Detection of Leishmania spp. in Chronic Dermatitis: Retrospective Study in Exposed Horse Populations

Leishmania infantum is a protozoan causing human zoonotic visceral leishmaniasis (ZVL) and visceral–cutaneous canine leishmaniosis (CanL) in the Mediterranean Basin. L. infantum is able to infect a large number of wild and domestic species, including cats, dogs, and horses. Since the 1990s, clinical cases of equine leishmaniasis (EL), typically characterized by cutaneous forms, have been increasingly diagnosed worldwide. The aim of the present study was to evaluate the presence of clinical forms of EL in CanL-endemic areas in Italy, where exposure of equine populations was ascertained from recent serological surveys. For this purpose, formalin-fixed and paraffin-embedded skin biopsies of 47 horses presenting chronic dermatitis compatible with EL were retrospectively selected for the study and subjected to conventional and q-PCR. A singular positivity for L. infantum was found; BLAST analysis of sequence amplicons revealed a 99–100% homology with L. infantum sequences. The histological examination revealed a nodular lymphoplasmacytic and histiocytic infil-trate; immunohistochemistry showed rare macrophages containing numerous positive amastigotes. The present retrospective study reports, for the first time, a case of a cutaneous lesion by L. infantum occurring in an Italian horse. Pathological and healthy skin samples should be investigated on a larger scale to provide information on the potential clinical impact of EL in the practice, and to define the role of horses in epidemiological ZVL and CanL scenarios

First Expert Elicitation of Knowledge on Drivers of Emergence of Bovine Besnoitiosis in Europe

Bovine besnoitiosis (BB) is a chronic and debilitating parasitic disease in cattle caused by the protozoan parasite Besnoitia besnoiti. South European countries are affected and have reported clinical cases of BB. However, BB is considered as emerging in other countries/regions of central, eastern and northern Europe. Yet, data on drivers of emergence of BB in Europe are scarce. In this study, fifty possible drivers of emergence of BB in cattle were identified. A scoring system was developed per driver. Then, the scoring was elicited from eleven recognized European experts to: (i) allocate a score to each driver, (ii) weight the score of drivers within each domain and (iii) weight the different domains among themselves. An overall weighted score was calculated per driver, and drivers were ranked in decreasing order of importance. Regression tree analysis was used to group drivers with comparable likelihoods to play a role in the emergence of BB in cattle in Europe. Finally, robustness testing of expert elicitation was performed for the seven drivers having the highest probability to play a key role in the emergence of BB: i.e., (i) legal/illegal movements of live animals from neighbouring/European Union member states or (ii) from third countries, (iii) risk of showing no clinical sign and silent spread during infection and post infection, (iv) as a consequence, difficulty to detect the emergence, (v) existence of vectors and their potential spread, (vi) European geographical proximity of the pathogen/disease to the country, and (vii) animal density of farms. Provided the limited scientific knowledge on the topic, expert elicitation of knowledge, multi-criteria decision analysis, cluster and sensitivity analyses are very important to prioritize future studies, e.g., the need for quantitative import risk assessment and estimation of the burden of BB to evidence and influence policymaking towards changing (or not) its status as a reportable disease, with prevention and control activities targeting, firstly, the top seven drivers. The present methodology could be applied to other emerging animal diseases

<i>Besnoitia besnoiti</i> among cattle in insular and northwestern Italy: endemic infection or isolated outbreaks?

Background: Bovine besnoitiosis, caused by the apicomplexan Besnoitia besnoiti, is a chronic and debilitating disease considered as emerging in Europe. In Spain, Portugal and France it is endemic and foci of infection were recorded in Germany, Switzerland, Hungary, Greece and Italy. In Italy, cases of bovine besnoitiosis were registered both in imported and autochthonous cattle, and mostly in central regions; high seroprevalence was also revealed by an epidemiological survey performed in the southern part of the country. Aiming to update information on the disease in northwestern and insular areas of Italy, where data on bovine besnoitiosis were missing, a serosurvey was designed for the present study. Methods: Three thousand one hundred and forty bovine blood samples from both dairy and beef farms (n = 126) were collected in northwestern regions (Lombardy, Piedmont and Liguria) and in the island of Sardinia. Samples were analyzed by a standardized in-house ELISA and those resulted positive were re-tested by Western Blot (WB) for confirmation. On results obtained by both ELISA and WB, apparent (AP) and true prevalence (TP) were calculated at individual and herd levels. Further, a panel of sera resulted positive to ELISA was analyzed by IFAT. Results: A total of 712 animals (AP = 22.7%; TP = 18.8%) and 109 farms (AP = 86.5%; TP = 88.2%) showed a positive reaction in ELISA. Only ten (AP = 0.3%; TP = 0%) specimens proceeding from five farms (AP = 3.9%; TP = 1.7%) from Lombardy were confirmed positive to the WB, corresponding to two Holstein Friesian cows and eight beef cattle. IFAT showed a low sensitivity (44.4%) scoring positive in only four samples out of 9 positive to WB. Conclusions: The survey demonstrated that bovine besnoitiosis cannot still be considered endemic in whole Italy. In fact, independent foci of infection were registered only in Lombardy region. Therefore, a sanitary strategy aimed to increase control measures and to organize monitoring plans, by adequate diagnostic tools is necessary to avoid overestimation of B. besnoiti in Italy.</br

