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Tapeworms are trophically-transmitted and multi-host parasites with a complex indirect life cycle, strictly depending on predator-prey interactions. Their presence in a free-living population, mainly definitive hosts, is arduous to study due to the complexity of collecting fecal samples. However, epidemiological studies on their frequency are crucial from a public health perspective, providing information on food habits and prey selection of predators. The present study aims to update the frequency of tapeworms detected in stool samples by molecular analysis in Italian wolf populations of Umbria and Marche regions collected from 2014 to 2022. Tapeworm's total frequency was 43.2%. In detail, Taenia serialis was detected in 27 samples (21.6%), T. hydatigena in 22 (17.6%), and Mesocestoides corti (syn. M. vogae) in 2 (1.6%). Three samples were identified as M. litteratus and E. granulosus s.s. (G3) and T. pisiformis, with a proportion of 0.8%, respectively. The low frequency of E. granulosus in a hyperendemic area is discussed. The results show for the first time a high frequency of Taenia serialis not comparable to other Italian studies conducted on wild Carnivora; thus, a new ecological niche is conceivable. These findings suggest a plausible wolf-roe deer cycle for T. serialisin the investigated area.

Radical cystectomy (RC) with pelvic lymph node dissection is the standard treatment for patients with limited-stage muscle-invasive bladder cancer. RC is associated with a complication rate of approximately 50–88%. Immunonutrition (IMN) refers to the administration of substrates, such as omega-3 fatty acids, arginine, glutamine, and nucleotides, that modulate the immune response. IMN has been associated with improved outcomes following surgery for esophagogastric, colorectal and pancreatic cancer. In this paper, we describe a study protocol for a multicentre, randomised, open-label clinical trial to evaluate the effect of IMN in patients undergoing RC for bladder cancer. A 7-day preoperative course of IMN is compared with a standard high-calorie high-protein oral nutritional supplement. The primary outcome of this study is the rate of complications (infectious, wound-related, gastrointestinal, and urinary complications) in the first 30 days after RC. Secondary outcomes include time to recovery of bowel function and postoperative mobilisation, changes in muscle strength and body weight, biochemical modifications, need for blood transfusion, length of stay, readmission rate, and mortality. The results of this study may provide new insights into the impact of IMN on postoperative outcomes after RC and may help improve IMN prescribing based on patient nutritional status parameters.

We have read with interest the comments and observations made regarding the paper “A terbinafine sensitive Trichophyton indotineae strain in Italy: the first clinical case of tinea corporis and onychomycosis” [...]

Background Malnutrition is a frequent problem in oncology and is associated with reduced response to cancer treatments, increased drug-related toxicity, higher rates of clinical complications, reduced quality of life (QoL) and worse prognosis. Guidelines on clinical nutrition in oncology emphasise the usefulness of early assessment of nutritional status for a prompt identification of malnutrition and the implementation of effective interventions, but no real-world clinical data are available on the adequate management of nutritional support in patients with cancer in Italy.Methods and analysis This is an observational, longitudinal, multicentre registry of patients with a new diagnosis of cancer or metastatic disease, candidates for active treatment. They will be identified in at least 15 Italian oncological centres, members of the Alliance Against Cancer Working Group ‘Survivorship Care and Nutritional Support’. At least 1500 patients with cancer are expected to be enrolled each year. Detailed clinical and nutritional data will be collected by oncologists and clinical nutritionists during the visits foreseen in the clinical practice, through an ad hoc developed digital platform (e-Nutracare). The effects of malnutrition and nutritional support—at diagnosis and during follow-up—on overall survival and progression-free survival, as well as on patients’ symptoms and QoL, will be investigated.Ethics and dissemination The study protocol was approved by the Ethics Committee of the Fondazione IRCCS Policlinico San Matteo, Pavia, Italy and from the Ethics Committees of all other participating centres. An informed consent will be obtained from each patient enrolled in the study. Study findings will be disseminated through peer-reviewed journals, conferences and patients with cancer or professional associations. The registry will allow a better monitoring of the nutritional status of patients with cancer, promoting adequate and sustainable nutritional support, with the ultimate goal of improving the care and prognosis of these patients.

