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Introduction: Gastrointestinal parasitic infestations are one of the major health concerns worldwide; hence, this study aimed to investigate the efficacy of a doramectin-based anti-parasitic drug. Methods: To achieve this goal, we collected fresh faecal samples (50-100 g) from 20 sheep and 20 cattle, which were subsequently examined through coproparasitic methods. Results: Pre-therapeutic coproscopic screening of samples from sheep and cattle led to the identification of digestive strongyles, Eimeria spp., Balantidium coli, Cestodes, and Toxocara spp. The prevalence rates for sheep were 100%, 35%, 35%, 30%, and 15%, respectively, and 80%, 30%, 30%, 10%, and 10% for cattle. Conclusion: The study led to the identification of parasites belonging to three different species: Sporozoa, Nematoda, and Cestoda. None of the investigated helminths proved resistant to Dectomax®, meaning that the average efficacy rate of Dectomax® in sheep and cattle naturally infected with digestive strongyles, Eimeria spp. or Toxocara spp. was 94% and 95%, respectively.

In the last few years, atopic dermatitis (AD) has become widespread both in humans and dogs in Romania. This skin disease impairs the quality of life (QoL) of affected patients. AD is usually evaluated using clinical approach and pruritus score. Several studies have been done recently to establish disease-specific questionnaires in order to assess both the QoL and health-related quality of life (HRQoL) in affected individuals (children with AD and their parents, owners of atopic dogs and their pets). In this respect, 100 questionnaires aiming to evaluate QoL and HRQoL both in owners and their dogs, affected by atopic dermatitis, were drafted and handed out to patients in Timișoara, Romania. A total of 54 (54%) valid questionnaires were received from owners. Answers were statistically analyzed to reveal the relationship between the owner and his/her pet. In this study, this relationship was not influenced by the age of the owner, and the higher the severity of the disease, the more severely the QoL is affected in both humans and dogs. Euthanasia is not an option for pet owners from Timișoara, and the costs of establishing and applying the treatment appear to be non-restrictive, thus disclosing a very close pet-owner relationship. This is the first study of this kind conducted in Romania.

Equine piroplasmosis (EP) is an endemic tick-borne disease found in most countries around the world. It affects all species of Equidae, and it is caused by

The anthropic impact, having various forms, negatively changes the parameters of water quality and disturbs the living environment of benthic macroinvertebrates. The purpose of the present study is to present how the main sources of pollution influence the way of life of the benthic macroinvertebrates community. 20 quantitative benthic samples were collected from different benthic areas of the Bega River water during from June 2014 to February 2017. In addition to identify all the changes that occur in the structure of macroinvertebrates community due to the activities performed by the human population. For the study to lead to conclusive results, samples were collected upstream and downstream from the main sources of pollution. Macroinvertebrates identified the macroinvertebrates and performed tests to determine the density, abundance and frequency of the samples. Based on these data, it is observed that, the presence of benthic macroinvertebrates that are sensitive to pollutants and higher-quality water in the upstream segment waters sector compared to the waters of the central segment where we identified benthic macroinvertebrates that prefer a polluted living environment. In the downstream segment, we could see that the river has a better quality, because naturally flowing on a long distance, allows an improvement in water quality through the self-purification process emphasized by the occurrence of some sensitive groups to pollution.

The lockdown period in Romania lasted for 60 days and had the purpose of limiting the spread of SARS-CoV-2 virus outbreak and manage its consequences through emergency measures on many important areas of activity. This study aimed to gather, assess, analyze and disseminate relevant social, economic, and medical aspects on the impact of COVID-19 pandemic during lockdown on the veterinary profession in Romania. A survey was created using an online questionnaire platform, and disseminated. The survey was completed by a total of 409 individuals. A high number of respondents (71.64%; n = 293) felt exposed to medium or high risk of infection with SARS-CoV-2 at their workplace and many (56.97%; n = 233) felt that their professional environment was more stressful than usual during lockdown. Almost all respondents (89.73%; n = 367) declared the implementation of several control measures recommended by FECAVA and FVE (e.g., social distancing, wearing protective equipment, hand washing), but few mentioned the opportunity of remote work or visiting restrictions. Overall, the results show that the impact of lockdown lies directly on four main categories of importance on veterinarian professional's life—human resource, activity management, relationship between veterinarian and authorities, and continuing education.

