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The prevalence of gastrointestinal helminth parasites in hunting dogs from the area of foothills of Olympus Mountain was investigated through fecal examination, using a sedimentation method. From a total of 317 fecal samples collected, the following parasites were identified: Toxocara canis (10.4%), Ancylostomidae (9.8%), Dipylidium caninum (8.8%), Trichuris vulpis (4.7%), Taeniidae (1.9%), Toxascaris leonina (1.3%), Spirocerca lupi (0.6%) and Alaria alata (0.6%). This study has recorded, with respect to the age of the infected dogs, a significantly higher prevalence in young dogs infected with Toxocara canis, compared to adults, and a significantly higher prevalence in adults of the Trichuris vulpis and Dipilidium caninum infection (p<0.05). As regards gender, a significantly higher prevalence was recorded in female rather than male dogs infected with the nematode Toxocara canis (p<0.05) [ABSTRACT FROM AUTHOR]

Abstract Background Hyalomma aegyptium is a hard-tick with a typical three-host life cycle. The main hosts are Palearctic tortoises of genus Testudo. However, other hosts can be used by immature ticks for feeding in natural conditions. Given this complex ecology and multiple host use, the circulation of pathogens by H. aegyptium between various hosts can be important from epidemiological point of view. The aim of this study was to evaluate the role of H. aegyptium as natural carrier of four important zoonotic pathogens. Methods From 2008 to 2011, 448 H. aegyptium ticks were collected from 45 Spur-thighed tortoises, Testudo graeca in Romania. DNA was extracted individually from each tick using a commercial kit. DNA was examined for the presence of specific sequences of Borrelia burgdorferi s.l., Anaplasma phagocytophilum, Ehrlichia canis and Coxiella burnetii by PCR, according to previously described protocols. Results PCR analysis of H. aegyptium revealed the presence of A. phagocytophilum (18.8%), E. canis (14.1%) and C. burnetii (10%). 32.4% of the ticks were infected with at least one pathogen and 9.8% had co-infections. The stages most frequently infected were nymphs (50%) followed by males (33.9%) and females (27%). The number of tortoises which harboured infected ticks was 27/45 examined (60%). From all tested T. graeca, 40% harboured ticks infected with A. phagocytophilum, 46.7% had ticks infected with E. canis and 33.3% had ticks with C. burnetii. This study reports for the first time the presence of A. phagocytophilum and E. canis in H. aegyptium. Conclusions The presence and relatively high prevalence of three important zoonotic pathogens in H. aegyptium raises the question of their epidemiologic importance in disease ecology. As tortoises are unlikely to be reservoir hosts for A. phagocytophilum and E. canis and both these pathogens are common in H. aegyptium, this is an important indication for (1) a possible increased host-switching behaviour of these ticks to competent reservoir hosts (i.e. hedgehogs) and (2) transstadial transmission. Furthermore, if we consider also the presence of C. burnetii, we conclude that T. graeca and its ticks should be evaluated more seriously when assessing the eco-epidemiology of zoonotic diseases.