Abstract Background Sarcopenia gained importance in the evaluation of patients with chronic respiratory diseases, including idiopathic pulmonary fibrosis (IPF), since it may impact negatively on clinical outcomes. Aim Aim of this study is to evaluate the prevalence and factors associated with sarcopenia, defined according to the European Working Group on Sarcopenia in Older People 2 (EWGSOP2) 2019 definition, and to evaluate the prevalence of the single criteria that define the EWGSOP2 definition (muscle strength, muscle quantity and physical performance), in a cohort of consecutive patients with IPF prospectively followed up in 9 hospitals in Northern Italy between December 2018 and May 2021. Methods Enrolled patients underwent an extensive pulmonary and nutritional assessment, including bioelectrical impedance analysis, dynamometry and 4-m gait speed test, both at IPF diagnosis and at 6-month follow-up. Results Out of the 83 patients (81% males, mean age 72.5 years) with IPF at disease diagnosis enrolled in the study, 19 (22.9%) showed sarcopenia, including 2 (2.4%) with severe sarcopenia, 5 (6.0%) with confirmed sarcopenia and 12 (14.5%) with probable sarcopenia. Sarcopenia was associated with a significantly higher severity of the disease and sedentary lifestyle, while no differences were observed in regards to body mass index, history of weight loss and comorbidities between patients with and without sarcopenia. Out of the 64 patients without sarcopenia at baseline, 16 cases showed alteration of muscle quantity and/or physical performance. In the 51 patients with complete data at 6-month follow-up, there were no cases of severe sarcopenia, 1 case (2.0%) showed confirmed sarcopenia, while the prevalence of probable sarcopenia was 19.6% (10 cases). No differences in regards to antifibrotic treatment received and onset of gastrointestinal side effects were observed between patients with and without sarcopenia at follow-up. Conclusions The prevalence of sarcopenia in patients with IPF both at diagnosis and at 6-month follow-up was low but not negligible and was associated with higher severity of the disease and sedentary lifestyle. In IPF patients, a comprehensive diagnostic work-up including all the criteria defining the EWGSOP2 definition might be more useful than a series testing for prompt recognition of nutritional and physical performance abnormalities.

Taeniids are multi-host parasites with an indirect life cycle that strictly depends on a predator-prey relationship. Parasites with a complex life cycle may exhibit different degrees of host-specificity at each life stage. Knowing the host breadth is a fundamental concept of the biology and epidemiology of these multi-host parasites. Morphological identification of tapeworms is challenging and occasionally may produce misdiagnosis. Thus, molecular investigations were carried out for the identification of parasitic cysts detected from muscle tissues in a male roe deer necropsied at the Istituto Zooprofilattico Sperimentale dell’Umbria e delle Marche “Togo Rosati” (Central Italy). Sanger sequencing showed 99% query cover, 2e-109 e-value, and 100% identity with Taenia serialis. The exact definitive host was not revealed in this report, but red foxes and Italian wolves may play a significant role as being widespread within the area. Wildlife surveillance is crucial to monitor for human and animal health since global distribution and flexibility in intermediate hosts of many and even more critical taeniids species may enlarge their host range.

Abstract Background Low muscle strength has been pointed out as a key characteristic of sarcopenia, but the prognostic significance of muscle function next to reduced skeletal muscle mass (SMM) in patients with cancer has been scantily investigated. Methods Data on muscle strength by handgrip (HG) dynamometry and total‐body SMM estimated by bioelectrical impedance analysis (BIA) of Italian and German patients with cancer observed prospectively until death or censoring were analysed (N = 1076). Patients were stratified in four risk categories based on low HG (60 months for both). After adjusting for sex, age, body mass index and percentage of weight loss, disease's stage, performance status and type of cancer, compared to reference category (normal HG and SMM; N = 210) the hazard ratios were: low SMM/normal HG (N = 342), 0.83 [95% confidence interval, CI, 0.67–1.02] (p = 0.073); normal SMM/low HG (N = 158), 1.19 [95% CI, 1.07–1.32] (p = 0.002); low SMM/low HG (N = 366), 1.39 [95% CI, 1.27–1.53] (p

Trichophyton indotineae is an emerging dermatophyte species that plays a relevant role in human healthcare. It has been associated with severe chronic skin infections and a high level of terbinafine resistance. T. indotineae is endemic to India, Iran, and Iraq but several cases have been reported in Europe, recently. In this manuscript, the authors report the first clinical description of a tinea corporis and onychomycosis due to T. indotineae. The patient was a 42-year-old female from India that has lived in Umbria (Central Italy) for the last two years. Firstly, a dermatological examination suggested dermatophytosis: mycology isolation from cultures and macro- and microscopical features identified the colonies as belonging to the T. mentagrophytes/T. interdigitale species complex. Subsequently, ITS1/ITS4 end-point PCR and Sanger sequencing identified the strain as T. indotineae. Lastly, a DermaGenius® Resistance Multiplex real-time PCR assay was carried out, targeting the mutations in the SQLE gene to establish terbinafine resistance or susceptibility of the strain. The melting curve observed was compatible with wild-type positive control, identifying the strain as T. indotineae terbinafine-sensitive. An oral terbinafine treatment was associated with a topical ciclopirox nail solution, resulting in remission in its clinical manifestation. On 3 July 2023, the local Prevention Service notified the case to the Ministry of Health that then reported the information at national and international levels.