Cat demodicosis is uncommon to rare, and is caused by Demodex cati, Demodex gatoi and another unnamed species. The investigated patient was a mix-breed, 10-year-old feline with no dermatological history. Alopecia, erythema, minor erosions and ulcerations and crusts, associated with pruritus and self-trauma, were observed on the head. Dark, agglutinated cerumen was also present in the external ear canal. The agent causing the skin condition in the feline patient was identified as being a Demodex genus mite, based on the specific, morphological characteristics noticed upon the microscopic examination of deep skin scrapes. Biological samples were collected from the patient with to perform a PCR assay for clear species-determination and morphological assessment. PCR amplification of DNA extracted from the Demodex mites produced a single band of ~330 bp, indicating the presence of the D. cati species. The acaricidal treatment consisted of topical treatment using a fluralaner and moxidectin-based spot-on. Upon follow-up appointments, scheduled three times at a monthly interval, the patient failed to provide a positive result upon deep skin scrapes. The negative scrapes were also accompanied by the complete resolution of the existing lesions. In conclusion, this is the first molecular study to highlight the presence of Demodex cati within the feline population of Romania, and the fluralaner-moxidectin spot-on therapy has led to a complete recovery of the feline patient affected by feline demodicosis.

Dirofilaria infections in dogs are recognized as widespread mosquito-borne diseases with zoonotic potential, that are caused by the filarioid nematode (Onchocercidae) species Dirofilaria immitis and Dirofilaria repens. The long-term survey conducted in western and south-western Romania was undertaken in order to provide valuable data on the occurrence of Dirofilaria infections in dogs. Blood samples from 1088 dogs, originating from 73 localities of 11 western and south-western counties in Romania, were collected and examined using the modified Knott test. Subsequently, all of the microscopically positive samples were subjected to molecular analysis for confirmation. Altogether, the data obtained showed the percentage of dogs with circulating microfilariae to be 21.42% (233/1088) of dogs tested in the investigated region. The identified species, in cases of monoinfections, were D. immitis, D. repens, and Acanthocheilonema reconditum in 106 (9.74%) samples, 102 (9.38%) samples, and 1 (0.09%) sample, respectively. Twenty-four (2.21%) samples were simultaneously positive for D. immitis and D. repens. There was no association (p &gt, 0.05) between infection status and breed, however, sex and lifestyle were positively associated (p &lt, 0.05) with the percentage of dogs with circulating microfilariae and might be regarded as risk factors for infection. The results of the present investigation indicate potential zoonotic risks for humans living in the screened area and support the imperative to increase awareness among both veterinarians and physicians, regarding the continuous spread of these zoonotic filariae.

Species of the genus Sarcocystis are recognized as protozoan parasites infecting a wide range of animals, including humans. This study aimed to provide data on the occurrence, genetic characterization, and epidemiological significance of Sarcocystis spp. in cattle destined for human consumption in Romania. A total of 117 heart samples from slaughtered cattle in three southwestern Romanian counties (Dolj, Timiș, and Gorj) were analyzed in order to detect sarcocysts, using fresh examination microscopic techniques. Subsequently, the isolated sarcocysts and/or cyst fragments (5–15 per sample) from each infected animal were molecularly characterized. Overall, 17.9% (21/117) of the tested animals were found to be Sarcocystis spp. positive by microscopy. Genetic characterization of Sarcocystis spp. isolates, based on sequence analysis of the 18S rRNA gene, showed the presence of a single species, namely S. cruzi. No correlation was found (p>0.05) between S. cruzi infection and the origin, age, breed, and gender of cattle, but the grazing farming system was positively associated (p=0.031) with the pathogen prevalence and can be considered a risk factor (OR = 3.6) in acquiring infection. To evaluate the possible public health risk, further investigation focused on the processing of other Sarcocystis-specific tissue matrices and evidence of human infections is recommended. This is the first study of bovine Sarcocystis infection in Romania.

Until now, in Romania, a country in southeastern Europe located on the Balkans Peninsula, information regarding the status on A. mellifera honey bee infection by Nosema spp. has not been published. The aims of the present study were to survey the occurrence and identification of Nosema spp. in A. mellifera honeybee colonies from 37 apiaries located in Arad, Caras-Severin and Timis Counties, Romania. Also, based on published literature an update on the distribution of N. ceranae infection among A. mellifera colonies worldwide was performed. Overall, a frequency of 55.1% (389/690) of Nosema infection was observed in the analyzed regions (ranging from 52% to 66%), by light microscopy. By PCR and DNA sequencing, N. ceranae was the only microsporidia identified. Nosema ceranae is clearly a novel, emergent pathogen of A.mellifera with potentially very serious effects on the individual and honeybee�s colonies in Romania. Data obtained provide new and important information on N. ceranae geographic prevalence and distribution, and on its impact at colony level and/or its role in colony losses. The present study intends to contribute to highlight the importance of implementing prevention, treatment and control measures of honeybee nosemosis, in Romania.

