The correct identification of mosquito vectors is often hampered by the presence of morphologically indiscernible sibling species. The Maculipennis complex is one of these groups that include both ...malaria vectors of primary importance and species of low/negligible epidemiological relevance, of which distribution data in Italy are outdated. Our study was aimed at providing an updated distribution of Maculipennis complex in Northern Italy through the sampling and morphological/molecular identification of specimens from five regions. The most abundant species was Anopheles messeae (2032), followed by Anopheles maculipennis s.s. (418), Anopheles atroparvus (28) and Anopheles melanoon (13). Taking advantage of ITS2 barcoding, we were able to finely characterize tested mosquitoes, classifying all the Anopheles messeae specimens as Anopheles daciae, a taxon with debated rank to which we referred as species inquirenda (sp. inq.). The distribution of species was characterized by Ecological Niche Models (ENMs), fed by recorded points of presence. ENMs provided clues on the ecological preferences of the detected species, with An. daciae sp. inq. linked to stable breeding sites and An. maculipennis s.s. more associated to ephemeral breeding sites. We demonstrate that historical Anopheles malaria vectors are still present in Northern Italy.
Abstract
Aedes koreicus
is an invasive mosquito species which has been introduced into several European countries. Compared to other invasive
Aedes
mosquitoes, little is known of its biology and ...ecology. To determine
Ae. koreicus
’ vectorial capacity, it is essential to establish its feeding patterns and level of anthropophagy. We report on the blood-feeding patterns of
Ae. koreicus
, examining the blood meal origin of engorged females and evaluating the influence of different biotic and abiotic factors on feeding behavior. Mosquitoes were collected in 23 sites in northern Italy by manual aspiration and BG-sentinel traps; host availability was estimated by survey. The source of blood meals was identified using a nested PCR and by targeting and sequencing the cytochrome c oxidase subunit I gene. In total, 352
Ae. koreicus
engorged females were collected between 2013 and 2020 and host blood meals were determined from 299 blood-fed mosquitoes (84.9%). Eleven host species were identified, with the highest prevalences being observed among roe deer (
Capreolus capreolus
) (
N
= 189, 63.2%) and humans (
N
= 46, 15.4%). Blood meals were mostly taken from roe deer in forested sites and from humans in urban areas, suggesting that this species can feed on different hosts according to local abundance. Two blood meals were identified from avian hosts and one from lizard.
Ae. koreicus’
mammalophilic feeding pattern suggests that it may be a potential vector of pathogens establishing transmission cycles among mammals, whereas its role as a bridge vector between mammals and birds could be negligible.
Recent climate and environmental changes have resulted in the geographical expansion of Mediterranean Leishmania infantum vectors towards northern latitudes and higher altitudes in different European ...countries, including Italy, where new foci of canine leishmaniasis have been observed in the northern part of the country. Northern Italy is also an endemic area for mosquito-borne diseases. During entomological surveillance for West Nile virus, mosquitoes and other hematophagous insects were collected, including Phlebotomine sand flies. In this study, we report the results of Phlebotomine sand fly identification during the entomological surveillance conducted from 2017 to 2019.
The northeastern plain of Italy was divided by a grid with a length of 15 km, and a CO
-CDC trap was placed in each geographical unit. The traps were placed ~ 15 km apart. For each sampling site, geographical coordinates were recorded. The traps were operated every two weeks, from May to November. Sand flies collected by CO
-CDC traps were identified by morphological and molecular analysis.
From 2017 to 2019, a total of 303 sand flies belonging to the species Phlebotomus perniciosus (n = 273), Sergentomyia minuta (n = 5), P. mascittii (n = 2) and P. perfiliewi (n = 2) were collected, along with 21 unidentified specimens. The trend for P. perniciosus collected during the entomological surveillance showed two peaks, one in July and a smaller one in September. Sand flies were collected at different altitudes, from -2 m above sea level (a.s.l.) to 145 m a.s.l. No correlation was observed between altitude and sand fly abundance.
Four Phlebotomine sand fly species are reported for the first time from the northeastern plain of Italy. Except for S. minuta, the sand fly species are competent vectors of Leishmania parasites and other arboviruses in the Mediterranean Basin. These findings demonstrate the ability of sand flies to colonize new environments previously considered unsuitable for these insects. Even though the density of the Phlebotomine sand fly population in the plain areas is consistently lower than that observed in hilly and low mountainous areas, the presence of these vectors could herald the onset of epidemic outbreaks of leishmaniasis and other arthropod-borne diseases in areas previously considered non-endemic.
