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Abstract 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.

West Nile virus (WNV) (Flaviviridae, Flavivirus) infection is a mosquito-borne zoonosis able of causing disease and death in humans and animals. Over the past decade, WNV infections have been a significant public health concern in Europe, and Italy has been among the most affected countries since 2008. The 2022 vector season has been characterized by an intense and early circulation of WNV. This report describes cases of co-circulation of WNV L1 and of WNV L2 occurring at the end of the 2022 vector season in Sicily and Tuscany, regions where no strains had ever been sequenced. The phylogenetic analysis of the detected strains confirmed the peculiar WNV scenario that has characterized the Italian West Nile disease (WND) epidemic since its appearance. The circulation observed in Tuscany was in fact a consequence of the spread of endemic strains to new areas while the Sicilian episodes were linked to new introductions of WNV L1 and L2 strains likely from other European countries.

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.

Abstract Background To date, there is a scarcity of information and literature on Macaca maura health status relative to viral diseases. The objectives of the present study were to investigate on the potential spread of enteric and non-enteric viruses shed in the environment through a wild macaque feces and to understand the possible interrelation in the spread of zoonotic viruses in a poorly studied geographical area, the Sulawesi Island. This study will also contribute providing useful information on potential threats to the health of this endangered species. Methods The sampling was conducted between 2014 and 2016 in the Bantimurung Bulusaraung National Park, in the south of the Sulawesi Island and non-invasive sampling methods were used to collect fresh stools of the M. maura, one of the seven macaque species endemic to the island of Sulawesi, Indonesia. The population under study consisted in two wild, neighboring social macaque groups with partially overlapping home ranges; twenty-four samples were collected and examined using negative staining electron microscopy and a panel of PCR protocols for the detection of ten RNA and two DNA viruses. Results Viral particles resembling parvovirus (5 samples), picornavirus (13 samples) and calicivirus (13 samples) were detected by electron microscopy whereas the PCR panel was negative for the 12 viruses investigated, except for one sample positive for a mosquito flavivirus. The results did not correlate with animal sex; furthermore, because all of the animals were clinically healthy, it was not possible to correlate feces consistency with viral presence. Conclusions As information on viral infections in wild moor macaques remains limited, further studies are yet required to identify the fecal–oral and blood transmitted potentially zoonotic viruses, which may infect the moor macaque and other macaque species endemic to the South Sulawesi Island.

Abstract Background The Asian tiger mosquito Aedes albopictus (Skuse 1894), which is native to Southeast Asia, is among the top 100 invasive species worldwide and one of the most troubling vector species. It has become established in more than 20 European countries. Since its arrival in Italy in the 1990s, the species has colonized all the regions of the country, up to an altitude of 600 m. Nevertheless, no thorough investigation has ever been performed to confirm or extend its elevation limit (EL) in Italy. Methods To define the EL of Ae. albopictus and analyse its phenology along an altitudinal gradient, we carried out an investigation by means of ovitraps placed in Lazio region, central Italy. Sampling was performed on a weekly basis in 13 villages within five 200-m altitudinal ranges [0–1000 m above sea level (asl)], with the addition of higher localities to the species range whenever the species was recorded in the highest range. Results Aedes albopictus has colonized sites well beyond its known EL, with established populations at 900 m asl and positive ovitraps recorded at 1193 m asl. The relationship between egg abundance and elevation was described by an exponential decay regression, which predicted an EL for oviposition at 1015 m asl. In the active season, egg-laying started earlier at low altitude and ended earlier within the highest altitudinal range. Aedes albopictus abundance and activity period (number of days active) decreased, respectively, by 95% and 34% from the lowest to the highest altitudinal range. Conclusions Using data from the present study, the altitudinal limit of Ae. albopictus in central Italy was updated from 600 to 900 m asl. In addition, established populations were predicted to exist up to 1015 m asl. Considering that up to 99.5% of Lazio region’s inhabitants could potentially be affected by Aedes-borne virus outbreaks, the surveillance area for Ae. albopictus should be expanded accordingly. However, our results also indicate that Ae. albopictus surveillance programs need to be revised in order to harmonize the resources earmarked for these with the altitudinal changes in the phenology of this species. Graphical abstract

