with the host. Finally, citizen sensitization should be implemented to promote responsible behaviors that could reduce human-related traumatic events.

IntroductionNeospora caninum is one of the main causes of abortion in cattle. In wildlife, the occurrence and relevance of vertical infection have not been yet clearly evaluated. The aim of this study was to verify the possibility of vertical transmission of N. caninum in three wild species extensively distributed in Europe, namely roe deer Capreolus capreolus, wild boar Sus scrofa and red fox Vulpes vulpes.MethodsA total of 190 fetuses (72 wild boars, 67 foxes and 51 roe deer) from 61 females were included in the study. All animals, which were either found dead or culled within selective control plans in North-western Italy, were tested, in parallel by PCR on central nervous system, skeletal muscle and kidney.Results and discussionThe efficiency of vertical transmission in the three target species was of 87.5% (95% CI 69.00–95.66).

Invasive species have a detrimental impact on native populations, particularly in island ecosystems, and they pose a potential zoonotic and wildlife threat. Black rats (Rattus rattus) are invasive species that disrupt native flora and fauna on islands and serve as potential competent reservoirs for various pathogens and parasites. Microparasites screening was conducted in rat populations from small islands in central Italy (the Pontine Islands and Pianosa) with the aim of assessing the role of rats in maintaining infections, particularly in cases where key reservoir hosts were scarce or absent. We focused on microparasites of zoonotic and veterinary relevance. A total of 53 rats was kill-trapped and target tissues were analysed with molecular techniques. We observed the absence or very low prevalence of Anaplasma spp., while Babesia was found in rats from all locations, marking the first recorded instance of Babesia divergens in wild rats. Data from Pianosa strongly suggest the presence of an autochthonous Leishmania infantum cycle in the Tuscan archipelago islands. Neospora caninum was absent from all islands, even in areas where dogs, the main reservoirs, were present. Toxoplasma gondii was only recorded on the Pontine Islands, where genotyping is needed to shed light on infection dynamics. This study confirms that invasive species, such as rats, may be responsible for maintaining an increased parasitological threat to fauna and human communities in certain ecosystems.

Abstract Background The present clinical field trial was conducted to assess the efficacy of a broad-spectrum parasiticide spot-on formulation containing eprinomectin (Broadline®) against Thelazia callipaeda eyeworm in naturally infected cats. Methods Fifteen privately owned cats harboring at least one live adult T. callipaeda were included in the study. Cats were randomly allocated to an untreated control group of seven cats or to a Broadline®-treated group of eight cats. Cats were treated on Day 0; ocular examinations were performed at inclusion and on Days 7 and 14; eyeworms were recovered and counted on Day 14. The primary efficacy assessment was based on group comparison of number of T. callipaeda on Day 14. Results Seven days after treatment, six of eight treated cats were negative for eyeworm infection per visual examination, and on Day 14 no eyeworms were found in the treated cats while the seven untreated cats were still infected (geometric mean: 1.97). All cats had inflammatory ocular signs at inclusion; on Day 14, five of eight treated cats had recovered while all untreated control cats were still symptomatic. All collected parasites were confirmed to be T. callipaeda by morphology and molecular characterization. Conclusions A single treatment with Broadline® provided 100% efficacy against feline thelaziosis and improved related ocular inflammation signs. Graphical abstract

Abstract Background Parasites that infect cats include protozoa, helminths and arthropods, many of which are transmissible to humans. Effective control relies on a good knowledge of parasite distribution and the risk factors for infection. The present study was aimed at evaluating the prevalence of major feline parasites in Italy and the risk factors associated with their occurrence. Methods Over a 12-month study period, feces, hair and ectoparasites from naturally infected cats from feral colonies, shelters and private households were analyzed at 13 study centers across Italy. Samples from these cats (n = 987) were analyzed at all centers using the same diagnostic methods. Prevalence values and risk factors were evaluated statistically for the identification of predictors of risk. Results The overall prevalence of gastro-intestinal and broncho-pulmonary (BP) nematodes was 35.9% (354/987). Toxocara cati was the most prevalent species (253/987; 25.6%), followed by Ancylostomatidae (98/987; 9.9%). Among BP nematodes, Aelurostrongylus abstrusus was the most common (76/987; 7.7%). Approximately 35.7% (352/987) of the study population was infested by ectoparasites, of which the most common were fleas (29.4%, 290/987), followed by ear mites Otodectes cynotis (9.8%, 97/987). Predictors of risk for parasite infection included age, a predominantly or exclusively outdoor lifestyle, geographic area and lack of antiparasitic treatment. Conclusions Both ecto- and endoparasites are still common in cats throughout Italy, many of them being of zoonotic concern and vectors of pathogens to humans. Given the presence of parasites throughout the entire study period, year-round treatment should be considered. Furthermore, data confirm the need to protect the human–animal bond using proper endo- and ectoparasiticides to reduce the risk of human infection, in application of the One-Health concept. Graphical abstract