IntroductionSeveral studies and meta-analyses suggested the role of vitamin D 25OH in preventing severe forms of coronavirus disease 2019 (COVID-19). However, the evidence on the clinical benefits of vitamin D 25OH adequacy in patients hospitalized for COVID-19 remain conflicting and speculative. We aimed to investigate the association between vitamin D 25OH serum levels and mortality in hospitalized patients with moderate to severe COVID-19.MethodThis prospective observational multicentre study included 361 consecutive patients with moderate to severe COVID-19 admitted to the Italian hospitals involved in the NUTRI-COVID19 trial from March to August 2020. For each patient, serum vitamin D 25OH levels were assessed 48 h since admission and classified as deficient (

Tapeworm infections are among the most relevant parasitic diseases in humans and animals. Tapeworms from the Genus Echinococcus are particularly important as they can cause cystic or alveolar echinococcosis. A molecular screening was performed on 279 fecal samples collected from carcasses of wild carnivores from Central Italy using PCR targeting diagnostic fragments of nad1, rrnS, and nad5 genes. Samples positive for either Taenia spp. or Echinococcus granulosus were sequenced to taxonomically identify the parasitic DNA. Of the 279 samples, 134 (48.0%) gave positive results in the multiplex PCR. Only one (0.4%) sample from an Apennine wolf tested positive for Echinococcus granulosus sensu stricto (genotype G3), whereas no sample tested positive for E. multilocularis. The most frequently detected tapeworms were: Mesocestoides corti (syn M. vogae) (12.9%), M. litteratus (10.8%), Taenia serialis (9.3%), and T. hydatigena (6.5%), other tapeworms were rarely detected. The results suggest that Echinococcus infections in Central Italy do not seem to be sustained by sylvatic cycles, confirming the absence of E. multilocularis in Central Italy. The survey corroborates, yet again, the importance of passive surveillance of wild animals that can serve as reservoirs for zoonotic pathogens, especially on wild canids that in other areas are strongly implicated in the transmission of E. granulosus and E. multilocularis.

Abstract Background Toxoplasmosis is one of the most common parasitic infections in both humans and animals. It is a frequent cause of abortion and stillbirth in intermediate hosts, especially sheep and goats but rarely causes fatal clinical form in adult animals. Case presentation In contrast, the study reports an unusual fatal case of toxoplasmosis in a young goat naturally infected with type II strain of Toxoplasma gondii. A three-month-old female goat was presented with dyspnea and died few days later. Grossly, lungs were firm, edematous and mottled with disseminated whitish areas. Generalized lymphadenopathy was found. The histopathological examination showed necrotic interstitial bronchopneumonia and necrotizing lymphadenitis with intralesional free and clustered within macrophages tachyzoites of T. gondii. DNA extracted from lungs and lymph nodes was positive for T. gondii by a fast qPCR. PCR-RFLP analysis and sequencing of GRA6 gene showed that the isolated strains belonged to type II genotype. Conclusions This is an unusual report of acute systemic toxoplasmosis caused by the type II strain of T. gondii with a fatal outcome in a young goat.

Background Nutritional status impacts quality of life and prognosis of patients with respiratory diseases, including idiopathic pulmonary fibrosis (IPF). However, there is a lack of studies performing an extensive nutritional assessment of IPF patients. This study aimed to investigate the nutritional status and to identify nutritional phenotypes in a cohort of IPF patients at diagnosis. Methods Patients underwent a thorough pulmonary and nutritional evaluation including questionnaires on nutritional status, and physical activity, anthropometry, body impedance, dynamometry, 4-m gait speed and blood tests. Results 90 IPF patients (78.9% males, mean age 72.7 years) were enrolled. The majority of patients were classified as Gender-Age-Physiology Index stage 2 (47, 52.2%) with an inactive lifestyle according to International Physical Activity Questionnaire score (39, 43.3%), and had mean forced vital capacity and diffusing capacity for carbon monoxide 86.5% and 54.2%, respectively. In regards to nutritional phenotypes, the majority of patients were normally nourished (67.8%, 95% CI 58.6–77.7%), followed by non-sarcopenic obese (25.3%, 95% CI 16.1–35.2%), sarcopenic (4.6%, 95% CI 0.0–14.5%) and sarcopenic obese (2.3%, 95% CI 0.0–12.2%). Among the normally nourished, 49.2% showed early signs of nutritional and physical performance alterations, including body mass index ≥30 kg·m−2 in 4.3%, history of weight loss ≥5% in 11.9%, and reduction of gait speed and hand grip strength in 11.9% and 35.6%, respectively. Low vitamin D values were observed in 56.3% of cases. Conclusions IPF patients at diagnosis are mainly normally nourished and obese, but early signs of nutritional and physical performance impairment can already be identified at this stage.

