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Paleoparasitological investigation in the archeological site of the Caspian Sea littoral revealed the presence of human Dioctophyma renale infection, a ubiquitous kidney-residing helminth of mammals, during the Parthian Empire of Iran (247 BC-224 AD). This study reports the oldest human infection with giant kidney worm in the Middle East, mainly in an area where the infection is reported nowadays. The paucity of records throughout the literature suggests that the same species of parasite occurred with low frequency in the past, as well. The cemetery in the archeological site of Kiasar represented a unique opportunity of studying human and animal parasites of the Parthian Empire in Iran.

Of the eleven species of shrews described in Iran, the Etruscan shrew (Suncus etruscus), the smallest known extant mammal by mass, is recorded in northern and southern provinces. During rodent control programs, a female S. etruscus was trapped and found naturally infected with a trematode which was morphologically identified as Brachylaima sp. This finding represents the first report of trematodiasis in this host species.

Although Joyeuxiella pasqualei is frequently detected in cats from Mediterranean Europe, information on its biology is still scarce. This cestode is relatively less frequently reported in dogs, possibly because it is often misdiagnosed with the better-known Dipylidium caninum. The occurrence of J. pasqualei proglottids in a dog living in a closed environment triggered us to delve into the biology of this cestode by collecting biological samples from lizards and a road-killed cat. Two reptile species, Podarcis siculus (Lacertidae), and Tarentola mauritanica (Geckonidae) were also collected in the garden and its surroundings. In addition, experimental infections with eggs obtained from gravid proglottids were performed in laboratory mice, and Tenebrio molitor (Coleoptera: Tenebrionidae) beetles. Proglottids from the dog's feces and adult cestodes detected at necroscopy of a cat were morphologically identified as J. pasqualei. Two out of 13 T. mauritanica collected in the garden had natural infections of J. pasqualei cysts in the liver and attached to the intestine. All P. siculus lizards and experimentally infected rodents and beetles were negative. DNA sequences obtained from J. pasqualei showed the highest nucleotide similarities with Versteria sp., Echinococcus sp., Raillietina sonini, Taenia polyacantha and D. caninum. Data herein provided show the inability of rodents to become infected by direct ingestion of gravid proglottids, suggesting a need for an invertebrate first intermediate host in the life cycle. Thus, more research study is advocated to better understand the biology of J. pasqualei such as its first intermediate host and its mechanism of transmission in reptiles and rodents.

Canine vector-borne pathogens (CVBPs) comprise a group of disease agents mainly transmitted by ticks, fleas, mosquitoes and sand flies. In this study, we assessed the presence of CVBPs in an Afro-descendent community (Quilombola) of northeastern, Brazil. Dog blood samples (n = 201) were collected and analyzed by rapid test for the detection of antibodies against Leishmania spp., Anaplasma spp., Ehrlichia spp. and Borrelia burgdorferi sensu lato (s.l.), and antigens of Dirofilaria immitis. In addition, polymerase chain reactions were performed for Anaplasmataceae, Babesia spp., Hepatozoon spp., Rickettsia spp. and B. burgdorferi s.l. Overall, 66.7% of the dogs scored positive to at least one pathogen at serological and/or molecular methods. Antibodies against Ehrlichia spp. were the most frequently detected (57.2%; n = 115/201), followed by Anaplasma spp. (8.5%; n = 17/201), Leishmania spp. (8.5%; n = 17/201) and B. burgdorferi s.l. (0.5%; n = 1/201). For D. immitis, 11 out of 201 (5.5%) animals scored positive. At the molecular analysis, 10.4% (n = 21/201) of the samples scored positive for Babesia spp./Hepatozoon spp., followed by Anaplasmataceae (5.0%; n = 10/201) and Rickettsia spp. (3.0%; n = 6/201). All samples were negative for B. burgdorferi s.l. Our data demonstrated the presence of CVBPs in the studied population, with a high seropositivity for Ehrlichia spp. In addition, considering the detection of zoonotic pathogens in dogs and their relationship with people from Quilombola communities, effective control strategies are advocated for minimizing the risk of infection in this socially vulnerable human population and their pets.

Dermatophagoides spp. (Acariformes; Pyroglyphidae), house dust-mite well known as the causative agent of atopic hypersensitivity and allergy could potentially cause severe dermatitis. Herein we report an uncommon case of scalp dermatitis associated with the presence of Dermatophagoides spp.A 17-year old male presented with patchy alopecia on the scalp without intense peeling or itching, surround by unchanged skin and hair. Initially, superficial fungal infection was suspected; however, parasitological examination revealed the presence of live mites.All the anatomical measurements and parameters from the specimens were compatible with Dermatophagoides spp.Dermatophagoides spp. are not yet confirmed as causative agents of parasitic infestation, but the presence of these mites could have caused an allergic reaction followed by dermatitis with mild-to-moderate clinical manifestations. However, true parasitism as well as phoresy could also be considered. The clinical manifestations caused by house-dust mite cannot be easily recognized and the lack of diagnostic tools is a hindrance that often leads to misdiagnosis and inadequate therapy.

