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. Between 2013–2014 and 2016–2018, CDC light traps and mouth aspirators were used to collect sand flies in 132 locations from Romania, indoors and around various animal species shelters. Species identification of collected specimens was done using morphological keys, genetic tools and MALDI-TOF protein profiling. Sand flies were present in seven localities (5.3%): Eibenthal, Baia Nouă, Gura Văii (south-western Romania, Mehedinţi County); Fundătura, Pâhneşti, Epureni (eastern Romania, Vaslui County); and Schitu (southern Romania, Giurgiu County). Of the total number of collected sand flies (n = 251), 209 (83.27%) were Phlebotomus neglectus, 39 (15.53%) P. perfiliewi, 1 (0.40%) P. papatasi, 1 (0.40%) P. balcanicus and 1 (0.40%) P. sergenti (sensu lato). We confirmed the presence of five sand fly species previously recorded in Romania. However, their updated distribution differs from historical data. The diversity of sand fly species in Romania and their presence in areas with Mediterranean climatic influences constitutes a threat for the reemergence of vector-borne diseases. In the context of CanL and VL reemergence in Romania, but also due to imported cases of the diseases in both humans and dogs, updates on vector distribution are imperative.

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.

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. 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. 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. 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.

African swine fever has a significant economic, social and environmental impact due to official regulation of the disease, namely the mass depopulation of all pigs in confirmed outbreaks. The main objective of the present study was to statistically analyze the possible correlation between the number of outbreaks and infected pigs from backyard farms with the altitude, seasonal average annual humidity, and average temperature during spring, summer, autumn and winter, as well as the distance from forests, rivers, and lakes in Romania. The study included all infected backyard pigs with African swine fever virus (n = 7764) and outbreaks (n = 404) that occurred in Romania between 6 February 2020 and 2 March 2021. The number of infected pigs and ASF outbreaks were significantly higher in localities at altitudes between 100 and 500 m, average annual humidity between 60% and 80%, average spring temperature between 10 and 14 °C, average summer temperature between 16 and 22 °C, average autumn temperature between 10 and 13 °C, average winter temperatures between −1 and 2 °C, and at distances of less than 5 km from the forests, less than 15 km from lakes and less than 5 km from the rivers. The number of affected pigs decreased significantly at summer temperatures below 16 °C.

Background: An entomological study was conducted in the Canaraua Fetii Special Protection Area in the Dobrogea region, South-Eastern Romania. Four sand fly species were recorded at this location between 1968 and 1970: Phlebotomus neglectus, Ph. balcanicus, Ph. sergenti and Sergentomyia minuta. The most abundant sand fly species recorded at that time were Ph. balcanicus and Se. minuta. In the context of a countrywide study to update the sand fly species diversity, we surveyed the same area, recording also a previously unknown Ph. (Transphlebotomus) sp., for which we provide a formal description here. Methods: Sand flies were collected between July and August in 2018 and 2019 in three sites from Canaraua Fetii, Dobrogea region, Romania. The general aspect of the landscape is of a canyon (vertical, narrow walls and deep valleys). Species identification was done using both morphological and molecular analyses. Results: Out of 645 collected sand flies, 644 (99.8%) were morphologically identified as Ph. neglectus, while one female specimen (0.2%) was assigned to a previously unknown species, belonging to the subgenus Transphlebotomus. The morphological and molecular examination of this specimen showed that it is a previously unknown species which we formally describe here as Phlebotomus (Transphlebotomus) simonahalepae n. sp. Also, Ph. balcanicus, Ph. sergenti, and Se. minuta (previously recorded in this location) were not present. Conclusions: The study revealed for the first time the presence of sand flies of the subgenus Transphlebotomus in Romania. Moreover, a new species, Ph. simonahalepae n. sp., was described based on a female specimen, raising the number of species in this subgenus to six. In the investigated natural habitat, the predominant species was Ph. neglectus instead of Ph. balcanicus and Se. minuta (recorded as the predominant species in 1968–1970). Graphical