Brucella canis has been isolated for the first time in Italy in a commercial breeding kennel. It was diagnosed after a deep investigation related to the onset of reproductive disorders. Animals were tested with direct and indirect techniques. The agent was first detected in two Chihuahua aborted foetuses by direct culture. Further, it was also isolated from blood samples of dogs hosted in the kennel, which also showed reaction to conventional serological tests (microplate serum agglutination test). The isolates were identified as B. canis by standard microbiological methods and a Bruce‑ladder multiplex PCR. To investigate the genomic diversity, whole genome sequencing was used, applying the core genome Multilocus Sequence Typing (cgMLST ). In a first round of serological testing performed on 598 animals, 269 (46.1%) tested positive. In the second round of laboratory testing carried out 4‑5 weeks apart, the number of serologically positive dogs was 241 out of 683 tested (35.3%), while the number of dogs positive to isolation was 68 out of 683 tested (10.0%). The PCR showed a lack of sensitivity when compared to direct isolation. The epidemiological investigation did not identify the source of the infection, given the time elapsed from the onset of abortions to the definitive diagnosis of B. canis infection in the kennel. The genomic analyses featured the strains as ST21 and, according to the cgMLST, revealed the presence of a tight cluster with a maximum diversity of four allelic differences. The observed limited genomic variation, largely within the known outbreak cut‑offs, suggests that the outbreak herein described was likely caused by a single introduction. Moreover, in a broader scale comparison using the public available genomes, we found that the closest genome, isolated in China, differed by more than 50 alleles making not possible to find out the likely origin of the outbreak. The lack of updated data on B. canis genome sequences in the public databases, together with the limited information retrieved from the epidemiological investigations on the outbreak, hampered identification of the source of B. canis infection.

Among the fungi responsible for deep mycosis, the genus Aspergillus plays a predominant role both in human and veterinary medicine. From a “One Health” perspective, infections by Aspergillus spp. often represent a public health problem linked to specific occupational categories that could have a greater risk of inhaling spores and developing any respiratory disease. This preliminary investigation allowed to acquire information about the spread of Aspergillus spp. in avian livestock of the Umbria region (Central Italy), their sensitivity to antifungals, and the presence of mycotoxins in the considered farms. Environmental, feed, animal, and human samples were collected for mycological investigations; chemical analyses were also performed in feed samples. Moreover, prevalence estimated of the fungal isolates were provided for each individual farm sampled. Direct fungal identification was possible in 298 out of the 559 total samples; 162 of the samples were positive for Aspergillus spp. Mycotoxins were detected in 5 out of the 21 feed samples collected. All the aspergilli tested for antifungal susceptibility were resistant to fluconazole. The results obtained show how much the genus Aspergillus is widespread in the investigated farms; therefore, the poultry livestock represents a favorable environment for the maintenance and spread of fungal spores and their potential transmission to animals and humans.

A direct sandwich enzyme-linked immunosorbent assay (sELISA) was developed for the detection of the atypical β2-toxin (CPB2) of Clostridium perfringens. Polyclonal (PAbs) and monoclonal (MAbs) antibodies were previously obtained employing recombinant CPB2 produced in the baculovirus system as antigen. In the current study, PAbs were used as capture molecules, while purified MAbs conjugated to horseradish peroxidase (MAbs-HRP) were used for the detection of atypical CPB2 toxin. MAbs 5C11E6 and 2G3G6 showed high reactivity, sensitivity and specificity when tested on 232 C. perfringens cell culture isolates. In addition, a reactivity variation among different strains producing atypical CPB2 toxin was observed using the conformation-dependent MAb 23E6E6, suggesting the hypothesis of high instability and/or the existence of different three-dimensional structures of this toxin. Results obtained by sELISA and Western blotting performed on experimentally CPB2-contaminated feces revealed a time-dependent proteolytic degradation as previously observed with the consensus allelic form of CPB2. Finally, the sELISA and an end-point PCR, specific for the atypical cpb2 gene, were used to test field samples (feces, rectal swabs and intestinal contents) from different dead animal species with suspected or confirmed clostridiosis. The comparison of sELISA data with those obtained with end-point PCR suggests this method as a promising tool for the detection of atypical CPB2 toxin.