Toxoplasma gondii and Neospora caninum are apicomplexan protozoa of major concern in livestock and T. gondii is also considered one of the major threats and a public health concern. These protozoa have a wide range of intermediate hosts, including birds. The present work aimed to assess the prevalence of these cyst-forming parasites in migratory and sedentary birds of prey. The skeletal muscle and myocardium of 159 birds of prey from Central Italy, belonging to 19 species and recovered across 6 Wildlife Recovery Centers/Care structures along the Italian migratory route, were collected specifically for molecular (PCR) and for histopathological analysis to detect T. gondii and N. caninum. For the molecular analysis, genomic DNA was extracted. The DNA was tested by sequence typing, targeting GRA6, 529 bp repeated element, B1, PK1, BTUB, SAG2, alt.SAG2, and APICO genes for T. gondii and to end-point PCR targeting NC5 gene for N. caninum. Thirty-seven out of the one hundred and fifty-nine analyzed samples tested positive for T. gondii with a prevalence of 23.27% and nine for N. caninum, with a prevalence of 5.66%. Thirty-two sequences were obtained from the thirty-seven isolates of T. gondii. Among these, 26 presented alleles compatible with type I strain in 1 or more loci, 4 with type II strain and 2 consisted of atypical strains. Toxoplasma gondii genetic variability in birds of prey confirms previous findings of wildlife as reservoirs of atypical strains. Results from the histology showed few protozoal tissue cysts in skeletal muscle (n. 4) and hearts (n. 2).

Abstract Background Ticks and tick-borne diseases are increasingly recognized as a cause of disease in dogs worldwide. The epidemiology of ticks and tick-transmitted protozoa and bacteria has changed due to the spread of ticks to urban and peri-urban areas and the movement of infected animals, posing new risks for animals and humans. This countrywide study reports information on distribution and prevalence of pathogens in ticks collected from privately-owned dogs in Italy. We analyzed 2681 Ixodidae ticks, collected from 1454 pet dogs from Italy. Specific PCR protocols were used to detect i) Piroplasms of the genera Babesia and Theileria, ii) Gram-negative cocci of the family Anaplasmataceae and iii) Borrelia burgdorferi sensu lato. Sequencing of positive amplicons allowed for species identification. Results Babesia/Theileria spp. DNA was detected in 435 homogeneous tick-pools (Minimum Infection Rate (MIR) = 27.6%; 95% confidence interval (CI) = 25.4–29.8%) with higher prevalence in Ixodes ricinus and Rhipicephalus sanguneus group. The zoonotic B. venatorum was the most prevalent species (MIR = 7.5%; 95% CI = 6.3–9.0%). Anaplasma and Ehrlichia species were detected in 165 tick-pools (MIR = 10.5%; 95% CI = 9.3–11.8%) and specifically, A. phagocytophilum was identified with MIR = 5.1% (95% CI = 4.1–6.3%). Borrelia burgdorferi s.l. and B. afzelii were detected with MIR = 0.4% (95% CI = 0.2–0.8%) and MIR = 0.3% (95% CI 0.1–0.7%) respectively. Conclusions Zoonotic pathogens B. venatorum and A. phagocytophilum were the most frequently detected in ticks collected from privately-owned dogs which might be used as markers of pathogens presence and distribution.

Although wildlife has progressively been recognized as a booster for the spillover of pathogens to humans and other wild and domestic animals, the interest of scientists in this topic has not been constant over time and uniform in its targets. Epidemiological outbreaks and technological development have contributed to this. Through the analysis of the number of publications from a citation database, we aimed to obtain an indicator of the scientific community’s interest towards wildlife diseases over the years. Using Web of Science, bibliographic searches were performed by adding to the basic search string “Wildlife AND Disease” terms detailing topics such as aspect investigated, pathogen type, aetiologic group and species group. For each host species group, the 100 most frequent Medical Subject Headings (MeSHs) related to specific diseases in each decade were extracted. The scientific production regarding wildlife diseases has increased 3.7 times the relative proportion of publications on diseases during the last 50 years, focusing mainly on zoonotic or epizootic pathogens, and with a sharp growth in contemporary hot topics. Wildlife disease scenarios are complex and challenging to approach. Knowing the trends in the scientific interest in the past decades may pose a guide to direct future steps and actions in several fields, from public health to ecosystem management.

