Background Ticks are increasingly acknowledged as significant vectors for a wide array of pathogens in urban environments with reports of abundant tick populations in recreational areas. The study aims to contribute to a better knowledge of the abiotic and biotic factors which impact the ecology of hard ticks in urban and peri-urban habitats in Romania. Methods Questing ticks were collected by flagging in seven recreational locations, from four types of habitats in Cluj-Napoca, Romania: parks; gardens; a cemetery; and peri-urban forests. Hedgehogs, birds and micromammals were also sampled and searched for ticks, using standard methods (i.e. torch-based searches, ornithological mist nets, snap-traps, etc.), while vegetation was evaluated on surveyed areas. Data on questing ticks were converted to abundance indices. Moodʼs median tests were used to assess the relationship between the abiotic and biotic factors and the abundance of questing ticks. Results Two species of questing ticks were found: Ixodes ricinus (96.8%) and Haemaphysalis punctata (3.2%). Ixodes ricinus was also the predominant engorged tick collected from urban wildlife. For I. ricinus the highest mean total abundance index/location (total no. of ticks/100 m2) was recorded in the urban gardens (3.79, 95% CI: ± 1.59) and parks (2.68, 95% CI: ± 0.75), whereas the lowest mean total abundance index was noted in the peri-urban forests (0.06, 95% CI: ± 0.03) and the urban cemetery (0.04, 95% CI: ± 0.02). The adults and nymphs of I. ricinus displayed a bimodal activity pattern, while the larvae showed a unimodal questing behaviour with an autumnal peak. Positive correlations were found between the mean total abundance index and the rise in the daily mean temperature and relative humidity, and between the global abundance of questing ticks and the presence of hedgehogs in the respective locations (P < 0.01). Conclusions Ticks were collected in all the recreational sites surveyed in Cluj-Napoca. Ixodes ricinus was the dominant questing and engorged tick species. Several abiotic and biotic factors shape the ecology of ticks in Cluj-Napoca city, with climate and the local presence of suitable hosts being the most important.

Despite the increasingly recognized eco-epidemiological importance of ticks as vectors for numerous zoonotic pathogens in urban areas, data regarding the pathogen diversity and co-infection rates in ticks and wildlife hosts in urban and peri-urban Romania are scanty. We aimed to establish the risk of human exposure to co-infected ticks in Cluj-Napoca, a major city in Romania. DNA was isolated from 151 questing ticks: Ixodes ricinus (n = 95), Haemaphysalis punctata (n = 53), Dermacentor reticulatus (n = 2), and Dermacentor marginatus (n = 1); 222 engorged ticks: I. ricinus (n = 164), I. hexagonus (n = 36), H. punctata (n = 16), H. concinna (n = 6), and 70 tissue samples collected from wildlife hosts during 2018 in five urban, and two peri-urban sites. Using a pre-designed Fluidigm real-time PCR dynamic array, all DNA samples were individually screened for the presence of 44 vector-borne pathogens. Subsequently, conventional PCRs were performed for a selection of samples to allow validation and sequencing. In total, 15 pathogens were identified to species and 6 to genus level. In questing ticks, single infections were more common than co-infections. Seven Borrelia spp. were detected in questing I. ricinus, and three in H. punctata ticks. An overall high prevalence 26.35% (95% CI: 19.46–34.22) and diversity of Borrelia burgdorferi sensu lato was seen in urban questing ticks. Other pathogens of the order Rickettsiales were present with variable prevalence. Co-infections occurred in 27.4% (95% CI: 18.72-37.48) of all infected questing ticks. In engorged ticks the overall Bo. burgdorferi sensu lato prevalence was 35.6% (95% CI: 29.29–42.27), with five species present. Pathogens of the order Rickettsiales were also frequently detected. We report for the first time in Romania the presence of Rickettsia aeschlimannii and Rickettsia felis. Overall, from the infected engorged ticks, 69.2% showcased co-infections. In Ixodes spp., dual co-infections, namely Borrelia spp. and Anaplasma phagocytophilum, and Rickettsia helvetica and A. phagocytophilum were the most prevalent. Given the outcome, we underline the need to establish proper tick-surveillance programs in cities and include co-infections in the management plan of tick-borne diseases in Romania.

