Prevalence and Genotyping of Water- and Food-Borne Parasitic Protozoans (Giardia duodenalis and Cryptosporidium spp.) in Hospitalized Patients from Northwestern Romania.

Giardia duodenalis and Cryptosporidium spp. are important zoonotic protozoan pathogens that infect the gastro-intestinal tract of numerous vertebrates, including humans, and both parasites are responsible for water- or food-borne outbreaks of disease worldwide. Although, globally, both parasites are highly prevalent, particularly in developing countries, epidemiological data from Romania are scarce, and genotyping has rarely been performed. The aims of the present study were to investigate the occurrence and genetic diversity of G. duodenalis and Cryptosporidium spp. in patients hospitalized in Northwestern Romania in relation to clinical and paraclinical presentation and to identify the relative frequency of non-specific symptoms and potential risk factors. Between June 2022 and January 2024, 426 fecal samples were screened for gastro-intestinal parasites by rapid tests and microscopical examination, further confirmed by PCR and sequencing. Giardia duodenalis was detected and characterized in 12 samples (2.82%), while Cryptosporidium parvum was confirmed in four samples (0.94%). A majority of positive patients were symptomatic and reported nausea and vomiting with a significantly higher frequency compared to negative ones. This study provides new insights into the epidemiological status and clinical implications of gastro-intestinal parasite species and genospecies in Romania that are necessary for an in-depth understanding of the potential zoonotic transmission and improvement of patient care.

Wildlife parasitology: sample collection and processing, diagnostic constraints, and methodological challenges in terrestrial carnivores.

Wild terrestrial carnivores play a crucial role as reservoir, maintenance, and spillover hosts for a wide parasite variety. They may harbor, shed, and transmit zoonotic parasites and parasites of veterinary importance for domestic hosts. Although wild carnivores are globally distributed and comprise many different species, some living in close proximity to human settlements, only a few studies have investigated parasites of wild terrestrial carnivores using non-specific techniques. Access to samples of wild carnivores may be challenging as some species are protected, and others are secretive, possibly explaining the data paucity. Considering the importance of wild carnivores' health and ecological role, combined with the lack of specific diagnostic methodologies, this review aims to offer an overview of the diagnostic methods for parasite investigation in wild terrestrial carnivores, providing the precise techniques for collection and analysis of fecal, blood, and tissue samples, the environmental impact on said samples, and the limitations researchers currently face in analyzing samples of wild terrestrial carnivores. In addition, this paper offers some crucial information on how different environmental factors affect parasite detection postmortem and how insects can be used to estimate the time of death with a specific highlight on insect larvae. The paper contains a literature review of available procedures and emphasizes the need for diagnostic method standardization in wild terrestrial carnivores.

Fleas of wild carnivores in Romania: diversity, distribution, and host-associations.

BACKGROUND: Fleas are important hematophagous insects, infesting mammals and birds with a worldwide distribution. Fleas of medical importance have been reported from various carnivores worldwide, such as felids, canids, or mustelids. Romania hosts a wide carnivore diversity, but very little is known about flea species that parasitize these animals in Romania. This study aimed to provide a better understanding of the fleas' diversity and their distribution in a relatively large and diverse number of wild carnivore hosts from Romania. METHODS: From 2013 to 2021, 282 carcasses of wild carnivores from different locations in Romania were collected and examined for the presence of ectoparasites. All collected fleas were morphologically identified using specific keys and descriptions. An analysis of the co-occurrence networks was performed. RESULTS: A total of 11 flea species were identified: Pulex irritans (41.09%), Paraceras melis (20.11%), Ctenocephalides felis (7.33%), Ctenocephalides canis (7.83%), Monopsyllus sciurorum (11.11%), Chaetopsylla trichosa (21.96%), Chaetopsylla homoea (5.5%), Chaetopsylla tuberculaticeps (100%), Chaetopsylla rothschildi (13.33%), Chaetopsylla sp. (14.34%), Chaetopsylla globiceps (5.12%), Echidnophaga gallinacea (10%). The statistical analyses showed a significant difference between the infestation of Martes foina with females being more frequently infected than males (66% versus 33%). Paraceras melis infesting Meles meles had a significantly higher prevalence in female badgers than in males (× 2 = 7.7977, P < 0.01) and higher intensities of infestations in males than in females (t = 1.871, P < 0.05). CONCLUSIONS: This is the first large-scale study investigating the distribution and diversity of flea species infesting wild carnivores in Romania. Three flea species were identified for the first time in Romania (E. gallinacea, C. homoea, and C. tuberculaticeps).