Dirofilaria immitis (the canine heartworm) is widespread in the tropics, with prevalence surpassing 30% in high-risk areas. In addition to the suitable climatic conditions that favour mosquito abundance and filarial larva development, there is low compliance with the recommended year-round use of preventives in these transmission hotspots. This represents a major concern, considering that melarsomine (first-line heartworm adulticide) is unavailable in several tropical countries, resulting in the so-called slow-kill protocol being the only available adulticide treatment option. In this article, the members of TroCCAP (Tropical Council for Companion Animal Parasites) review the current distribution of heartworm in the tropics and the availability of melarsomine, and discuss alternatives for the management of heartworm infections in dogs. Graphical Abstract

Background The Asian elephant (Elephas maximus), which is an endangered species, harbors several parasites. Among the ectoparasites that it harbors, ear mites of the genus Loxanoetus have the potential to cause external otitis, an inflammation that may also be associated with the presence of other microorganisms. We assessed the relationships between ear mites, nematodes, yeast, bacterial rods, and cocci sampled from the ears of captive Asian elephants in Thailand. In addition, we discuss the possibility that dust-bathing behavior may be triggered by ear mite infestation, and that this in turn may lead to contamination of the ears with soil microorganisms. Methods Legally owned captive Asian elephants (n = 64) were sampled. Ear swabs were individually collected from both ears and microscopically examined for the presence of mites, nematodes, yeast, bacterial rods, cocci, and host cells. Mites and nematodes were identified to species level using morphological and molecular methods. Results Loxanoetus lenae mites were present in 43.8% (n = 28/64) of the animals (19 animals with mites in one ear and nine animals with mites in both ears). Nematodes of the genus Panagrolaimus were detected in 23.4% (n = 15/64) of the animals (10 with nematodes in one ear and five with nematodes in both ears). In adult elephants (Fisher’s exact test, P = 0.0278) and female elephants (Fisher’s exact test, P = 0.0107), the presence of nematodes in both ears was significantly associated with the presence of mites. In addition, higher categorical burdens of nematodes were also significantly associated with the presence of mites (Fisher’s exact test, P = 0.0234) and epithelial cells (Fisher’s exact test, P = 0.0108), and marginally significantly associated with bacterial cocci (Fisher’s exact test, P = 0.0499). Conclusions The presence of L. lenae mites in the ear canals of the Asian elephants was significantly associated with the occurrence of other microorganisms, such as soil nematodes, bacteria and yeasts. The presence of mites in their ears may increase the dust-bathing behavior of elephants which, if confirmed, represents a further paradigmatic example of a parasitic infestation affecting animal behavior. Graphical Abstract

One of the most common behaviors of cats that have an indoor/outdoor lifestyle is to bring hunted “gifts” to their owners, represented by small mammals, reptiles and birds. Access to the outdoors by dogs and cats may represent a problematic issue, since they may be at risk of diseases, traffic accidents and ingestion of toxins. Yet, the impact of this population of roaming dogs and cats predating wildlife is another concerning issue that receives less attention. Despite these risks, most owners still prefer to give outdoor access to their pets to allow them to express their “natural instincts,” such as hunting. Therefore, with a growing population of > 470 million dogs and 373 million cats worldwide, predation not only represents a threat to wildlife, but also a door of transmission for parasitic diseases, some of them of zoonotic concern. In this review, the role played by dogs, and especially cats, in the perpetuation of the biological life cycle of zoonotic parasites through the predation of rodents, reptiles and birds is discussed. Feral and domestics dogs and cats have contributed to the population collapse or extinction of > 63 species of reptiles, mammals and birds. Although the ecological impact of predation on wild populations is well documented, the zoonotic risk of transmission of parasitic diseases has not received significant attention. The parasitic diseases associated to predation vary from protozoan agents, such as toxoplasmosis, to cestodes like sparganosis and even nematodes such as toxocariasis. Raising awareness about predation as a risk of zoonotic parasitic infections in dogs and cats will aid to create responsible ownership and proper actions for controlling feral and free-roaming cat and dog populations worldwide. Graphical Abstract

Culex pipiens (Linnaeus), one of the most abundant mosquito species in Europe, plays a crucial role in the endemic transmission of West Nile virus and caused the large outbreak with1600 human cases in 2018. Although evidence of resistance to pyrethroids has been reported for Cx. pipiens populations from Spain and Greece, resistance monitoring has been largely neglected in Italy. Herein, we investigate susceptibility of Italian Cx. pipiens populations to the pyrethroids permethrin and deltamethrin. Results from WHO-tube-bioassays revealed mortalities ranging from 14-54%, indicating high levels of resistance, in four out of 10 populations exposed to permethrin (0.75%) and of 63% in one of three populations exposed to deltamethrin (0.05%). Reduced susceptibility (mortality98%) was detected in almost all other populations. A clear association is shown between the resistant phenotype and the presence of kdr-alleles in position 1014 of the VSSC, strongly suggesting its role in reducing susceptibility. The study provides the first evidence of pyrethroid-resistance in Italian Cx. pipiens populations and reports levels of resistance paralleled in the European region only in Turkey. This highlights the urgent need to implement insecticide-resistance management plans to restore the efficacy of the nowadays only chemical weapon available to control arbovirus transmission in Europe.

Parasites of the genus Leishmania are unusual unicellular microorganisms in that they are characterized by the capability to subvert in their favor the immune response of mammalian phagocytes, including dendritic cells. Thus, in overt leishmaniasis, dendritic cells and macrophages are converted into a niche for Leishmania spp. in which the parasite, rather than being inactivated and disassembled, survives and replicates. In addition, Leishmania parasites hitchhike onto phagocytic cells, exploiting them as a mode of transport to lymphoid tissues where other phagocytic cells are potentially amenable to parasite colonization. This propensity of Leishmania spp. to target dendritic cells has led some researchers to consider the possibility that the non-pathogenic, reptile-associated Leishmania tarentolae could be exploited as a vaccine platform and vehicle for the production of antigens from different viruses and for the delivery of the antigens to dendritic cells and lymph nodes. In addition, as L. tarentolae can also be regarded as a surrogate of pathogenic Leishmania parasites, this parasite of reptiles could possibly be developed into a vaccine against human and canine leishmaniases, exploiting its immunological cross-reactivity with other Leishmania species, or, after its engineering, for the expression of antigens from pathogenic species. In this article we review published studies on the use of L. tarentolae as a vaccine platform and vehicle, mainly in the areas of leishmaniases and viral infections. In addition, a short summary of available knowledge on the biology of L. tarentolae is presented, together with information on the use of this microorganism as a micro-factory to produce antigens suitable for the serodiagnosis of viral and parasitic infections. Graphical Abstract