Neospora caninum is an apicomplexan (family: Sarcocystidae) protozoan parasite with a global distribution. In the N. caninum life cycle, dogs and other related canids are the definitive hosts, while other animals such as water buffaloes (Bubalus bubalis) constitute the intermediate host for this parasite. In many countries, the water buffalo is of high economic importance, providing valuable high-quality products for human needs. Although knowledge concerning the prevalence of this parasite in intermediate animal host populations is crucial, data from water buffalo are scarce. Keeping this in mind, the aim of this study was to examine the presence and assess the prevalence rates of N. caninum infection in water buffaloes in Northern Greece, where this animal husbandry sector started to raise, as well as to determine associated risk factors, with the application of a commercially available Neospora ISCOM ELISA test kit, developed for the detection of antibodies against N. caninum in milk samples The study was conducted during January-June 2023 in a total of 172 individual raw milk samples, collected from dairy buffaloes, reared under a semi extensive system, in three different farms. Information on the status of N. caninum infection in buffaloes from Greece was so far unknown with a lack of epidemiological or risk factors associated. For the detection of N. caninum, the commercially available Neospora ISCOM ELISA test kit (SANOVIR ®, Svanova Biotech AB, Uppsala, Sweden) was utilized. Results demonstrated the presence of N. caninum in water buffaloes from Greece for the first time. All positive N. caninum animal were asymptomatic, with absence of any disease signs. The overall prevalence of infection was 22.10%, whereas the main risk factors include the presence of dogs as well as the low biosecurity measures., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this study., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Multi-host pathogens that infect various animal species and humans are considered of great importance for public and animal health. Leishmania spp. parasites are a characteristic example of such pathogens. Although leishmaniosis in humans is endemic for about 100 countries around the world it is classified as a neglected tropical disease. There are three main forms of leishmaniosis in humans: cutaneous (CL), visceral (VL) and mucocutaneous leishmaniosis (MCL). Each year, about 30,000 new cases of VL and more than 1 million new cases of CL are recorded. In Europe L. infantum is the dominant species with dogs being reservoir hosts. Apart from dogs, infection has been recorded in various animals, which suggests that other species could play a role in the maintenance of the parasite in nature. Herein we provide an in-depth review of the literature with respect to studies that deal with Leishmania infantum infections in domestic and wild animal species in Europe. Given the fact that domesticated and wild animals could contribute to the incidences of leishmaniosis in humans, the aim of this paper is to provide a comprehensive review which could potentially be used for the development of measures when it comes to the control of the Leishmania infantum parasite., (© 2024. The Author(s).)

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., (© 2022. The Author(s).)

Remote sensing of inland waters is challenging, but also important, due to the need to monitor the ever-increasing harmful algal blooms (HABs), which have serious effects on water quality. The Ocean and Land Color Instrument (OLCI) of the Sentinel-3 satellites program is capable of providing images for the monitoring of such waters. Atmospheric correction is a necessary process in order to retrieve the desired surface-leaving radiance signal and several atmospheric correction methods have been developed through the years. However, many of these correction methods require programming language skills, or function as commercial software plugins, limiting their possibility of use by end users. Accordingly, in this study, the free SNAP software provided by the European Space Agency (ESA) was used to evaluate the possible differences between a partial atmospheric correction method accounting for Rayleigh scattering and a full atmospheric correction method (iCOR), applied on Sentinel-3 OLCI images of a shallow, highly eutrophic water reservoir. For the complete evaluation of the two methods, in addition to the comparison of the band reflectance values, chlorophyll (CHL) and cyanobacteria (CI) indices were also calculated and their values were intercompared. The results showed, that although the absolute values between the two correction methods did not coincide, there was a very good correlation between the two methods for both bands' reflectance ( r  > 0.73) and the CHL and CI indices values ( r  > 0.95). Therefore, since iCOR correction image processing time is 25 times longer than Rayleigh correction, it is proposed that the Rayleigh partial correction method may be alternatively used for seasonal water monitoring, especially in cases of long time-series, enhancing time and resources use efficiency. Further comparisons of the two methods in other inland water bodies and evaluation with in situ chlorophyll and cyanobacteria measurements will enhance the applicability of the methodology., Competing Interests: Konstantinos A. Kormas is an Academic Editor for PeerJ., (©2022 Katsoulis-Dimitriou et al.)