Pneumonia is the most frequent cause of death for Alpine ibex (Capra ibex) in Gran Paradiso National Park, (Italy). The etiology of this form of pneumonia is currently unknown and the identification of the primary etiological agent remains difficult due to biological and logistic constraints. Uncovering individual differences in Protostrongylid prevalence and intensity is important to further investigate the epidemiology of respiratory diseases and their relationship to heterozygosity and inbreeding in a once almost extinct population like C. ibex. In a group of 21 individually recognizable adult male we monitored monthly prevalence and intensity of Protostrongylid first-stage larvae using Baerman's technique from June to September 2019. First-stage larvae of 5 genera were detected. Muellerius (P = 100%, CI95% = 84–100) and Protostrongylus (P = 86%, CI95%:71–100) were two dominant genera according to Bush's importance index. Neostrongylus (P = 38%,CI95%: 17–59), Cystocaulus (P = 33%,CI95% = 13–53) were classified as co-dominant genera while Dictyocaulus filaria (P = 0.05%, CI95% = 0.04–0.13) was detected, for the first time in Alpine ibex, in one subject. Protostrongylidae larval excretion varied significantly over time, with minimum L1 excretion in July. Individual median larval intensity ranged from 4.4 lpg to 82.2 lpg with Poulin's discrepancy index showing highly aggregated distribution patterns for Muellerius spp. (D = 0.283, CI95% = 0.760–0.895) and Protostrongylus spp. (D = 0.635, CI95% = 0.580–0.705). Presented data provide the necessary base point to further investigate how lungworm infection account for the different rates of progression of pneumonia in C. ibex. Individual aggregation of larval intensity must be further evaluated to determine whether these differences mirror different levels of parasitic infection related to individual differences in immune response, hormonal-states or genetic fitness.

Tick-borne diseases and especially protozoa of the genus Babesia, are gaining increasing attention as emerging zoonotic pathogens. Zoonotic species like B. venatorum and B. microti have wild animals as main reservoir hosts. We propose a habitat suitability model for Babesia spp., as tool for institutions and policy makes to better understand the entity of Babesia presence, to improve diagnostic awareness and to optimize screening and preventive actions. The probability of presence of Babesia spp. was estimated using as presence data, wild ruminants positive by PCR to Babesia spp. which were correlated to environmental factors that can favor or limit vector and host availability. We developed three separate models to discriminate the different roles of Red deer and Roe deer and Alpine chamois in Babesia spp. epidemiology. A comprehensive model using all presence data from all ungulates species was also developed. The overall suitable area for Babesia spp. in this simulation is of 3723 km2, which correspond to 15.51% of the background regional territory. The model developed was empirically validated assessing tick abundance in randomly chosen areas classified by the model as moderately or highly suitable for Babesia spp. Collected ticks were tested by PCR for Babesia spp. to confirm model predictions as infection prevalence with Babesia spp. was significantly higher in areas predicted as highly suitable compared to those classified by the model as moderately suitable for Babesia spp. (X2=5.05 p

Abstract Background The geographical distribution of ticks on companion animals needs to be monitored to develop and plan effective control measures, as suggested by the European Scientific Counsel on Companion Animal Parasites. The aim of this study was to conduct the first Italian national survey of tick distribution on privately owned dogs. Methods The study was performed over 20 months (February 2016 - September 2017) and involved 153 veterinary practices in 64 different provinces covering 17/20 (85%) Italian regions. Participating practitioners were asked to examine five different dogs per month at random and complete a questionnaire for each dog. Differences in tick infestation associated with: sex, age and hair length (long and short); the dog’s habitat (indoor or outdoor/kennel); and the dog’s environment (urban or rural/sylvatic) were evaluated. The attachment site of ticks on the dog was also recorded. Acaricide efficacy was evaluated for the subset of dogs for which complete information on product used, date of sampling and date of last ectoparasiticide treatment was available. Results Of the 3026 dogs examined, 1383 (45.7%) were carrying at least one tick. Overall, 2439 tick samples were collected and a total of 14 tick species identified. Rhipicephalus sanguineus group were the most predominant ticks (63.6%), followed by Ixodes ricinus (30.6%) and I. hexagonus (5.6%). Twenty-four dogs had mixed tick infestations. Long-haired dogs had a higher tick infestation risk as did dogs with outdoor and rural/sylvatic lifestyles. Ticks were located on the head (37.4%), the neck (28.8%), the muzzle (15.5%) and the back (15.3%). A higher prevalence of Rhipicephalus was found in the interdigital spaces (10.8%) compared to Ixodes (0.2%). Finally, ectoparasiticide treatments were found significantly protective against tick infestation, especially orally administered formulations. Conclusions Privately owned dogs in Italy have a high prevalence (45.7%) of infestation with ixodid ticks and this risk varies by dog phenotype and lifestyle.