Molecular survey of flea-borne pathogens in fleas associated with carnivores from Algeria and an Artificial Neural Network-based risk analysis of flea-borne diseases.

As ectoparasites and efficient vectors of pathogens fleas constitute a source of nuisance for animals as well as a major issue for public health in Algeria. In this study, a molecular survey has been conducted to investigate the presence of pathogens in fleas infesting domestic and wild carnivores in the central north and eastern north and south of Algeria. The molecular screening that targeted Acanthocheilonema reconditum, Bartonella spp.,and Dipylidium caninum, was supplemented by a comprehensive analysis of risk factors related to flea-borne pathogens, drawing data from all documentation across multiple languages and sources from Morocco, Algeria, and Tunisia. In the current study, several Bartonella spp. 56/430 (13.02%) and Dipylidium caninum 3/430 (0.7%) were identified. The sequencing results revealed 5/23 (21.74%) B. clarridgeiae, 3/23 (13.04%) B. henselae, and 3/23 (13.04%) B. vinsonii. The two haplotypes, H1 and H2, of D. caninum were identified for the first time in North Africa. The results of the Artificial Neural Network risk analyses unveiled that the prevalence of pathogens and the presence of host generalist fleas as well as the vectorial competence are the most determinant risk factors of flea-borne diseases in Maghreb.

ParSCo: celebrating 10 years of a unique parasitology summer course.

ParSCo (Parasitology Summer Course) is an intense, 1-week-long summer course organized by the Parasitology Unit of the Department of Veterinary Medicine, University of Bari, Italy, with the support of the World Association for the Advancement of Veterinary Parasitology (WAAVP), the European Veterinary Parasitology College (EVPC) and Parasites and Vectors. The course, which is conducted in southern Italy, is planned for parasitologists and post-graduate students working in the field of parasitology. The course consists of theoretical and practical lessons, which include the collection, identification and diagnosis of parasites of pets, livestock and wildlife. The participants in ParSCo are afforded the opportunity to be involved in clinical examination and sample collection for the diagnosis of parasitic diseases (e.g. leishmaniosis, thelaziosis and many tick-borne diseases) present in the Mediterranean Basin. The course is conducted at Casa di Caccia, a hunting lodge situated in the Gallipoli Cognato Forest near the Basento River in the Basilicata region in southern Italy. In addition to the training purpose, ParSCo is a great opportunity for sharing knowledge and expertise while becoming part of the parasitology community in a pleasant environment. In this editorial, we share some information and celebrate 10 years of ParSCo, looking forward to forthcoming sessions of this unique parasitology summer course.

On the efficacy of preventive toltrazuril treatments and the diagnosis of Cystoisospora suis infections in intensively raised piglets in farms from southeast Spain.

This study investigated the presence and abundance of Cystoisospora suis oocysts in faecal samples from 131 one- to three-week-old pig litters belonging to eight intensively raised, indoor herds in Spain. Seven herds used preventive anticoccidial toltrazuril treatments administered orally or by intramuscular injection, and one did not use preventive anticoccidial treatments. The diagnosis was performed using two oocyst flotation-concentration methods, Bailenger's method in every herd and the more recent Joachim's method in four herds. Oocysts were detected in every farm, the proportion of oocyst-positive samples was higher with Bailenger's technique, and the estimated overall prevalence (95% confidence interval) was 40 (32-49)%, including 47 (29-65)% in non-medicated litters, 52 (38-67)% in orally medicated litters and 28 (16-40)% in intramuscularly medicated litters (p < 0.05). However, mixed logistic regression models indicated that the risk of infection was not significantly associated with preventive anticoccidial treatments (p > 0.05), and increased with age, was higher in herds with partially compared to fully slatted dung floors in farrowing pens and in litters with pigs with diarrhoea (p < 0.05). The median (range) oocysts per gram of faeces (OpG) in infected litters by Bailenger's method was 623 (35-49048) and mixed negative binomial models revealed no significant association between infection intensity in positive litters and pen's floor type and piglets age, faecal consistency and treatment status (p > 0.05). The apparent low efficacy of Toltrazuril suggests treatment administration failures, reduced residual efficacy or low susceptibility of C. suis strains in study farms and needs further investigation.