Abstract Baylisascaris procyonis is a nematode parasite of the raccoon (Procyon lotor), and it can be responsible for a severe form of larva migrans in humans. This parasite has been reported from many countries all over the world, after translocation of its natural host outside its native geographic range, North America. In the period between January and August 2021, 21 raccoons were cage-trapped and euthanized in Tuscany (Central Italy), in the context of a plan aimed at eradicating a reproductive population of this non-native species. All the animals were submitted for necroscopic examination. Adult ascariids were found in the small intestine of seven raccoons (prevalence 33.3%). Parasites have been identified as B. procyonis based on both morphometric and molecular approaches. The aim of the present article is to report the first finding of this zoonotic parasite from Italy, highlighting the sanitary risks linked to the introduction of alien vertebrate species in new areas. Graphical Abstract

The Asian tiger mosquito, Aedes albopictus, competent vector of several arboviruses, poses significant impact on human health worldwide. Although global warming is a driver of A. albopictus range expansion, few studies focused on its effects on homodynamicity (i.e. the ability to breed all-year-round), a key factor of vectorial capacity and a primary condition for an Aedes-borne disease to become endemic in temperate areas. Data from a 4-year monitoring network set in Central Italy and records from weather stations were used to assess winter adult activity and weekly minimum temperatures. Winter oviposition occurred in 38 localities with a seasonal mean photoperiod of 9.7 : 14.3 (L : D) h. Positive collections (87) occurred with an average minimum temperature of the two and three weeks before sampling of approximately 4°C. According to these evidences and considering the climate projections of three global climate models and three shared socio-economic pathways for the next three 20-year periods (from 2021 to 2080), the minimum temperature of January will increase enough to allow an all-year-round oviposition of A. albopictus in several areas of the Mediterranean Basin. Due to vector homodynamicity, Aedes-borne diseases could become endemic in Southern Europe by the end of the twenty-first century, worsening the burden on human health.

Abstract Background West Nile (WNV) and Usutu (USUV) are emerging vector-borne zoonotic flaviviruses. They are antigenically very similar, sharing the same life cycle with birds as amplification host, Culicidae as vector, and man/horse as dead-end host. They can co-circulate in an overlapping geographic range. In Europe, surveillance plans annually detect several outbreaks. Methods In Italy, a WNV/USUV surveillance plan is in place through passive and active surveillance. After a 2018 WNV outbreak, a reinforced integrated risk-based surveillance was performed in four municipalities through clinical and serological surveillance in horses, Culicidae catches, and testing on human blood-based products for transfusion. Results Eight WNV cases in eight equine holdings were detected. Twenty-three mosquitoe catches were performed and 2367 specimens of Culex pipiens caught; 17 pools were USUV positive. A total of 8889 human blood donations were tested, and two asymptomatic donors were USUV positive. Conclusions Different surveillance components simultaneously detected WNV only in horses and USUV only in humans and mosquitoes. While in endemic areas (i.e. northern Italy) entomological surveillance is successfully used as an early detection warning, this method in central Italy seems ineffective. To achieve a high level of sensitivity, the entomological trapping effort should probably exceed a reasonable balance between cost and performance. Besides, WNV/USUV early detection can be addressed by horses and birds. Further research is needed to adapt the surveillance components in different epidemiological contexts.

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 implications.

Abstract Background Scalp Eschar and Neck LymphAdenopathy after Tick bite is a zoonotic non-pathogen-specific disease most commonly due to Rickettsia slovaca and Rickettsia raoultii. Diagnosis is mostly based only on epidemiological and clinical findings, without serological or molecular corroboration. We presented a clinical case in which diagnosis was supported by entomological identification and by R. slovaca DNA amplifications from the tick vector. Case presentation A 6-year-old child presented with asthenia, scalp eschar and supraclavicular and lateral-cervical lymphadenopathy. Scalp Eschar and Neck LymphAdenopathy After Tick bite syndrome following a Dermacentor marginatus bite was diagnosed. Serological test on serum revealed an IgG titer of 1:1024 against spotted fever group rickettsiae, polymerase chain reaction assays on tick identified Rickettsia slovaca. Patient was successfully treated with doxycycline for 10 days. Conclusions A multidisciplinary approach including epidemiological information, clinical evaluations, entomological identification and molecular investigations on tick, enabled proper diagnosis and therapy.

