Local terrestrial snails as natural intermediate hosts of the zoonotic parasite Angiostrongylus cantonensis in the new European endemic area of Valencia, Spain.

AIM: The rat lungworm, Angiostrongylus cantonensis, has recently been found in the city of Valencia, parasitizing rats, Rattus norvegicus and Rattus rattus, its natural definitive hosts. This is the first finding of this zoonotic nematode in continental Europe. After informing local and national health authorities, the collection of local terrestrial snails took place with the aim of elucidating their potential role as intermediate hosts of A. cantonensis. METHODS AND RESULTS: A total of 145 terrestrial snails, belonging to the species Cernuella virgata, Cornu aspersum, Eobania vermiculata, Otala punctata, Pseudotachea splendida, Rumina decollata and Theba pisana, were randomly collected between May and December 2022 in public gardens, parks and orchards in six districts of Valencia, in five of which A. cantonensis had been reported previously in rats. Once collected and identified, the snails were frozen at -20°C. Subsequently, the DNA was isolated and screened by PCR using specific primers targeting the A. cantonensis COI gene. Seven individual snails, belonging to the species C. virgata, C. aspersum and T. pisana, were positive, for an overall prevalence of 4.8%. The PCR product from one of them was sequenced by Sanger sequencing. CONCLUSIONS: The three positive terrestrial snail species are among the edible species that are frequently included in various dishes in Spain. C. virgata is reported as a previously unrecorded intermediate host and should be added to the list of more than 200 species of terrestrial snails that have been reported worldwide as intermediate hosts of the rat lungworm. Considering that these terrestrial snails may release infective larvae of A. cantonensis on leafy green vegetables on which they feed and during their handling and preparation for consumption, prophylactic measures to prevent human neuroangiostrongyliasis in Valencia and other regions to which this zoonotic parasite may spread are recommended.

A Geomedical Survey: Is There an Association Between Climatic Conditions and Leishmania Species Distribution in Iran During the Years 1999-2021?

PURPOSE: Iran is among the high-risk leishmaniasis regions in the world. WHO recommends the use of GIS as an ideal tool for healthcare authorities to predict the evolution of a disease, delimit the risk of outbreaks and identify critical areas. The aim of this research is to find the association between the main species of Leishmania (L. major, L. tropica, L. infantum) dispersion and climatic variables in Iran. METHODS: All molecular-based reports of leishmaniasis from Iran between 1999 and 2021 were gathered from reliable medical sources. Meteorological data (air and soil temperatures, annual rainfall and humidity) of the country along the study period were obtained from the Iranian Climatological Research Centre. The data concerning species distribution and climatic conditions during this period were moved to a base-map through raster layers using ArcGIS 10.4.1 software. The relationship between parasitological and climatic models was examined using ANOVA. RESULTS: High risk area maps, based on the cut-off thresholds, were generated for Leishmania major, L. tropica and L. infantum. According to the molecular-based reports, the L. major distribution was significantly related to all climatic variables, while L. tropica was merely related to rainfall and humidity, and the L. infantum distribution was significantly associated with rainfall, soil and air temperatures. CONCLUSION: The association between climatic conditions and Leishmania species distribution in Iran has been confirmed. Consequently, both, the relationship between climatic conditions and the geographical distribution of Leishmania species, and the use of GIS to better understand the spatial epidemiology of leishmaniasis, have been reaffirmed.

Update on the First Finding of the Rat Lungworm, Angiostrongylus cantonensis, in Rattus spp. in Continental Europe, Valencia, Spain, 2022.

The rat lungworm, Angiostrongylus cantonensis, is an emerging parasite that can cause eosinophilic meningitis in humans. Over the past 60 years, the worm has greatly expanded its original Asian distribution to most tropical and subtropical areas of the world, mostly by traveling on ships with its definitive hosts, rats. Angiostrongylus cantonensis was recently found for the first time in Continental Europe, specifically in 3 (2 Rattus norvegicus and 1 Rattus rattus) out of 27 rats trapped in the sewer system of the city of Valencia, Spain. Updating the investigation, the parasite has subsequently been found in 8 (5 R. norvegicus and 3 R. rattus) out of 94 rats analyzed. The highest prevalence of infection (20%) was obtained in rats trapped in the orchards that surround the city, where both snails and slugs (intermediate hosts) abound, and where vegetables consumed in Valencia, other parts of Spain, and abroad, are produced. The presence of the parasite in rats does not necessarily mean that the disease it causes becomes a relevant public health concern since it strongly depends on the food habits of the population at risk. If proper precautions are taken, the risk of acquiring neuroangiostrongylosis should be minimal.