Parasitic diseases cause significant economic losses in swine, including free-range swine farms, the number of which in Romania has increased in the last decades. The current study aimed to identify the parasitic profile of swine raised on two free-range (low-input) farms from Transylvania. Nine hundred sixty samples collected from weaners, fatteners, and sows were investigated by flotation, centrifugal sedimentation, modified Ziehl-Neelsen stained fecal smear, modified Blagg technique, and oocyst/egg cultures. The number of oocysts (OPG), cysts (CPG), and eggs (EPG) were counted per gram of fecal matter. The examination revealed parasitic infections with Balantidium coli, Eimeria spp., Ascaris suum, Trichuris suis, Oesophagostomum spp., Strongyloides ransomi and Cryptosporidium spp. Prevalence (P) and the mean intensity (MI) of the infections varied according to age, swine category, farm, and season. The overall prevalence in both free-range farms according to the age category was 63.2%—Eimeria spp., 70.31%—B. coli, 9.38%—Oesophagostomum spp., 3.75% S. ransomi, and 18.12% Cryptosporidium spp. in weaners. In fatteners Eimeria spp. revealed a prevalence of 50.93%, B. coli—72.5 %, A. suum—63.13%, T. suis—39.06%, and in sows Eimeria spp.—39.06%, B. coli—62.19%, A. suum—34.06%, Oesophagostomum spp.—27.19%, S. ransomi—1.56% and Cryptosporidium spp.—9.38%. The study revealed statistically significant (p < 0.05) differences between age groups, seasons, and farms for all diagnosed parasites. Further research is required to better understand the epidemiology of these infections in swine from Transylvania.

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 10 5 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.

Nematodes of genus Trichinella are wide-spread zoonotic parasites, able to infect a large variety of vertebrates. Animal hosts are usually regarded as asymptomatic carriers. However, there is little data regarding the functional consequences that T. spiralis infection renders on muscle cells. The aim of the present study was to assess the effect of T. spiralis on the effort capacity of experimentally infected mice. Overall, 60 mice, divided into three groups were used: M (uninfected), L200 and L1000, infected with 200 or 1000 larvae/mouse respectively. The mice were periodically weighed and their effort capacity was evaluated (days 0, 7, 15, 35 and 60). From each group, two randomly selected mice were euthanized after evaluation carcasses were artificially digested in order to establish the number of larvae per gram (LPG). On day 0, there were no significant differences among groups. Starting with day 7, the effort capacity of infected groups decreased, with significant differences between group M and the infected groups. From day 15, the differences between the infected groups also became significant. The LPG gradually increased and the differences between groups were always significant. A strong correlation between the LPG and decreased effort capacity was noted. The present study demonstrates the reduction of muscular capacity in mice experimentally infected with Trichinella spiralis, in correlation with the infective dose, providing new insights in this parasite's transmission strategy., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Thelazia callipaeda (Spirurida, Thelaziidae) is a vector-borne zoonotic eye worm with a broad host spectrum. In Europe, it is an emerging threat, having greatly expanded its geographical distribution during the past two decades. In Romania, T. callipaeda has been previously reported in domestic and wild canids and felids. The aim of the present study was to assess the occurrence of T. callipaeda in mustelids in the country., Methods: Between March 2015 and April 2019, 77 road-killed mustelids (3 pine martens, Martes martes; 6 European polecats, Mustela putorius; 13 beech martens, Martes foina; and 55 European badgers, Meles meles) were examined by necropsy. If present, all ocular nematodes were collected and stored in absolute ethanol, for subsequent morphological and molecular identification., Results: Two animals were found to be infected with T. callipaeda: one European badger and one beech marten. The molecular analysis revealed a 100% nucleotide similarity to T. callipaeda haplotype h1 for all the sequenced specimens., Conclusions: To our knowledge, the present study demonstrates for the first time the occurrence of T. callipaeda in mustelids from Romania, records the easternmost locality of the parasite in Europe, and represents the first report of T. callipaeda in the European badger, Meles meles, extending the known host range for this parasite in Europe.