The raccoon (Procyon lotor) is a carnivore native to North and Central America, gradually introduced into Asia and Europe, including Italy. It is an important carrier of multiple endoparasites, both Protozoa and Helminths, some of them being zoonotic. The aim of this study was to investigate the endoparasites of the non-native raccoon population of Central Italy. Sixty-two raccoons were collected by local competent authorities (sixty trapped and euthanized, two found dead) and subjected to necroscopic examination. Carcasses underwent a broad parasitological investigation, including coprological techniques (macroscopic examination of the gastrointestinal tract, lungs, trachea, and heart, Flotac®, Baermann test, and immunofluorescence for Giardia duodenalis and Cryptosporidium spp.), research on respiratory/urinary capillariosis and artificial digestion for Trichinella spp. larvae, and a histopathological examination of the ileum. Ascarid parasites were further identified at the species level using a next-generation sequencing-based amplicon sequencing approach. The results showed the presence of different Protozoa and Nematodes: Baylisascaris procyonis (26/62; 41.9%), Pearsonema sp. (6/62; 9.6%), Capillariidae (6/62; 9.6%), Eimeria sp. (2/62; 3.2%), Cryptosporidium sp. (2/62; 3.2%), and Ancylostomatidae (2/62; 3.2%). B. procyonis is an emerging helminthic zoonotic agent considered a serious concern for public and animal health, given the possibility of its transmission to paratenic hosts, including humans and pets. The demonstrated role of the raccoon as a multi-parasite carrier should be an incentive to continuing the eradication/control of this alien species, and supports the need to implement related disease surveillance programs.

Abstract Background Pediculus humanus, the human body louse, is widespread where overcrowding and lack of hygiene are present, in areas of the world affected by poverty, war, famine and presence of refugees. It has recently been considered re-emerging among homeless populations in developed countries. In Italy, it was last reported in 1945. Pediculus humanus is a vector of highly relevant human pathogens. Methods In October 2018, a woman found small insects on a T-shirt bought second-hand in a local street market in a village 35 km south of Rome (central Italy). Insects were identified both morphologically and by molecular analysis. Moreover, they were analyzed molecularly for the presence of Rickettsia prowazekii, Borrelia recurrentis, Bartonella quintana, Coxiella burnetii and Yersinia pestis. Results Morphological and molecular analyses of the insects identified them as 26 lice (12 females, 10 males and 4 nymphs) of the species P. humanus. Many nits were found on the T-shirt seams. DNA of the investigated pathogens was not detected in any of the lice. Conclusions The exceptionality of the described case lies both in the report of P. humanus from a country where it had not been reported since 1945, and in its finding from second-hand clothes for sale in a market, constituting a potential source of infection for people buying this type of goods. The question arises, how did adults and nits of P. humanus infest clothes for sale on a market stall in a country where it had not been reported for decades. Given that the body louse requires frequent blood meals to survive and develop, its arrival on clothes imported from abroad is highly improbable. Hence, it must be presumed that people infected with the human body louse are present in Italy. This report points out a serious regulatory problem regarding the management of second-hand clothes prior to sale and, more generally, of controls in street markets.

This paper reported a case of a metastrongyloid nematode Angiostrongylus vasorum infection in a fennec (Vulpes zerda) kept in a zoo in central Italy. The fennec had shown paralysis of the hind limbs, anorexia, weakness and respiratory signs before death. Cardiomegaly and granulomatous pneumonia were the major anatomopathological findings. Inflammatory lesions associated with parasitic larvae were observed in the lungs, brain, liver, heart, spinal cord and kidney of the fennec at histology. A. vasorum diagnosis was confirmed by both morphological and molecular identification of adult worms recovered at necropsy. Fennecs are active predators and maintain their hunting behaviour in captivity. Hence, it is likely that the animal was exposed to infection by preying on parasitised gastropods, intermediate hosts of A. vasorum, entering zoo enclosures from the surrounding environment. This is the first report of A. vasorum systemic infection in a captive fennec (V. zerda) in a zoo in Italy.