Ticks are hematophagous parasites that can transmit a variety of human pathogens, and their life cycle is dependent on several climatic factors for development and survival. We conducted a study in Piedmont and Aosta Valley, Italy, between 2009 and 2018. The study matched human sample serologies for Borrelia spp. with publicly available climatic and meteorological data. A total of 12,928 serological immunofluorescence assays (IFA) and Western blot (WB) tests were analysed. The median number of IFA and WB tests per year was 1236 (range 700–1997), with the highest demand in autumn 2018 (N = 289). In the study period, positive WB showed an increasing trend, peaking in 2018 for both IgM (N = 97) and IgG (N = 61). These results were consistent with a regional climatic variation trending towards an increase in both temperature and humidity. Our results suggest that coupling data from epidemiology and the environment, and the use of a “one health” approach, may provide a powerful tool in understanding disease transmission and strengthen collaboration between specialists in the era of climate instability.

Leishmaniasis is a heterogeneous group of neglected tropical diseases with various clinical syndromes, which is caused by obligate intracellular protozoa of the genus Leishmania and transmitted by the bite of a female phlebotomine sandfly. Humans and several animal species are considered as reservoirs of the disease. Among other animal species, dogs are the most important reservoirs in a domestic environment, maintaining the endemic focus of the parasite. The behavior of the disease progression and the clinical symptoms of the disease in the infected dog is mainly associated with depressed cellular immunity and strong humoral response. This study aimed to assess the role of Western blotting in the analysis of the idiotype expression of the two main immunoglobulins (IgG1 and IgG2) in dogs that are naturally infected with Leishmania infantum (L. infantum) and treated with N-methyl meglumine antimoniate. Interestingly, for the first time, our study identified several L. infantum antigen polypeptides (14, 31, 33, 49, 64, 66, 99, and 169 kDa) that more frequently stimulate an immune reaction in recovered dogs after treatment, whereas in the non-recovered group of dogs, four antigen polypeptides of L. infantum with molecular weights of 31, 49, 66, and 115 kDa with unfavorable prognosis were identified. Clearly, these interesting findings confirm the strong association between the detected immunodominant bands and the successful recovery in treated dogs that can be used for differentiating the treated dogs from the untreated dogs, as well as the markers of a favorable or unfavorable prognosis and, as a consequence, the prediction of the clinical outcome of the disease. Likewise, these data could be helpful in the implementation of novel vaccines from the detected antigens.

Leishmaniasis remains one of the major neglected tropical diseases. The epidemiological profile of the disease comprises a wide range of hosts, including dogs and cats. Despite several studies about feline Leishmaniosis, the role of cats in disease epidemiology and its clinical impact is still debated. The present study raises awareness about the impact of leishmaniasis in cats from an endemic region in of Northwestern Italy (Liguria). A total number of 250 serum and 282 blood samples were collected from cats, then assessed for Leishmania infantum (L. infantum) serologically using western blot (WB) and molecularly using polymerase chain reaction (PCR). We also tested the association of Leishmania infection with some infectious agents like haemotropic Mycoplasma, Feline immunodeficiency virus (FIV) and Feline leukemia virus (FeLV) together with the hematobiochemical status of the examined animals. Interestingly, all tested animals were asymptomatic and out of 250 examined serum samples, 33 (13.20%) samples (confidence interval (CI) 95% 9.56–17.96%) were positive at WB for L. infantum, whereas of the 282 blood samples, 80 (28.36%) returned a positive PCR (CI 95% 23.43–33.89%). Furthermore, there was a statistical association between PCR positivity for L. infantum and some hematological parameters besides FIV infection as well as a direct significant correlation between Mycoplasma infection and WB positivity. Taken together, the present findings report high prevalence of L. infantum among cats, which reinforces the significance of such positive asymptomatic animals and confirms the very low humoral response in this species. In addition, the laboratory values provide evidence that infection by the parasite is linked to alteration of some hematological parameters and is correlated to some infectious agents. These data are of interest and suggest future research for accurate diagnosis of such zoonosis.