Canine granulocytic anaplasmosis (CGA) is an important tick-borne disease with worldwide distribution. The importance of this disease resides in the ability of Anaplasma phagocytophilum to infect humans and several animal species. The aim of the study was to evaluate the prevalence rate of CGA in different altitudinal areas of Romania. A total of 357 canine blood samples were collected during 2010-2013 from eight counties. To assess the influence of the altitude on A. phagocytophilum prevalence, the samples were collected from four different altitude areas (coastal 0-5 meters; lowland 6-100 meters; hilly areas 200-300 meters; low mountain areas >500 meters). These samples were evaluated for the presence of A. phagocytophilum DNA by amplifying part of the Ankyrin repeat protein (AnkA) gene. A higher prevalence was obtained for coastal compared with remaining areas, suggesting an influence of altitude on the CGA. Moreover, the results suggest an influence of climate and rainfall. In the present research work, we highlight the risk of granulocytic anaplasmosis in Central and Southern Romania, with a greater risk associated to Southern lowland region, especially in coastal areas. The importance of these results resides in the zoonotic potential of the canine A. phagocytophilum strains. In conclusion, the altitude and precipitation level may be risk factors for A. phagocytophilum infection in dogs and other hosts, including humans.

Introduction: Toxoplasmosis is a worldwide reported zoonotic infection caused by the protozoon Toxoplasma gondii. Pigs may become infected by ingesting feed or water contaminated with cat faeces, by cannibalism, and/or by eating infected rodents. T. gondii infected edible tissues of pigs are a source of infection for humans. This study was undertaken to detect seropositivity of pigs against T. gondii and identify possible risk factors as well as to compare two serological methods., Methods: A total of 609 blood samples were collected from 10% of the sows from 65 pig farms located in mainland Greece. Information about the geographical location, size, and biosecurity level of the herd was recorded. Samples were tested for the presence of antibodies against T. gondii employing the indirect fluorescent antibody test and the enzyme-linked immunosorbent assay., Results: Twenty-six positive samples were detected (4.3%) in 17 out of the 65 farms tested (26.2%). The average seroprevalence in affected herds was 26.8% (95% confidence interval 21.0-32.6%). Among the categorical data evaluated, the biosecurity level (odds ratio 0.17, p < 0.01) and the geographical characteristics (odds ratio 13.55, p < 0.05) significantly affected the presence of toxoplasmosis in the herd. Toxoplasmosis was recorded in significantly higher percentages in mountainous farms as compared to lowlands (p < 0.05) and in those with poor biosecurity levels than in those with very good (p < 0.05). A very good agreement (κ = 0.958) was observed between the two serological methods., Conclusion: The presence of antibodies in pigs is indirect information on the risk of the infection and an indication of the necessity of biosecurity measures to be taken in order to control infection at least in the areas at highest risk.

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.

In June 2005, a free-ranging grass snake, Natrix natrix infected with Rhabdias fuscovenosa was collected for histologic examination of the lungs. Gross lesions were not noted; however, histologic examination revealed vacuolar degeneration of the respiratory epithelium, hemorrhage, smooth muscle degeneration, faveolae necrosis and obstruction, and intralesional nematodes. Eosinophils were the most common inflammatory cell observed in the areas of necrosis and interstitium. Herein we describe histopathologic changes due to R. fuscovenosa in a free-ranging grass snake.

From May 2007 to May 2008 we have examined by otoscopy a total number of 214 cats, aged between 0 and 6 months, brought in for their first veterinary examination to a private veterinary clinic. All cats were of urban origin. In all positive cats we performed a washing of the ear using warm paraffin oil in order to determine intensity of infection. Statistical analysis was performed using the Chi-square test. A total number of 30 cats were found to be infected with Otodectes cynotis (prevalence 14.02%). Prevalence of infection was significantly (p<0.05) higher in cats aged between 3 and 6 months (17.58%) than in cats aged below 3 months (11.38). Intensity of infection ranged between 7 and 85 (mean intensity 35.60) mites per infected cat. The same age group of cats between 3 and 6 months had significantly (p<0.05) higher mean intensity (47.19) compared to cats aged under 3 months (22.36). No statistical significance was found between males and females for neither prevalence nor intensity. Differences and similarities with other studies are discussed.