BACKGROUND: Over the recent decades, container-breeding mosquito species belonging to the genus Aedes have frequently been recorded far from their place of origin. Aedes koreicus was first reported ...in north-eastern Italy in 2011, in a region endemic for Dirofilaria immitis, the agent of canine heartworm disease. The vector competence of Ae. koreicus for D. immitis was here tested under laboratory conditions, by infecting mosquitoes with a local strain of D. immitis. METHODS: Blood containing 3000 microfilariae/ml was offered to 54 mosquitoes (T group) while 29 were left as a control (C group). Mosquitoes killed at scheduled days post infection (dpi) and naturally dead were divided in head, thorax and abdomen and examined for D. immitis larval stages by dissection under a microscope and molecularly. RESULTS: Of the 45 engorged mosquitoes in T, 32 (71.1%) scored positive for D. immitis larval stages. L3 were found as early as 8 dpi in the Malpighian tubules and then in the thorax, salivary glands, palp and proboscis. At the end of the study a total of 18 mosquitoes developed L3 giving an estimated infection rate at 12 dpi of 68.2% and a vector efficiency index of 25.2%. The rate of mortality in T group within the first 9 days post infection was significantly higher in T group (47.6%) than in C group (8.3%) (p < 0.01). The concordance between microscopy and PCR was high (0.8-0.9), however, a positivity for D. immitis in the head was found molecularly at 13 dpi, three days before microscopy. CONCLUSIONS: Aedes koreicus, a new invasive species for Europe, is most likely a competent vector of D. immitis being of potential relevance in the natural cycle of the parasite. This poses a new threat for animal and human health in endemic areas for dirofilariosis and enhances the risk of spreading the infection in previously non-endemic areas. These results stress the importance of active surveillance and control strategies to minimize the risk of introduction of invasive alien species.
BACKGROUND: Ticks may transmit a large variety of pathogens, which cause illnesses in animals and humans, commonly referred to as to tick-borne diseases (TBDs). The incidence of human TBDs in Italy ...is underestimated because of poor surveillance and the scant amount of studies available. METHODS: Samples (n = 561) were collected from humans in four main geographical areas of Italy (i.e., northwestern, northeastern, southern Italy, and Sicily), which represent a variety of environments. After being morphologically identified, ticks were molecularly tested with selected protocols for the presence of pathogens of the genera Rickettsia, Babesia, Theileria, Candidatus Neoehrlichia mikurensis, Borrelia and Anaplasma. RESULTS: Ticks belonged to 16 species of the genera Argas, Dermacentor, Haemaphysalis, Hyalomma, Ixodes and Rhipicephalus, with Ixodes ricinus (59.5%) being the species most frequently retrieved, followed by Rhipicephalus sanguineus sensu lato (21.4%). Nymphs were the life stage most frequently retrieved (41%), followed by adult females (34.6%). The overall positivity to any pathogen detected was 18%. Detected microorganisms were Rickettsia spp. (17.0%), Anaplasma phagocytophilum (0.8%), Borrelia afzelii (0.5%), Borrelia valaisiana (0.3%), C. N. mikurensis (0.5%) and Babesia venatorum (0.6%). CONCLUSIONS: Results indicate that people living in the Italian peninsula are at risk of being bitten by different tick species, which may transmit a plethora of TBD causing pathogens and that co-infections may also occur.
