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

PREVALENCE OF CANINE DISTEMPER VIRUS IN CANIDS IN CENTRAL ITALY AND FIRST IDENTIFICATION OF ARCTIC LINEAGE IN MARCHE AND UMBRIA REGIONS: A PRELIMINARY STUDY

Canine distemper virus (CDV) is one of the most commonly virus implicated in outbreaks in wild and domestic carnivores. CDV causes severe systemic diseases which normally involves the respiratory, gastrointestinal and nervous systems. To our knowledge the literature about the real incidence of such disease is scarce, particularly in wild animals population. Recently, outbreaks of CDV have been documented in Italian grey wolves (Canis lupus italicus) [1], a least concerned species in IUCN Red List. Therefore, the surveillance of CDV is a priority for the conservation of the wolves and, more generally, for the protection of wild carnivores which are widespread in Central Italy, especially in the National Parks. In total, 215 samples, belonging to 148 canids for CDV presence, were analysed from November 2012 to December 2016 in the laboratory of IZSUM. Of these, 37.2% were dogs, 33% wolves and 29.8% foxes. Animals were collected in 12 different provinces of 6 Regions: Umbria, Marche, Emilia Romagna, Tuscany, Lazio and Apulia. All samples were collected from dead animals which were sent to the Diagnostic Units of Istituto Zoprofilattico Sperimentale Umbria e Marche and subjected to autopsy. The RNA was extracted from organ pools and swabs with a commercial kit, retrotranscripted to cDNA and amplified by the real time PCR with QuantiFast SYBR Green RT PCR kit (Qiagen GmbH, Hilden, Germany) using primers for a fragment of 278bp in CDV nucleoprotein (NP) gene [2]. Samples having a melting temperature (TM) value ±0.5°C versus TM value of positive control were considered positive. Moreover, samples were visualized by UV rays with GelRed TM (Biotium Inc.) after electrophoresis in agarose gels and bands of appropriate sizes were excised, extracted and sequenced. Sequences obtained (n=11) were aligned with NP gene sequences of CDV available in GenBank by MUSCLE. Molecular phylogenetic analysis (MEGA 7.0) was carried out by using Maximum Likelihood method based on the Tamura 3-parameter model. The CDV RNA was identified in 20.3% of the analysed animals. A high positivity rate was identified in dogs with 10.1% of 148 sample tested positive followed by wolves (6.08%) and foxes (5.11%). The Artic Lineage of CDV was identified in 9 out of 11 sequenced samples, in both wild and domestic canids. This strain was identified in 3 different provinces (PU, AP, PG), raising concerns given the vastness of the affected area. Two Onderstepoort strains were also identified. In conclusion, this study shows a wide CDV circulation involving different ecotypes and species in the investigated area. Further studies, based on epidemiological and genetic analysis, will be carried out in order to assess the phylogenetic correlation among the identified strains. This follow up will be important in order to highlight potential risk factors associated with the introduction of this new genotype and to better understand the role played by domestic and wild carnivores interactions in virus spreading. These additional studies should be carried out as soon as possible in order to prevent virus dissemination and to perform ad hoc vaccination campains

Ultrastructural morphometrical and immunocytochemical analyses of hepatocyte nuclei from mice fed on genetically modified soybean

No direct evidence that genetically modified (GM) food may represent a possible danger for health has been reported so far; however, the scientific literature in this field is still quite poor. Therefore, we carried out an ultrastructural morphometrical and immunocytochemical study on hepatocytes from mice fed on GM soybean, in order to investigate eventual modifications of nuclear components of these cells involved in multiple metabolic pathways related to food processing. Our observations demonstrate significant modifications of some nuclear features in GM-fed mice. In particular, GM fed-mice show irregularly shaped nuclei, which generally represents an index of high metabolic rate, and a higher number of nuclear pores, suggestive of intense molecular trafficking. Moreover, the roundish nucleoli of control animals change in more irregular nucleoli with numerous small fibrillar centres and abundant dense fibrillar component in GM-fed mice, modifications typical of increased metabolic rate. Accordingly, nucleoplasmic (snRNPs and SC-35) and nucleolar (fibrillarin) splicing factors are more abundant in hepatocyte nuclei of GM-fed than in control mice. In conclusion, our data suggest that GM soybean intake can influence hepatocyte nuclear features in young and adult mice; however, the mechanisms responsible for such alterations remain unknown