A severe case of hyperinfection by Strongyloides stercoralis in a pet dog from Romania.

Strongyloides stercoralis is a zoonotic soil-transmitted nematode affecting mainly humans and dogs but identified also in non-human primates, cats and wild carnivores. It has a cosmopolitan distribution being endemic in tropical and subtropical areas. In Romania, the infection was reported on several occasions in dogs with low prevalence (3.5% -3.8%), assessed by coproscopy and it was confirmed in human patients with no travel history. A 2-year-old male Boston Terrier dog presented to a private clinic due to severe digestive problems, in July 2022. The animal had a long history of health problems. The dog was in a very bad clinical condition with severe abdominal pain, bloody diarrhea, and weight loss. Coproparasitological examinations using the saline flotation method and the modified Baermann's technique were done, both being negative. In addition, an intestinal biopsy was performed during the second endoscopy. Nematodes were collected and identified morphologically and molecularly confirmed. Histology revealed severe inflammation of the duodenal mucosa with areas of edema, necrosis, and hemorrhage, and in the intestinal glands, there were numerous nematodes suggesting a parasitic infection by Strongyloides spp. PCR followed by sequencing confirmed the infection with S. stercoralis. The dog was treated with a combination of oral fenbendazole and milbemycin oxime for 5 months. No relapse was observed 3 months after negativity was attained. This case describes a severe clinical infection by Strongyloides stercoralis in a domestic dog from Romania and the recovery after long-term treatment.

Pets, Wildlife and Parasites.

In our dynamic world, borders of different sorts are being rapidly altered or even erased [...].

An overview of fleas (Siphonaptera) in wild and domestic mammals from Algeria with new data from the central north and south of the country.

Fleas (Siphonaptera) are medically and veterinary important ectoparasites known to infest a wide range of host species. This study presents a systematic review complemented by new data on the diversity of flea species and host associations in Algeria. 198 mammals were examined, including previously understudied hosts such as sheep, red foxes, fennec foxes, and golden African wolves. Of these animals, 108 (54.55%) were infested with fleas. Overall, 1.906 fleas belonging to seven species (Archaeopsylla erinacei s.l., Ctenocephalides felis, C. canis, Nosopsyllus fasciatus, Pariodontis riggenbachi, Pulex irritans, Xenopsylla cheopis) were identified, originating from seven provinces across Algeria. Statistical analysis techniques were employed to process the data, including descriptive statistics, statistical tests, and principal component analysis (PCA). This study reveals the diverse flea fauna in Algeria and their association with various host species, including wild and domestic animals. This comprehensive survey aimed to combine literature reviews, fieldwork, and statistical analysis to understand fleas' diversity and host associations. The findings contribute to the growing knowledge of flea ecology and host-parasite interactions, with implications for public health and veterinary practices in Algeria and potentially other regions with similar ecological characteristics.

Is the Angiostrongylus vasorum infection in domestic dogs underestimated or misdiagnosed? A comprehensive presentation of four lethal cases.

Introduction: Angiostrongylus vasorum (A. vasorum) is a widely distributed gastropod-borne nematode, causing severe cardio-pulmonary disorders in dogs. In Romania, A. vasorum was detected in foxes and serologically confirmed in domestic dogs, but no clinical cases are currently diagnosed. Methods: Four dogs with clinical history of respiratory distress, originating from different geographical regions of Romania, were included in the study. One dog (case 1) was clinically evaluated using thoracic radiology and cardiac ultrasound; examination of feces and PCR were also performed for the etiologic diagnosis. The postmortem exam was performed in the other three cases, followed by parasitological and molecular analyses. Results: In the first case, parasitic pneumonia was suspected based on the radiographic examination of the thorax and the infection with A. vasorum was confirmed by L1 morphological identification and PCR. The main postmortem changes included large, coalescing, dark red areas of pulmonary consolidation (n = 3) and numerous adult nematodes in the pulmonary arteries (n = 2). The histopathological examination of the lungs showed necrotizing and granulomatous pneumonia with severe hemorrhages and chronic pulmonary arterial changes. Intralesional nematodes were seen in all necropsied cases. Additional inflammatory changes related to A. vasorum infection were identified in the brain and tracheobronchial and mediastinal lymph nodes (n = 2). Identification of larvae, histopathology and PCR confirmed the infection with A. vasorum. Conclusions: This study describes the first cases of canine cardiopulmonary angiostrongylosis in domestic dogs in Romania, and focuses on clinical presentation, pathological findings and molecular analysis. Angiostrongylosis should be included on the list of differential diagnoses of canine cardiopulmonary distress and/or haemorrhagic diathesis in Romania and awareness should be raised among clinicians to avoid post-mortem diagnosis in the future.