The introduction of the Eastern grey squirrel (Sciurus carolinensis) in Europe is one of the best-known cases of invasive alien species (IAS) colonisation, that poses a severe risk to the conservation of biodiversity. In 2003, it was released in a private wildlife park near the city of Perugia (Italy), where it is replacing the native Eurasian red squirrel (Sciurus vulgaris). The LIFE13 BIO/IT/000204 Project (U-SAVEREDS) was set up for the Sciurus vulgaris conservation in Umbria through an eradication campaign of grey squirrels. One hundred and fifty-four animals were analysed for bacteriological, mycological, virological, and serological investigations (C4 action). Sanitary screening showed that Sciurus carolinensis is a dermatophyte carrier, and therefore, it could cause public health issues for humans, considering its confident behaviour. Moreover, it has been marginally responsible for the spreading of Candida albicans, Coxiella burnetii, and Borrelia lusitaniae. Health status evaluation conducted on the Sciurus carolinensis population indicated that it is necessary to raise awareness of its impacts on biodiversity and human health. Moreover, the health status and behaviours of the IAS must be considered when control or eradication campaigns are planned.

Background: Nutritional support, including nutritional counseling and oral nutritional supplements (ONSs), has been recommended at the earliest opportunity in head and neck (H&N) cancer patients. The limited available evidence on the efficacy of immunonutrition during chemoradiotherapy (CT-RT) in H&N cancer patients is positive with regard to some secondary endpoints, but is still scanty, particularly with regard to toxicity and treatment tolerance. We hypothesize that early systematic provision of ONSs with a high-protein–high-calorie mixture containing immunonutrients (Impact) compared to standard high-calorie–high-protein nutritional blends, in addition to nutritional counseling, may be beneficial to patients with H&N cancer during CT-RT. Hence, we designed the present study to evaluate the efficacy, in terms of treatment tolerance, toxicity and response, body weight, body composition, protein-calorie intake, quality of life (QoL), fatigue, muscle strength and immunological profile of the early systematic provision of ONSs enriched in immunonutrients compared to isonitrogenous standard blends, in H&N cancer patients undergoing CT-RT. Methods: This is a pragmatic, bicentric, randomized (1:1), parallel-group, open label, controlled, pilot clinical trial. Discussion: Many efforts are still to be taken to improve the efficacy of nutritional support in oncology. Immunonutrition represents a promising approach also in H&N cancer patients, but the evidence on its efficacy in improving clinical outcomes during CT-RT is still inconclusive. The present pilot study, which guarantees the early provision of nutritional assessment and support to all the enrolled patients in accordance with the recent guidelines and recommendations, could represent one of the first proofs of the clinical effectiveness of early oral immunonutrition in cancer patients undergoing CT-RT and could stimulate further large randomized trials, potentially resulting in the improvement of supportive care quality. Trial registration: This study is registered on ClinicalTrials.gov Identifier: NCT04611113.

Nannizzia gypsea (formerly Microsporum gypseum) belongs to geophilic dermatophytes, fungi involved in keratin degradation in the soil; however, they are also found in dogs and cats. Transmission to humans can occur directly by contact with soil, but indirect transmission via domestic animals is reported too. The exact source of the infection is usually difficult to assess and in most cases only hypothesised and rarely investigated. This case report describes a kerion caused by N. gypsea in a 2-year-old boy, where the contagion was probably secondary to domestic healthy carrier dogs. A “One-Health” approach involving human dermatologists and veterinarians, combined with the use of conventional and molecular-based techniques, allowed tracing of the epidemiological chain and managing of not only the treatment but also the prevention of a recurrence. The child's lesion began to regress after about 8 weeks of treatment with both systemic and topical therapy, while the dogs were given chlorhexidine and miconazole baths. No recurrences nor new infections occurred, demonstrating the effectiveness of the strategies used.

Nowadays, food fortification with bioactive compounds deriving from agri-food waste is of great interest all over the world. In this work, apple pomace (AP), the most abundant by-product of apple juice manufacturing, was characterised by chemical, chromatographic and spectrophotometric analyses. AP showed valuable antioxidant activity, due to the presence of phenolic compounds (8.56 mg gallic acid equivalents/g), including quercetin-3-O-galactoside, quercetin-3-O-arabinofuranoside, and phloridzin. Dried AP, at 7% and 14%, was added to pork meat to produce Italian salami, then subjected to 25 days of ripening. Physicochemical, colorimetric and microbiological analyses were carried out at days 0, 5, 11, 19 and 25, while nutritional and sensory evaluations were performed at the end of the ripening. The overall acceptability was slightly higher for 7% AP compared to 14% AP sample, and generally the replacement of a percentage of meat with apple pomace allowed the production of salami with sensory properties comparable to those obtained with classic recipes. The improved fibre and phenol content, together with the lower fat and calories, represent the most interesting characteristics of fortified salami. The results confirm that the addition of AP represents a valid approach to adding healthy compounds to salami.