In Europe, free-ranging wildlife has been linked to the emergence of several vector-borne diseases such as rodents for Borrelia burgdorferi s.l. In particular, wild carnivorans are one of the most important sources of emerging zoonotic pathogens worldwide, although little information is available regarding the epidemiology of vector-borne parasites in these animals. Thus, the aim of this paper was to investigate the prevalence of Babesia spp., Anaplasma spp., Ehrlichia spp., Hepatozoon spp. and Leishmania infantum in alpine wild canids and mustelids from Italy. For this study, spleen samples of 157 foxes (Vulpes vulpes), 45 badgers (Meles meles), and 33 wolves (Canis lupus) collected between 2009 and 2017 in Northwest Italy were examined by using conventional PCR. Logistic regression was used to identify possible risk factors for pathogen infections. DNA of any of the tested pathogens was found in more than 90% of the analyzed animals. In particular, Babesia spp. showed significantly higher prevalence in foxes (89.7%) and badgers (89.6%) than in wolves, while the latter were considerably more infected with Hepatozoon canis (75.8%) than foxes (5.1%). None of the badger tested positive for Hepatozoon spp., although they showed high prevalence of Leishmania infantum (53.3%). Sequencing results revealed the presence, among others, of Babesia vulpes, Babesia sp. isolate badger type A and B, and Anaplasma phagocytophilum. Moreover, previously unreported pathogen/host associations were observed, such as Babesia capreoli in wolves and badgers. The prevalence of vector-borne pathogens observed in the present study is one of the highest reported so far, suggesting the importance of free-ranging carnivorans in the epidemiology and maintenance of the sylvatic cycle of the pathogens. Moreover, several of these pathogens are of particular importance regarding human (A. phagocytophilum, L. infantum) and pet health (L. infantum, B. vulpes).

The aim of this study was to evaluate the effect of Hermetia illucens (HI) and Tenebrio molitor (TM) live larvae as environmental enrichment on the mucin composition, local immune response and microbiota of broilers. A total of 180 four-day-old male broiler chickens (Ross 308) were randomly allotted to three dietary treatments (six replicates/treatment; ten animals/replicate): (i) control (C); (ii) C+HI; (iii) C+TM. Live larvae were distributed based on 5% of the expected daily feed intake. At slaughter (39 days of age), samples of duodenum, jejunum and ileum (twelve animals/diet) were submitted to mucin histochemical evaluation. Expression of MUC-2 and cytokines was evaluated by rt-qPCR in jejunum. Mucin staining intensity was not influenced by diet (p > 0.05); however, this varied depending on the intestinal segment (p < 0.001). No significant differences were recorded for IL-4, IL-6 TNF-α, MUC-2 and INF-γ gene expression in jejunum, while IL-2 was lower in the TM group compared to HI and C (p = 0.044). Caecal microbiota showed higher abundance of Clostridium, Saccharibacteria and Victivallaceae in the HI group, while Collinsella was higher in the TM group. The results suggested that live insect larvae did not impair mucin composition or local immune response, and can slightly improve caecal microbiota by enhancing a minor fraction of short chain fatty acid-producing taxa.

Nematodes are an important cause of disease and loss of performance in horses. Changes in the parasitic fauna of horses have occurred in the past few decades, making cyathostomins the major parasites in adult horses, while large strongyles have become less prevalent. Parascaris spp. remains the most important parasite infecting foals and weanlings. Anthelmintic resistance is highly prevalent in cyathostomins and Parascaris spp. worldwide and it must be factored into treatment decisions. To assess anthelmintic efficacy in Northern Italy, we sampled 215 horses from 17 sport and horse-breeding farms. Fecal egg count reduction tests (FECRT) were used to assess anthelmintic efficacy. Copromicroscopic analysis was performed using MiniFLOTAC before treatment with fenbendazole, pyrantel pamoate or ivermectin, and repeated 14 days post-treatment. Strongyle-type eggs were detected in 66.91% of horses (CI95% 61.40–73.79%), while Parascaris spp. was detected in 2.79% (CI95% 1.94–5.95%). Reduced efficacy against cyathostomins was observed for fenbendazole in 55.56% of the treated animals (CI95% 41.18–69.06%), and for pyrantel pamoate in 75% of animals (CI95% 30.06–95.44%). Ground-based actions must be set in place to promote the uptake of state-of-the-art worm control plans that will prevent clinical disease while minimizing the selection pressure of resistant parasites.