Diversity of Crenosoma species in mustelids with the first molecular characterization of C. melesi and C. petrowi.

Species of genus Crenosoma have a wide distribution and are reported in Europe, the Americas, and Asia. Currently, the genus includes 14 nominal species, out of which 9 are parasitic in mustelids. Two species are mostly reported in mustelids from Europe, namely C. melesi and C. petrowi. Up to now, no genetic sequences are deposited in GenBank for any of the two. The aims of this study were to investigate the distribution, prevalence, and diversity of Crenosoma spp. infecting mustelids in Romania and to genetically characterize the species. Mustelids (n = 247) were collected over a period of 7 years from different locations in Romania and the respiratory tract was removed and examined for nematodes. Detected nematodes were morphologically identified and fragments of two genes were sequenced. Sampled mustelids included Eurasian badger, Meles meles (n = 102), Eurasian otter, Lutra lutra (n = 20), beech marten, Martes foina (n = 36), European pine marten, Martes martes (n = 5), steppe polecat, Mustela eversmanii (n = 1), European mink, Mustela lutreola (n = 1), least weasel, Mustela nivalis (n = 2), European polecat, Mustela putorius (n = 78), and marbled polecat, Vormela peregusna (n = 1). Nematodes from Eurasian badgers were morphologically identified as C. melesi (n = 13, 12.74%) and C. petrowi (n = 3, 2.94%). Nematodes from the beech martens were identified as C. petrowi (n = 6, 16.66%), C. vulpis (n = 1, 2.78%) and Crenosoma spp. (n = 3, 8.33%). Co-infections with two Crenosoma species were detected in one beech marten (C. petrowi + C. vulpis, n = 1, 2.77%) and in one European pine marten [C. petrowi + C. vulpis (n = 1, 20%)]. Two genes of Crenosoma melesi and C. petrowi were partly sequenced for the first time. We report new host-parasite associations for M. martes and C. vulpis. However, further studies are needed in order to determine the host-parasite associations and to improve the understanding of the epidemiology of Crenosoma nematodes.

First report of Thelazia callipaeda Railliet & Henry, 1910 in a zoo lion (Panthera leo) (Linnaeus, 1758) from Romania.

Thelazia callipaeda, the zoonotic "oriental eye worm", is an emerging nematode known to infect a wide range of hosts, particularly carnivores (wild and domestic canids and felids, mustelids, and ursids), but also other mammal groups (suids, lagomorphs, monkeys, and humans), across a wide geographical area. New host-parasite associations and human cases have mostly been reported in endemic areas. A less studied group of hosts is represented by zoo animals that may harbor T. callipaeda.A 23-year old male lion (Panthera leo) that was born in a public zoo in Reșița (Caraș-Severin County, Romania) was submitted to the University of Agricultural Sciences and Veterinary Medicine of Cluj-Napoca for investigations after death from unknown reasons in November 2022. Four nematodes were collected from the right eye during the necropsy and submitted to morphological and molecular characterization.Collected nematodes were morphologically identified as three females and one male T. callipaeda. The BLAST analysis revealed 100% nucleotide identity to numerous isolates of T. callipaeda haplotype 1.The present paper presents a new host-parasite association between T. callipaeda and P. leo from a zoo in Romania.

First report of Crenosoma vulpis in Africa and Eucoleus aerophilus in Algeria.

Introduction: Crenosoma vulpis and Eucoleus aerophilus are widely distributed lungworms infecting carnivores, mainly red foxes, and are localized in the trachea, bronchi and bronchioles. There are no reports from Africa regarding the presence of C. vulpis. The aim of the present study was to report a co-infection with C. vulpis and E. aerophilus in a red fox from Algeria. Materials and methods: In January 2022, a road-killed male juvenile red fox (Vulpes vulpes) was collected from Bouhadjar-Tarf locality and was submitted for a complete parasitological necropsy. Detected nematodes were collected and preserved in ethanol for morphological and molecular identification. Tissue samples were also collected and analyzed by histopathological methods. Results: Collected nematodes were identified as a male C. vulpis and several Eucoleus aerophilus. The histological techniques of the lung tissue did not reveal the presence of any larvae, or lung inflammation. Conclusion: To the best of our knowledge, this is the first report of C. vulpis infecting a carnivore in this continent, highlighting the importance of further studies to update the geographical distribution of C. vulpis. E. aerophilus was first reported in Algeria. Red foxes are important spreaders of parasitic diseases. Further studies are required for a better understanding of its epidemiology across North Africa and other areas overlapping the range of the red fox.