The spotted fever group of Rickettsiae is a heterogeneous group of Rickettsiae transmitted by ticks, causing similar diseases in humans (spotted fever). Until recently, it was supposed that a single pathogenic tick-borne SFG Rickettsia circulated in each different geographic area and that R. conorii subsp. conorii was the SFG Rickettsiae circulating in Italy, but in the last decade, thanks to molecular diagnostic, several different Rickettsia species, previously not considered pathogenic for decades, have been isolated from ticks and definitively associated to human disease, also in Italy. The present survey was carried out with the aim of investigating the presence of different SFG Rickettsia species in a geographic area where no information was available. Ticks collected from animals submitted to necropsy, removed from humans in local hospitals and collected from the environment were identified and tested by PCR for Rickettsia spp. based on the gltA gene, and positive PCR products were sequenced. A total of 3286 ticks were collected. Fifteen tick species were recognized, the most represented (79.52%) species in the collection was Ixodes ricinus, followed by Rhipicephalus sanguineus (9.13%). The overall prevalence of Rickettsia infection was 7.58%. Eight species of Rickettsia were identified, the most frequent was R. monacensis (56%), followed by R. helvetica (25.50%). Noteworthy, is the detection in the present study of Rrhipicephali, detected only twice in Italy. These are the first data available on SFG Rickettsiae circulation in the study area and they can be considered as starting point to assess the possible risk for humans.

Clostridium (C.) perfringens is the causative agent of several diseases and enteric infections in animals and humans. The pathogenicity of the bacterium is largely mediated by the production of a wide range of toxins. Individual C. perfringens strains produce only subsets of this toxin repertoire, which permits the classification in seven toxinotypes (A–G). In addition, a variety of minor toxins further characterizes the single strains. The aim of this work was to evaluate, using Polymerase Chain Reaction (PCR) assays, the diversity of 632 C. perfringens strains isolated in Italy over 15 years. The genotyped strains were analyzed to determine the presence of major and minor toxins (cpe, consensus, and atypical cpb2), their geographical origins, and the source of isolation (animal species or food). Our study shows that toxinotype A had the greatest representation (93%) and correlated mainly with consensus cpb2 in a variety of animal species, as well as with atypical cpb2 in the five food samples. Type D, associated with cpe and atypical cpb2 minor toxins, was identified in 3% of the cases, and type F was identified in 2.5%. Seven type C isolates (1.1%) were detected in cattle, whereas the only type B atypical cpb2 isolated in Italy was detected in a goat, and one type E cpe+atypical cpb2 was detected in a sheep. Type G was not detected.

The grey squirrel is an invasive alien species that seriously threatens the conservation of the native red squirrel species. With the aim of characterizing the reproductive physiology of this species due to its great reproductive success, the function of the ovarian nerve growth factor (NGF) system was analyzed in a grey squirrel population living in central Italy. During the breeding and nonbreeding seasons, the ovarian presence, distribution, and gene expression of NGF, neurotrophic tyrosine kinase receptor 1 (NTRK1), and nerve growth factor receptor (NGFR), as well as NGF plasma concentrations, were evaluated in female grey squirrels. NGF was found in the luteal cells and in the thecal and granulosa cells of follicles, while NTRK1 and NGFR were only observed in follicular thecal and granulosa cells. NGF and NGFR transcripts were almost two-fold greater during the breeding season, while no seasonal differences were observed in NTRK1 gene expression. During the breeding season, NGFR was more expressed than NTRK1. Moreover, no changes were observed in NGF plasma levels during the reproductive cycle. The NGF system seems to be involved in regulating the ovarian cycle mainly via local modulation of NGF/NGFR, thus playing a role in the reproductive physiology of this grey squirrel population.

The reproductive cycle of an invasive alien Italian grey squirrel population was studied to understand its adaptation and limit its spread, in order to conserve the autochthonous red squirrel. Female and male genital traits were evaluated throughout the reproductive cycle, including the ovary, uterus, testicle, epididymis, seminiferous tubule morphometry, and germinative epithelium histology. Moreover, individual female fecundity was determined by counting uterine scars. Ovary width and uterus weight, length, and width reached their highest values in the luteal and pregnancy phases. On conducting a histological evaluation of the testicular germinal epithelium, four morphotypes related to the different reproductive phases of the male squirrels were identified: immature, pubertal, spermatogenesis, and regressive. Testicle and epididymis weights and seminiferous tubule diameters reached their largest values during spermatogenesis. Uterine scar analysis showed that 69% of the females had given birth to one or two litters, while 31% had no uterine scars. Litters were larger in the first breeding period than in the second; annual fecundity was 4.52 ± 1.88 uterine scars/female. Umbrian grey squirrels have adapted to their non-native range, showing two annual mating periods at times similar to those in their native range, and high reproductive success.

