Background: Phlebotomine sand flies (Diptera: Psychodidae) are haematophagous insects that transmit the protozoan parasite Leishmania infantum (Kinetoplastida: Trypanosomatidae), the main causative agent of both zoonotic visceral leishmaniasis (VL) and canine leishmaniasis (CanL) in the Mediterranean basin. Eight species of sand flies have been previously recorded in Romania: Phlebotomus papatasi, Phlebotomus alexandri, Phlebotomus sergenti, Phlebotomus perfiliewi, Phlebotomus neglectus, Phlebotomus longiductus, Phlebotomus balcanicus and Sergentomyia minuta. Three of them (P. perfiliewi, P. neglectus and P. balcanicus) were incriminated as vectors of L. infantum. Recent reports of autochthonous CanL in Romania require updates on sand fly distribution and diversity in this country.

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BackgroundCanine leishmaniasis is a severe, potentially life-threatening, systemic vector-borne disease of dogs caused by protozoan parasites of the genus Leishmania. Romania has been traditionally regarded as a non-endemic country for leishmaniasis with sporadic human disease cases. However, the recent report of an autochthonous canine leishmaniasis case (the first in the last 80 years) suggested the presence of an infection focus in the area of Râmnicu Vâlcea. The present study describes a survey of canine leishmaniasis in this geographical area with comparison to a georeferenced dataset of sand fly distribution based on historical literature records.MethodsThe study was carried out in Râmnicu Vâlcea and included samples (serum, blood and conjunctival swabs) collected from 80 dogs including client-owned dogs from two local practices and dogs from two public shelters. Serum anti-leishmanial antibodies were assessed by ELISA. All blood and conjunctival samples were assessed by real-time quantitative PCR, targeting the leishmanial kinetoplast minicircle DNA.ResultsThree dogs (3.7 %) were seropositive and another four (5.0 %) showed borderline results indicative of exposure or infection. TaqMan PCR was performed for all dogs, on both blood and conjunctival swabs. Seven dogs (8.7 %) were positive by conjunctival swab PCR and one dog (1.2 %) by blood PCR. None of the positive dogs presented clinical signs compatible with canine leishmaniasis.ConclusionsThis is the first study evaluating canine leishmaniasis in a dog population in Romania by both highly sensitive PCR and serology. Although the prevalence was relatively low compared to other endemic regions, our results clearly demonstrate the presence of a canine leishmaniasis focus in Romania.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1583-5) contains supplementary material, which is available to authorized users.

Background Angiostrongylus vasorum is a metastrongyloid nematode localized in the right heart and the pulmonary arteries of domestic dogs. The number of reports in Europe has recently increased, presumably as a consequence of a growing awareness among clinicians, animal owners and researchers, but also due to a growing incidence and territorial spread. So far, no studies have been conducted to assess the prevalence and distribution of A. vasorum in domestic dogs in Romania, and the awareness among veterinarians is limited or absent. The aim of the present study was to evaluate the countrywide seroprevalence of circulating antigens of A. vasorum and specific antibodies against A. vasorum in domestic dogs from Romania. Methods Between November 2016 and July 2017, blood was sampled from a total of 1545 domestic dogs from 23 counties of Romania. Details about their gender, age, breed, housing, use and origin were collected. All serum samples were tested for the presence of A. vasorum circulating antigens (AG) using monoclonal and polyclonal antibodies in a sandwich ELISA. Additionally, a sandwich ELISA using A. vasorum adult somatic antigen purified by monoclonal antibodies was used for specific antibody (AB) detection. Results A total of 33 dogs (2.14%, 95% CI: 1.82–3.56%) were seropositive for A. vasorum antigen or antibodies against the parasite. Three dogs were positive for antigen only (0.19%, 95% CI: 0.07–0.57%) and 30 dogs (1.94%, 95% CI: 1.36–2.76%) were positive for antibodies only. No dog was positive for both tests. The overall prevalence (AB or AG) and the AB prevalence were significantly higher in pure breed dogs compared to mixed breeds and mongrel dogs ( P < 0.05) and in shepherd dogs compared to other groups ( P < 0.05). There was no significant difference between males and females, between urban and rural dogs, between dogs with unrestricted access and with restricted access to the environment, and between dogs living outdoors and indoors. Conclusions Our data suggest that the disease is present in Romania in dogs, as it was previously demonstrated in foxes. However, so far, no clinical case has been reported in the country and this may be related to a low awareness among vets. Electronic supplementary material The online version of this article (10.1186/s13071-019-3481-0) contains supplementary material, which is available to authorized users.