Background Ophionyssus natricis is the main species of mite that infests captive reptiles. High infestations may result in the host experiencing general discomfort and deleterious effects, even death. Moreover, O. natricis is an important vector of reptile vector-borne diseases and is considered to be the putative vector of the Reptarenavirus, the causal agent of the inclusion body disease. Despite the cosmopolitan distribution of O. natricis in captive reptiles, treatment options are limited. The aim of the present study was to assess the efficacy of afoxolaner (NexGard®; Boehringer Ingelheim, Ingelheim, Germany) in heavily infested, privately owned snakes, evaluate the prevalence of mites and drug availability in the plasma of treated snakes (pharmacokinetics) and perform a clinical examination of animals. Methods The study was conducted in two snake breeding facilities, where many snakes were infested with mites. Each animal was clinically examined and weighed, and mite infestations were assessed on the animals and in their enclosures (environment). Animals were treated with a dose of 2.5 mg afoxolaner per kilogram body weight (2.5 mg/kg) administered orally. All animals were examined pre-treatment (T0) and at various time points post-treatment (T1, 6 h; T2, 24 h; T3, 14 days; T4, 28 days). The collected mites were morphologically identified at the species level and the species identity also confirmed molecularly. Results Overall, 81 snakes from the two participating facilities (i.e. 70 from site 1 and 11 from site 2) were screened, and 31 (38.3%) snakes were found to have at least one mite. All mites were identified morphologically and molecularly as O. natricis. Lampropeltis was the genus of snakes with highest number of infested individuals. Mites were found to be alive on snakes at T1, but at T2 only dead mites were observed, and at T3 and T4 mites were no longer present on the animals or in their environment. No side effects were observed in the treated snakes. Conclusions A single oral administration of afoxolaner at 2.5 mg/kg was a safe treatment for snakes and 100% effective for the eradication of natural O. natricis infestation without the need to treat the environment of the snake. Graphical Abstract

In spite of the increasing medical interest in Malassezia yeasts, the virulence factors of Malassezia furfur causing bloodstream infections (BSI) were never investigated. Therefore, phospholipase (Pz), lipase (Lz), hemolysin (Hz), biofilm production, and in vitro antifungal susceptibility profiles were evaluated in M. furfur strains, isolated from both pityriasis versicolor (PV) patients (n = 18; Group 1) or from preterm infants BSI (n = 21; Group 2). All the test stains exhibited Pz activity, whereas 92.3% and 97.4% of strains exhibited Lz and Hz activities, respectively. Pz, Lz, and Hz activities were higher (i.e., lower values) within Group 1 strains (i.e., 0.48, 0.40, and 0.77) than those within Group 2 (i.e., 0.54, 0.54, and 0.81). The biofilm production was higher within Malassezia isolates from Group 2 (0.95 ± 0.3) than from Group 1 (0.72 ± 0.4). Itraconazole and posaconazole were the most active drugs against M. furfur, followed by amphotericin B and fluconazole. The minimum inhibitory concentrations (MIC) values varied according to the origin of M. furfur strains being statistically lower in M. furfur from Group 1 than from Group 2. This study suggests that M. furfur strains produce hydrolytic enzymes and biofilm when causing PV and BSI. Data show that the phospholipase activity, biofilm production, and a reduced antifungal susceptibility profile might favor M. furfur BSI, whereas lipase and hemolytic activities might display a synergic role in skin infection.

In Europe wildlife animals such as the red fox (Vulpes vulpes) are considered the main reservoir for Angiostrongylus vasorum as well as a potential threat for domestic dog infection. Though this parasite is endemic in fox populations, data on A. vasorum infection in wolves (Canis lupus italicus) are still scant, having only recently been described in Northwestern Spain, in Italy, in Croatia and in Slovakia. Based on the rising number of cases of canine lungworm infection in Central Italy (Abruzzo region), the aim of the present study was to investigate the infection by A. vasorum in fox and wolf populations sharing the same geographical area of dogs. From October 2008 to November 2019, A. vasorum specimens were collected, through routine post-mortem examination, from 56 carcasses (44 foxes and 12 wolves). Adult parasites were searched for in the right side of the heart and in pulmonary artery of all carcasses. First stage of larvae (L1) was searched in faeces using the Baermann technique and in lungs by tissue impressions. Overall, 230 adult specimens were collected and identified on a morphological basis. To confirm the morphological identification, 4 adult specimens (n = 3 from fox, n = 1 from wolf) were molecularly identified as A. vasorum by amplification of partial fragment of nuclear 18S rRNA (~1700 bp) genes. The anatomo-pathological and parasitological examinations indicated the presence of A. vasorum in 33 foxes (75%) and in 8 wolves (66.7%). The level of prevalence of infested wolves was higher than the previous one reported in other European countries. Interestingly, the prevalence of infection in foxes herein recorded was higher than that described in dogs (8.9%) living in the same geographical area. This result may confirm the hypothesis that the spread of canine angiostrongylosis is linked to fox populations infection., Graphical abstract Image 1, Highlights • Angiostrongylus vasorum was found in dead foxes and wolves in Chieti province, Italy. • The prevalence in foxes was higher than that described in dogs. • The prevalence in wolves was higher than the previous one reported in European countries. • The spread of canine angiostrongylosis is linked to fox populations infection. • The province of Chieti appears to be an enzootic focus for canine angiostrongylosis.