Wild Mesocarnivores as Reservoirs of Endoparasites Causing Important Zoonoses and Emerging Bridging Infections across Europe.

Mesocarnivores are small- or mid-sized carnivore species that display a variety of ecologies and behaviours. In Europe, wild mesocarnivores are represented by the red fox (Vulpes vulpes), the golden jackal (Canis aureus), the European wildcat (Felis silvestris), the Mustelidae of the genera Meles, Martes, Mustela, Lutra, the invasive species of raccoon dog (Nyctereutes procyonoides), raccoons (Procyon lotor), and American mink (Neogale vison). These abundant animals thrive in various habitats and often develop their activity close to human settlements. Thus, they may play an important role in the introduction, maintenance, and transmission of major parasitic zoonoses and promote bridging infections with domestic animals. Against this background, this article reports and discusses some of the most important endoparasites of wild mesocarnivores living in Europe, on the basis of their actual role as reservoirs, spreaders, or sentinels. The data derived from epizootiological studies in different European countries, and the proven or speculated implications of the detected endoparasites in human and domestic animals' health, are discussed. Through older and recent literature review, the state-of-the-art knowledge on the occurrence and prevalence of the parasites under consideration is presented, showing further, warranted investigations and the need for surveillance and vigilance.

Trichinella britovi in wild boar meat from Italy, 2015-2021: A citizen science approach to surveillance.

As a result of the increase of game meat intended for human consumption through Europe, a plethora of food-borne diseases, including trichinellosis, may occur in consumers, posing a relevant public health threat. Thus, this study aims to a citizen science approach to monitor the occurrence of Trichinella spp. in wild boar meat intended for human consumption, evaluating the risk of infection for consumers. Following the European Regulation 2015/1375 (laying down specific rules on official controls for Trichinella in meat), from 2015 to 2021, hunters (n = 478) were involved to collect diaphragm pillar samples of wild boars from mainland southern Italy, which were tested for Trichinella spp. L1 larvae via HCl-pepsin digestion and Multiplex PCR. Overall, 139,160 animals were collected (average of 19,880 per year), being 14 (i.e., 0.01%) tested positive to Trichinella britovi by the combined biochemical and molecular approach. An average larval burden of 28.4 L1 per gram of meat was found (minimum 3.2 - maximum 132.6). A statistically significant difference was found in the prevalence according to hunting seasons (p < 0.01, with higher values in 2016 and 2021) and regions of the study area (p < 0.01). No statistically significant decrease in the prevalence of T. britovi throughout the study period was found (p = 0.51), except in Apulia region (p < 0.01). These findings revealed a stable prevalence of T. britovi in wild boar meat intended for human consumption, suggesting a risk of infection for consumers, especially hunters and local markets users. Citizen science surveillance models could be promoted to improve trichinellosis control and prevention in a One Health perspective.

Efficacy of a topical formulation containing esafoxolaner, eprinomectin and praziquantel (NexGard Combo®) against natural infestations with the cat louse, Felicola subrostratus under field conditions.

Felicola subrostratus is the only species of louse affecting domestic cats. Although it is considered a rare ectoparasite of pet cats, it occurs mainly in stray or shelter animals and is sometimes associated with intense pruritus and secondary bacterial infections. The aim of the present study was to evaluate the efficacy of a single dose of the topical formulation of esafoxolaner, eprinomectin and praziquantel (NexGard Combo®, Boehringer Ingelheim) in cats for the treatment of naturally acquired chewing lice infestation in a multi-site, positive-control, blinded clinical field study. Thirty-one domestic cats presenting natural F. subrostratus infestation were included in the study. The animals had not been treated with any ectoparasiticide within the previous 60 days. After inclusion, each cat was randomly assigned to one of the two groups: group 1, 14 cats treated with NexGard Combo® or group 2, 17 cats treated with Frontline Combo®. A clinical evaluation was performed at days 0 (inclusion), 14 and 30 and consisted in scoring the skin lesions and symptoms and scoring the presence of lice. On day 30, all cats from both groups scored 0 for the presence of live lice and no dead lice were found, demonstrating 100% efficacy. The clinical scores significantly improved from day 0 to day 30 in both groups. During the 30 days of surveillance, no reinfestations due to the hatching of eggs were observed and none of the cats had any adverse reactions. Esafoxolaner demonstrated high efficacy for the treatment of F. subrostratus infestation.

