A case of mortality in a re-introduced European bison associated with severe pneumonia caused by Dictyocaulus viviparus.

BACKGROUND: Dictyocaulosis is a parasitic disease caused by pulmonary nematodes from genus Dictyocaulus affecting various ungulate hosts. It can cause verminous bronchopneumonia and for heavily infected individuals, fatal outcomes can occur. CASE PRESENTATION: The study describes the case of a male European bison which died three months after relocation from Slovakia to one of the reintroduction areas in Nucșoara village, Fagaraș Mountains, Romania. Necropsy revealed the presence of pulmonary nematodes in the respiratory tract. Morphology and molecular diagnosis was performed and Dictyocaulus viviparus was identified. CONCLUSION: Rigorous health monitoring is essential to assure the success of reintroduction programs, understanding and preventing infectious diseases and limiting their impact on population health.

Diversity of lice and flea- and lice-borne pathogens in free-ranging dogs in Uzbekistan.

Vector-borne diseases pose a significant threat to human and animal health worldwide, with arthropods, including fleas and lice, acting as key vectors for transmitting various pathogens. In Uzbekistan, where millions of domestic dogs coexist with humans, the diversity of vector-borne pathogens carried by ectoparasites remains largely unexplored. This study aimed to investigate the diversity and prevalence of lice and pathogens carried by fleas and lice collected from domestic dogs in Uzbekistan. Ectoparasites were collected from 77 dogs across five regions and subjected to morphological identification and molecular analysis. The 47 lice collected were identified as Trichodectes canis Fabricius, 1780 (Psocodea: Trichodectidae), Linognathus africanus Kellogg and Paine, 1911 (Psocodea: Linognathidae) and Bovicola bovis Linnaeus, 1758 (Phthiraptera: Trichodectidae), alongside with 197 fleas. Notably, L. africanus was detected for the first time on dogs worldwide. Molecular screening detected Rickettsia spp. in 27.9% fleas and 44.7% lice, Dipylidium caninum Linnaeus, 1758 (Cyclophyllidea: Dipylidiidae) in 1.5% of fleas and Bartonella spp. in 0.5% of fleas. Sequencing confirmed Rickettsia asembonensisMaina et al., 2016 (Rickettsiales: Rickettsiaceae), Rickettsia helvetica Beati et al., 1993 (Rickettsiales: Rickettsiaceae), Rickettsia monacensis Simser et al., 2019 (Rickettsiales: Rickettsiaceae), D. caninum and Bartonella clarridgeiae Lawson and Collins, 1996 (Hyphomicrobiales: Bartonellaceae). These findings highlight the potential role of dogs as reservoirs for vector-borne pathogens and emphasises the need for further research to inform effective disease control strategies.

Toxoplasma gondii in small mammals in Romania: the influence of host, season and sampling location.

BACKGROUND: Toxoplasma gondii is a protozoan parasite that infects a large spectrum of warm-blooded animals, including humans. Small rodents and insectivores play an important role in the epidemiology of T. gondii and may serve as a source of infection for both, domestic and wild definitive felid hosts. Factors influencing the occurrence of T. gondii in wild small mammals are unknown, despite the fact that many intermediate host species are identified. We have used small mammals (Rodentia and Lipotyphla) captured over two years in various habitats, both in urbanised and in natural landscapes. We assessed the importance of land-use, season and host ecology on T. gondii infection. RESULTS: We examined 471 individuals belonging to 20 small mammal species, collected at 63 locations spread over wide altitude, habitat and land-use ranges from Romania. Heart tissue samples were individually analysed by PCR targeting the 529 bp repetitive DNA fragment of T. gondii. The overall prevalence of infection was 7.3%, with nine species of rodents and two species of shrews being found to carry T. gondii DNA. Five species showed high frequency of infection, with the highest prevalence found in Myodes glareolus (35.5%), followed by Spermophilus citellus (33.3%), Sorex minutus (23.1%), S. araneus (21.7%) and Micromys minutus (11.1%). Adults seemed more often infected than young, however when controlling for season, the difference was not significant, as in spring both adults and young showed higher infection rates, but more adults were sampled. Contrary to our expectations, urban/rural areas (with their implicit high density of domestic feline presence) had no effect on infection prevalence. In addition, neither habitat, nor land-use at sampling sites was important as only geographical location and host species were contributing factors to the infection risk. CONCLUSIONS: High prevalence of T. gondii infection showed a highly localised, patchy occurrence, with long living and higher mobility host species being the most common carriers, especially during autumn.