Hedgehogs are synanthropic mammals, reservoirs of several vector-borne pathogens and hosts of ectoparasites. Arthropod-borne pathogens (i.e., Rickettsia spp., Borrelia spp., and Anaplasmataceae) were molecularly investigated in ectoparasites collected on hedgehogs (n = 213) from Iran (161 Hemiechinus auritus, 5 Erinaceus concolor) and Italy (47 Erinaceus europaeus). In Iran, most animals examined (n = 153; 92.2%) were infested by ticks (Rhipicephalus turanicus, Hyalomma dromedarii), and 7 (4.2%) by fleas (Archeopsylla erinacei, Ctenocephalides felis). Of the hedgehogs infested by arthropods in Italy (i.e., 44.7%), 18 (38.3%) were infested by fleas (Ar. erinacei), 7 (14.9%) by ticks (Haemaphysalis erinacei, Rh. turanicus, Rhipicephalus sanguineus sensu lato), and 6 (12.8%) by mites (Caparinia tripilis, Acarus nidicolous, Ornithonyssus spp.). Phoretic behavior of C. tripilis on Ar. erinacei was detected in two flea specimens from Italy. At the molecular analysis Rickettsia spp. was detected in 93.3% of the fleas of Italy. In Iran, Rickettsia spp. was detected in 8.0% out of 212 Rh. turanicus ticks, and in 85.7% of the Ar. erinacei fleas examined. The 16S rRNA gene for Ehrlichia/Anaplasma spp. was amplified in 4.2% of the 212 Rh. turanicus ticks. All sequences of Rickettsia spp. from fleas presented 100% nucleotide identity with Rickettsia asembonensis, whereas Rickettsia spp. from Rh. turanicus presented 99.84%–100% nucleotide identity with Rickettsia slovaca, except for one sequence, identical to Rickettsia massiliae. The sequences of the 16S rRNA gene revealed 99.57%–100% nucleotide identity with Anaplasma spp., except for one, identical to Ehrlichia spp. A new phoretic association between C. tripilis mites and Ar. erinacei fleas has been herein reported, which could be an important route for the spreading of this mite through hedgehog populations. Additionally, spotted fever group rickettsiae were herein detected in ticks and fleas, and Anaplasma/Ehrlichia spp. in ticks, suggesting that hedgehogs play a role as reservoirs for these vector-borne pathogens.

Background For a long time known as the oriental eyeworm, Thelazia callipaeda is a zoonotic nematode that infects the eyes of a wide range of vertebrate hosts including dogs, cats, wildlife carnivores, lagomorphs, and humans. The high occurrence of this infection in Europe and the first cases in the United States have increased scientific interest in the parasite, as it also represents a risk for people living in endemic areas. Therefore, treatment and prevention of thelaziosis in canine population are advocated to reduce the risk of human infection as well. Here, we assessed the efficacy of a formulation containing sarolaner/moxidectin/pyrantel (Simparica Trio®) administered orally at monthly intervals, for the prevention of establishment of infection with T. callipaeda in naturally infected dogs. In this formulation, moxidectin is expected to have efficacy against eyeworms, whereas sarolaner and pyrantel are not. Methods The study was conducted in eyeworm endemic areas of Italy and France, where dogs (n = 125) were assigned into two groups consisting of a negative control group (G1; n = 62), in which animals were treated monthly with a control product (sarolaner; Simparica®), and a treatment group (G2; n = 63) in which animals were treated monthly with Simparica Trio (sarolaner/moxidectin/pyrantel) from day 0 to day 150. In total, nine animals were withdrawn from the study (two animals became positive at day 30, and seven for reasons unrelated to eyeworm infection), resulting in 116 animals (n = 58 for G1; n = 58 for G2). Results In G1, 16 out of 58 animals (27.6%) were observed with eyeworms during the study, and none of the animals from G2 were ever observed with eyeworms, resulting in 100% efficacy (P T. callipaeda infection. Adult nematodes and fourth-instar (L4)-stage larvae were recovered from the eyes of positive animals, counted, and morphologically identified as T. callipaeda. In addition, specimens from Italy were molecularly confirmed as belonging to the haplotype 1 (i.e., the only one circulating in Europe so far). Conclusions Data presented herein demonstrated 100% efficacy of Simparica Trio for the prevention of T. callipaeda eyeworm infection in dogs from highly endemic areas of France and Italy. The use of this formulation is advantageous, as it is a licensed product in Europe with a wide efficacy spectrum against other nematodes, multiple tick species, and fleas. In addition, preventing the development of infection in dogs could also be a prophylaxis measure for zoonotic T. callipaeda infection in humans inhabiting endemic areas. Graphical Abstract

Although

Canine leishmaniosis (CanL) by Leishmania infantum (L.i.) and heartworm disease by Dirofilaria immitis (D.i.) are common zoonotic vector-borne diseases (VBDs) characterized by a variety of pathological and clinical signs. The immunopathology in both VBDs is extremely complex, and their clinical manifestations are strongly dependent on the type of immune response elicited by the parasites. In particular, the formation of circulating immune complexes (CICs) plays an important role in the pathogenesis of these VBDs. Based on the international guidelines, dogs with high anti-L. infantum antibody titres and one or more clinical and/or laboratory signs related to CanL require anti-Leishmania treatment. Consequently, the CICs measurement could be used for improving the clinical staging process of CanL. The aim of the study was to assess the CICs level by a competitive inhibition enzyme immunoassay, in healthy or sick dogs seropositive to L.i. and in healthy dogs positive to D.i.. Out of 51 enrolled dogs, 11 were included in Group A (seronegative to L.i., D.i. negative and healthy), 15 in Group B (exposed to L.i., D.i. negative and healthy), 12 in Group C (seropositive to L.i., D.i. negative and sick) and 13 in Group D (seronegative to L.i, D.i. positive and healthy). The comparison of CIC level in canine sera revealed a significant difference among groups (P 0.001), with the highest concentration (i.e., median = 104.6 µg/mL) in dogs with CanL. The findings of the study highlight the CICs measurement as a useful tool in the clinical staging of CanL for avoiding misclassification of dogs as leishmaniotic, thus not requiring anti-Leishmania therapy, as well as the possibility of results misuse in geographical areas where both leishmaniosis and heart-worm disease are endemic.