West Nile virus lineage 2 (WNV-2) was mainly confined to sub-Saharan Africa until the early 2000s, when it was identified for the first time in Central Europe causing outbreaks of human and animal ...infection. The aim of this study was to reconstruct the origin and dispersion of WNV-2 in Central Europe and Italy on a phylodynamic and phylogeographical basis. To this aim, discrete and continuous space phylogeographical models were applied to a total of 33 newly characterised full-length viral genomes obtained from mosquitoes, birds and humans in Northern Italy in the years 2013-2015 aligned with 64 complete sequences isolated mainly in Europe. The European isolates segregated into two highly significant clades: a small one including three sequences and a large clade including the majority of isolates obtained in Central Europe since 2004. Discrete phylogeographical analysis showed that the most probable location of the root of the largest European clade was in Hungary a mean 12.78 years ago. The European clade bifurcated into two highly supported subclades: one including most of the Central/East European isolates and the other encompassing all of the isolates obtained in Greece. The continuous space phylogeographical analysis of the Italian clade showed that WNV-2 entered Italy in about 2008, probably by crossing the Adriatic sea and reaching a central area of the Po Valley. The epidemic then spread simultaneously eastward, to reach the region of the Po delta in 2013, and westward to the border area between Lombardy and Piedmont in 2014; later, the western strain changed direction southward, and reached the central area of the Po valley once again in 2015. Over a period of about seven years, the virus spread all over an area of northern Italy by following the Po river and its main tributaries.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
In Northern Italy, a hastening of hare population decline was noticeable from 2008. In the same year hunters reported a sudden increase of hares infected by Taenia sp. larvae, whose morphology was ...consistent with T. pisiformis cysticerci. The aim of the survey was: i)to identify the parasites through morphological features and molecular techniques; ii)to quantify the prevalence and abundance of cysticerci in hunted hares; iii)to describe pathological aspects of parasite-induced lesions; iv)to evaluate the short-term trend of the infection comparing two different hunting seasons; v)to highlight possible relationship between T. pisiformis infection and hare-related variables.
In 2013, 2015 the viscera of 54 and 61 hares legally hunted in agro-ecosystems of the Po Plain were collected.
Peritoneum, liver and lungs were examined for cysticercosis; abundance was estimated counting superficial parasites in liver; parasites were microscopically identified by shape and measure of both large and small hooks. One cysticercus from each hare was analized by a PCR targeting Taeniid species and then sequenced. Frozen liver, lungs and gastrointestinal peritoneum were macroscopically observed and, after thawing, representative samples from the available organs were collected for histologic examination to verify parasitic cysts and the subsequent damage of the involved organs. Sex, weight and age class of the animals were recorded. Generalized linear models were used for statistical analysis.
T. pisiformis was isolated in 8 hares in 2013 (prevalence 14.8%; abundance range: 0–400; mean abundance 17.8) and in 2 hares in 2015 (prevalence 3.28%; abundance range: 0–180; mean abundance 3.22). Identification was confirmed morphologically and by PCR. The DNA sequencing confirmed T. pisiformis in all samples. The sequences were all identical each-other. Infection was significantly related with adult age class, sampling year and low full-weight.
Epidemiological and pathological pattern suggest both a possible role on host population health and a tendency toward host-parasite equilibrium.
•Hastening of hare population decline was noticeable in Northern Italy.•Hunters reported a sudden increase of hares infected by Taenia sp. larvae.•T. pisiformis was isolated.•Observed pattern suggest both a possible role on host population health and a tendency toward host-parasite equilibrium.
Surveillance of Echinococcus multilocularis at the edge of its range is hindered by fragmented distributional patterns and low prevalence in definitive hosts. Thus, tests with adequate levels of ...sensitivity are especially important for discriminating between infected and non-infected areas. In this study we reassessed the prevalence of E. multilocularis at the southern border of its distribution in Province of Bolzano (Alto Adige, northeastern Alps, Italy), to improve surveillance in wildlife and provide more accurate estimates of exposure risk. We compared the diagnostic test currently implemented for surveillance based on coproscopy and multiplex PCR (CMPCR) to a real-time quantitative PCR (qPCR) in 235 fox faeces collected in 2019 and 2020. The performances of the two tests were estimated using a scraping technique (SFCT) applied to the small intestines of a subsample (n = 123) of the same foxes as the reference standard. True prevalence was calculated and the sample size required by each faecal test for the detection of the parasite was then estimated. True prevalence of E. multilocularis in foxes (14.3%) was markedly higher than reported in the last decade, which was never more than 5% from 2012 to 2018 in the same area. In addition, qPCR showed a much higher sensitivity (83%) compared to CMPCR (21%) and agreement with the reference standard was far higher for qPCR (0.816) than CMPCR (0.298) meaning that for the latter protocol, a smaller sample size would be required to detect the disease. Alto Adige should be considered a highly endemic area. Routine surveillance on definitive hosts at the edges of the E. multilocularis distribution should be applied to smaller geographic areas, and rapid, sensitive diagnostic tools using directly host faeces, such as qPCR, should be adopted.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Feline vector-borne pathogens (FeVBPs) have been increasingly investigated for their impact on cat health and their zoonotic potential. The aim of the present study was to assess the prevalence of ...FeVBPs and haemoplasmas in cats across Italy and to identify potential risk factors linked to their occurrence.