VectorNet: collaborative mapping of arthropod disease vectors in Europe and surrounding areas since 2010.

BackgroundArthropod vectors such as ticks, mosquitoes, sandflies and biting midges are of public and veterinary health significance because of the pathogens they can transmit. Understanding their distributions is a key means of assessing risk. VectorNet maps their distribution in the EU and surrounding areas.AimWe aim to describe the methodology underlying VectorNet maps, encourage standardisation and evaluate output.Methods: Vector distribution and surveillance activity data have been collected since 2010 from a combination of literature searches, field-survey data by entomologist volunteers via a network facilitated for each participating country and expert validation. Data were collated by VectorNet members and extensively validated during data entry and mapping processes.ResultsAs of 2021, the VectorNet archive consisted of ca 475,000 records relating to > 330 species. Maps for 42 species are routinely produced online at subnational administrative unit resolution. On VectorNet maps, there are relatively few areas where surveillance has been recorded but there are no distribution data. Comparison with other continental databases, namely the Global Biodiversity Information Facility and VectorBase show that VectorNet has 5-10 times as many records overall, although three species are better represented in the other databases. In addition, VectorNet maps show where species are absent. VectorNet's impact as assessed by citations (ca 60 per year) and web statistics (58,000 views) is substantial and its maps are widely used as reference material by professionals and the public.ConclusionVectorNet maps are the pre-eminent source of rigorously validated arthropod vector maps for Europe and its surrounding areas.

The current situation of Angiostrongylus vasorum in Romania: a national questionnaire-based survey.

BACKGROUND: Angiostrongylus vasorum (Nematoda, Metastrongyloidea) is a vascular nematode that resides in the pulmonary arteries and the right side of the heart of a wide variety of carnivores, with an indirect life cycle using coprophagic gastropods as intermediate hosts. For domestic dogs, the infection with A. vasorum can be asymptomatic, but more frequently, it is associated with a wide range of clinical manifestations like cardio-respiratory signs, bleedings, neurological signs, and ocular problems which can lead to death when not treated accordingly. Angiostrongylosis was confirmed for the first time in Romania in red foxes (Vulpes vulpes) in 2017 and two years later a seroepidemiologic study was conducted among domestic dogs. However, to this date, no clinical canine angiostrongylosis cases were published in Romania. The aim of the present paper was to evaluate the knowledge about canine angiostrongylosis among veterinarians in Romania and to update the distribution of this disease using a national wide anonymous questionnaire. RESULTS: Overall, 147 unique responses were submitted, from 31 out of 42 counties. Twelve veterinarians (8%) from 8 counties (26%) acknowledged diagnosing a case of angiostrongylosis including 5 from the Bucharest and 1 from each of the remaining seven counties. All affected dogs had respiratory distress, 75% suffered cardiopathy, 16% coagulopathies and 8% neurological signs. Case diagnosis was based mostly on larval detection by coprology (67%) and serological antigen detection test (42%). CONCLUSIONS: Romanian veterinarians are aware of canine angiostrongylosis and a significant number have clinical experience with the disease. Epidemiological studies are now needed to assess its distribution in the country, and further efforts are required to improve understanding of the disease, its diagnostic and treatment methods among veterinarians.

First report of Cytauxzoon sp. infection in Germany: organism description and molecular confirmation in a domestic cat.