Background The hemotropic mycoplasmas (hemoplasmas) of the genus Mycoplasma are recognized as important bacteria that parasitize red blood cells, causing hemolytic anemia in many mammalian species, including cats. No information is available concerning the presence of feline hemoplasma infections in cats in Romania. Thus, the objective of the present study was to provide data on the occurrence and molecular characterization of hemotropic mycoplasmas in client-owned cats in Romania. Methods Blood samples from 51 unhealthy cats, originating from Timişoara Municipality, Romania, were screened for the presence of hemoplasmas using conventional polymerase chain reaction (PCR) targeting the 16S rRNA gene and sequencing assays. PCR-positive samples were subsequently analyzed by phylogenetic and population genetic analysis. Results Molecular analysis revealed 11 (21.6%) positive samples, consisting of 8 (72.7%) Candidatus Mycoplasma haemominutum and 3 (27.3%) Mycoplasma haemofelis confirmed positives. Candidatus Mycoplasma turicensis was not detected, and no co-infections were registered. No significant associations (p > 0.05) were found between the hemoplasma infection status and age, gender, breed, presence of ectoparasites, feline leukemia virus/feline immunodeficiency virus positivity of cats, or the sampling season. However, outdoor access was positively associated (p = 0.049) with infection and could be considered a risk factor (OR = 4.1) in acquiring feline hemotropic mycoplasmas. Phylogenetic analysis revealed that our sequences clustered with those selected from the GenBank database in two distinct clades. The registered population genetic indices were strongly supportive of the great variance in sequences between the recorded Mycoplasma species. Conclusions The findings support the occurrence of feline hemoplasma infections in previously uninvestigated territories of Europe, providing useful information for small animal practitioners. To our knowledge, the present survey is the first reported molecular evidence of feline hemoplasma infections in Romania.

Background: The hemotropic mycoplasmas (hemoplasmas) of the genus Mycoplasma are recognized as important bacteria that parasitize red blood cells, causing hemolytic anemia in many mammalian species, including cats. No information is available concerning the presence of feline hemoplasma infections in cats in Romania. Thus, the objective of the present study was to provide data on the occurrence and molecular characterization of hemothropic mycoplasmas in client owned cats in Romania. Methods : Blood samples from 51 unhealthy cats, originating from Timişoara Municipality, Romania, were screened for the presence of hemoplasmas using conventional polymerase chain reaction (PCR) targeting the 16S rRNA gene and sequencing assays. Results: Molecular analysis revealed 11 (21.6%) positive samples, consisting of 8 (72.7%) Candidatus Mycoplasma haemominutum and 3 (27.3%) Mycoplasma haemofelis confirmed positives. Candidatus Mycoplasma turicensis was not detected, and no co-infections were registered. No significant associations ( p > 0 . 05) were found between the hemoplasma infection status and age, gender, breed, presence of ectoparasites, FeLV/FIV positivity of cats or the sampling season. However, outdoor access was positively associated ( p =0.049) with infection and could be considered a risk factor (OR=4.1) in acquiring feline hemotropic mycoplasmas. Conclusions : The findings support the emergence of feline hemoplasma infections in previously uninvestigated territories of Europe, providing useful information for small animal practitioners. To our knowledge, the present survey is the first reported molecular evidence of feline hemoplasma infections in Romania.

Background The hemotropic mycoplasmas (hemoplasmas) of the genus Mycoplasma are recognized as important bacteria that parasitize red blood cells, causing hemolytic anemia in many mammalian species, including cats. No information is available concerning the presence of feline hemoplasma infections in cats in Romania. Thus, the objective of the present study was to provide data on the occurrence and molecular characterization of hemothropic mycoplasmas in client owned cats in Romania. Methods Blood samples from 51 unhealthy cats, originating from Timişoara Municipality, Romania, were screened for the presence of hemoplasmas using conventional polymerase chain reaction (PCR) targeting the 16S rRNA gene and sequencing assays. Results Molecular analysis revealed 11 (21.6%) positive samples, consisting of 8 (72.7%) Candidatus Mycoplasma haemominutum and 3 (27.3%) Mycoplasma haemofelis confirmed positives. Candidatus Mycoplasma turicensis was not detected, and no co-infections were registered. No significant associations ( p > 0.05) were found between the hemoplasma infection status and age, gender, breed, presence of ectoparasites, FeLV/FIV positivity of cats or the sampling season. However, outdoor access was positively associated ( p =0.049) with infection and could be considered a risk factor (OR=4.1) in acquiring feline hemotropic mycoplasmas. Conclusions The findings support the emergence of feline hemoplasma infections in previously uninvestigated territories of Europe, providing useful information for small animal practitioners. To our knowledge, the present survey is the first reported molecular evidence of feline hemoplasma infections in Romania.