With over 300 terrestrial and aquatic mammalian species, Iran is considered a country with an ample mastofauna. Although many studies have assessed the distribution of gastrointestinal helminth parasites in animals and humans in Iran, lungworms have not received adequate attention. Following a previous article in which we reviewed the diversity and prevalence of lungworm infections in pastoral and wild ruminants of Iran, this report compiles the available scientific information about the occurrence of lungworms in non-ruminant mammals and humans from 1980 to 2022 to provide insights into the epidemiology of these infections. International and national scientific databases were searched, and twenty-six articles in peer-reviewed journals, one conference paper, and one D.V.M. thesis were included in the study. In total, 10 species belonging to seven genera, including Dictyocaulus, Deraiophoronema, Protostrongylus, Crenosoma, Eucoleus, Aelurostrongylus, and Metastrongylus, were reported in the respiratory tract or feces of humans, domestic animals (i.e., camels, equids, dogs, and cats), and wildlife species (i.e., hedgehogs, wild boars, and hares). Most of the studies (22/28) were performed using post-mortem examinations. The overall prevalence of respiratory nematode infection varied according to animal species in camels (14.83%), equids (13.31%), dogs (5%), wild boars (45.66%), hedgehogs (42.57%), and hares (1.6%). In addition, pulmonary capillariasis caused by Eucoleus aerophilus was reported in a 9 year old child. The prevalence of lungworm species in domestic camels, equids, and dogs, combined with a lack of labeled anthelmintic products, supports the need to improve our understanding of these important nematode parasites and inform the development of sustainable control strategies. From a zoo and wildlife medicine point of view, there is a shortage of information about the presence and prevalence of lungworm infections in the majority of mammalian species, pending epidemiological studies that integrate classical parasitology and molecular methods.

Species of Ixodiphagus (Hymenoptera: Encyrtidae) are parasitoid wasps whose immature forms develop inside ixodid and argasid ticks (Acari: Ixodida). Following oviposition by adult female wasps into the idiosoma of ticks, larvae hatch and start feeding on their internal contents, eventually emerging as adult wasps from the body of the dead ticks. Species of Ixodiphagus have been reported as parasitoids of 21 species of ticks distributed across 7 genera. There are at least ten species described in the genus, with Ixodiphagus hookeri being the most studied as an agent for biological control of ticks. Although attempts of tick control by means of this parasitoid largely failed, in a small-scale study 150,000 specimens of I. hookeri were released over a 1-year period in a pasture where a small cattle population was kept, resulting in an overall reduction in the number of Amblyomma variegatum ticks per animal. This review discusses current scientific information about Ixodiphagus spp., focusing on the role of this parasitoid in the control of ticks. The interactions between these wasps and the ticks’ population are also discussed, focusing on the many biological and logistical challenges, with limitations of this control method for reducing tick populations under natural conditions.

Wolbachia, a maternally transmitted Gram-negative endosymbiont of onchocercid nematodes and arthropods, has a role in the biology of their host; thus it has been exploited for the filariasis treatment in humans. To assess the presence and prevalence of this endosymbiont in reptiles and their ectoparasites, blood and tail tissue as well as ticks and mites collected from them were molecularly screened for Wolbachia DNA using two sets of primers targeting partial 16S rRNA and Wolbachia surface protein (wsp) genes. Positive samples were screened for the partial 12S rRNA and cytochrome c oxidase subunit 1 (cox1) genes for filarioids. Of the different species of lizards (Podarcis siculus, Podarcis muralis and Lacerta bilineata) and snakes (Elaphe quatuorlineata and Boa constrictor constrictor) screened from three collection sites, only P. siculus scored positive for Wolbachia 16S rRNA. Among ectoparasites collected from reptiles (Ixodes ricinus ticks and Neotrombicula autumnalis, Ophionyssus sauracum and Ophionyssus natricis mites), I. ricinus (n = 4; 2.8%; 95% CI, 0.9–7) from P. siculus, N. autumnalis (n = 2 each; 2.8%; 95% CI, 0.9–6.5) from P. siculus and P. muralis and O. natricis (n = 1; 14.3%; 95% CI, 0.7–55.4) from Boa constrictor constrictor scored positive for Wolbachia DNA. None of the positive Wolbachia samples scored positive for filarioids. This represents the first report of Wolbachia in reptilian hosts and their ectoparasites, which follows a single identification in the intestinal cells of a filarioid associated with a gecko. This data could contribute to better understand the reptile filarioid-Wolbachia association and to unveil the evolutionary pattern of Wolbachia in its filarial host.