Cytauxzoonosis is described as an emerging tick-borne disease of domestic and wild felids caused by protozoans of the genus Cytauxzoon. While in the Americas the condition is described as a fatal disease, in Europe, reports on the clinical expression of the infection are scarce. This study describes the first case of Cytauxzoon sp. infection in Germany, in a domestic cat. A 6-year-old male domestic cat living in Saarlouis (Saarland) was presented with anorexia, lethargy and weight loss. The cat had an outdoor lifestyle and had not travelled abroad. Serum clinical chemistry analysis revealed azotaemia with markedly increased symmetric dimethylarginine, hypercreatinemia, hyperphosphatemia and hypoalbuminemia. Moreover, a mild non-regenerative anaemia was present. Approximately 1 year prior to these findings, the domestic cat was diagnosed with a feline immunodeficiency virus (FIV) infection. These results pointed toward a decreased glomerular filtration rate, presumably as a result of kidney dysfunction. Round to oval signet ring-shaped intraerythrocytic organisms, morphologically suggestive for a piroplasm, were revealed during blood smear evaluation with a degree of parasitaemia of 33.0%. PCR analyses and sequencing of a region of the 18S rRNA gene confirmed the presence of a Cytauxzoon sp. infection, with 99-100% nucleotide sequence identity with previously published Cytauxzoon sp. isolates. As this is the first molecularly confirmed Cytauxzoon sp. infection in a domestic cat in Germany, these findings suggest that cytauxzoonosis should be considered as a differential diagnosis in cases of anaemia in outdoor domestic cats, particularly in areas where wild felid populations are present.

The invasive Asian tiger mosquito Aedes albopictus in Romania: towards a country-wide colonization?

Currently, five invasive Aedes mosquito species are of concern in Europe according to the European Centre for Disease Prevention and Control: Aedes aegypti, Ae. albopictus, Ae. atropalpus, Ae. japonicus, and Ae. koreicus. Among these, only Ae. albopictus was reported to occur in Romania, in Bucharest. The aim of this study was to update the knowledge on the distribution of this invasive mosquito species in Romania, by investigating new potential locations. Monitoring of Ae. albopictus was carried out between May 2017 and October 2018. Three types of traps (CDC-Gravid Traps, CDC miniature Light Traps, ovitraps) were placed in 53 localities in 13 counties at sites suitable for container-breeding mosquitoes. Collected adult mosquitoes were counted and identified according to morphological criteria. Larvae were found present in domestic containers and rain catch basins. Aedes albopictus adults and eggs were collected in 10 localities in eight counties across Romania. Our study confirms nine new localities and seven counties where Ae. albopictus became established in Romania, highlighting the need for surveillance to further assess the species' distribution and abundance, as well as the pathogen transmission risk related to that vector species.

Seasonal dynamics of a population of Phlebotomus (Larroussius) perfiliewi Parrot, 1930 (Diptera: Psychodidae) in North-Eastern Romania.

Sand flies were collected in a location from Romania in order to estimate their abundance and seasonal variation in correlation with environmental and anthropic factors. From May to October 2017, eight premises with different animal species were sampled for sand flies in a household from Fundatura village, Vaslui County, in North-Eastern Romania. Animal-related data, shelter-related data, and climatic parameters were recorded. All (n = 150) collected sand flies were Phlebotomus perfiliewi. A mono-modal type of abundance trend has been recorded (a single peak at the beginning of August). The first day of capture was in mid-July. The total number of females during the peak season was significantly higher than the total number of males. The highest percentage of males was recorded at the beginning and at the end of the sand fly activity. Only the traps placed in the poultry enclosure built from clay and wood were positive. A strong positive correlation was recorded between the total number of collected sand flies and the minimum and the maximum temperature. The analysis of the climatic data shows that the first presence of sand flies was registered only after the average minimum temperature for the previous 7 days was above 15 °C.

A new Borrelia on the block: Borrelia miyamotoi - a human health risk?

BackgroundBorrelia miyamotoi clusters phylogenetically among relapsing fever borreliae, but is transmitted by hard ticks. Recent recognition as a human pathogen has intensified research into its ecology and pathogenic potential.AimsWe aimed to provide a timely critical integrative evaluation of our knowledge on B. miyamotoi, to assess its public health relevance and guide future research.MethodsThis narrative review used peer-reviewed literature in English from January 1994 to December 2018.ResultsBorrelia miyamotoi occurs in the world's northern hemisphere where it co-circulates with B. burgdorferi sensu lato, which causes Lyme disease. The two borreliae have overlapping vertebrate and tick hosts. While ticks serve as vectors for both species, they are also reservoirs for B. miyamotoi. Three B. miyamotoi genotypes are described, but further diversity is being recognised. The lack of sufficient cultivable isolates and vertebrate models compromise investigation of human infection and its consequences. Our understanding mainly originates from limited case series. In these, human infections mostly present as influenza-like illness, with relapsing fever in sporadic cases and neurological disease reported in immunocompromised patients. Unspecific clinical presentation, also occasionally resulting from Lyme- or other co-infections, complicates diagnosis, likely contributing to under-reporting. Diagnostics mainly employ PCR and serology. Borrelia miyamotoi infections are treated with antimicrobials according to regimes used for Lyme disease.ConclusionsWith co-infection of tick-borne pathogens being commonplace, diagnostic improvements remain important. Developing in vivo models might allow more insight into human pathogenesis. Continued ecological and human case studies are key to better epidemiological understanding, guiding intervention strategies.

