First Worldwide Evidence of Bronchopulmonary Strongyle Nematodes and the First Report on Italy of Cryptosporidium sp. in Allochthonous Nutria (Myocastor coypus)

Nutria (or coypu, Myocastor coypus), is a semi-aquatic rodent that is native to South America and has been introduced almost all over the world since the end of the 19th century. In Europe, this rodent is considered an invasive species. In this report, we analyzed nutria fecal samples in a small coastal wetland of Central Italy, using different techniques (fresh smear, direct immunofluorescence, Baermann technique, flotation, ethyl acetate sedimentation) to obtain an arrangement of eukaryote endoparasites (Protozoa and Helminths) and compare them with data available in the literature for both Italy and worldwide. We recorded five taxa, with a dominant occurrence (>70%) of nematodes of the genus Strongyloides. Moreover, we reported for the first time in nutria a bronchopulmonary strongyle nematode (Muellerius vel. Angiostrongylus) and, for the first time in Italy, protozoans of the genus Cryptosporidium. Since nutria co-occurs with humans and domestic animals in the study area, we highlighted possible sanitary and management implication

Giardia duodenalis assemblages and Entamoeba species infecting non-human primates in an Italian zoological garden: zoonotic potential and management traits

<p>Abstract</p> <p>Background</p> <p><it>Giardia duodenalis </it>and <it>Entamoeba </it>spp. are among the most common intestinal human protozoan parasites worldwide and they are frequently reported in captive non-human primates (NHP). From a public health point of view, infected animals in zoos constitute a risk for animal caretakers and visitors. In this study we carried out the molecular identification of <it>G. duodenalis </it>and <it>Entamoeba </it>spp. from nine species of primates housed in the zoological garden of Rome, to better ascertain their occurrence and zoonotic potential.</p> <p>Results</p> <p><it>G. duodenalis </it>was found only in <it>Lemur catta </it>(47.0%). <it>Entamoeba </it>spp. were detected in all species studied, with the exception of <it>Eulemur macaco </it>and <it>Varecia rubra</it>. The number of positive pools ranged from 5.9% in <it>L. catta </it>to 81.2% in <it>Mandrillus sphinx; </it>in <it>Pan troglodytes </it>the observed prevalence was 53.6%. A mixed <it>Entamoeba</it>-<it>Giardia </it>infection was recorded only in one sample of <it>L. catta</it>. All <it>G. duodenalis </it>isolates belonged to the zoonotic assemblage B, sub assemblage BIV. Three <it>Entamoeba </it>species were identified: <it>E. hartmanni</it>, <it>E. coli </it>and <it>E. dispar</it>.</p> <p>Conclusions</p> <p>Our results highlight the importance of regularly testing animals kept in zoos for the diagnosis of zoonotic parasites, in order to evaluate their pathogenic role in the housed animals and the zoonotic risk linked to their presence. A quick detection of the arrival of pathogens into the enclosures could also be a prerequisite to limit their spread into the structure via the introduction of specific control strategies. The need for molecular identification of some parasite species/genotype in order to better define the zoonotic risk is also highlighted.</p