Trypanosomes are haemoflagellate protozoa transmitted by blood-feeding arthropods causing infections in a wide range of mammals, including humans. Adult badgers (Meles meles, n = 2), displaying severe paralysis, ataxia and severe ectoparasite infestation, were rescued from a peri-urban area of Bari (southern Italy). Blood samples and ectoparasites were screened for Trypanosoma spp. by the combined PCR/sequencing approach, targeting a fragment of 18S rRNA gene. Smears of haemolymph, guts and salivary glands of the alive ticks were microscopically observed. No haematological alterations, except thrombocytopenia, were found. Trypomastigotes and epimastigotes were observed in the blood smears of both badgers and Trypanosoma pestanai was molecularly identified. Out of 33 ticks (i.e. n = 31 Ixodes canisuga, n = 2 Ixodes ricinus) and two fleas (Ctenocephalides felis), 11 specimens (n = 5 I. canisuga engorged nymphs, n = 4 engorged females and n = 2 I. ricinus engorged females) tested positive only for T. pestanai DNA. All smears from ticks were negative. The present study firstly revealed T. pestanai in Ixodidae and badgers from Italy, demonstrating the occurrence of the protozoan on the peninsula. Further studies are needed to clarify the occurrence of the only known vector of this parasite, Paraceras melis flea, as well as other putative arthropods involved in the transmission of T. pestanai.

To investigate the role of dipterans in the transmission of Onchocerca lupi and other zoonotic filarioids, samples were collected from different sites in Algarve, southern Portugal, morphologically identified and molecularly tested for filarioids. Culex sp. (72.8%) represented the predominant genus followed by Culicoides sp. (11.8%), Ochlerotatus sp. (9.7%), Culiseta sp. (4.5%), Aedes sp. (0.9%) and Anopheles sp. (0.3%). Nineteen (2.8%) specimens scored positive for filarioids, with Culex pipiens quinquefasciatus (2%) positive for Dirofilaria immitis (1.4%), Dirofilaria repens, Acanthocheilonema reconditum, Onchocerca lupi, unidentified species of Filarioidea (0.2%, each) and Onchocercidae (0.6%). Additionally, Culiseta longiareolata (6.5%), Ochlerotatus caspius (3%) and Culex laticinctus (0.2%) scored positive for unidentified Onchocercidae, A. reconditum and for O. lupi, respectively. This is the first report of the occurrence of DNA of O. lupi, D. repens and A. reconditum in Culex spp. in Portugal. Information regarding the vectors and the pathogens they transmit may help to adopt proper prophylactic and control measures., First report of the occurrence of the DNA of Onchocerca lupi, Dirofilaria repens and Acanthocheilonema reconditum in Culex spp. from Portugal.Unidentified species of Filarioidea and Onchocercidae were detected in mosquitoes from Portugal.The detection of Wolbachia supergroup E in Culex laticinctus positive for O. lupi is new to science and deserves further investigation.

For a long time known as the oriental eyeworm, Thelazia callipaeda is a zoonotic nematode that infects the eyes of a wide range of vertebrate hosts including dogs, cats, wildlife carnivores, lagomorphs, and humans. The high occurrence of this infection in Europe and the first cases in the United States have increased scientific interest in the parasite, as it also represents a risk for people living in endemic areas. Therefore, treatment and prevention of thelaziosis in canine population are advocated to reduce the risk of human infection as well. Here, we assessed the efficacy of a formulation containing sarolaner/moxidectin/pyrantel (Simparica TrioThe study was conducted in eyeworm endemic areas of Italy and France, where dogs (n = 125) were assigned into two groups consisting of a negative control group (G1; n = 62), in which animals were treated monthly with a control product (sarolaner; SimparicaIn G1, 16 out of 58 animals (27.6%) were observed with eyeworms during the study, and none of the animals from G2 were ever observed with eyeworms, resulting in 100% efficacy (P 0.0001) in the prevention of establishment of T. callipaeda infection. Adult nematodes and fourth-instar (L4)-stage larvae were recovered from the eyes of positive animals, counted, and morphologically identified as T. callipaeda. In addition, specimens from Italy were molecularly confirmed as belonging to the haplotype 1 (i.e., the only one circulating in Europe so far).Data presented herein demonstrated 100% efficacy of Simparica Trio for the prevention of T. callipaeda eyeworm infection in dogs from highly endemic areas of France and Italy. The use of this formulation is advantageous, as it is a licensed product in Europe with a wide efficacy spectrum against other nematodes, multiple tick species, and fleas. In addition, preventing the development of infection in dogs could also be a prophylaxis measure for zoonotic T. callipaeda infection in humans inhabiting endemic areas.

A scarcity of information on the occurrence of zoonotic vector-borne pathogens (VBPs), alongside a lack of human and animal health authorities' awareness of pre-existing data, augment the risk of VBP infection for local people and limit our ability to establish control programs. This holds especially true in low-middle income countries such as Bosnia and Herzegovina (BiH). This dearth of information on zoonotic VBPs is bolstered by the inability of previously used diagnostic tests, including conventional molecular diagnostic methods, to detect the full spectrum of relevant pathogens. Considering this, we set out to apply a microfluidic qPCR assay capable of detecting 43 bacterial and protozoan pathogens from blood to accrue critical baseline data for VBPs occurrence in BiH. A total of 408 dogs were tested of which half were infected with at least one VBP of zoonotic or veterinary importance. Leishmania infantum was found in 18% of dogs, reaching a prevalence as high as 38% in urbanized areas of Sarajevo. These data highlight substantially higher levels of L. infantum prevalence when compared to that previously reported using conventional methods using the same samples. Additionally, this high-throughput microfluidic qPCR assay was able to detect pathogens rarely or never reported in canines in BiH, including Anaplasma phagocytophilum (3%), Anaplasma platys (0.2%), haemotropic Mycoplasma (1%) and Hepatozoon canis (26%). Our report of the endemicity of important zoonotic pathogens and those of clinical significance to dogs emphasizes the need for urgent implementation of surveillance and control for VBPs in BiH, targeting both animal and human infections within the country.