An updated meta-analysis of the distribution and prevalence of Borrelia burgdorferi s.l. in ticks in Europe.

BACKGROUND: The bacteria of the group Borrelia burgdorferi s.l. are the etiological agents of Lyme borreliosis in humans, transmitted by bites of ticks. Improvement of control measures requires a solid framework of the environmental traits driving its prevalence in ticks. METHODS: We updated a previous meta-analysis of the reported prevalence of Borrelia burgdorferi s.l. in questing nymphs of Ixodes ricinus with a literature search from January 2010-June 2017. This resulted in 195 new papers providing the prevalence of Bb for 926 geo-referenced records. Previously obtained data (878 records, years 2000-2010) were appended for modelling. The complete dataset contains data from 82,004 questing nymphs, resulting in 558 records of B. afzelii, 404 of B. burgdorferi s.s. (only 80 after the year 2010), 552 of B. garinii, 78 of B. lusitaniae, 61 of B. spielmanii, and 373 of B. valaisiana. We associated the records with explicit coordinates to environmental conditions and to a categorical definition of European landscapes (LANMAP2) looking for a precise definition of the environmental niche of the most reported species of the pathogen, using models based on different classification methods. RESULTS: The most commonly reported species are B. afzelii, B. garinii and B. valaisiana largely overlapping across Europe. Prevalence in ticks is associated with portions of the environmental niche. Highest prevalence occurs in areas of 280°-290° (Kelvin) of mean annual temperature experiencing a small amplitude, steady spring slope, together with high mean values and a moderate spring rise of vegetation vigor. Low prevalence occurs in sites with low and a noteworthy annual amplitude of temperature and the Normalized Difference Vegetation Index (colder areas with abrupt annual changes of vegetation). Models based on support vector machines provided a correct classification rate of the habitat and prevalence of 89.5%. These results confirm the association of prevalence of the three most commonly reported species of B. burgdorferi s.l. in Europe to parts of the environmental niche and provide a statistically tractable framework for analyzing trends under scenarios of climate change.

Ixodid ticks parasitizing wild carnivores in Romania.

In Romania, data regarding hard-tick diversity and tick-host associations in wild carnivores are scarce. We aimed to identify tick species in wild carnivores and to establish reliable data on tick-host associations. The study was conducted in various Romanian localities from all five ecoregions found in the country. Fourteen species of wild carnivores were examined. Immature and adult ticks were collected and identified using the morphological keys. The frequency and mean intensity of tick infestation, overall and differentiated by species, developmental stage and host were calculated. Of 202 wild carnivores, 68 were parasitized by seven tick species (predominantly Ixodes ricinus, I. hexagonus and Dermacentor reticulatus). The mean intensity of tick infestation was similar in males (6.97, BCa 95% CI 5.15-9.88) and females (5.76, BCa 95% CI 4.15-9.17). The highest prevalence of infested animals was recorded in the pannonian and steppic ecoregions, 66.7 and 52.7%, respectively. In the continental ecoregion the prevalence was 26.7%, whereas in the pontic ecoregion it was 28%. The lowest value, 16.7% was recorded in the alpine ecoregion. In total 430 ticks were collected, and 24.8% (n = 50) of the animals were infested with more than one tick species. Fourteen new tick-host associations were recorded. Our results suggest that anthropogenic changes of the environment lead to the diminishing of the boundaries, between wild and domestic animals, increasing the exposure for both animals and humans, to infective agents, including tick-borne pathogens.

Angiostrongylus chabaudi (Biocca, 1957) in wildcat (Felis silvestris silvestris, S) from Romania.