Avian malaria is a parasitic disease of birds caused by protozoa belonging to the genus Plasmodium, within the order Haemosporida. Penguins are considered particularly susceptible, and outbreaks in captive populations can lead to high mortality. We used a multidisciplinary approach to investigate the death due to avian malaria, occurred between 2015 and 2019, in eight African penguins (Spheniscus demersus) kept in two Italian zoos located in central Italy, and situated about 30 km apart. We also provided information about the presence and circulation of Plasmodium spp. in mosquitoes in central Italy by sampling mosquitoes in both zoos where penguin mortalities occurred. In the eight dead penguins, gross and histopathological lesions were consistent with those previously observed by other authors in avian malaria outbreaks. Organs from dead penguins and mosquitoes collected in both zoos were tested for avian malaria parasites by using a PCR assay targeting the partial mitochondrial conserved region of the cytochrome b gene. Identification at species level was performed by sequencing analysis. Plasmodium matutinum was detected in both dead penguins and in mosquitoes (Culex pipiens), while Plasmodium vaughani in Culex pipiens only. Parasites were not found in any of the PCR tested Aedes albopictus samples. Based on our phylogenetic analysis, we detected three previously characterized lineages: Plasmodium matutinum LINN1 and AFTRU5, P. vaughani SYAT05. In Culex pipiens we also identified two novel lineages, CXPIP32 (inferred morphospecies Plasmodium matutinum) and CXPIP33 (inferred morphospecies P. vaughani). Significantly, LINN1 and AFTRU5 were found to be associated to penguin deaths, although only LINN1 was detected both in penguins (along the years of the study) and in Culex pipiens, while AFTRU5 was detected in a single penguin dead in 2017. In conclusion, in our study Plasmodium matutinum was found to cause avian malaria in captive penguins kept in Europe, with Culex pipiens being its most probable vector. Our results are in agreement with previous studies suggesting that Culex pipiens is one of the main vectors of Plasmodium spp. in Europe and the Northern Hemisphere. Zoos maintaining captive penguins in temperate areas where Culex pipiens is abundant should be well aware of the risks of avian malaria, and should put every effort to prevent outbreaks, in particular during the periods when the number of vectors is higher.

Wildlife is frequently infected by intestinal protozoa, which may threaten their fitness and health. A diverse community of Eimeria species is known to occur in the digestive tract of mountain-dwelling ungulates, including chamois (genus Rupicapra). However, available data on Eimeria diversity in these taxa is at times inconsistent and mostly dated. In the present study, we aimed to revisit the occurrence of Eimeria spp. in the Alpine subspecies of the Northern chamois (Rupicapra rupicapra rupicapra) and the Apennine subspecies of the Southern chamois (Rupicapra pyrenaica ornata) in Italy, using an integrated approach based on a hierarchical cluster analysis (HCPC) applied to oocyst morphology and morphometry. A total of 352 fecal samples were collected from R. r. rupicapra (n = 262) and R. p. ornata (n = 90). Overall, 85.3% (300/352) of the animals tested microscopically positive to Eimeria spp. Based on morphological analysis, we identified all the eimerian species described in chamois. Through the HCPC method, five clusters were generated, corresponding to E. suppereri, E. yakimoffmatschoulskyi, E. riedmuelleri (two different clusters), and E. rupicaprae morphotypes. The well-defined clusters within E. riedmuelleri support the existence of two distinct morphological groups, possibly referable to different taxonomic units. This study suggests that combining a morphometrical approach with a powerful statistical method may be helpful to disentangle uncertainties in the morphology of Eimeria oocysts and to address taxonomic studies of eimeriid protozoa at a specific host taxon level.

Fragmented data are so far available on genotype diversity of G. duodenalis in wildlife in different countries in Europe, in particular, in Italy. In the present study, G. duodenalis sequences obtained from different Italian wild animals [12 porcupines (Hystrix cristata), 4 wild boars (Sus scrofa), 1 wolf (Canis lupus italicus), 6 Alpine chamois (Rupicapra rupicapra rupicapra)] were compared with those available from wild host species in Europe to add new data on the geographic distribution of Giardia assemblages/sub-assemblages and their transmission patterns among natural hosts. Thirty-eight sequences were obtained by MLG analysis (SSU-rRNA, bg, gdh, and tpi genes) and subsequently compared by phylogenetic and network analyses with those from wild species monitored in the last decades in Europe. The results revealed the presence of potentially zoonotic (A-AI, A-AII from wild boar; B from porcupine) and host-adapted (D from wolf; E, A-AIII from chamois) assemblages and sub-assemblages and represent the first report for Italian wild boar. The analysis did not find any evidence of spatial or host segregation for specific genetic variants, mostly shared between different hosts from different European countries. However, conflicting evidence was found in genotypic assignment, advocating for data improvement and new genomic approaches.