CRESS DNA viruses are significant pathogens of birds and pigs and have been detected repeatedly in human samples (stools, serum, and cerebrospinal fluid), both from healthy individuals and from patients with neurological disease, eliciting in 2013 a risk assessment by the European Centre for Disease Prevention and Control (ECDC). Sequences of CRESS DNA viruses previously reported in humans (TN9, TN12, and TN25), and detected in different animal species (e.g., birds, dogs, and bats) were herein detected in fecal samples of synanthropic squamates (geckos and lizards).

Background Some species of drosophilid flies belonging to the genus Phortica feed on ocular secretions of mammals, acting as biological vectors of the zoonotic eyeworm Thelazia callipaeda. This study describes an effective breeding protocol of Phortica variegata and Phortica oldenbergi in insectary conditions. Methods Alive gravid flies of P. oldenbergi, P. variegata and Phortica semivirgo were field collected in wooded areas of Lazio region (Italy) and allowed to oviposit singularly to obtain isofamilies. Flies were maintained in ovipots (200 ml) with a plaster-covered bottom to maintain high humidity level inside. Adult feeding was guaranteed by fresh apples and a liquid dietary supplement containing sodium chloride and mucin proteins, while larval development was obtained by Drosophila-like agar feeding medium. The breeding performances of two media were compared: a standard one based on cornmeal flour and an enriched medium based on chestnut flour. All conditions were kept in a climatic chamber with a photoperiod of 14:10 h light:dark, 26 ± 2 °C and 80 ± 10% RH. Results From a total of 130 field-collected Phortica spp., three generations (i.e. F1 = 783, F2 = 109, F3 = 6) were obtained. Phortica oldenbergi was the species with highest breeding performance, being the only species reaching F3. Chestnut-based feeding medium allowed higher adult production and survival probability in both P. oldenbergi and P. variegata. Adult production/female was promising in both species (P. oldenbergi: 13.5 F1/f; P. variegata: 4.5 F1/f). Conclusions This standardized breeding protocol, based on controlled climatic parameters and fly densities, together with the introduction of an enriched chestnut-based feeding medium, allowed to investigate aspects of life history traits of Phortica spp. involved in the transmission of T. callipaeda. Obtaining F3 generation of these species for the first time paved the road for the establishment of stable colonies, an essential requirement for future studies on these vectors in controlled conditions. Graphical abstract

Insects, ticks, and mites represent a threat to animal health globally, mainly due to their role as vectors of pathogens. Among the most important diseases, those transmitted by mosquitoes (e.g., malaria and arboviral infections) and ticks (e.g., Lyme borreliosis, babesiosis, and viral haemorrhagic fever) have a huge impact on human health. The principal methods available for reducing the public health burden of most vector-borne diseases are vector-based intervention relying to insecticides and acaricides. However, the use of these products is challenged by the introduction of invasive species, the quick development of physiological insecticide and acaricide resistance, and their non-target effects on human health and environment. In this scenario, insecticide/acaricide-free control approaches based on the employment of entomopathogenic fungi (EPFs) are currently considered a promising tool in Integrated Pest/Vector Management, even if their large-scale use is still limited. In this article, we provide an overview on current knowledge about the role of EPFs for mosquito and tick management to assess solutions improving the delivery and efficacy of EPFs in the field. Laboratory research provided solid evidence that EPFs represent a next-generation control tool to manage mosquito and tick populations. However, the viability, infectivity, and persistence of fungal spores under field conditions are still inadequate. Herein we also discuss the development and optimization of EPF-based lure and kill approaches through biopolymers to improve cost-competitive, safety and eco-friendly pest and vector control tools.

Essential oils (EOs) of

Pest management is looking for green and cost-effective innovative solutions to control tiger mosquitoes and other pests. By using biomimetic principles and biocompatible/biodegradable biopolymers, it could be possible to develop a new approach based on substrates that selectively attract insects by reproducing specific natural environmental conditions and then kill them by hosting and delivering a natural biopesticide or through mechanical action (biomimetic lure and kill approach, BLK). Such an approach can be theoretically specialized against tiger mosquitoes (BLK-TM) by designing hydrogels to imitate the natural oviposition site's conditions to employ them inside a lure and kill ovitraps as a biomimetic oviposition substrate. In this work, the hydrogels have been prepared to prove the concept. The study compares lab/on-field oviposition between standard substrates (absorbing paper/masonite) and a physical and chemically crosslinked hydrogel composition panel. Then the best performing is characterized to evaluate a correlation between the hydrogel's properties and oviposition. Tests identify a 2-Hydroxyethylcellulose (HEC)-based physical hydrogel preparation as five times more attractive than the control in a lab oviposition assay. When employed on the field in a low-cost cardboard trap, the same substrate is seven times more capturing than a standard masonite ovitrap, with a duration four times longer.

Dirofilaria immitis and Dirofilaria repens are mosquito-borne filarioids of zoonotic concern, which primarily cause canine heartworm disease (HWD) and subcutaneous dirofilariosis, respectively. The geographical distribution of these nematodes is constantly changing, due to many factors such as the colonization by new invasive mosquito species, the increased movement of animals and the absence of chemoprophylactic strategies in non-endemic regions. Although HWD has always been considered endemic in northern Italy, an increase in the prevalence of this disease has been recorded in the last decades in central and southern regions. We describe the southernmost hyperendemic European focus of heartworm disease in the Pelagie archipelagos. From June to November 2020, 157 dogs and 46 cats were sampled in Linosa and Lampedusa islands for the detection of Dirofilaria immitis and Dirofilaria repens by modified Knott's, rapid antigen and molecular tests. Mosquitoes were collected using CDC light and BG sentinel-2 traps and aspirators, and tested for host blood meal and Dirofilaria spp. DNA. Out of 56 dogs in Linosa and 101 in Lampedusa, 33 (i.e. 58.9%) and 8 (i.e. 7.9%) were positive to D. immitis and D. repens, respectively. Three cats scored positive to D. immitis (i.e. 17.6%) in Linosa. Six mosquito species were identified, and the abdomen of a non-engorged Aedes albopictus was positive for D. immitis and human DNA. The results suggest that D. immitis infection could spread to new previously non-endemic territories in southern Europe, representing a real threat to animal and human health.