Intestinal parasites infecting captive non-human primates in Italy

Non-human primates (NHPs) living in captive conditions are susceptible to intestinal parasites that can contribute to mortality and morbidity, and cause zoonotic infections. Thus, parasite surveys on NHP populations under human care are relevant as part of the evaluation of NHPs welfare and in the zoonotic disease risk assessment, as well as in the exploration of parasite transmission pathways, according to the One-Health concept. This study aimed to identify intestinal parasites infecting NHPs living in two wildlife recovery centers and in a zoological garden, in Italy. Ninety-three fecal samples from Macaca tonkeana, Macaca fascicularis, Sapajus apella, Chlorocebus aethiops, Macaca fuscata, Macaca sylvanus, and Cebus capucinus were collected at Piano dell’Abatino Park (Lazio), and fecal smears and flotation were performed in order to identify parasites according to morphological keys. Additionally, one carcass of M. fuscata from the Bioparco Zoological Garden of Rome (Lazio) and one of M. fascicularis from the Center for the Recovery of Exotic and Maremma Wild Animals (Tuscany) were necropsied and intestinal adult nematodes were collected and characterized at morphological and molecular level, using the mitochondrial cox1 and rrnL markers. Protozoans (Entamoeba coli, Iodamoeba bütschlii, Dientamoeba fragilis-like, Giardia sp.), chromists (Balantidium/ Buxtonella sp.) and nematodes (Capillaria sp., Trichuris sp., strongyliform larvae and Oesophagostomum sp.) were found through fecal smears and flotation. The collected adult nematodes from dead NHPs were morphologically identified as whipworms (genus Trichuris). Phylogenetic analyses grouped Trichuris specimens into the Trichuris trichiura complex of species, with specimens from M. fuscata clustering into a host-specific branch, and whipworms from M. fascicularis clustering within a clade formed by Trichuris infecting several primate species, including humans. The results here collected revealed the presence of potentially zoonotic parasites circulating in captive primates in Italy, providing useful information for the formulation of management and care plans for captive NHPs, and for the elaboration of safety measures for visitors and animal keepers

Nuclear and mitochondrial data on Trichuris from Macaca fuscata support evidence of host specificity

Whipworms are parasitic intestinal nematodes infecting mammals, and traditionally humans and other primates that have so far been considered infected by Trichuris trichiura. Recent molecular studies report a more complex scenario suggesting the presence of a species complex with several Trichuris taxa specifically infecting only one primate species as well as taxa able to infect a range of primate species. The systematics of the group is important for taxonomic inference, to estimate the relative zoonotic potential, and for conservation purposes. In fact, captive animals living in zoological gardens are usually infected by persistent monoxenous intestinal parasites. Here, two Japanese macaques living in the Bioparco Zoological Garden of Rome were found infected by Trichuris sp. Nematodes were characterized at the molecular level using nuclear (btub and 18S) and mitochondrial (16S and cytb) markers and then compared to Trichuris collected previously in the same location, and to other Trichuris infecting primates. Evidences from mitochondrial and nuclear markers allowed for the identification of Trichuris sp. specific to Macaca fuscata. Results obtained here also described a uniform taxonomic unit of Trichuris, separated but closely related to Trichuris trichiura, thus, emphasizing its zoonotic potential for workers and visitors

Intestinal parasites in Rupicapra spp. populations: study in the framework of the relevant italian project (PRIN)

Intestinal parasites can seriously threaten the performances and well-being of wild ungulates. In this study, we investigated the occurrence and parasitic burden of protozoans and gastro-intestinal helminths (GIH) in Rupicapra spp. From September 2013 to January 2016, 352 fresh fecal samples were collected from Rupicapra rupicapra rupicapra in the Alps (N=262) and from Rupicapra pyrenaica ornata in the Apennines (N=90). Samples were examined using standard copro-parasitological methods for Eimeria and GIH and an immunofluorescence test for Cryptosporidium and Giardia duodenalis. Parts of gp60 and ssRNA/gdh/βgiardin genes were used to identify these protozoa species/genotypes. In R.r.rupicapra and in R.p.ornata, 7 and 6 parasite taxa were identified, respectively, with a mean number of 1.7 species/host (min-max 0-5) and 2.05 (min-max: 0-4), respectively. Overall, 85.3% (95%, C.I.=81.5-89.1) of the animals investigated scored microscopically positive to Eimeria spp. with a mean intensity of emission (m.i.e.) of up to 776 o.p.g.; 5.4% (95%, C.I.=3.07.7) were positive to G. duodenalis and 82% (95%, C.I. 77.91-86.15) to GIH with a m.i.e. of up to 147 e.p.g. Prevalence in R.r.rupicapra was 81.2% with a m.i.e. of 380 o.p.g. for Eimeria, 6.87% for Giardia, and 77.45% for GIH with a m.i.e. of 142 e.p.g. Prevalence in R.p.ornata was 94.4% with m.i.e. of 1,093 o.p.g. for Eimeria, 1.1% for G.duodenalis, and 94.4% for GIH with a m.i.e. of 151 e.p.g. Assemblages A/AI and E were identified in R.r.rupicapra and assemblage A/AIII in R.p.ornata. None of the animals tested positive for Cryptosporidium. The results show that the prevalence of Eimeria, G.duodenalis and GIH in both host species is nonnegligible, with a significantly higher parasitic burden in R.p.ornata. The detection of G. duodenalis in Rupicapra spp. is noteworthy. This study updates the data on parasitic fauna of these wild bovids. The impact of these parasites on chamois population dynamics will be inferred from the results/variables obtained throughout the entire interdisciplinary project