The prevalence and intensity of infection with digestive, liver, and pulmonary parasites in wild boars in Romania was determined by examination of 280 cadavers from 26 hunting grounds during the period 2012–2016. Eleven genera of parasites were recovered: nine within the digestive system (Eimeria, Ascaris, Globocephalus, Gongylonema, Hyostrongylus, Oesophagostomum, Physocephalus, Trichuris, and Macracanthorinchus); and two (Dicrocoelium, Metastrongylus ) located in the hepatic and pulmonary systems. The overall prevalence of infection was 80.7% (n = 280). Polyparasitism was found in 82.8% of positive cases. The mean intensity of parasitism was highest for pulmonary parasites (Metastrongylus salmi, 25.95). Regarding gastrointestinal parasites, the highest mean intensity occurred in the case of Oesophagastomum dentatum infections (22.14), whilst the lowest was that of Macracanthorhynchus hirudinaceus (1.66). Wild boars are an important source of infection for domestic pigs in Romania and neighboring countries where extensive breeding systems occur.

Forty seven working horses from Romania were post-mortem examined for small strongyles (Cyathostominae) infections. All horses were found infected. The overall cyathostomins intensity ranged from 390 to 13,010 and horses were infected by 8-24 species. The intensity was higher in ventral colon (1531) and dorsal colon (824), the lowest in the caecum (524). Twenty four species were identified. Cyathostomum catinatum, Cylicocyclus insigne, and C. Nassatus had 100% of prevalence. Over 50% of horses were infected by Coronocyclus coronatus, Cylicostephanus calicatus, C. goldi, and C. longibursatus. Other prevalent species (34%-45%) were Cyathostomum tetracanthum, Cylicostephanus minutus and Gyalocephalus capitatus. Coronocyclus labiatus, Parapoteriostomum mettami, Poteriostomum imparidentatum and P. ratzii had the lowest prevalence. Most species showed high organ preference with a niche breadth value between 1 and 1.96 while only 7 species (Coronocyclus labiatus, Cyathostomum tetracanthum, C. brevicapsulatus, Cylicocyclus elongatus, C. insigne, C. leptostomum and C. radiatus) showed a more generalist selection. The niche breadth of 10 species was significantly (p0.05) influenced by itself intensity (Coronocyclus labratus, Cyathostomum pateratum, C. tetracanthum, Cylicocyclus elongatus, C. radiatus, C. ultrajectinus, C. leptostomum, Cylicodontophorus euproctus, Poteriostomum imparidentatum, P. ratzii). The niche breadth of Cylicocyclus nassatus was positively (p0.05) influenced by the summed intensity of the other species while that of Cylicocyclus elongatus was negatively (p0.05) influenced by the intensity of the other species. The cluster analysis of the Cyathostominae community composition showed a major cluster composed by the three dominant species, followed by a cluster composed by Coronocyclus coronatus, while all the other species presented a tree like structure.

Thelazia callipaeda is an emerging vector-borne zoonotic helminth parasitizing the conjunctival sac of a broad spectrum of definitive hosts, such as dogs, cats, rabbits, wild carnivores, and humans. Its presence is associated with mild to severe ocular disease. Here, we report two new clinical cases in dogs originating from western and southern Romania, with no travel history. On clinical examination, the nematodes were retrieved from the conjunctival sac and identified using morphological keys and molecular tools. Twenty-two adult nematodes (8 males, 14 females) were collected and were identified as T. callipaeda by morphology. The molecular analysis revealed a 100% identity with haplotype h1 of T. callipaeda. This study describes the occurrence of new autochthonous cases of thelaziosis in Romania, reinforcing the spreading trend of this zoonotic eyeworm and highlighting the need for increased awareness among medical and veterinary practitioners. Moreover, we provide additional molecular evidence for the exclusive distribution of haplotype 1 of T. callipaeda in Europe.

