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Nematodes are abundant and ubiquitous animals which are poorly known at intraspecific level. This work represents the first attempt to fill the gap on basic knowledge of genetic variability and differentiation in Protostrongylus oryctolagi , a nematode parasite of lagomorphs. 68 cox1 sequences were obtained from brown hares collected in five locations in Northern and Central Italy, highlighting the presence of a high amount of genetic variation inside this species. The eleven haplotypes identified (Haplotype diversity equal to 0.702) were split into two lineages: lineage A (comprising six different haplotypes, A1-A6) and lineage B (B1-B5). The mean intra-lineage amount of genetic variation was 0.3%, whereas the inter-lineage percentage of variation was ten-fold higher (3%). These two lineages were non-randomly distributed in the investigated areas. Lineage A showed a preference for Central Italy (Tuscany) even if it was sporadically found also in northern territories (Emilia-Romagna), while B-haplotypes were present exclusively in Emilia-Romagna. The analysis of molecular variance identified two main barriers to gene flow: (i) a strong major one which separate samples of Central Italy (PIA and GR7) from the northern ones (RE1, RE3 and MO1; Φ ST = 0.750, P = 0.00); (ii) a secondary faint barrier which separates Pianosa island from Grosseto (Φ ST = 0.133, P = 0.00). Any difference was found among northern samples (Φ ST = 0.009, P = 0.00). The observed data may be explained by several factors ranging from the parasite’s biology (presence of a narrow host spectrum), the final host’s behaviour (small home range), the natural dispersion of the host-parasite dyad occurred in past or the recent passive men-mediated migration. Finally, the presence of unconventional shortened amplicons revealed the presence of NUMTs (nuclear copy of mitochondrial genes) in the P . oryctolagi nuclear genome, suggesting caution when using DNA barcode as unique marker for the identification of species belonging to this genus. “ In short , if all the matter in the universe except the nematodes were swept away , our world would still be dimly recognizable ”. Nathan Augustus Cobb, from \"Nematodes and Their Relationships\", 1915

Gastrointestinal nematodes (GIN) represent a major constraint to sheep production, and sustainable alternatives to routine anthelmintic use are increasingly required. This study compared two parasite control strategies in Zerasca sheep: routine albendazole treatment administered twice yearly (T) and a long-term non-chemical approach based on rotational grazing (relocation to a new pasture when grass height fell below 10 cm) combined with quarterly veterinary monitoring (NT). Twenty-four adult ewes (n = 12 per group) were monitored over an eight-month period. Mean faecal egg counts (EPG) differed significantly between groups (T: 210 ± 78; NT: 529 ± 89; p = 0.0007). In group T, EPG values decreased markedly 7 days after treatment but increased again by 150 days, resulting in no persistent differences between groups over time. Despite higher parasite burdens, NT sheep showed significantly higher body condition scores compared with treated animals (3.00 ± 0.61 vs. 2.51 ± 0.53; p = 0.0014). Haematological parameters were largely comparable between groups, although mild reductions in RBC, HGB, and HCT were observed in both. Treated sheep exhibited higher AST activity (p < 0.0001) and transient increases in ALT and BUN following treatment. Hair cortisol concentrations did not differ significantly between groups. Overall, these findings suggest that a non-chemical parasite management strategy, when combined with controlled grazing and veterinary monitoring, may maintain acceptable parasite levels while supporting body condition and stable welfare indicators, potentially reducing reliance on routine anthelmintic treatments.

Background This study investigates the prevalence of leishmaniasis and American trypanosomiasis (Chagas disease) – two neglected vector-borne diseases – in humans and dogs in the Bolivian Chaco region, where high poverty levels increase population vulnerability. Leishmaniasis, which affects millions globally, is widespread in Bolivia, a country reporting some of the highest rates of cutaneous and mucocutaneous cases in Latin America. Chagas disease is endemic across the region, with an estimated 4.5 million individuals affected. Methods Blood and serum samples were collected from 189 dogs living in rural communities near the cities of Camiri and Villa Montes. Samples were tested using serological assays and quantitative PCR (qPCR) to detect Leishmania spp. and Trypanosoma cruzi . Additionally, serum samples from 151 school-aged children from both areas were screened for anti- Leishmania antibodies. Results The seroprevalence of Leishmania in dogs was significantly higher in Villa Montes (46%) compared with Camiri (26%). Moreover, Leishmania DNA was detected by qPCR in 9 out of 125 dogs tested (7.2%). Among children, 13.2% from Villa Montes tested positive for Leishmania antibodies, while no positive cases were found in Camiri. Serological evidence of previous T . cruzi infection was identified in 17.7% of dogs, although all qPCR results for T . cruzi were negative. Conclusions These findings highlight the relevance of a One Health approach, as dogs may serve as reservoirs for both parasites, potentially increasing the risk of human transmission. Integrated control measures – including vector management and ongoing surveillance – are essential to reduce transmission and protect public health. Future research should prioritise mapping infection patterns and exploring ecological factors influencing disease dynamics. Graphical Abstract