Wild rodents are reservoirs of several Bartonella species that cause human bartonellosis. The aim of this study was to assess the presence of Bartonella spp. DNA in wild rodents in Pianosa island, Italy. Rats (Rattus spp.; n = 15) and field mice (Apodemus spp.; n = 16) were captured and spleen DNA tested for the presence of Bartonella spp. by means of an initial screening using a qPCR amplifying a short segment of the 16S-23S rRNA gene intergenic transcribed spacer region (ITS, ~200 bp) followed by conventional PCR amplification of a longer ITS fragment (~600 bp) and of a citrate synthase (gltA, ~340 bp) gene segment. A total of 25 spleen DNA samples obtained from 31 rodent carcasses (81%) yielded positive qPCR results. Bartonella genus was confirmed by amplicon sequencing. By conventional PCR, eight out of 25 samples (32%) yielded bands on gels consistent with ITS segment, and 6/25 (24%) yielded bands consistent with the gltA locus. Amplicon sequencing identified B. henselae and B. coopersplainsensis in 1/25 (4%), and 4/25 (16%) samples, respectively. Moreover, 5/25 (20%) of Bartonella spp. positive samples showed gltA sequences with about 97% identity to B. grahamii. These results provide support to recently published observations suggesting that B. henselae circulates in wild rodent populations.

Sarcocystis spp. are protozoan parasites with an obligatory two-host life cycle, with herbivores as intermediate hosts and carnivores as definitive hosts. Cattle are intermediate hosts for several species of Sarcocystis: indeed, in addition to S. cruzi, S. hirsuta and S. hominis, at least four new species were recently identified in bovine muscle: S. bovifelis, S. rommeli, S. bovini and S. heydorni. Since is not possible to unambiguously discriminate between S. hominis and the new species either morphologically or by the analysis of the 18S ribosomial (rRNA) gene, the aim of the present study was to use molecular techniques to discriminate cattle Sarcocystis species, taking advantage of the higher discriminative power of the Cytochrome C Oxidase subunit I mitochondrial (mtDNA COI) gene. Therefore, 119 bovine muscle samples were tested to identify S. hominis-like sarcocystis using a multiplex PCR of the 18S rRNA gene; later, positive samples were tested using a newly designed primer set for the PCR amplification of COI gene. Species identification was achieved by sequencing the amplified products: 16 sequences were confirmed to belong to S. bovifelis, while 12 sequences didn’t constitute the best BLAST match of any of the published sequences, allowing to speculate the possible presence of S. hominis. This study confirms the higher discriminatory power of COI mitochondrial gene; besides, our work provides the first report of S. bovifelis in Italy.

Introduction: A parasite community is usually well adapted and specific to the host species they co-evolved with. Although exotic pathogens infecting autochthonous species have been documented, the infection of an alien species with native parasites is rare in lagomorphs. Trichostrongylus retortaeformis is a nematode parasite infecting the small intestine of domestic and wild lagomorphs in Europe. Methods: Thirty-two Eastern cottontails from a naturalized population in Italy were processed to describe the gastrointestinal parasite community. Results and discussions: T. retortaeformis is reported for the first time in the Eastern cottontail Sylvilagus floridanus introduced to Europe. The Eastern cottontail is an invasive lagomorph, living in sympatry with the autochthonous European brown hare in certain areas of Italy. This study provides new insights into the dynamics of parasite communities of native and alien lagomorph species in sympatric areas.