Fluorescent Asymmetrically Cobalt-Tipped CdSe@CdS Core@Shell Nanorod Heterostructures Exhibiting Room-Temperature Ferromagnetic Behavior

A colloidal two-step seeded-growth approach has been devised to selectively synthesize three-component magnetic/semiconductor hybrid nanocrystals (HNCs) with a matchstick-like profile and tunable geometric parameters. The newly developed heterostructures individually comprise a single metallic Co head connected to either apexes of one rod-shaped section made of a CdSe core eccentrically embedded in a CdS shell. The specific topological arrangement realized arises from the peculiar anisotropic reactivity of the noncentrosymmetric CdSe@CdS core@shell nanorods that have been used as substrates to seed heterogeneous nucleation of Co in a surfactant-free environment from an organometallic precursor. The HNCs retain appreciable fluorescent emission in spite of photoexcited charge transfer from the semiconductor to the metal domain and exhibit unusual ferromagnetic-like behavior at room temperature

Giardia duodenalis in colony stray cats from Italy

Giardia duodenalis is the most common intestinal protozoan in humans and animals worldwide, including eight morphologically identical assemblages, infecting pets, livestock, wildlife and human beings. Assemblages A and B are those with the higher zoonotic potential, and they have been detected in several mammals other than humans; the others (C to H) show a higher host specificity. Cats can harbour both the specific Assemblage F and the zoonotic ones A and B. Several studies have been carried out on G. duodenalis genotypes in cats; however, the role of this species in the epidemiology of giardiasis is still poorly understood. In this scenario, the present study carried out the detection and genetic characterization at sub-assemblage level of G. duodenalis from colony stray cats in central Italy. In the period 2018-2019, 133 cat faecal samples were analysed for the presence of G. duodenalis cysts by a direct immunofluorescence assay. Positive samples were subsequently subjected to molecular analyses for assemblage/sub-assemblage identification. Forty-seven samples (35.3%) were positive for G. duodenalis cysts by immunofluorescence. G. duodenalis DNA was amplified at SSU-rDNA locus from 39 isolates: 37 were positive for zoonotic Assemblage A and 2 showed a mixed infection (A + B). Positive results for the beta-giardin gene were achieved for 25 isolates. Sequence analysis revealed 16 isolates belonging to Sub-assemblage AII and 8 to Sub-assemblage AIII. One isolate resulted as ambiguous AI/AIII. Large sequence variability at the sub-assemblage level was detected, with several double peaks and mutations, making complex a proper isolate allocation. When compared with previous studies, the 35.3% prevalence of G. duodenalis in cats reported in the present article was surprisingly high. Moreover, all positive cats resulted to be infected with zoonotic assemblages/sub-assemblages, thus indicating stray cats as a possible source of human giardiasis and highlighting the sanitary relevance of cat colonies in the study area

A novel allele specific polymerase chain reaction (As-pcr) assay to detect the v1016g knockdown resistance mutation confirms its widespread presence in aedes albopictus populations from Italy