The gut microbiota plays a basic role in maintaining the psychophysical health and well-being of horses. Understanding the complex interactions among microbial communities in relation to age, management, and health is a topic of growing interest. The equine microbiota, given its complexity, is subject to variations caused by internal or external stressors that can lead to metabolic problems and pathologies, i.e., obesity and laminitis. The aim of this study was to characterize the fecal microbiota of 12 purebred Arabian horses and to compare the microbial communities in two distinct management systems (Group 1 = box 22 h/day + paddock 2 h/day and Group 2 = paddock 24 h). Fecal samples were analyzed using high-throughput sequencing of 16S rRNA V3-V4 amplicons. The microbiota was predominantly composed of Firmicutes (32–53%) and Bacteroidetes (32–47.8%), with Ruminococcaceae emerging as the most prevalent bacterial family (10.9–24%). Comparisons of alpha and beta diversity revealed no statistically significant differences between the groups based on different management systems. This preliminary characterization contributes valuable data for understanding the equine fecal microbiota and lays the groundwork for future studies on the relationship among intestinal microbiota and equine health, performance, and management strategies.

This study reports the first case of bronchial myiasis in a dog caused by Oestrus ovis, a parasite known to infest sheep, and occasionally other mammals. A 9‐year‐old dog, from a rural area near Ancona (Italy), exhibited persistent coughing despite antibiotic and anti‐inflammatory treatments. Bronchoscopy revealed a live first‐instar larva of O. ovis, confirmed through morphological and molecular analyses. Larval removal and treatment with isoxazolines resulted in a rapid resolution of symptoms. This case highlights the critical importance of including atypical parasitic infestations in the differential diagnosis of respiratory symptoms in dogs. It underscores the importance of heightened vigilance, particularly in areas with sheep flocks, to prevent and manage such infestation effectively. Oestrus ovis bronchial myasis is described in a dog for the first time. Bronchoscopy proved essential for larval visualization and removal. Isoxazoline compounds should be considered both as a treatment and as a preventive option.

The Italian wolf population in human-modified landscapes has increased greatly in the last few decades. Anthropisation increases the risk of transmission of many zoonotic infections and in this context, control of taeniid cestode species needs to be addressed from a One Health perspective. Predator-prey interactions are at the root of taeniid cestode transmission, and the wolf plays a key role in the maintenance and transmission of taeniids. To date, all available data on the taeniids of wolves in Italy refer to populations living in a wild habitat. Between 2018 and 2019, we investigated taeniids in a wolf pack living in a highly anthropic hilly agro-ecosystem. Thirty-eight faecal samples were collected and analysed, 4 of which were also genetically characterised for individual wolves and belonged to three different animals. Samples collected were analysed microscopically and by molecular analysis in order to identify the taeniid species. Taeniid eggs were detected in 34.2% (13/38) of samples. Within samples positive to taeniid eggs only Echinococcus granulosus s.s. and Taenia hydatigena were identified in 26.3% and 10.5% of the samples, respectively. On microscopic examination, Capillaria spp., Ancylostomatidae and Toxocara canis eggs, Crenosoma vulpis larvae, and coccidian oocysts were also found. The combination of low biodiversity of taeniid species with a high occurrence of E. granulosus s.s. recorded in this study could be the consequence of a deeper link occurring between wolves and livestock in human-modified landscapes than in wild settings. La population de loups italiens dans les paysages modifiés par l’homme a considérablement augmenté au cours des dernières décennies. L’anthropisation augmente le risque de transmission de nombreuses infections zoonotiques et, dans ce contexte, le contrôle des espèces de cestodes Taeniidae doit être abordé dans une perspective One Health. Les interactions prédateurs-proies sont à la base de la transmission des Cestodes Taeniidae, et le loup joue un rôle clé dans le maintien et la transmission des Taeniidae. À ce jour, toutes les données disponibles sur les Taeniidae du loup en Italie se réfèrent aux populations vivant dans un habitat sauvage. Entre 2018 et 2019, les Taeniidae d’une meute de loups vivant dans un agro-écosystème vallonné hautement anthropique ont été étudiés. Trente-huit échantillons fécaux ont été collectés et analysés, dont quatre ont également été génétiquement caractérisés pour des loups individuels et appartenaient à trois animaux différents. Les échantillons prélevés ont été analysés au microscope et par analyse moléculaire afin d’identifier les espèces de Taeniidae. Des œufs de Taeniidae ont été détectés dans 34,2 % (13/38) des échantillons. Dans les échantillons positifs aux œufs de Taeniidae, seul Echinococcus granulosus s.s. et Taenia hydatigena ont été identifiés dans 26,3 % et 10,5 % des échantillons, respectivement. Lors de l’examen microscopique, des œufs de Capillaria spp., d’Ancylostomatidae et de Toxocara canis , des larves de Crenosoma vulpis et des oocystes de coccidie ont également été trouvés. La combinaison d’une faible biodiversité d’espèces de Taeniidae avec une forte occurrence d’ E. granulosus s.s. rapportées dans cette étude pourrait être la conséquence d’un lien plus profond entre le loup et le bétail dans les paysages humains que dans les paysages sauvages.