Parasite Fauna and Coinfections in Urban Rats Naturally Infected by the Zoonotic Parasite Angiostrongylus cantonensis.

When the zoonotic parasite of rodents that can cause human neuroangiostrongyliasis, i.e., Angiostrongylus cantonensis, is found in its natural definitive hosts, it is usually reported in isolation, as if the rat lungworm were the only component of its parasite community. In this study, we report the coinfections found in rats naturally infected by A. cantonensis in urban populations of Rattus norvegicus and Rattus rattus in Valencia, Spain. In addition to the rat lungworms, which were found in 14 of the 125 rats studied (a prevalence of 11.20%), 18 other parasite species (intestinal and tissular protists, microsporidia and helminths) were found, some of them with high burdens. Fourteen of these nineteen species found are potential zoonotic parasites, namely Blastocystis, Giardia duodenalis, Cryptosporidium spp., Enterocytozoon bieneusi, Encephalitozoon hellem, Toxoplasma gondii, Brachylaima spp., Hydatigera taeniaeformis s.l. larvae, Hymenolepis nana, Hymenolepis diminuta, Angiostrongylus cantonensis, Calodium hepaticum, Gongylonema neoplasticum and Moniliformis moniliformis. The total predominance of coinfected rats as well as their high parasite loads seem to indicate a trend towards parasite tolerance.

Autochthonous Angiostrongylus cantonensis Lungworms in Urban Rats, Valencia, Spain, 2021.

To determine the role of rats as potential reservoirs of zoonotic parasites, we examined rats trapped in urban sewers of Valencia, Spain, in 2021. Morphologic and molecular identification and sequencing identified autochthonous Angiostrongylus cantonensis nematodes, the most common cause of human eosinophilic meningitis, in pulmonary arteries of Rattus norvegicus and R. rattus rats.

Spatio-temporal Prediction of the Malaria Transmission Risk in Minab District (Hormozgan Province, Southern Iran).

INTRODUCTION: Malaria is the most important parasitic disease in tropical and subtropical regions, with more than 240 million cases reported annually. In Iran, indigenous cases occur in its south-eastern region. The aim of this study is to assess the environmental risk of malaria transmission in an endemic area of southern Iran. METHODS: The study was carried out in Minab district (Hormozgan province, southern Iran), with the aim to assess the environmental risk of malaria, based on a spatio-temporal study, using Growing Degree Days (GDD)-based predictions, larval habitat ecology, MaxEnt spatial predictions and malaria transmission data. RESULTS: The Gradient Model Risk index showed the highest malaria transmission risk period to be during January-April and October-December. The ecological conditions of water bodies of larval habitats of the four vector species (Anopheles culicifacies, A. dthali, A. fluviatilis and A. stephensi) were assessed, with A. stephensi being the most prevalent and the most widely distributed species. CONCLUSION: These findings, together with the MaxEnt Anopheles predictive distribution models, allowed identifying villages in danger of malaria transmission in Minab district. This spatio-temporal prediction of malaria transmission risk should be incorporated in the design of malaria control initiatives towards a local malaria early warning system. Moreover, the proposed transmission risk model can be extrapolated, at local scale, to other malaria endemic areas of tropical and subtropical regions.

Nematode Parasites of the European Pilchard, Sardina pilchardus (Walbaum, 1792): A Genuine Human Hazard?

The European pilchard is one of the most frequently consumed fish species in Mediterranean countries, especially in Italy and Spain, and has been reported as the cause of at least eight human anisakidosis cases in Spain. With the aim to shed light on the potential human parasitosis risk posed by nematode larvae belonging to families Anisakidae or Raphidascarididae, a total of 350 sardines captured in the Atlantic Ocean (175 specimens) and the Mediterranean Sea (175 specimens), acquired in various Spanish nationwide supermarket chains, were helminthologically analyzed. The statistical analysis of some helminth parameters revealed a higher presence of nematodes belonging to the genus Hysterothylacium (prevalence 24.29%; mean abundance of 2.36), usually considered non-parasitic for humans (only three cases reported worldwide), when compared to nematodes of the genus Anisakis (5.71%; 0.16). The human anisakidosis risk after the consumption of raw or undercooked sardines and the role of Hysterothylacium, the most frequent nematode, is discussed, providing information to consumers. To avoid human infection by anisakid larval nematodes, the established preventive measures are confirmed and new ones are proposed, such as the consumption of sardines preferably caught in the Mediterranean and of small-sized specimens available, and the immediate evisceration after fishing.

Anisakid Nematodes and Potential Risk of Human Anisakiasis through the Consumption of Hake, Merluccius spp., Sold Fresh in Spanish Supermarkets.