Introduction: West Nile virus (WNV), Usutu virus (USUV), and the tick-borne encephalitis virus (TBEV) are all arboviruses belonging to Flaviviridae family. All are characterized by vectorial transmission and sometimes associated with neuroinvasive infections. The circulation of these viruses is considered endemic in parts of Europe, with human cases reported in many countries. Among hosts, the viruses are vectored by hematophagous arthropods, such as mosquitoes (WNV, USUV) and ticks (TBEV). Considering the currently outdated knowledge regarding the epidemiology of these viruses in Romania, the aim of our study was to assess the seroprevalence rates of WNV, USUV, and TBEV among healthy blood donors in north-western Romania. Methods: Human blood samples from healthy donors were collected between November 2019 and February 2020 in six counties from the north-western region of Romania. The samples were serologically tested by ELISA and serum neutralization test. Results: Overall, we obtained a seroprevalence of 3.17% for WNV, 0.08% for TBEV, and 0% for USUV. Conclusion: Despite the low seroprevalence of WNV, USUV, and TBEV in our study, we highlight the need for continuous nationwide vector and disease surveillance and implementation of control measures. Further research is required for an optimal overview of the epidemiological status of the Romanian population regarding these flaviviruses together with countrywide awareness campaigns.

Nematodes of the genus Trichinella are among the most widespread parasites of domestic and wild omnivores and predatory animals. The present study aimed to evaluate the antiparasitic effect of Lactobacillus casei ATCC 393 (original) and L. paracasei CNCM in CD-1 mice experimentally infected with Trichinella britovi. Four groups of 20 mice (10 females and 10 males/group) were used, with two control (C) groups and two experimental (E) groups, in which each animal received a daily oral dose of 100 µl of 105 CFU/ml probiotics in Ringer’s solution. On day 7, all mice (except the negative control group) were infected orally with Trichinella (100 larvae/animal) as well as the two probiotics. On day 9 post-infection (p.i.), 10 mice/group were euthanized, and the presence of adult parasites in the intestinal content and wall was tested. On day 32 p.i., 10 mice/group were euthanized, then trichinoscopy and artificial digestion were performed to assess the muscle infection with T. britovi. On day 9 p.i., the experimental group pretreated with L. casei ATCC 393 (6.3 ± 3.03) showed a significantly lower number of adult parasites in the intestinal wall compared with the positive control group (24.6 ± 4.78). Additionally, a significantly lower adult parasite count in the intestinal wall was registered in female mice pretreated with L. paracasei CNCM (7.4 ± 4.71) compared to female mice from the positive control (29.0 ± 5.17). No statistically relevant results were obtained concerning the male mice or the data obtained at 32 days p.i., irrespective of mice gender.

Background Gaining insight into the risk perceptions and the knowledge evolution of the public about emerging or changing health risks is vital for the improvement of health promotion activities. Currently, scientific evidence regarding the attitudes of the Romanian public towards ticks is scanty. This study aimed to identify how the lockdown enforced by the COVID-19 pandemic in Romania impacted the recreational behaviour, risk perceptions, and protective practices of the Romanian population regarding ticks. Methods A cross-sectional, nationwide web-based questionnaire was designed and distributed via social media to evaluate if, and how, the COVID-19 lockdown impacted the behaviour of the Romanian public concerning ticks. The survey was available online from 6 May until 15 May 2020, which marked the last day of the travel ban in Romania. The collected data were processed by applying both uni- and multivariate methods. Results Respondents reported a higher frequency of finding ticks on themselves and their dogs during the lockdown. Bathing/showering and checking the body for ticks were the two most used protective behaviours both before and during the lockdown. Nevertheless, an overall lower usage rate of protective measures was registered during the lockdown. Almost all dog owners used a form of ectoparasite control for their dogs, and only three stopped due to lockdown-associated reasons. Respondent characteristics that were found to be positively associated with risk perceptions were being female and living in peri-urban/suburban/rural environments. Conclusions Despite spending less time outdoors during the lockdown, more respondents reported finding ticks on themselves or their dogs. Changes in the preferences for recreational locations, rates of protective practices usage, amount of time spent in specific areas, or tick seasonal activity might have contributed to this outcome. Concerning risk groups, men of all ages, senior citizens, and rural inhabitants should be targeted by the relevant Romanian authorities when promoting local or nationwide tick awareness campaigns. Graphical abstract