Adult specimens of Trichuris sp. collected from crested porcupines (Hystrix cristata) from Italy were characterized using an integrative taxonomic approach involving morphological and molecular tools. The morphological features of this Trichuris sp. were compared to data already available for Trichuris spp. from Hystrix sp., revealing diagnostic traits, such as spicule length in males or vulva shape in females, which distinguish this Trichuris sp. from the other species. Evidence from sequences analysis of the partial mitochondrial COX1 region indicated that the taxon under study is a distinct lineage. Biometrical and genetic data suggested this Trichuris sp. to be a valid and separated taxon. However, since molecular data from other Trichuris spp. infecting Hystrix, such as T. infundibulus, T. hystricis, T. javanica, T.landak and T. lenkorani, are missing in public repositories, the number and identity of distinct lineages able to infect porcupines remain only partially defined.

Abstract Background Angiostrongylus vasorum is a nematode residing in the heart and pulmonary vessels of dogs and wild carnivores. In Europe the red fox is its reservoir, while only three records from wolves have been published. Angiostrongylus vasorum has a worldwide distribution, and many pieces of evidence demonstrate that it is spreading from endemic areas to new ones. In Italy, A. vasorum was reported with increasing frequency in dogs and foxes in the last decades, and now it is considered endemic throughout the country. Angiostrongylus vasorum can be asymptomatic or cause respiratory and circulatory disorders, at times causing severe disseminated infections. Methods Between February 2012 and December 2016, 25 wolves found dead in central Italy were submitted to the Istituto Zooprofilattico del Lazio e della Toscana for post-mortem examination. Samples of lungs, heart, liver, spleen, kidneys, mediastinic lymph nodes and brain were collected from each animal for histological examination. When adult and larval nematodes were microscopically seen in lungs, the other organs were processed, and five histological sections for each organ were examined. To confirm parasite identification, lung samples were submitted to a PCR-sequencing protocol targeting the ITS2 region of A. vasorum. Results Seven wolves (28.0%) harboured nematode larvae in lung sections. In two of the positive wolves, adult nematodes were visible in pulmonary arteries, in four animals larvae were also detected in other organs. DNA sequencing reactions confirmed parasite identification as A. vasorum in all the cases. Conclusions As a result of the high prevalence of A. vasorum reported in wolves in the present study, a focus of high circulation could be hypothesised in central Italy. Nevertheless, the similarly high prevalence in foxes originating from the same areas were reported in previous papers. Histopathological evidence highlights the pathogenic potential of A. vasorum in the wolf, especially in juvenile animals.

Intestinal and respiratory parasites are among the most common pathogens in dogs and some of them are recognized as zoonotic agents. In Italy, various taxa have been reported, with variable prevalence estimates depending on study area, dog category and coprological exam technique. In this paper, we report the results of six years of passive surveillance. In the period January 2006-December 2012, 2,775 dog faecal samples from Lazio Region (Central Italy), were examined for parasites, 1,156 from household and 1,619 from shelter dogs, respectively. The following parasites were detected: Giardia duodenalis, Cystoisospora sp., Ancylostomatidae, Toxocara canis, Toxascaris leonina, Trichuris vulpis, Eucoleus böhmi, Eucoleus aerophilus, Mesocestoides sp., Taeniidae and Dipylidium caninum. Helminths were more frequent than protozoa, with total prevalences of 29.1% and 10.7%, respectively. T. vulpis and Ancylostomatidae were the most common parasites, with prevalences of 9.9% and 9.6% respectively. T. vulpis and Ancylostomatidae were significantly more prevalent in shelter dogs than in household ones. T. canis and Cystoisospora sp. were significantly more frequent in household dogs.The assessment of the prevalence in sheltered and in household dogs is useful to infer the occurrence of different parasites in the origin population and to plan possible control intervention. Keywords: Ancylostomatidae, Cystoisospora sp., Dog, Shelter, T. Canis, T. vulpis, Zoonosis