Funding Information: Funding: The study received support from the project ”Sapienza 2017” and JP received support from FCT-Fundação para a Ciência e a Tecnologia, I.P., through national funds in the framework of the project PTDC/BIA-OUT/29477/2017. Funding Information: The study received support from the project ?Sapienza 2017? and JP received support from FCT-Funda??o para a Ci?ncia e a Tecnologia, I.P., through national funds in the framework of the project PTDC/BIA-OUT/29477/2017. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Polymerase chain reaction (PCR)-based genotyping of mutations in the voltage-sensitive sodium channel (vssc) associated with resistance to pyrethroid insecticides is widely used and represents a potential early warning and monitoring system for insecticide resistance arising in mosquito populations, which are vectors of different human pathogens. In the secondary vector Aedes albopictus—an Asian species that has invaded and colonized the whole world, including temperate regions—sequencing of domain II of the vssc gene is still needed to detect the V1016G mutation associated with pyrethroid resistance. In this study we developed and tested a novel allele-specific PCR (AS-PCR) assay to genotype the V1016G mutation in this species and applied it to the analysis of wild populations from Italy. The results confirm the high accuracy of the novel AS-PCR and highlight frequencies of the V1016G allele as >5% in most sampling sites, with peaks of 20–45% in coastal touristic sites where pyrethroid treatments are extensively implemented, mostly for mosquito nuisance reduction. The high frequency of this mutation observed in Italian Ae. albopictus populations should serve as a warning bell, advocating for increased monitoring and management of a phenomenon which risks neutralizing the only weapon today available to counteract (risks of) arbovirus outbreaks.publishersversionpublishe

Unbiased sequence analysis of vgsc gene reveals circulation of novel and known knock-down resistance mutations in Culex pipiens, challenging vector control measures

Pyrethroids, targeting the voltage gated sodium channel (VGSC), are fundamental for the control of arboviral disease circulation. The spread of pyrethroid resistance among vector species represents thus a major public health concern. Culex pipiens is one of the most abundant European mosquito species and main vector of West Nile virus, leading cause of arboviral encephalitis worldwide. Despite this, monitoring of its resistance status and the understanding of underlying mechanisms are widely neglected. Herein, we performed an oligo-hybridization capture approach on 82 Cx. pipiens specimens from Italy and Greece to investigate the whole coding sequence of the vgsc gene for the presence of known and potential knock-down resistance (kdr) mutations associated with target-site resistance to pyrethroids in insects. Among the 26 non-synonymous substitutions revealed by the analysis, the super-kdr haplotype—i.e. the association of kdr-alleles 918T and 1014F, known for conferring a strongly enhanced resistance phenotype in Musca domestica – was revealed for the first time in mosquitoes. Three more potential kdr alleles were detected for the first time in Cx. pipiens and multiple kdr variants were observed for locus 1014, with allele 1014F, reaching frequencies &gt; 80%. Overall, results depict a worrisome situation that could affect the ability to control West Nile virus outbreaks in southern Europe. To avoid this, resistance monitoring needs to be intensified and an enhancement of the diagnostic tool box for the easy detection of different kdr-variants (including in particular the super-kdr haplotype) and for subsequent functional studies on the resistance phenotype of detected variants, is require

VectAbundance: a spatio-temporal database of Aedes mosquitoes observations

Modelling approaches play a crucial role in supporting local public health agencies by estimating and forecasting vector abundance and seasonality. However, the reliability of these models is contingent on the availability of standardized, high-quality data. Addressing this need, our study focuses on collecting and harmonizing egg count observations of the mosquito Aedes albopictus, obtained through ovitraps in monitoring and surveillance efforts across Albania, France, Italy, and Switzerland from 2010 to 2022. We processed the raw observations to obtain a continuous time series of ovitraps observations allowing for an extensive geographical and temporal coverage of Ae. albopictus population dynamics. The resulting post-processed observations are stored in the open-access database VectAbundance.This initiative addresses the critical need for accessible, high-quality data, enhancing the reliability of modelling efforts and bolstering public health preparedness