Angiostrongylus chabaudi is a rare cardio-pulmonary nematode infecting felids. Although almost 60 years have passed since the original description of the species in Italy, this parasite has been seldom found in domestic and wildcats in southern Europe. The present study aims to report a new case of patent A. chabaudi infection in a road-killed wildcat from Maramureș County in Northern Romania. The necropsy revealed the presence of parasites in the pulmonary arteries and the right ventricle, and the fecal examination showed the presence of L1 larvae. Parasites were morphologically and morphometrically characterized as A. chabaudi, showing 100 % nucleotide similarity to an Angiostrongylus sp. originating from a wildcat from Germany and 99 % to A. chabaudi from Italy. This study reports A. chabaudi for the first time in Eastern Europe, expanding knowledge about the distribution range of this species.

Autochthonous Hepatozoon infection in hunting dogs and foxes from the Czech Republic.

Blood samples from 21 red foxes (Vulpes vulpes) and 8 hunting dogs from the same locality in the Czech Republic were examined for presence of Hepatozoon canis/Hepatozoon sp. The dogs were selected based on their close contact with foxes during fox bolting and because they had not traveled into known endemic areas. Using diagnostic PCR amplifying partial 18S rDNA fragment, Hepatozoon DNA was detected in 20 red foxes (95 %) and 4 dogs (50 %). From 8 positive foxes and 2 positive dogs, we obtained nearly complete 18S rDNA sequences. Phylogenetic analyses of these sequences revealed very low variability. Buffy coat smears from positive dogs were prepared and examined. No Hepatozoon gamonts were found. This study provides the first report of autochthonous infection of H. canis/Hepatozoon in dogs and foxes from the Czech Republic. Our study indirectly demonstrates cross infection between red foxes and dogs and confirms autochthonous infection of Hepatozoon canis in dogs living in a geographic area well outside the range of Rhipicephalus sanguineus sensu lato, which is so far the only known vector of H. canis in Europe.

Current surveys on the prevalence and distribution of Dirofilaria spp. and Acanthocheilonema reconditum infections in dogs in Romania.

During the last decades, Dirofilaria spp. infection in European dogs has rapidly spread from historically endemic areas towards eastern and northeastern countries, but little or no information is available from these geographical regions. The present study provides a picture of filarial infections in dogs from Romania and compares two tests for the diagnosis of Dirofilaria immitis. From July 2010 to March 2011, blood samples were collected from 390 dogs from nine counties of Romania and serological SNAP tests were performed for the detection of D. immitis antigen. The remaining blood clots were subsequently used for DNA extraction followed by multiplex PCR for assessing filarioid species diversity (i.e. D. immitis, Dirofilaria repens and Acanthocheilonema reconditum). Based on molecular detection, an overall prevalence of 6.92 % (n = 27; 95 % confidence interval (CI) 4.70-10.03 %) for D. repens, 6.15 % (n = 24; 95 % CI 4.07-9.14 %) for D. immitis and 2.05 % (n = 8; 95 % CI 0.96-4.16 %) for A. reconditum was recorded, with significant variations according to sampling areas. Coinfections of D. immitis and D. repens were recorded in 23.91 % (n = 11) positive dogs. A slightly higher prevalence for D. immitis was detected at the SNAP test (n = 28, 7.17 %; 95 % CI 4.91-10.33 %), but this difference was not statistically significant (p = 0.66). However, only 53.57 % (n = 15) of antigen-positive dogs were confirmed by PCR, while other dogs (n = 9) PCR positive for D. immitis were negative at the serology. The present study shows that Dirofilaria species are endemic in the southern and southeastern areas of Romania, This article also provides, for the first time, an epidemiological picture of the distribution of A. reconditum in Romania.

Laboratory development of Dermacentor marginatus ticks (Acari: Ixodidae) at two temperatures.

The influence of two temperatures on the development of Dermacentor marginatus evolutive cycle was studied. Tests performed under controlled laboratory conditions at 21 °C, 80 % RH and 27 °C, RH 80 %, on ten fully engorged female ticks collected from naturally infested goats, in Cluj County, Romania. Hatched larvae were fed on white mice and the nymphs and adults on guinea pigs. The following parameters were evaluated: egg incubation; pre-feeding, feeding and pre-moulting for larvae and nymphs; pre-feeding, feeding, pre-oviposition and oviposition for females. Significant differences (p < 0.001) were observed in: egg incubation period (29.4 ± 1.07 at 21 °C; 5.9 ± 0.73 at 27 °C) and pre-moulting duration of larvae (18.9 ± 1.02 at 21 °C; 6.1 ± 0.58 at 27 °C) and nymphs (21.3 ± 0.87 at 21 °C; 19.9 ± 0.71 at 27 °C). The average duration of the developmental cycle was 133.9 days (range 122-154 days) at 21 °C, and 94.2 days (range 83-111 days) at 27 °C.