Molecular epidemiology of Blastocystis sp. in dogs housed in Italian rescue shelters

Blastocystis is a ubiquitous protozoan with a wide range of hosts. In humans, its presence has been associated with gastrointestinal disorders, although its role as a pathogen still needs to be elucidated. Until now, 17 Blastocystis subtypes (STs) have been identified, with ST1\u2013ST4 the most commonly found in humans. Among domestic animals, the same STs reported in humans have been detected in dogs. An epidemiological survey on dog kennels was carried out to evaluate the prevalence of Blastocystis and the STs involved. Overall, 99 faecal samples were collected from the rescue shelters. Blastocystis detection was performed through conventional barcoding PCR targeting the 1800-bp SSU-rDNA, followed by sequencing and phylogenetic analysis. Blastocystis DNA was found in 21 faecal samples (21.2%), and all samples were successfully sequenced and identified as ST3 in a unique monophyletic group. The presence of Blastocystis was reported for the first time in dogs from Italy, with the identification of ST3, the subtype most commonly found in humans

Epidemiology and genetic diversity of Blastocystis sp. in dogs housed in sanitary and rescue shelters

INTRODUCTION. Blastocystis is an intestinal protist associated with gastrointestinal disorders and affecting worldwide both humans and a wide range of animals. Up-to-now, 17 Blastocystis subtypes (STs) have been recognized on the basis of subunit ribosomal RNA gene (SSU-rDNA) analysis, being ST1-ST4 the most commonly found in humans. Among domestic animals, dogs seem to be involved in the spread and maintenance of the infection, naturally harboring STs in common with humans. With the aim of investigate on dogs as a potential source of Blastocystis infection to humans, an epidemiological survey on kennels’ dogs in Lombardy was planned, to evaluate the prevalence of Blastocystis infection and the STs involved. MATERIALS AND METHODS. Ninety-nine individual fecal samples were collected in six kennels, with an average of 19 animals per surveyed kennel. Samples included eight pure breeds and cross-breeds, and animals of varying age from three months to 14 years. Genomic DNA was extracted from each sample using a commercial kit; the DNA samples were subjected to PCRs in order to amplify a fragment of about 600 base pair within the 1800 bp SSU-rDNA of Blastocystis (Scicluna et al., 2006. Protist. 157: 77–85) and the fragments obtained purified for sequencing. STs were identified aligning obtained sequences with available published ST1-ST10 sequences. To infer the phylogenetic relationships among the sequences, a phylogenetic tree was constructed by using the neighbor-joining (NJ) method in MEGA v.6.0.6 software. RESULTS AND CONCLUSIONS. Twenty-one fecal samples proved to be positive by PCR (21.2%), with prevalence values ranging from 18.2% to 37.5% according to the kennels; all dogs from one kennel scored negative for Blastocystis. Sequences obtained showed a high identity (98–100%) to homologous sequences of Blastocystis isolates previously reported in GenBank. The phylogenetic analysis showed that all sequences clustered with ST3 in a monophyletic group with a high bootstrap value (>95), being the first report in Italy of this ST in dogs. Previous surveys carried out in Italy showed the occurrence of ST3 as the most prevalent in human fecal samples (Mattiucci et al., 2016, Epidemiol Infect. 144:635-646. Meloni et al., 2011. Parasitol Res. 109:613-619); the results obtained in the present survey suggest dogs as possible zoonotic reservoirs for the parasite

Infestazione da Angiostrongylus vasorum nel panda rosso (Ailurus fulgens): review della letteratura e presentazione di un caso clinico = Angiostrongylus vasorum infestation in the red panda (Ailurus fulgens): review of the literature and presentation of a case report

L\u2019infestazione da Angiostrongylus vasorum nel panda rosso (Ailurus fulgens) determina una polmonite parassitaria ad andamento cronico potenzialmente letale. Gli Autori presentano una revisione della letteratura e un caso clinico. Un panda rosso \ue8 stato riferito all\u2019Ospedale Veterinario dell\u2019Universit\ue0 degli Studi di Milano a Lodi con una diagnosi di infestazione da A. vasorum che era stata formulata a maggio del 2015, non appena trasferito in un parco faunistico del Nord Italia. La diagnosi \ue8 stata emessa sulla base del riscontro di larve di prima et\ue0 (L1) nelle feci con tecnica di Baermann e della presenza dell\u2019antigene tramite tecnica di immunomigrazione eseguita presso un laboratorio di riferimento. Il soggetto \ue8 stato successivamente trattato con successo mediante somministrazione per via orale di milbemicina ossima e praziquantel rispettivamente alla dose settimanale di 12,5 mg/soggetto e 125 mg/soggetto per 3 settimane consecutive alternate a 20 giorni di sospensione. Il trattamento \ue8 continuato con lo stesso schema fino al controllo eseguito a Lodi a dicembre 2018. Grazie alla tecnica del rinforzo positivo e dell\u2019animal training, l\u2019animale ha assunto il farmaco regolarmente e direttamente dalle mani del Keeper nascosto in acini d\u2019uva, alimento molto gradito dai panda rossi. Non \ue8 stato riscontrato alcun effetto collaterale