The improvement of rabbit gut microbiota by modifying nutritional components of the feed or favoring its early intake of feed has been previously investigated. The early administration of dehydrated alfalfa (A) or fresh grass (G) for rabbits, during the peri-weaning period (30 and 45 days of age), and their effect on performance and caecal microbiota compared to a standard diet (C) were evaluated. Until 15 days of age, nine litters/group were housed in the maternal cage and milked once per day. From 15 to 30 days, the young rabbits could consume both milk and solid feed (pelleted for C or supplemental feed for A and G). At 30 days of age, the rabbits were weaned and, until 45 days, were kept in single cages following the same dietary protocol. No significant changes were found in the milk intake or the individual weight of young rabbits at 30 and 45 days. The caecal Firmicutes/Bacteroidetes (bacterial phyla ratio) increased with age (from 2.43 to 6.05 on average, at 30 and 45 days). The Ruminococcaceae/Lachinospiraceae (bacterial family ratio) was highest in the A group at both ages, followed by G then C. The early administration of dehydrated alfalfa is a promising solution to improve health status by favoring an appropriate digestive microbiota.

Dientamoeba fragilis (D. fragilis) is a worldwide distributed protozoan parasite; it is pathogenic for humans. A wide spectrum of gastrointestinal symptoms has been described in infected patients: diarrhoea, flatulence, abdominal pains, colic and weight loss. However, asymptomatic infection has been also described. D. fragilis is still not well known; no cystic stage has been demonstrated and only the trophozoites are detected in stool samples. For identifying this typically more often binucleate protozoan, is necessary to perform permanent stain (eg. Giemsa) on fresh stool specimens. This protozoan is extremely difficult to cultivate but molecular techniques such as the Polymerase Chain Reaction offer promise as a means of diagnosing infection. In five years time (2006-2010), faecal samples were collected from pigs housed in farrow-to-finish herds (494 samples, splitted in three different categories: sows, growers, finishing pigs) and from hunted or slaughtered wild boars (87 samples). Simultaneously, the study was undertaken on human faeces (17 samples) to evaluate the presence of D. fragilis in pig breeders. All samples were collected directly from the rectum, cooled and sent to the laboratory where they were examined for D. fragilis by direct microscopic examination. The fresh faecal smears were stained with a 10% Giemsa solution in distilled water for 30 min. Biomolecular investigations (TaqMan real-time PCR which targets the 5.8S rRNA, nested PCR for the 18S rRNA, nested PCR for the internal transcribed spacer 1 region) were carried out on 38 pigs and 17 pig breeders specimens. The microscopic examination of the fresh fecal smears revealed positivity in 277 domestic pigs, corresponding to 56.07%. In particular higher positivity was observed on youngest animal (76.57%), while oldest or mature pigs recorded an important decreasing of positivity according the age (Table 1). Concerning wild boars, we revealed positivity in 35 animals (40.22%). Among humans, the positivity was 76.47% and these positive specimens came from people with a close contact with pigs. Biomolecular investigations carried out on human and animals amplified positive products revealed 100% homology with the 5.8S rRNA gene of D. fragilis, genotype 1 (e.g., Genbank DQ233451). During a five years research project we demonstrated the presence of D. fragilis in domestic pigs populations as well as in hunted or slaughtered wild boars. Due to the high percentage of positivity we could assume the domestic and/or wild pigs can play a role as natural reservoir of the parasite. In this scenario, outdoor pig farms and/or “confined” wild boars rearing can act as important link of exchange of this parasite. The demonstrated homology of D. fragilis sequences obtained from both humans and animals suggests the potential role of this parasite as zoonotic agent. If an environmentally resistant and infective stage of D. fragilis exists, we suppose the environmental contamination with domestic/wild pigs feces could be as an important factor in the transmission of this parasite to other hosts, including humans.

All helminths parasitosis transmitted to humans trough ingestion of infested fleshes, where man is definitive host too, are represented by four groups of helminths: the cestodes Dyphyllobothrium spp and Spirometra spp. (Sparganum proliferum is the name of the immature plerocercoid larva), the trematodes Opisthorchis Clonorchis “group” (many could be the genera and species involved), and the nematode Capillaria philippinensis. So, for fishes humans foods (fresh or salted water) the control and prevention in veterinary health must be directed to investigation regarding intermediate stages of these parasites in fishes for human alimentation; if present, they must be eliminated. The helminths parasitosis transmitted to humans trough ingestion of infected mammals meats, are represented by taeniasis (Taenia saginata, T. solium and T. saginata asiatica), where man id definitive host and the infection is caused by ingestion of bovine or swine meat, containing larvae of these cestodes, and by trichinellosis, where humans represent a intermediate stage, and the eventual pathology is caused as by adult (acute infection) as by larvae (chronic infection) of this nematode: usually the meats responsible are infected pork, wild pork or horse (Trichinella spp. Is inside the meats of these animals). So the veterinary control and prophylaxis are necessary to avoid this disease and preventing the infection that could be severe.