Nematode parasite species belonging to the Anisakis simplex complex are the most important cause of human anisakiasis through the consumption of (mainly) undercooked, previously not frozen, or conveniently treated fish. In Spain, the consumption of hake has been recognized as an important source of this parasitosis. With the aim of shedding light on the risk factors that can influence the potential risk of human anisakiasis in Spain through the consumption of fresh hake sold by nationwide supermarket chains, a total of 536 small hake specimens belonging to the species Merluccius bilinearis caught off the Northeast American coasts and Merluccius merluccius caught in the Northeast Atlantic and Mediterranean waters was analysed. Anisakids morphologically identified as Anisakis type I were found as the most prevalent and the most abundant species and were considered the main potential cause of human anisakiasis. Intrinsic and extrinsic factors concerning the hake, such as its origin and season of capture, its size, as well as the days passed between its capture and consumption, should be taken into account to avoid this human parasitosis. It is essential that consumers have access to fish label information which should include, as regulated by the European Commission, traceability data.

Molecular detection of Leishmania infantum in rats and sand flies in the urban sewers of Barcelona, Spain.

BACKGROUND: Classically, dogs have been considered to be the only reservoir of leishmaniasis in urban areas. However, in a previous study, we found a 33.3% prevalence of Leishmania infantum in the spleens of Norway rats (Rattus norvegicus) sampled in the underground sewer system of the city of Barcelona (Spain). The aim of the present study was to verify, using molecular methods, the potential reservoir role of these rats in the same sewer system. METHODS: A sensitive real-time PCR (qPCR) assay, DNA sequencing and phylogenetic analysis were carried out to identify and quantify the presence of L. infantum DNA in sand fly individuals captured in the same underground sewer system of Barcelona as in our previous study and in the spleens and ears of rats captured in the same sewer system. RESULTS: Leishmania infantum DNA was found in 14 of the 27 (51.9%) sand flies identified as Phlebotomus perniciosus, and 10 of the 24 (41.7%) rats studied were infected. Leishmania infantum was found in the spleens (70%) and in the ears (40%) of the infected rats. Quantitative results revealed the presence of high loads of L. infantum in the rats studied (> 3 × 106 parasites/g ear tissue) and among the sand flies (> 34 × 106 parasites in 1 individual). CONCLUSIONS: The molecular methods used in this study demonstrated a high prevalence of L. infantum in the underground sewer populations of both R. norvegicus and P. perniciosus. These results suggest that sewer rats, in addition to dogs, are likely to act as reservoirs of leishmaniasis in cities, where sewer systems seem to offer the ideal scenario for the transmission of leishmaniasis. Therefore, to achieve the WHO 2030 target on the elimination of leishmaniasis as a public health problem successfully, an efficient control strategy against leishmaniasis in rats and sand flies should be implemented, particularly in the sewer systems of urban areas of endemic countries.

First Data on the Helminth Community of the Smallest Living Mammal on Earth, the Etruscan Pygmy Shrew, Suncus etruscus (Savi, 1822) (Eulipotyphla: Soricidae).

Suncus etruscus is the smallest living mammal on Earth by mass. Most adults weigh 1.8-3 g with a body length of 35-48 mm. Catching it in small mammal traps in nature is extremely difficult due to its minute size, and therefore special trapping methods must be used. We had the unique opportunity of studying, for the first time, the helminth parasites of 166 individuals of S. etruscus, part of the largest collection in the world, which belonged to the French scientist Dr Roger Fons (1942-2016). A total of 150 individuals were captured in the Banyuls-Cerbère area (France) and 16 in the island of Corsica (France). We found seven helminth species, specifically, the cestodes Joyeuxiella pasqualei larvae, Mesocestoides sp. larvae, Staphylocystis claudevaucheri, S. banyulsensis, S. cerberensis, and Pseudhymenolepis sp., and the nematodes Aonchotheca sp. and Nematoda gen. sp. larvae. Neither trematodes nor acanthocephalans were detected. We provide prevalences, infracommunity compositions, and helminth associations. The adult helminth community of S. etruscus seems to be highly specific, i.e., oioxenous, and linked to its insectivore diet. Due to its small size, S. etruscus has undergone numerous physiological adaptations that have probably influenced its helminth spectrum as well as its helminth specificity.

Absence of Toxoplasma gondii in 100% Iberian products from experimentally infected pigs cured following a specific traditional process.