Medium sized opossums (Didelphisspp.) are among the most fascinating mammals of the Americas, playing important ecological roles (e.g., dispersal of seeds and control of insect populations) in the environment they inhabit. Nevertheless, as synanthropic animals, they are well adapted to human dwellings, occupying shelters within the cities, peripheral areas, and rural settings. These marsupials can harbor numerous pathogens, which may affect people, pets, and livestock. Among those, some protozoa (e.g.,Leishmania infantum,Trypanosoma cruzi,Toxoplasma gondii), helminths (e.g.,Ancylostoma caninum,Trichinella spiralis,Alaria marcianae,Paragonimusspp.) and arthropods (e.g., ticks, fleas) present substantial public health and veterinary importance, due to their capacity to cause disease in humans, domestic animals, and wildlife. Here, we reviewed the role played by opossums on the spreading of zoonotic parasites, vectors, and vector-borne pathogens, highlighting the risks of pathogens transmission due to the direct and indirect interaction of humans and domestic animals withDidelphisspp. in the Americas.

Two cats infected by Dirofilaria immitis and Dirofilaria repens, respectively, were taken to two different private practitioners for a clinical examination. The analyses conducted on the first cat revealed a microfilaraemia due to D. repens of 66 mfs/mL by a modified Knott’s test. No clinical signs of D. repens infection were observed in the cat. The animal was euthanised because of a lymphoma condition, and two adult females of D. repens were found in the subcutaneous tissue of the lumbar and left scapular regions at the post-mortem examination. The second cat showed severe abnormalities in the white blood cells, including eosinophil count. Microfilariae of D. immitis were detected in the blood smear, with an average length (n = 2) of 296.2 μm. These clinical cases represent the first reports of feline dirofilarioses in southern Italy and are indicative of a common occurrence of dirofilarial infection in the local canine population.

Cladosporium infections have a poor prognosis in animals, most likely due to a lack of knowledge about diagnosis and treatment. In this study, we described a case of a lethal Cladosporium allicinum infection in a captive bullfrog (Pyxicephalus adspersus) in Europe. One adult male bullfrog was referred with clinical signs of lethargy and a cutaneous nodule. Fungal infection was suspected on cytology and confirmed by histology and cultural isolation. The mold was identified by molecular methods using partial sequencing of the TEF1α gene and the ITS region of rDNA. Climbazole antifungal treatment was started but the frog died after 30 days, and necropsy was done. Pigmented hyphae and structures consistent with muriform bodies were found on a background of diffuse granulomatous inflammation at cytological and histopathological examinations. Fungal culture revealed the presence of pigmented fungi identified as Cladosporium allicinum only by partial sequencing of the TEF1α gene. A focally extensive granuloma with intralesional hyphae and muriform bodies effacing the architecture of head, liver, kidneys, lungs, and large intestine were retrieved after necropsy. This study is the first Italian report of the occurrence of lethal C. allicinum infection in a frog and highlights the role of this Cladosporium sp. in chromoblastomycosis.

Pigeons are common birds around the world and may act as intermediate hosts of the tissue cyst-forming apicomplexan parasites Toxoplasma gondii, Neospora caninum and Sacrocystis spp. This study aimed to provide an overview on the prevalence of and exposure to these parasites in Iranian domestic rock pigeon (Columba livia domestica) through molecular, serological and histopathological examination. Blood and tissue samples (i.e., brain, heart, gizzard, neck, thigh, and pectoral muscles) were taken from 100 pigeons. Sera were screened by agglutination tests for detection of anti- T. gondii and N. caninum antibodies, genomic DNA from tissue samples were assessed by respective species-specific PCRs, and histopathological examination of tissues was carried out. A seroprevalence of 45 % to anti-T. gondii and 35 % to anti-N. caninum IgG was recorded. PCR detected T. gondii DNA in 28 pigeons. Sacrocystis spp. was detected in one animal, but sequencing of the 28 S rRNA gene product did not reveal the identity of the species. Histopathology revealed myocarditis, myositis, and gliosis in the heart, skeletal muscles, and brain, respectively. No Sarcocystis tissue-cysts were detected, but T. gondii tissue cyst-like structures in the brain (i.e., 4 %) and heart (i.e., 3 %) were found by histology. Data reported herein demonstrate that pigeons from Iran are infected with tissue cyst-forming apicomplexans, particularly T. gondii. Since domestic pigeons are in close contact with human populations, and consumption of their meat and egg is popular in different societies, control strategies for minimizing the risk of infection in both pigeons and humans are suggested.

Global changes have influenced our societies in several ways with both positive (e.g., technology, transportation, and food security), and negative impacts (e.g., mental health problems, spread of diseases, and pandemics). Overall, these changes have affected the distribution patterns of parasites and arthropod vectors with the introduction and spreading of alien species in new geographical areas, eventually posing new challenges in public health. In this framework, the Acta Tropica Special Issue "Emerging parasites and vectors in a rapidly changing world: from ecology to management" provides a focus on the biology, ecology and management of emerging parasites and vectors of human and veterinary importance. Herein we review and discuss novel studies dealing with interactions of parasites and vectors with animals in changing environmental settings. In our opinion, a special focus on the implementation of management strategies of parasitic diseases to face anthropogenic environmental changes still represent a priority for public health. In the final section, key research challenges in this rapidly changing scenario are outlined.