Ecological niche modelling of Culicoides imicola and future range shifts under climate change scenarios in Italy.

Culicoides imicola is the main vector of viral diseases of livestock in Europe such as bluetongue (BT), African horse sickness and epizootic haemorrhagic disease. Climatic factors are the main drivers of C. imicola occurrence and its distribution might be subject to rapid shifts due to climate change. Entomological data, collected during BT surveillance, and climatic/environmental variables were used to analyse ecological niche and to model C. imicola distribution and possible future range shifts in Italy. An ensemble technique was used to weigh the performance of machine learning, linear and profile methods. Updated future climate projections from the latest phase of the Climate Model Intercomparison Project were used to generate future distributions for the next three 20-year periods, according to combinations of general circulation models and shared socioeconomic pathways and considering different climate change scenarios. Results indicated the minimum temperature of the coldest month (BIO 6) and precipitation of the driest-warmest months (BIO 14) as the main limiting climatic factors. Indeed, BIO 6 and BIO 14 reported the two highest values of variable importance, respectively, 9.16% (confidence interval [CI] = 7.99%-10.32%), and 2.01% (CI = 1.57%-2.44%). Under the worst-case scenario of climate change, C. imicola range is expected to expand northward and shift away from the coasts of central Italy, while in some areas of southern Italy, environmental suitability will decrease. Our results provide predictions of C. imicola distribution according to the most up-to-date future climate projections and should be of great use to surveillance management at regional and national scales.

Is Asian tiger mosquito (Aedes albopictus) going to become homodynamic in Southern Europe in the next decades due to climate change?

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.

Blackflies (Simulium spp.) attacks on humans and animals in Rome and surrounding areas (Central Italy).

BACKGROUND: Due to their abundance in some environments, the blood-sucking habit and the involvement in the transmission of several diseases, blackflies of the genus Simulium (Diptera: Simuliidae) can be considered among the most annoying biting arthropods. METHODS: Following repeated attacks to humans and animals, entomological investigations were carried out in green areas of Rome and surroundings. Site and period of attacks were reported, together with the human and animal reactions to the bites. RESULTS: Four Simulium species have been identified through morphological or molecular analysis: Simulium intermedium, Simulium lineatum, Simulium lundstromi and Simulium ornatum (complex). Larval breeding sites were identified in small moats, where a high density of blackflies larvae and pupae was revealed. CONCLUSIONS: Being able to thrive in highly polluted water, even in few mm depth watercourses, some blackfly species are widely distributed in the area and their bites caused mild to severe reactions to humans and animals.

Delusional parasitosis: an entomological perspective after a 20-years-experience in two public medical and veterinary entomology laboratories.

Delusional parasitosis (DP) is psychiatric disorder characterized by the unshakeable belief of being infested by endo- or ectoparasites, without any evidence of infestations. Hence, DP sufferers often consult medical entomologists or dermatologists, rather than seeking help from a mental health practitioner. Here we present 39 cases of suspected DP occurred in twenty years in two Italian public medical entomology laboratories, to highlight their common features and peculiarities, based on the interviews and material brought to our laboratories. Our results confirmed that DP affects both sexes, but is more frequent in females over the age of 50 years. Cases of psychological contagion, mostly affecting elderly people, were reported within seven family units. Some rare or unique features of DP sufferers involved: the fixed belief to be infested by endoparasites, olfactory hallucinations of unpleasant smell, self-harm to get rid of the imaginary parasite and suicidal thoughts. We suggest the term "Digital pics sign" to describe a modern derivative of the "Matchbox sign", where the DP sufferers showed pictures and videos, in person or sent by email, as proofs of being infested. Medical entomologists, being the most frequently contacted professionals by DP sufferers, may be crucial to raise awareness about this psychological disorder and to encourage a multidisciplinary approach amongst medical professionals and pest control operators.

Aedes albopictus abundance and phenology along an altitudinal gradient in Lazio region (central Italy).

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.

First Report of Culicoides Biting Midges (Diptera: Ceratopogonidae) Attacking People in Italy, With the Description of Extreme Larval Breeding Sites and Diurnal Activity of Culicoides riethi.

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) play a paramount role in medical and veterinary entomology worldwide, particularly as vectors of pathogens which cause animal diseases. Biting midges are also infamous for the nuisance they provoke to people involved in outdoor activities. Nonetheless, attacks to man by midges from any Culicoides species have not been reported in Italy. An entomological investigation was performed following repeated attacks to man in a nature park near Rome (central Italy). The study area is a natural degassing zone, characterized by widespread hazardous gas emissions of CO2 and H2S, with several water bodies including permanent lakes, ponds, and pools. The biting midge C. riethi Kieffer, 1914 was very active during daytime in the period April-June. The species has been identified as responsible for attacks on people in the area. An in-depth analysis of the extreme environmental conditions revealed the ability of larvae to thrive in several water bodies, characterized by an extremely low pH and a high concentration of sulfates.