Title: Efficacité d'une formulation topique contenant de l'esafoxolaner, de l'éprinomectine et du praziquantel (NexGard Combo®) contre les infestations naturelles par le pou du chat, Felicola subrostratus, dans des conditions de terrain. Abstract: Felicola subrostratus est la seule espèce de pou affectant les chats domestiques. Bien que ce pou soit rare comme ectoparasite chez les chats de compagnie, il infeste principalement les animaux errants ou en refuge et est parfois associé à un prurit intense et à des infections bactériennes secondaires. Le but de la présente étude était d'évaluer l'efficacité d'une dose unique de la formulation topique d'esafoxolaner, d'éprinomectine et de praziquantel (NexGard Combo®, Boehringer Ingelheim) chez le chat pour le traitement de l'infestation naturelle par les poux broyeurs dans une étude de terrain multi-sites, randomisée, avec un contrôle positif. Trente et un chats domestiques présentant une infestation naturelle par F. subrostratus ont été inclus dans l'étude. Les animaux n'avaient été traités avec aucun ectoparasiticide au cours des 60 jours précédents. Après inclusion, chaque chat a été assigné au hasard à l'un des deux groupes : groupe 1, 14 chats traités avec NexGard Combo®, ou groupe 2, 17 chats traités avec Frontline Combo®. Une évaluation clinique a été réalisée aux jours 0 (inclusion), 14 et 30 et a consisté à un score clinique des lésions et symptômes cutanés et à noter la présence de poux. Au jour 30, tous les chats des deux groupes ont obtenu un score de 0 pour la présence de poux vivants et aucun pou mort n'a été trouvé, démontrant une efficacité de 100 %. Les scores cliniques se sont significativement améliorés du jour 0 au jour 30 dans les deux groupes. Au cours des 30 jours de surveillance, aucune réinfestation due à l'éclosion des œufs n'a été observée et aucun des chats n'a présenté d'effet indésirable. L'esafoxolaner a démontré une efficacité élevée pour le traitement de l'infestation par F. subrostratus.

A Survey of Helminths of Dogs in Rural and Urban Areas of Uzbekistan and the Zoonotic Risk to Human Population.

Dogs are very popular pets that can be infected with a wide diversity of endo- and ectoparasites, some of which have zoonotic potential. The aim of the present study was to determine the diversity and prevalence of helminths in rural and urban dogs in Tashkent, Samarkand and Karakalpakstan regions of Uzbekistan. A total of 399 dogs from rural and urban areas were examined by necropsy between November 2016 and March 2022. All helminth species were morphologically identified. A total of 31 species belonging to the classes Trematoda (3), Cestoda (9), Nematoda (18) and Acanthocephala (1) were identified in 378 dogs (94.7%). Twenty-one species are indicated for the helminth fauna of urban dogs and 31 species for rural dog populations. From the 31 species of helminths identified 18 species are zoonotic and four of them (Echinicoccus granulosus, Dipylidium cani-num, Toxocara canis, Dirofilaria repens) have a significant epidemiological importance. The study showed that the prevalence and diversity of helminths in dogs in rural areas of Uzbekistan is higher than in urban dogs.

Efficacy of afoxolaner (NexGard®) against natural infestations with Trichodectes canis in dogs under field conditions.

BACKGROUND: Trichodectes canis is a small chewing louse found globally that primarily infests dogs. Limited information is available on the efficacy of isoxazolines against infestation with the chewing louse. In the present study, we evaluated the efficacy of afoxolaner, an isoxazoline class compound, in naturally infested domestic dogs. METHODS: The field study was carried out in Romania. Between September 2021 and December 2021, 43 dogs with confirmed T. canis infestation were included in the study. On the day of the inclusion (day 0), each animal was clinically examined and randomly treated with a control product labeled for use against lice [fipronil-(S)-methoprene combination (Frontline Combo®; Boehringer Ingelheim)] or with the investigational product [chewable tablets containing afoxolaner (NexGard®; isoxazoline)]. Each animal was evaluated for the presence of lice at 15 and 30 days post-inclusion. RESULTS: Of the 48 dogs initially included in the study, 43 completed the treatment period [18 in the control group (CG) and 25 in the investigational group (IG)]. At day 14, no living T. canis lice were detected on the dogs in either group. At day 14, dead lice were detected in four dogs in the IG, while eggs were present in two dogs in the IG and in one dog in the CG. At day 30, no lice were detected in either group, while eggs were still present in one dog in the CG. CONCLUSION: These results suggest that afoxolaner is a feasible treatment option against chewing lice in dogs, providing 100% curative efficacy.