Blood samples from 958 owned cats living in the North (n = 556), Centre (n = 173) and South (n = 229) of Italy were tested for Babesia spp., Hepatozoon spp., Ehrlichia spp., Anaplasma spp. and filarioids by conventional PCR (cPCR) and for haemoplasmas and Bartonella spp. by SYBR green real-time PCR. Cats included in the study represent a sub-sample from a larger number of animals enrolled in a previous study, which were selected based on the geographical origin. Data on cats' positivity for Leishmania infantum, feline leukaemia virus (FeLV) and for feline immunodeficiency virus (FIV), available from the previous study, were included and examined. Potential risk factors for pathogen infection were assessed in relationship to categorical variables including sex, geographical origin, breed, neutering status and age of cats.
Out of the 958 cats, 194 (20.2%) were positive for at least one of the tested pathogens, 89 (16%) from the North, 32 (18.5%) from the Centre and 73 (31.9%) from the South of Italy. A high prevalence of FeVBPs was detected in male cats (n = 125, 27.8%), living in the southern part of the country (n = 73, 31.9%), younger than 18 months of age (n = 24, 22.4%) and not neutered (n = 39; 27.5%). In particular, 24 cats (2.5%) tested PCR-positive for Bartonella spp., of which 1.6% for B. henselae and 0.9% for B. clarridgeiae. A total of 111 cats scored PCR-positive for haemoplasmas (11.6%), specifically "Candidatus Mycoplasma haemominutum" (n = 95, 9.9%), M. haemofelis (n = 14, 1.5%) and "Candidatus Mycoplasma turicensis" (n = 2, 0.2%). Moreover, 39, 31 and 8 cats were positive for FeLV (4.1%), L. infantum (3.2%) and FIV (0.8%), respectively. Co-infections were registered for 19 (9.8%) cats.
These results confirm the occurrence of haemoplasmas and FeVBPs throughout Italy. Preventive measures to protect both animal and human health should be carried out also for owned cats, even if no health status of animals has been assessed in this study.
Echinococcus multilocularis is a small tapeworm affecting wild and domestic carnivores and voles in a typical prey-predator life cycle. In Italy, there has been a focus of E. multilocularis since ...1997 in the northern Italian Alps, later confirmed in red foxes collected from 2001 to 2005. In this study, we report the results of seven years of monitoring on E. multilocularis and other cestodes in foxes and describe the changes that occurred over time and among areas (eco-regions) showing different environmental and ecological features on a large scale.
Eggs of cestodes were isolated from feces of 2872 foxes with a sedimentation/filtration technique. The cestode species was determined through multiplex PCR, targeting and sequencing ND1 and 12S genes. Analyses were aimed to highlight variations among different eco-regions and trends in prevalence across the study years.
Out of 2872 foxes, 217 (7.55%) samples resulted positive for cestode eggs at coproscopy, with differences of prevalence according to year, sampling area and age class. Eight species of cestodes were identified, with Taenia crassiceps (2.65%), Taenia polyacantha (1.98%) and E. multilocularis (1.04%) as the most represented. The other species, Mesocestoides litteratus, Taenia krabbei, T. serialis, T. taeniaeformis and Dipylidium caninum, accounted for < 1% altogether. Echinococcus multilocularis was identified in foxes from two out of six eco-regions, in 30 fecal samples, accounting for 1.04% within the cestode positives at coproscopy. All E. multilocularis isolates came from Bolzano province. Prevalence of cestodes, both collectively and for each of the three most represented species (T. crassiceps, T. polyacantha and E. multilocularis), varied based on the sampling year, and for E. multilocularis an apparent increasing trend across the last few years was evidenced.
Our study confirms the presence of a focus of E. multilocularis in red foxes of northeast Italy. Although this focus seems still spatially limited, given its persistence and apparent increasing prevalence through the years, we recommend research to be conducted in the future on the ecological factors that, on a smaller scale, allow this zoonotic species to persist. On the same scale, we recommend a health education campaign to inform on the measures to prevent this zoonosis, targeted at people living in the area, especially hunters, dog owners, forestry workers and other potentially exposed categories.
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