Infection with Toxoplasma gondii in humans has usually been related to the consumption of raw, undercooked or cured meat. Our study is based on the detection of T. gondii in cured legs and shoulders made from 100% Iberian sows fed mainly with acorn and raised as outdoor livestock in Aracena (Spain), which having been elaborated following a specific curing process (time period and location). An outdoor farm with a total of 636 animals was studied, showing a seroprevalence of 10% for the parasite T. gondii. Twenty individuals were chosen to be experimentally infected and slaughtered 60 days post-infection. Their legs and shoulders were processed to make 100% Iberian ham legs and shoulders. The meat ready to be eaten was analyzed by quantification and viability assays using magnetic capture real-time qPCR and bioassay techniques proving that this specific traditional "Cinco Jotas" curing process 100% Iberian ham is strong enough to eliminate the parasite T. gondii, resulting in a safe product for consumers.

Leishmaniasis in Norway Rats in Sewers, Barcelona, Spain.

We detected Leishmania infantum in 98 Norway rats (Rattus norvegicus) trapped in parks and sewers of Barcelona, Spain. The 84 rats from the sewers showed a prevalence of 33.3% and up to 2,272 estimated parasites. These results, in the most abundant potential reservoir in cities, is of public health concern.

First survey on zoonotic helminthosis in urban brown rats (Rattus norvegicus) in Spain and associated public health considerations.

The brown rat, Rattus norvegicus, with a worldwide distribution, is the most commensal species among synanthropic rodents, since its main habitat, in urban as well as in rural areas, is always linked to humans. Therefore, people living in close proximity to rodent populations can be exposed to infection. Whereas bacteria and viruses are the best known rat-associated zoonoses in urban environments, the role of brown rats as reservoirs for helminth parasites and the associated risk for humans are less well known. Specifically, this role has not been analyzed in Spain to date. A total of 100 R. norvegicus trapped in the sewage system (n = 85), and parks (n = 15) of Barcelona was examined. The overall prevalence of helminth infection was 85%. The helminths found were Hymenolepis nana (17%), H. diminuta (33%) (Cestoda), Calodium hepaticum (17%), Eucoleus gastricus (28%), Aonchotheca annulosa (12%), Trichosomoides crassicauda (7%), Nippostrongylus brasiliensis (46%), Heterakis spumosa (62%), Gongylonema neoplasticum (20%) (Nematoda) and Moniliformis moniliformis (6%) (Acanthocephala). Five of the ten helminth species are considered zoonotic parasites, with rats acting as reservoirs for human infection, i.e. H. nana, H. diminuta, C. hepaticum, G. neoplasticum and M. moniliformis. G. neoplasticum and M. moniliformis are reported for the first time in urban rats in Europe. H. nana, H. diminuta and C. hepaticum are the most widespread species in European cities. For H. nana and C. hepaticum, rats act as effective spreaders of the human infective stage (eggs). For H. diminuta, G. neoplasticum and M. moniliformis, rats act as indirect reservoirs of the zoonoses since the eggs shed by the rats are infective for their insect intermediate hosts only. Medical practitioners need to be made aware of the range of parasites carried by rats, as there is a realistic likelihood that ill health currently caused by rat infestations may be misdiagnosed.

The helminth community component species of the wood mouse as biological tags of a ten post-fire-year regeneration process in a Mediterranean ecosystem.

Serra Calderona Natural Park, a Mediterranean ecosystem, has been in post-fire regeneration for 10 years. To elucidate which helminth community component species of the wood mouse, Apodemus sylvaticus, can be considered biological tags of this process, the influence of intrinsic (host density; host sex and age) and extrinsic factors (site, year, and period of capture; vegetation recovery) on their prevalence and abundance has been analysed, comparing a burned and an unburned area. A total of 564 wood mice (408 from the burned and 156 from the unburned area), from the 2nd to the10th post-fire year, was included in this helminthoecological study. The results suggest that the area in post-fire regeneration is still more vulnerable to periodic environmental changes than the unburned area as deduced from the analysis of the helminth populations of Pseudocatenotaenia matovi, Skrjabinotaenia lobata, Trichuris muris, Eucoleus bacillatus and Aonchotheca annulosa. The intermediate and definitive host populations presented a greater variability to these environmental changes in the burned area (Taenia parva, P. matovi, S. lobata, A. annulosa, Syphacia stroma and S. frederici). In the regenerating area, some behavioural changes in certain populations determined by the host sex are taking place (T. parva, Helgimosomoides polygyrus and S. frederici). During the last years studied, a greater similarity in the populational development of some component species between both areas can be appreciated (H. polygyrus and S. stroma). The role of the wood mouse and its helminth parasites as biological tags of the post-fire regeneration process in Mediterranean ecosystems has been confirmed.

Pufferfish nests vs. parasite hooks: A bizarre resemblance.

Image 1.