Seasonal monitoring of Aedes albopictus: practical applications and outcomes

The introduction of the Asian tiger mosquito Aedes (Stegomya) albopictus (Diptera: Culicidae) into temperate regions poses serious concerns for the risk of the spreading of arboviral epidemics, as confirmed by the Chikungunya fever outbreak in Italy. This article describes the implementation and the results of a strategy for the pest management implemented over 4 years in Pesaro (a city in the Marche region, Italy). The strategy used 60 integrated wide- sized ovitraps for monitoring purposes. Twenty-day larvicide-based treatment cycles were implemented for the manholes of the urban area and also the inhabitants were involved in pest control relating to their own properties. It was observed that the weekly median of eggs laid decreased consistently from 2008 to 2011, indicating the good performance of the vector control and a reduction in the related epidemics risk

Are two coincidences a proof ? The European roe deer (Capreolus capreolus) is a suitable intermediate host for Taenia serialis

INTRODUCTION: Taeniids are multi-host parasites with an indirect life cycle that strictly depends on a predator-prey relationship. This is referred to as a multi-host trophically-transmitted parasite system, which involves mammalian species in both adult and larval stage. Parasites with a complex life cycle may exhibit different degrees of host-specificity at each life stage. To measure host-specificity it is not sufficient the number of the species that a parasite can infect but also the relatedness to each other. Trophically-transmitted parasites necessitate high predation rates assuring high transmission levels within the food chain. A generalist parasite that uses several intermediate host species has more likelihood to infect its definitive host if this feeds broadly among many prey species. Thus, knowing the host breadth at the different life cycle stage is a fundamental concept of the biology and epidemiology of these multi-host parasites. MATERIALS AND METHODS: A free-ranging adult female European roe deer (Capreolus capreolus) died after a car crash in February 2022 in Bolognola (MC), within Monti Sibillini National Park. The carcass was brought in to the IZSUM of Tolentino (MC), where a necropsy was performed. Cyst-like lesions were recovered from the epicardial and endocardial surfaces and the intercostal muscles. Histological and molecular investigations were carried out for the cysts identification. RESULTS AND CONCLUSIONS: M-PCR gave a ̴270-bp amplicon, referable to Taenia spp. (267 bp). Sanger sequencing showed 99% query cover, 2e-109 e-value, and 100% identity with T. serialis. Histology of the parasitized heart revealed the presence of a metacestode larval stage (coenurus) compatible with taeniid larval stage of T. serialis. A large and more tissue-compressing cyst, which contains only a single protoscolex was observed, surrounded by dystrophic, vacuolated and compressed cardiomyocytes. No inflammatory cells were observed around the cyst that means, as for Echinococcus multilocularis, immune tolerance mediated by specialized regulatory T cells and related cytokines such as IL-10 and TGF-β. Wildlife plays a more prominent role in the T. serialis epidemiology. Taenia serialis-coenurosis is already diagnosed in lagomorphs, rodents, and several cases in primates rarely detected also in cats, sheep, and marsupials. Few diagnoses have been confirmed by using molecular analysis and no one by hystopathological approach. Thus, T. serialis may be more flexible in the selection of intermediate hosts than previously hypothesized. To our knowledge, few published articles report the presence of T. serialis either in definitive or intermediate hosts in Italy. The interesting finding we describe here is that in a relatively short period of time and close to each other, two roe deer were detected positive to T. serialis-coenurosis (Morandi et al., 2022. Int J Parasitol: Parasites Wildl, 17:110-13). Studies on the T. serialis-dynamic will be conducted. Wildlife surveillance is crucial to monitor for human and animal health as intermediate hosts breadth of critical taeniids may suddenly change

Seasonal monitoring of Aedes albopictus: practical applications and outcomes

The introduction of the Asian tiger mosquito Aedes (Stegomya) albopictus (Skuse, 1897) (Diptera: Culicidae) into temperate regions poses serious concerns for the risk of the spreading of arboviral epidemics, as confirmed by the chikungunya fever outbreak, in Italy. Integrated wide-sized ovitrap monitoring is a helpful tool in any good pest-control strategy. The Ae. albopictus population dynamics were monitored over a four-year period in the town of Pesaro (Marche, Italy), using 60 ovitraps. Twenty-day larvicide-based treatment cycles were implemented for the manholes of the urban area and also the inhabitants were involved in pest control relating to their own properties. The weekly median of eggs laid was reduced from 2008 to 2011, which indicated the good performance of the vector control and a reduction in the related epidemics risk. The monitoring system adopted is described here, and proposed as a cost-effective and available system for extensive vector surveillance