Avian malaria is a worldwide distributed, vector-born disease of birds caused by parasites of the order Haemosporida. There is a lack of knowledge about the presence and pathogenetic role of Haemosporida in Psittacidae. Here we report a case of avian malaria infection in lovebirds (Agapornis roseicollis), with the genetic characterization of the Plasmodium species involved. The birds were hosted in a zoo located in Italy, where avian malaria cases in African penguins (Spheniscus demersus) were already reported. Animals (n = 11) were submitted for necropsy after sudden death and were subjected to further analyses including histopathology, bacteriology, and PCR for the research of haemosporidians. Clinical history, gross lesions and histopathological observation of schizonts, together with positive PCR results for Plasmodium spp., demonstrated that avian malaria was the cause of death for one animal and the possible cause of death for the other nine. The sequences obtained were compared using BLAST and analyzed for similarity to sequences available at the MalAvi database. Genetic analyses demonstrated a 100% nucleotide identity to Plasmodium matutinum LINN1 for all the obtained sequences. To our knowledge, this is the first report describing avian malaria in lovebirds.

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.

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.

Abstract Background Although chamois Rupicapra spp. are the most abundant mountain ungulates in Europe, no data are available on the presence of Giardia duodenalis infecting these species. Methods A total of 157 fecal samples from Alpine Rupicapra rupicapra rupicapra and Apennine Rupicapra pyrenaica ornata chamois were tested for the presence of G. duodenalis by immunofluorescence test, quantitative Real Time PCR and end-point PCR for genotype characterization. Results G. duodenalis was detected in R. r. rupicapra and R. p. ornata, with a percentage value of 4.45 (5.82 and 1.85 %, respectively), and a cyst burden of up to 31,800 cysts/g of feces. Assemblages A/AI and E were identified in R. r. rupicapra and assemblage A/AIII in R. p. ornata. Conclusions The present study represents the first record of Giardia duodenalis in Rupicapra spp., suggesting that these wild bovids can play an epidemiological role in environmental contamination and transmission of both zoonotic and non-zoonotic genotypes.

The present study was carried out in Circeo National Park (Lazio region, Central Italy), in order to collect data about mosquito (Diptera, Culicidae) fauna in a protected area for biodiversity. From 2003 to 2004 seasonal surveys allowed to collect and to identify 380 larvae and 713 adult mosquitoes in 6 sites. A total of 15 mosquito species belonging to 6 genera were recorded; the most abundant species were Culex pipens Linnaeus, 1758 known as the main West Nile virus vector, Ochlerotatus detritus (Haliday, 1933) and Culiseta annulata (Dhrank, 1776). Present data show a noteworthy number of other mosquito species, even if less abundant, reflecting the considerable environmental richness. Respect to the past collections of Anophelinae mosquitoes carried out in the same area once affected by malaria, the present research represents the first monitoring of the whole Culicidae Family in Circeo National Park, up to now. This paper reports the collected data as a first base for a future checklist in this protected area.

The distribution of Bluetongue virus (BTV) in Europe can be represented by two distinct and interconnected epidemiological systems (episystems), each characterized by different ecological characteristics and vector species. This study investigated the vector competence of Italian populations of Culicoides imicola and Culicoides obsoletus/scoticus to some representative BTV strains after artificial oral infection. The BTV strains were selected according to their ability to spread to one or both episystems and included BTV-4 ITA, responsible of the recent Italian and French BTV-4 outbreaks; the BTV-2 strain which caused the first BTV incursion in Italy, Corsica, and Balearic Islands; BTV-4 MOR, responsible for the epidemic in Morocco; and BTV-8, the strain which spread through Europe between 2006 and 2008. Blood-soaked cotton pledgets and Hemotek membrane feeder using Parafilm® membrane were used to artificially feed midges. For each population/strain, recovery rates (positive/tested heads) were evaluated using serogroup- and serotype-specific RT-PCR. The trial demonstrated that, except for the Abruzzo population of C. obsoletus/C. scoticus, which was refractory to BTV-4 MOR infection, all the investigated Culicoides populations are susceptible to the selected BTV strains and that, if prompt vaccination programs and restriction measures had not been implemented, BTV-2 and BTV-4 MOR could have spread all over Europe.