A survey was carried out to assess the prevalence and the intensity of helminth infections in the roe deer (Capreolus capreolus L.), red deer (Cervus elaphus L.) and fallow deer (Dama dama L.) from 16 hunting areas in western Romania. Overall, 122 deer shot during the hunting season 2013-2015 were examined. Haemonchus contortus, Nematodirus filicollis, Oesophagostomum venulosum and Dictyocalus spp. were found in all the deer species. Trichuris spp. and Dicrocoelium dendriticum were found in the roe deer and red deer and Moniezia expansa in roe deer, only. Overall, the prevalences (35%) and the mean intensities (20) were quite low in the abomasums and intestine, but H. contortus in fallow deer (50%). No2 or 3 helminth species were found in the abomasum, small and large intestine. Lung helminthes in the roe deer and follow deer accounted for 57.1% and 71.4%, respectively. The only specie with a prevalence50% was in the red deer. D. dendriticum was found in the liver of roe deer and fallow deer.

During 2003 the blowflies of Lucilia, Calliphora and Protophormia genera were monitored between April 20 and October 30. For Calliphora genus 6 population peaks were recorded. First capture was achieved on April 20, at a 12.1ºC mean air temperature, 15.9ºC mean soil temperature and 50% relative humidity, and the last one on October 21st, at a 15.1ºC mean air temperature, 14.7ºC mean soil temperature and 84% relative humidity, respectively. For Lucilia genus 7 population peaks were recorded, with May 01st the first capture date, and October 07 the last capture date. For Protophormia genus were also 6 population peaks recorded, as for Calliphora genus. First capture was performed on May 02nd at a mean air temperature of 22.5ºC, a mean soil temperature of 24.3ºC and 54% relative humidity, and the last one on October 07, at a mean air temperature of 10.4ºC, a mean soil temperature of 10.8ºC and 73% relative humidity.

Traumatic myiasis is a serious threat for worldwide sheep flocks. It is known that the blowflies’ abundance and occurrence of strikes could be diminished by traps use. The aim of this study was to ascertain the role of olfaction in blowflies’ orientation towards the odorous traps. Three genera of blowflies: Calliphora, Lucilia and Protophormia were monitored. For the 50 m distance all three genera had a positive response to detect the odorous bait, while for the 100 m distance its perception was decreased, but the structure in re-trapped blowflies remained the same, better for Calliphora (18%) and diminished for Protophormia (2%). For the 200 m distance only blowflies of Calliphora (10%) and Lucilia (2%) genera were recovered, respectively.

Serum samples from 197 clinically healthy dogs residing in the Banat Region, the western historical part of Romania, were assayed by an indirect fluorescent antibody test for the presence of anti-Babesia canis antibodies. Overall, the seroprevalence was 19.8% (39/197). The percent of seropositive dogs in rural areas (28.4%; 19/67) was significantly higher (P0.05) compared to dogs living in urban areas (15.4%; 20/130). Seroprevalence of B. canis infection in hunting dogs was also found to be significantly higher (P0.05) compared to canines with other lifestyles, but no significant difference was found between companion and kennel dogs. The statistical analysis showed that no significant differences (P0.05) were present between the seroprevalence of infected animals associated with age, gender, or breed. The hunting lifestyle was the only factor (OR = 4.57; 95% CI = 2.1-10.2; P = 0.002) positively associated with seroprevalence in dogs and can be considered the risk factor in the acquisition of infection. Also, the results of the present survey indicate that infection with B. canis in dogs is common in the sampling area and that it is an important pathogen for the local canine population.

Serum samples from 376 randomly selected adult cattle, from 25 farms located in 3 counties (Arad, Bihor, and Timiş) from western Romania, were sampled for Neospora caninum antibodies using a commercial ELISA-kit. Seroprevalence values and risk factors for neosporosis (cow age, breed, herd size, farming system, previous abortion, and number of farm dogs) were examined using a generalized linear mixed model with a binomial distribution. Overall, the seroprevalence of N. caninum was 27.7% (104/376) with a prevalence of 27.9% (24/86) in Arad, 26.9% (25/93) in Bihor, and 27.9% (55/197) in Timiş. Of 25 cattle herds, 23 were seropositive with a prevalence ranging from 10.0 to 52.2%. No correlation was found between N. caninum seropositivity and age, breed, herd size, breeding system, and previous abortion. The number of farm dogs was the only factor (P(Wald) = 0.03) positively associated with seroprevalence in cows and can be considered the risk factor in the acquiring of infection. The present work is the first regarding serological evidence of N. caninum infection in cattle from western Romania.