Dientamoeba fragilis, atypical protozoon because “flagellate without flagella” but amoeba-like, of whom we know only the trophozoitic stage (an so its brittleness outside intestinal tract), is a frequent responsible of intestinal human infections, worldwide, and some authors relate that D. fragilis is the most frequent protozoon and parasite that can infects humans. Actually we don’t know a sure potential “reservoir” in animals who are strictly in contact with humans, and it is difficult to understand its epidemiological chain, otherwise the transmissions to humans and from humans to humans. For all these reasons we performed another study on subjects of swine breedings, and among people who work in these breedings, that are in direct contact or not with pigs. Using standardized methodologies, we analyzed 224 faecal specimens of swine and 15 human specimens.We use for identification of D. fragilis the Giemsa stain.These were the results: D. fragilis was observed in 50.9% of pigs and 20% among humans (30% in workers strictly in contact with breedings and pigs, 0% in familiars or other without a closed contact with swines). Other commensal protozoa were observed with variable associations, but in this article we want to analyze the possible transmission from this pigs to humans (and for us this protozoon is undoubtedly a “reservoir” of D. fragilis for humans), and underline two aspects: for the research of this protozoon, standard procedures area mandatory, with a permanent stain, as Giemsa stain, is necessary, and in all humans with various intestinal infections or troubles, particularly “irritable bowel syndrome” (or similar ones), the specimens must be analyzed for D. fragilis. At least we think that in the near future molecular studies are important for confirming this our observations, and for verifying eventual and probable differences inside genotypes of this very suggestive protozoon, that until now present not rarely phenotypic diversities.

Over the past five years have been described, in Umbria, the first human cases of infestation with Opistorchis felineus, as well as detect the presence of this fluke eggs in the feces of Digenea stray seeds or stray cats on the island of Maggiore Lake Trasimeno (Umbria) in central Italy. Previous research metacercariae in fish caught in this lake and the recovery of the shellfish B. leach into the lake area have been rather insignificant.This work confirms the presence of eggs of O. felineus in 33.3% of cat litter and, most importantly, it shows active infestation in two cats found dead on that island, anatomical lesions - histopathological opisthorchiasi and consistent with the observation of the presence of adult worm with eggs is in intrahepatic bile home that when pancreatic O. felineus.

Dientamoeba fragilis is a worldwide distributed protozoon which is now estimated responsible of enteric diseases in humans. A wide spectrum of intestinal symptoms has been described: diarrhoea (acute or prolonged), flatulence, abdominal pains, and not specific bowel syndrome; however, asymptomatic infection has been also reported. D. fragilis is still not well know because only the delicate trophozoite stage has been demonstrated in stool samples and definitive identification is based on permanent stains, usually not performed in ova and parasites (O&P) examinations, in Italy.D. fragilis is difficult to cultivate but molecular techniques, such as PCR, appear promising for a sure diagnosis of this protozoan infection.The epidemiology of dientamoebiasis is not clear too, and this paper presents preliminary results concerning the presence of D. fragilis in swine faeces in order to evaluate the possible role of pigs as reservoir of the parasite. So, 121 stool specimens of swine population were examined using Giemsa permanent stain and D. fragilis was observed in 53 stool specimens (43.8%). Other consideratioona are reported too.

The Authors describe two small and apparently asymptomatic epidemics of Opisthorchiasis caused by Opisthorchis felineus, which were observed in provence of Perugia, Umbrie region, Italy.The first one was observed in a couple of young adults during 2003 because both ones ate pickled (at 2-8°C) tenches fished in Trasimeno Lake in province of Perugia.The second episode was observed during 2006 among eight young adults that ate pickled fishes (tenches, carps and perches, always at 2-8°C); source of these fishes was the same lake.The request for a coproparasitological examination of stools in one of the two subjects of the first episode and in one of the eight subjects of the second episode was the casual observation of an increased count of periferical eosinophils. During January 2006, ten faecal samples of cats, living on “Isola Maggiore” of the Trasimeno Lake, were collected for a parasitological test; 40% was positive for ova of O. felineus.The Authors describe the diagnostical items for this parasitosis, relate the epidemiological features of this trematode infection, and suggest the importance of a good approach to this zoonotic parasitosis, which interest public health too and never were before observed in Italy among humans.