Fleas from the Silk Road in Central Asia: identification of Ctenocephalides canis and Ctenocephalides orientis on owned dogs in Uzbekistan using molecular identification and geometric morphometrics.

BACKGROUND: The Silk Road connected the East and West for over 1500 years. Countries in Central Asia are valuable in addressing the hypothesis that parasites on domestic animals were introduced along the Silk Road. Adult fleas are obligate parasites, having worldwide distribution. In dogs, Ctenocephalides canis, C. felis and C. orientis are the most common species identified. The distribution of the Oriental cat flea, C. orientis, is restricted to southeast Asia. The purpose of this study was to determine the diversity of dog fleas from Uzbekistan, a country in Central Asia, with particular reference to C. orientis. METHODS: Fleas were collected from 77 dogs from 5 locations in Uzbekistan. The cox1 gene sequences from Ctenocephalides spp. were compared to global collection of Ctenocephalides cox1 haplotypes. Landmark-based geometric morphometrics have been applied to the head and curvature to compare C. canis and C. canis using canonical variate analysis and discriminant function analysis. RESULTS: Overall, 199 fleas were collected and identified as C. canis (n = 115, 58%), C. orientis (n = 53, 27%) and Pulex irritans (n = 22, 11%). None of the fleas were C. felis. All Ctenocephalides spp. fleas were subject to cox1 amplification and 95% (166/175) yielded DNA sequence. There were 25 cox1 haplotypes; 14 (22/25, 88%) were C. canis cox1 haplotypes and 3 (3/25, 12%) were C. orientis cox1 haplotypes. Molecular analysis confirmed the absence of C. felis. Four (4/22) and one (1/3) cox1 haplotypes were identical to cox1 haplotypes belonging to C. canis and C. orientis cox1 haplotypes identified elsewhere, respectively. Overall morphometric analysis confirmed significant differences between the head shape of C. canis and C. orientis and improved four-fivefold the species identification compared to traditional morphological key. CONCLUSION: We report for the first time the presence of C. orientis in Uzbekistan. Differentiation of C. orientis from C. canis and C. felis remains difficult in regions where these species coexist. Studies in Central and Southeast Asia should confirm species identity using cox1 locus to enable retracing of the distribution of the Ctenocephalides in Asia. The presence of C. orientis suggests that this species may have been introduced from the east along the ancient Silk Road.

Synopsis of the ticks of Algeria with new hosts and localities records.

BACKGROUND: Ticks are obligate hematophagous arthropods with a world-wide distribution that are extremely important not only in terms of human and animal health but also economically. In Algeria, information on tick species is scarce. METHODS: A systematic literature review was performed using online databases. The information extracted from the databases was was supplemented by information from an original study. Ticks were collected from various hosts and by flagging from January 2018 to December 2019. RESULTS: To date, in Algeria a total of 36 valid tick species belonging to two families have been recorded: (1) family Argasidae, with three Argas species and nine Ornithodoros species recorded; and (ii) family Ixodidae, with one Dermacentor species, three Haemaphysalis species, 10 Hyalomma species, four Ixodes species and six Rhipicephalus species recorded. The geographical distribution for each species was determined and listed. Eight new tick-host associations were recorded: four for Ixodes inopinatus sensu Estrada-Peña et al. 2014, one for Rhipicephalus bursa, one for R. turanicus, one for Hyalomma marginatum and one for Hy. lusitanicum. To our best knowledge, this study is the first to report the presence of I. inopinatus sensu Estrada-Peña et al. 2014 in Algeria. We also report here for the first time all tick species (Argasidae and Ixodidae) known to be present in Algeria. CONCLUSION: This article represents a tool for students and scientists who work in the field of ticks and provides important new data on the distribution of ticks in Algeria.