West Nile and Usutu viruses co-circulation in central Italy: outcomes of the 2018 integrated surveillance.

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.

Plasmodium matutinum Transmitted by Culex pipiens as a Cause of Avian Malaria in Captive African Penguins (Spheniscus demersus) 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.

First evidence of blaNDM-1 and blaOXA-23 carbapenemase genes in human body lice infesting a second-hand T-shirt in a street market in Italy.

BACKGROUND: The spread of carbapenems resistance is a public health concern. The main group of carbapenemases encoding the ß-lactamases activity (bla genes) is the Metallo-ß-lactamases (MBLs). METHODS: The presence of carbapenemase blaOXA-23-like, blaOXA-40-like, blaOXA-51-like, blaOXA-58-like, and blaNDM-1 genes was screened by real time PCR in 26 Pediculus humanus insects identified from second-hand clothes in a local market in Central Italy. Bacteria diversity was also characterized through shotgun metagenomic amplification for a deep sequencing of the host-associated bacterial microbiomes. RESULTS: The blaOXA-23 and blaNDM-1 carbapenemases genes were found and metagenomic analysis showed a great presence of Acinetobacter species. CONCLUSIONS: These results suggest a new potential transmission path for carbapenemase gene spread through bacteria ingested by insects infesting humans.

Scalp eschar and neck lymphadenopathy by Rickettsia slovaca after Dermacentor marginatus tick bite case report: multidisciplinary approach to a tick-borne disease.

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.

Aedes albopictus (Diptera: Culicidae) Monitoring in the Lazio Region (Central Italy).

The Asian tiger mosquito Aedes albopictus (Skuse 1894) is assuming an ever-increasing importance as invasive species in Europe and consequently as human health and nuisance concern. In Central Italy, the species has been recently involved in a chikungunya outbreak. A 3 yr Ae. albopictus monitoring was carried out in 21 municipalities of the Lazio region (Central Italy), belonging to three provinces. Samplings were performed on a weekly basis using ovitraps, in order to investigate climatic and spatial variables driving egg abundance and Ae. albopictus period of activity. A temperature of 10.4°C was indicated as lower threshold for the onset of egg-laying activity, together with a photoperiod of 13:11 (L:D) h. The whole oviposition activity lasted 8 mo (May-December), with 95% of eggs laid between early June and mid-November and a peak at the end of August. Egg abundance was positively influenced by accumulated temperature (AT) of the 4 wk preceding sampling and negatively by precipitation during the week before. Egg-laying activity dropped with decreasing AT, increasing rainfall, and with a photoperiod below 10:14 (L:D) h. Our results pinpointed the importance of fine-scaled spatial features on egg abundance. Some of these fine-scaled characteristics have been highlighted, such as the presence of vegetation and human footprint index. Our model estimated an almost doubled maximum number of laid eggs for the maximum value of human footprint. Compelling evidence of the relevance of fine-scaled characteristics was reported, describing cases where human-made breeding sites driven the abundance of Ae. albopictus.

Investigation of Ixodid ticks as vectors of Babesia caballi and Theileria equi (Protozoa: Apicomplexa) in central Italy.

Babesia caballi and Theileria equi are widely recognized as causative agents of equine pirolasmosis (EP), an acute, sub-acute, and chronic disease of equines, with relevant economic impact on horse trade worldwide. Although several studies on EP prevalence from central Italy have been published, data on ticks responsible for its transmission are still lacking. In this study, we identified a potential competent vector, investigating main features of its ecology together with EP infection rates. A two-year sampling of questing ticks was carried out for the first time in Italy in an area known for high EP prevalence in horse sera, detecting the association between Rhipicephalus bursa and causative agents of EP. Most of the positive pools harbored a single infection (91.1%); mixed infections were also detected (8.9%). The infection rate for T. equi slightly decreased among years; B. caballi showed a lower, but increasing, infection rate. Tick phenology, climate variables, and peaks of EP prevalence indicated late May and second half of June as periods with the highest risk of new infections, especially during warm and dry days.

Report of the human body louse (Pediculus humanus) from clothes sold in a market in central Italy.

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.

Outbreak of swimmer's itch in Central Italy: Description, causative agent and preventive measures.