Given its numerous biologically active components, Euphorbiaceae has been found to be a large plant family and polyvalent with quite interesting therapeutic activity that can be studied.The ixodicidal activity of Euphorbia cyparissias extracts was studied in vitro and in vivo. Tested concentrations were 10, 5, 2, 1, 0.5 and 0.25%.For the in vitro study, conducted on field-collected female specimens of Dermacentor marginatus and Haemaphysalis punctata, the efficacy results showed that the ticks died after exposure in the case of 10, 5, and 2% tincture concentrations. The effects appeared after 30 minutes and became more visible 120 minutes after each exposure. The statistical differences regarding the used concentrations were found to be: F = 6.51, df = 5, P0.001. The in vivo study of the efficacy of E. cyparissias concentrations was performed on 35 naturally infested sheep and on 30 bovines parasitized with Ixodes ricinus, sprayed with tincture and glycerinate dilutions (bovines) on days 0 and 7. The results revealed detrimental effects on the survivability of female ticks, the most prominent being the reduction of their movement capacity. In sheep in vivo efficiency observed within 24 hrs varied, between 1 and 23% for D. marginatus and between 7 and 27% for H. punctata and respectively between 2 and 53% after 24 hrs, for I. ricinus, comparable effects being also found 72 hrs after the second administration of Euphorbia extracts.Extracts from E. cyparissias may be used, with results, as an ecologic alternative tick control management method, being a cheap solution, with a sizeable role in reducing the use of synthetic and/or other harming and resistance source ixodicidal conditionings.

The larvae of blowflies from the Calliphoridae family cause fly strikes in sheep and other species of economic importance. Impaired wool, decrease of ewe fertility, and even death can occur in heavy infestations. This paper describes the Calliphora spp. blowflies’ behavior on and around a trap baited with liver and ammonia before they entered in. More than half of Calliphora spp. blowflies (50.88%) stayed a medium time (eight to fourteen seconds) on the standard trap, while only 1.79% of them spent a longer time (26 to 30 seconds) before entering the trap.

Blood samples of 119 red foxes, originating from 44 hunting grounds of 3 western counties (Arad, Hunedoara, and Timiş) of Romania, have been examined for the presence of Hepatozoon canis infection using the conventional polymerase chain reaction (PCR) of the fragment of 18S rRNA gene. Overall, 15 (12.6%) samples were found to be PCR-positive. Of the sampled hunting grounds, 29.5% (13/44) were found positive. Positive samples were recorded in all screened counties with the prevalence of 14.8% (9/61) in Arad, 9.8% (5/51) in Timiş, and 14.3% (1/7) in Hunedoara, respectively. No correlation was found (P0.05) between H. canis positivity and gender or territorial distribution of the infection. To confirm PCR results, 9 randomly selected amplicons were sequenced. The obtained sequences were identical to each other, confirmed the results of the conventional PCR, and showed 98-100% homology to other H. canis sequences. The results of the current survey support the role of red foxes as sylvatic reservoirs of H. canis in Romania.

This study was undertaken to investigate the occurrence and public health significance of Cryptosporidium species/genotypes and subtypes in a newborn lambs. A total of 175 diarrheic fecal samples from lambs (younger than 21 days) were collected in seven sheep flocks located in western Romania, and were microscopically examined for the presence of Cryptosporidium oocysts after staining with modified Ziehl–Neelsen technique. Twenty-four (13.7%) fecal samples were tested Cryptosporidium positive by microscopy and were subjected for molecular characterization. All positive samples were successfully amplified through a nested polymerase chain reaction (PCR) of the small subunit (SSU) rRNA gene (18S). Cryptosporidium species were determined by restriction fragment length polymorphism (RFLP) analysis of the secondary PCR products using the conventional Ssp I and Vsp I restriction enzymes. The identified species were: Cryptosporidium parvum (20/24), C. ubiquitum (2/24) and C. xiaoi (2/24), respectively. PCR-RFLP results for C. ubiquitum and C. xiaoi isolates were confirmed by DNA sequencing. Subsequently, subtyping of seven randomly selected C. parvum isolates, based on sequence analysis of the GP60 gene, revealed the presence of five different subtypes (IIaA17G1R1, IIaA16G1R1, IIdA20G1, IIdA24G1 and IIdA22G2R1) belonging in two zoonotic subtype families (IIa and IId). These findings may suggest the potential role of the newborn lambs as a source for human cryptosporidiosis. This is the first published report about the presence of C. ubiquitum and C. xiaoi in lambs from Romania.