Studies on short- and long-term memory and their decline with aging in horses are still limited. This research aimed to assess the learning and short- and long-term memory abilities of adult and senior horses. A total of 21 adult horses (5–15 years) and 23 senior horses (16 years and older) were subjected to a behavioral test (Target Touch Test), which was repeated three times each for the initial test (T1) and again after 10 days (T10). Statistical analysis revealed no significant differences between adult and senior horses in the three tests conducted at T1. However, a significant difference was observed at T10 (χ2 = 42.1; p < 0.001). In fact, senior horses took longer to complete the test than adults. A significant difference between males and females, adults and seniors, at T1 and T10 (χ2 = 56.3; p < 0.001) was found, but no difference between male and female adult and senior horses in completing the three single tests at T1 and T10 was evidenced. In conclusion, even senior horses are capable of associative learning, maintaining the memory of this learning even after 10 days. However, they show slower recovery times for recalling memorized information compared to animals under 16 years of age.

A cross-sectional study on Trypanosoma cruzi was carried out in 2013 to evaluate the role of dogs as possible source of infection for humans in two rural communities of the highly endemic Bolivian Chaco (Bartolo, Chuquisaca Department, n = 57 dogs; and Ivamirapinta, Santa Cruz Department, n = 48 dogs). Giemsa-stained thick and thin smears, rapid immunochromatographic test (ICT) (Chagas Quick test, Cypress Diagnostic, Belgium) and polymerase chain reaction for T. cruzi on dried blood spots were performed. All smears proved negative by microscopic examination, whereas 23/103 (22%) were positive by ICT and 5/105 (5%) blood samples contained T. cruzi DNA, evidencing the potential role of dogs in the domestic transmission of the parasite.

Introduction: Chagas disease (CD) remains a public health concern in several Latin American countries. At global level, Bolivia has the highest CD burden and the Chaco region, in the southeast of the country, is the most affected area. We report the results of four serosurveys for Trypanosoma cruzi antibodies, carried out approximately ten years apart from each other, during the lapse 1987-2013, in different localities of the Bolivian Chaco. Methodology: Four cross-sectional surveys were conducted in various localities, mostly rural, of the Bolivian Chaco, during the period 1987-2013. Results: Although a reliable analysis of CD epidemiological trend is challenging, a partial reduction of anti-T. cruzi seroprevalence over the past four decades in the Bolivian Chaco may be assumed. In particular, in 1987 the exposure to T. cruzi in rural setting was universal since the first years of life, while it resulted gradually lower and age-dependent thereafter. Moreover, T. cruzi seroprevalence among women of reproductive age (15-45 years) has been persistently high in rural areas. Conclusions: T. cruzi transmission is still active and CD remains a concern throughout the Bolivian Chaco. More efforts are needed in order to achieve a sustainable interruption of vector-borne CD transmission in this area.

Introduction: In 2013 a coproparasitological survey was carried out in two rural communities of the Bolivian Chaco to determine the prevalence of intestinal parasitic infections (IPIs) and to investigate on possible infection drivers through a questionnaire interview. Methodology: Faecal samples were examined by microscopy. Samples positive for Entamoeba histolytica complex and Blastocystis were molecularly examined to identify the species/subtypes involved. Results: The overall infection rate was 86%, identical in both communities and mostly due to protozoa. Soil-transmitted helminths were detected in <3% of children and adults. Discussion: The protozoa detected, including Blastocystis subtypes, indicate faecal contamination of the environment by both humans (as confirmed by the presence of Hymenolepis nana) and animals. Nested-PCR identified E. histolytica, thus signalling the possible occurrence of invasive amoebosis. Lack of safe water, environmental contamination, poor sanitation and hygiene, shared by both communities, are the main drivers of IPIs. In addition, unlike gender and socioeconomic factors, childhood (only for some species), crowding and cohabitation with animals proved to be further significant protozoon infection risk factors. Conclusions: These results highlight the need for the promotion of access to clean water, improved sanitation and better hygiene, thus reducing the frequency of preventive chemotherapy for STHs while continuing to monitor the population for possible recrudescence.