Abstract Background Toxoplasma gondii is an apicomplexan parasite that is able to infect almost all warm blooded animals. In Europe, the domestic cat is the main definitive host. Worldwide, 6 billion people are infected with this parasite. The goal of our research is to evaluate the prevalence of T. gondii infection in wild animals from a previously unsampled area in Northern Italy where 0.1% of women seroconvert during pregnancy each year. Methods We sampled and tested skeletal muscle and central nervous system tissue of 355 wild animals by PCR (n = 121 roe deer Capreolus capreolus, n = 105 wild boar Sus scrofa, n = 94 red fox Vulpes vulpes, n = 22 alpine chamois Rupicapra rupicapra, n = 13 red deer Cervus elaphus). Results The overall prevalence of infection with T. gondii was 10.99% (confidence interval (CI) 95% 8.14%-14.67%). A higher rate of infection was recorded in carnivores and omnivores (red fox 20.21%, CI 95% 13.34%-29.43%; wild boar 16.19%, CI 95% 10.36%-24.41%) compared to ruminants (2.48%, CI 95% 0.85%-7.04% in roe deer; 0.00%, CI 95% 0.00%-22.81% in red deer, and 0.00% alpine chamois (CI 95% 0.00%-14.87%) confirming the importance of tissue cysts in transmitting infection. Conclusions The relatively high prevalence of T. gondii DNA in highly consumed game species (wild boar and roe deer) gives valuable insights into T. gondii epidemiology and may contribute to improve prevention and control of foodborne toxoplasmosis in humans.

Abstract Background Piroplasmosis are among the most relevant diseases of domestic animals. Babesia is emerging as cause of tick-borne zoonosis worldwide and free-living animals are reservoir hosts of several zoonotic Babesia species. We investigated the epidemiology of Babesia spp. and Theileria spp. in wild ungulates and carnivores from Northern Italy to determine which of these apicomplexan species circulate in wildlife and their prevalence of infection. Methods PCR amplification of the V4 hyper-variable region of the 18S rDNA of Babesia sp./Theileria sp was carried out on spleen samples of 1036 wild animals: Roe deer Capreolus capreolus (n = 462), Red deer Cervus elaphus (n = 52), Alpine Chamois Rupicapra rupicapra (n = 36), Fallow deer Dama dama (n = 17), Wild boar Sus scrofa (n = 257), Red fox Vulpes vulpes (n = 205) and Wolf Canis lupus (n = 7). Selected positive samples were sequenced to determine the species of amplified Babesia/Theileria DNA. Results Babesia/Theileria DNA was found with a mean prevalence of 9.94% (IC95% 8.27-11.91). The only piroplasms found in carnivores was Theileria annae, which was detected in two foxes (0.98%; IC95% 0.27-3.49). Red deer showed the highest prevalence of infection (44.23%; IC95% 31.6-57.66), followed by Alpine chamois (22.22%; IC95% 11.71-38.08), Roe deer (12.55%; IC95% 9.84-15.89), and Wild boar (4.67%; IC95% 2.69-7.98). Genetic analysis identified Babesia capreoli as the most prevalent piroplasmid found in Alpine chamois, Roe deer and Red deer, followed by Babesia bigemina (found in Roe deer, Red deer and Wild boar), and the zoonotic Babesia venatorum (formerly Babesia sp. EU1) isolated from 2 Roe deer. Piroplasmids of the genus Theileria were identified in Wild boar and Red deer. Conclusions The present study offers novel insights into the role of wildlife in Babesia/Theileria epidemiology, as well as relevant information on genetic variability of piroplasmids infecting wild ungulates and carnivores.

Alpine marmots Marmota marmota occupy a narrow altitudinal niche within high elevation alpine environments. For animals living at such high elevations where resources are limited, parasitism represents a potential major cost in life history. Using occupancy models, we tested if marmots living at higher elevation have a reduced risk of being infected with gastro-intestinal helminths, possibly compensating the lower availability of resources (shorter feeding season, longer snow cover and lower temperature) than marmots inhabiting lower elevations. Detection probability of eggs and oncospheres of two gastro-intestinal helminthic parasites, Ascaris laevis and Ctenotaenia marmotae, sampled in marmot feces, was used as a proxy of parasite abundance. As predicted, the models showed a negative relationship between elevation and parasite detectability (i.e. abundance) for both species, while there appeared to be a negative effect of solar radiance only for C. marmotae. Site-occupancy models are used here for the first time to model the constrains of gastrointestinal parasitism on a wild species and the relationship existing between endoparasites and environmental factors in a population of free-living animals. The results of this study suggest the future use of site-occupancy models as a viable tool to account for parasite imperfect detection in eco-parasitological studies, and give useful insights to further investigate the hypothesis of the contribution of parasite infection in constraining the altitudinal niche of Alpine marmots.