First report of Cercopithifilaria spp. in dogs from Eastern Europe with an overview of their geographic distribution in Europe.

Dogs in Europe may be parasitized by a variety of species of filarioids. While some species have been studied for decades, others have been only recently reported, and their range of distribution is still unknown. The present study was aimed to investigate the occurrence of Cercopithifilaria spp., whose microfilariae reside in the dermis and are transmitted by Rhipicephalus sanguineus s.l. ticks, in a dog population from Romania and to present the current knowledge of their distribution in Europe. Sampling was performed in a rural locality from Danube Delta region of Romania, location chosen due to the presence of the only known vector for these species. Skin samples were collected from 39 dogs. The samples were tested for the presence of Cercopithifilaria spp. by PCR methods. One animal (2.56%) was positive, and the amplified sequence showed a 100% similarity to Cercopithifilaria bainae. This study reports C. bainae for the first time in Eastern Europe, extending the known range of the genus in Europe.

A new canid host for subcutaneous ticks: A report from Algeria in an African golden wolf (Canis lupaster).

BACKGROUND: Ticks are hematophagous ectoparasites that feed on the skin surface of their host. Subcutaneous localizations of ticks have predominantly been associated with red foxes, documenting new cases still brings new and potentially valuable data for tackling this unusual situation. The objective of this study is to report the presence of subcutaneous ticks in a new host and on a new continent. METHODS: Between February 2022 and September 2023, 47 road-killed animal carcasses belonging mostly to carnivorous species were subjected to a comprehensive parasitological necropsy. Upon detection of subcutaneous tick, the protuberance was excised, dissected, and preserved in labeled vials containing ethanol. Identification of the subcutaneous tick was carried out at the species level. RESULTS: A single nodule of subcutaneous ticks was found in an African golden wolf (Canis lupaster) collected from Northern Algeria; no other cases of unusual tick locations were observed. Based on morphological criteria the tick was identified as Ixodes ricinus / I. inopinatus. CONCLUSION: This study represents the first documentation of subcutaneous tick in an African golden wolf (Canis lupaster), thereby broadening the host spectrum for this atypical location. Furthermore, it underscores North Africa as a new geographical area where this phenomenon occurs.

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.

Comparison of Five Serological Methods for the Detection of West Nile Virus Antibodies.

The West Nile Virus (WNV), a member of the family Flaviviridae, is an emerging mosquito-borne flavivirus causing potentially severe infections in humans and animals involving the central nervous system (CNS). Due to its emerging tendency, WNV now occurs in many areas where other flaviviruses are co-occurring. Cross-reactive antibodies with flavivirus infections or vaccination (e.g., tick-borne encephalitis virus (TBEV), Usutu virus (USUV), yellow fever virus (YFV), dengue virus (DENV), Japanese encephalitis virus (JEV)) therefore remain a major challenge in diagnosing flavivirus infections. Virus neutralization tests are considered as reference tests for the detection of specific flavivirus antibodies, but are elaborate, time-consuming and need biosafety level 3 facilities. A simple and straightforward assay for the differentiation and detection of specific WNV IgG antibodies for the routine laboratory is urgently needed. In this study, we compared two commercially available enzyme-linked immunosorbent assays (anti-IgG WNV ELISA and anti-NS1-IgG WNV), a commercially available indirect immunofluorescence assay, and a newly developed in-house ELISA for the detection of WNV-NS1-IgG antibodies. All four tests were compared to an in-house NT to determine both the sensitivity and specificity of the four test systems. None of the assays could match the specificity of the NT, although the two NS1-IgG based ELISAs were very close to the specificity of the NT at 97.3% and 94.6%. The in-house WNV-NS1-IgG ELISA had the best performance regarding sensitivity and specificity. The specificities of the ELISA assays and the indirect immunofluorescence assays could not meet the necessary specificity and/or sensitivity.

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.