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Background A study conducted at the International Airport of Cluj-Napoca, Romania, with the aim of investigating the presence/absence of invasive Aedes mosquito species resulted in finding Aedes japonicus japonicus (Theobald 1901) eggs in one of the ovitraps placed on site. Methods The study was carried out between 30 June and 29 September 2020. On 24 August, 26 eggs were collected and later hatched at the University of Agricultural Sciences and Veterinary Medicine of Cluj-Napoca’s insectary. On 15 October another adult female Ae. japonicus was caught entering a building in the center of the city, about 7 km from the first sampling spot. Results The mosquitoes were identified morphologically and confirmed by molecular analysis, based on the genetic analysis of the mitochondrial gene cytochrome c oxidase subunit 1 (COI). Conclusion This is the first report of the species in Romania, highlighting the need for surveillance and implemented control methods. However, in Romania to our knowledge only Aedes albopictus has been established; further studies are required to learn about this new invasive species' status in Romania.

Canine vector-borne diseases are caused by pathogens transmitted by arthropods including ticks, mosquitoes and sand flies. Many canine vector-borne diseases are of zoonotic importance. This study aimed to assess the prevalence of vector-borne infections caused by Dirofilaria immitis , Ehrlichia canis , Borrelia burgdorferi sensu lato, Anaplasma spp. and Leishmania infantum in a dog kennel in Argeș County, Romania. Dog kennels are shelters for stray dogs with no officially registered owners that are gathered to be neutered and/or boarded for national/international adoptions by various public or private organizations. The international dog adoptions might represent a risk in the transmission of pathogens into new regions. In this context, a total number of 149 blood samples and 149 conjunctival swabs from asymptomatic kennel dogs were assessed using serology and quantitative real-time polymerase chain reaction. Antibodies against B. burgdorferi s.l. were detected in one dog (0.6%), anti- Anaplasma antibodies were found in five dogs (3.3%), while ten dogs (6.7%) tested positive for D. immitis antigen. Overall, 20.1% (30/149) of dogs were positive for L. infantum DNA. All samples were seronegative for anti- Leishmania antibodies. When adopting dogs from this region of Romania, owners should be aware of possible infection with especially L. infantum . The travel of infected dogs may introduce the infection to areas where leishmaniasis is not present.

Background Equine thelaziosis is a neglected vector-borne parasitic disease in modern veterinary medicine, lacking recent reports. It is transmitted by Musca autumnalis, and potentially other Muscidae species, by ingesting the lachrymal secretions of its equine host. The distribution of both Thelazia lacrymalis and its intermediate hosts remains largely unknown throughout Europe, with most studies dating back 20 years. The aim of this study was to assess the presence, prevalence and distribution of T. lacrymalis in horses from Romania. Methods The eyes of 273 horses, slaughtered at two abattoirs from the Northwestern and Western regions of Romania, were examined for the presence of T. lacrymalis between March and November 2021. Upon detection, the nematodes were collected and morphologically identified using the keys from literature. Following identification, one specimen from each animal was selected for molecular analysis while the rest underwent detailed morphometric measurements. Mapping and distribution, according to ecoregions, was done using the QGis 3.20 software, while sequences obtained were compared to those available in GenBank through BLAST analysis using the MEGA X software. Results Of the 273 animals sampled, 12 (4.39%) were positive for Thelazia spp. infection. Eighty-seven nematodes were recovered, all morphologically identified as T. lacrymalis. The intensity of infestation varied between one and 33 nematodes/animal while five animals presented a bilateral infestation and seven a unilateral one. The highest prevalence was encountered in Pannonian ecoregion (12.12%) while the lowest was in the Alpine ecoregion (0%). Seventy-five intact specimens underwent detailed morphometric analysis, of the 18–20 parameters, resulting in notable differences in striation lengths compared to the data available in other reports. BLAST analysis identified a 96.46–98.60% similarity to the only other COI gene sequence available for T. lacrymalis. Conclusions The current study represents the first report of T. lacrymalis in horses in Romania. The low prevalence rates are probably linked to the wide use of macrocyclic lactones. Graphic Abstract