Swimmer's itch is caused by the penetration of free-swimming larvae of trematodes of the family Schistosomatidae in human skin. It is usually reported in people engaged in recreational water activities in freshwater bodies and in most of cases, it is provoked by bird schistosomes of the genus Trichobilharzia. In the summer 2017, many cases of dermatitis were recorded in people bathing in the waters of the Albano Lake (Rome, Italy) and a parasitological investigation was carried out in order to ascertain the causative agent of these cases. Snails of the family Lymnaeidae, natural intermediate hosts of bird schistosomes, were collected from lake shallow waters to detect the presence of trematodes of the genus Trichobilharzia. Pools of maximum 10 snails were placed in Petri dishes, and cercarial emergence was stimulated exposing snails to strong artificial light intensity at 25°C. Three hundred and thirty-seven snails were collected and screened for the shedding of cercariae. Furcocercariae of the family Schistosomatidae, with a morphology overlapping that of the genus Trichobilharzia, were detected in seven Petri dishes. Assuming that in each positive Petri dish just one snail was shedding furcocercariae, the minimum infectious rate was 2.1%. Molecular analysis of furcocercariae allowed ascribing them to the species Trichobilharzia franki. Snails of the species Radix auricularia were identified as intermediate hosts of the parasite. This is the second record of T. franki causing cercarial dermatitis in Central Italy, the third in Italy. The 2017 was in Italy exceptionally warm and dry. Trematodes are sensitive to changes in temperature, being cercarial production and emission rates temperature dependent. Small increases in water temperature would speed up parasite development and transmission, leading to a manifold increase in cercarial emergence. Moreover, high temperatures raise chances to acquire the infection, due to increased time spent in water by people.

Monitoring for the possible introduction of Crimean-Congo haemorrhagic fever virus in Italy based on tick sampling on migratory birds and serological survey of sheep flocks.

Crimean-Congo haemorrhagic fever (CCHF), endemic in Africa, Asia, Eastern Europe and the Middle East, is caused by a tibovirus (CCHFV) transmitted in particular by the Hyalomma genus of the Ixodidae family that can remain attached to the host for up to 26days, which in case of migratory birds allows long distance carriage. Although CCHF in domestic ruminants is usually subclinical, they may become reservoirs and act as sentinels for the introduction and/or circulation of CCHFV. In this study, possible CCHFV introduction and circulation in Italy were monitored by tick sampling on migratory birds and by a serosurvey conducted on sheep. While bird tick sampling was conducted in thirteen ringing sites of Central and Southern Italy, the serosurvey was performed on flocks grazing in coastal provinces of Central Italy that are stop over areas for birds flying from Africa, where Hyalomma ticks and CCHFV are endemic, to Central and Northern Europe. A total of 282 ticks (80.8% were Hyalomma spp.) were collected from 139 (0.28%) migratory birds of the 50,325 birds checked with 0.22% infested by Hyalomma spp., involving 22 avian species with a mean number of 1.6 Hyalomma spp. per infested bird. For the serosurvey, 540 sheep sera were randomly collected that resulted all negative when examined by an indirect IgG ELISA, employing a recombinant antigen coded by the CCHFV S gene. While the present study confirmed the introduction of CCHFV potential vectors in Central Italy, transported by migratory birds arriving from endemic areas, the serosurvey results did not put in evidence the concomitant arrival of the virus in the study area during the survey period. In general, in areas potentially at risk of CCHFV introduction and circulation, structured serological monitoring of susceptible domestic animals represents a rational system for an early detection of virus circulation.

Caged young pigeons mortality by Coleoptera larvae.

Dermestidae and Tenebrionidae are well known inhabitants of bird's nests and poultry farms, under favourable conditions they can be very abundant under favourable conditions. At times, their larvae shift from a scavenging behaviour to a parasitic/predatory one, entering nestling's plumage and feeding on skin and feathers, and nally provoking skin damages and blood losses. These episodes mainly involve species of the genus Dermestes, but the tenebrionid Alphitobius diaperinus h also been reported to be responsible of similar cases. In June 2014, a mortality of caged young pigeons occurred in a family farm in Central Italy. Post-mortem examination of 1 of the dead nestlings revealed the presence, near the cloacal ori ce, of a triangular shaped hole of about 1 cm side, with rounded edges facing inward and with bleeding from the cavity. Five coleoptera larvae 0.5-2 cm long were collected from the edges of the hole. Bacteriological examination of liver, intestine, and lungs revealed the presence of Escherichia coli in the lung samples. The 5 larvae were morphologically identi ed as Dermestes bicolor (4) and Alphitobius diaperinus (1). This is the rst reported case of pigeon nestling's mortality caused by Dermestidae and Tenebrionidae larvae acting as parasites/predators in Italy.

Recent entomological enquiry on mosquito fauna in Circeo National Park.

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.