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Background: Neosporosis is a parasitic disease caused by Neospora caninum. This parasite is an obligate intracellular coccidia similar to Toxoplasma gondii with a global distribution. With regard to the experimental studies, vertical transmission of the parasite in the monkey (non-human primates) has increased the concern about the zoonotic potential of this disease. The principal aim of the current research was to perform a mini-review on investigations regarding the Neospora infection in humans on a global scale for the first time. Methods: All peer-reviewed articles (published until April 2021) on the Neospora infection in humans were searched in English databases such as Google Scholar, ScienceDirect, Scopus, PubMed, and ProQuest. Results: Based on data in the available articles, the presence of antibodies against the Neospora infection was between 0 and 37.7% in people from different countries. The seroprevalence rate of this infection in HIV-positive individuals was higher (26.6% and 37.7%) compared to other cases. Finally, the genomic DNA of Neospora was detected up to 1% using molecular biology techniques. Conclusions: Overall, the detection of anti-Neospora antibodies in humans indicated that people have been exposed to the parasite. Comprehensive research studies are essential for clarifying the risk factors associated with the Neospora infection in humans. This report provides the baseline information for future researchers. Molecular investigations and genotypic works on N. caninum isolates are highly recommended as well.

Gastrointestinal nematodes are leading causes of loss in livestock and are the primary restriction to its profitable production, worldwide. This study was carried out to determine the prevalence and diversity of sheep abomasum nematode species in Urmia (Iran) and Soran (Iraq) slaughterhouses from October 2019 to January 2021. A total of 280 abomasa (each city 140 samples) were randomly collected from the slaughtered sheep. The abomasal content and mucosa were removed and washed. The collected nematodes were morphologically identified. Genomic DNA was extracted from identified nematodes and a fragment from the internal transcribed spacer 2 ribosomal ribonucleic acid (ITS2-rDNA) gene was amplified. In Urmia city, two species including Teladorsagia circumcincta (40.7%), and two morphotypes of Marshallagia species; Marshallagia marshalli (35.0%) and M. trifida (4.3%) were identified. In Urmia city, 52.9% of the examined sheep were infected with at least one species of nematodes. The overall prevalence of abomasa infection with nematodes in Soran city was 91.4%. In the examined sheep abomasa in Soran city, four species of nematodes were identified, including Marshallagia species with two morphotypes, M. marshalli (85.0%) and M. trifida (20.7%), Teladorsagia circumcincta (32.1%), Parabronema skrjabini (1.4%), and Haemonchus contortus (0.7%). Except for H. contortus, all the other identified nematode species were confirmed using molecular techniques. It was concluded that abomasal nematode infections are widespread in sheep particularly in Soran city. Marshallagia marshalli and T. circumcincta were most prevalent nematodes in both regions. In addition, further molecular studies are recommended to understand the intra-specific variations in the genus Marshallagia and more accurate identification of morphotypes in these regions.

Sarcocystis species in animals and humans cause a zoonotic disease called Sarcocystosis. This study was aimed to investigate the morphology and pathology of Sarcocystisin naturally infected sheep. The carcasses of slaughtered sheep at Urmia slaughterhouse were inspected for evidence of infection with Sarcocystis macrocysts. Histopathological sections were prepared and stained routinely by Hematoxylin-Eosin (H&E) staining. A total of 1372 out of 4121 (33.3%) removed macrocysts were full of bradyzoites (54.3×223.66μm) ranged from 5-10 mm. Histologically, the reaction of the muscle tissue varied from degenerative to inflammatory around the macrocysts. The infected muscle demonstrated evidence of myocytolysis with infiltration of inflammatory cells in focal pockets, which were mainly comprised of lymphocytes and macrophages. The other findings were arterial wall hyperplasia, hyperplastic proliferation, and giant cell presence around the macrocysts. The results indicated that morphologic features of macrocysts of Sarcocystis in naturally infected sheep were informative and causing pathologic changes in muscle tissue.

The aim of this study was to determine molecularly the species of larvae of trichostrongylid nematodes in arrested development in the abomasum of sheep from Iran and their seasonality. A total of 240 sheep abomasa were randomly collected at different seasons from Ilam abattoir in 2017–2018. The adult nematodes were isolated and identified. The mucosa of the abomasum was removed and digested using a pepsin solution. DNA from different parts of the abomasum was extracted using a SinaPureTM kit. Polymerase chain reaction (PCR) amplification was performed; the ITS2-rRNA gene was amplified using specific primers. Amplified PCR products were purified and sequenced by Bioneer Corporation (South Korea). The obtained ITS2 sequences were aligned using BLAST analysis and submitted to GenBank. Morphologically, five species of adult nematodes of four genera including Haemonchus contortus, Marshallagia marshalli, Marshallagia occidentalis, Parabronema skrjabini, and Teladorsagia circumcincta were identified. Marshallagia marshalli (65.4%, 120.2 ± 28.5) had the highest prevalence during autumn. Molecularly, arrested development of larvae of M. marshalli and T. circumcincta were demonstrated in mucosa of the abomasum in autumn and early winter using PCR of the ITS-2 region of rRNA. The results evidenced the first molecular characterization of the larvae of trichostrongylid nematodes from abomasum of sheep from Iran.

Knowledge on neosporosis and associated risk factors in different species of animals are so important for designing the control programs and reduce the economic losses globally. This literature review targeted for evaluating the infection rate of Neospora caninum in animals in Iran. Until April 2020, all of published documents in the main English and Persian-language databases were searched. A total number of 110 documents (English = 85 and Persian = 25) were extracted. Most of reports were sero-epidemiological studies using ELISA in Iranian cattle population. The range of Neospora infection was 3.8–76.2% in cattle, 0–54.6% in dogs, 0.9–9.9% in sheep, 6.2% in goats, 19.2–55.9% in buffaloes, 20–42.2% in horses, 52% in donkeys, 3.2–27% in camels, 14% and 19% in cats, and 0–20.4% in rodents. This rate in birds was 17.3% in chicken, 9.8% and 30.4% in pigeons, 2.8% and 3.7% in sparrows, and 9.9% in hooded crows. This is a comprehensive literature review on Neospora infection in Iran at the first time. The infection of N. caninum is widespread in Iran especially in dogs and cattle population. This review can provide baseline information for future research. Study on other hosts especially on wild and exotic animals is recommended for exact estimate of neosporosis in Iran. Investigations into molecular diagnosis and genotyping of N. caninum strains are also needed; this will be helpful for developing vaccines and finding the connection among wild and domestic cycles of disease. Education on the risk factors associated with N. caninum infection for is suggested farmers and rural public.

Background: Bovine theileriosis is an important disease in Iran and throughout the world with economic losses in Iranian cattle husbandry. The aim of the current study was to determine prevalence and geographic distribution of Theileria annulata infection in cattle and ixodid ticks species diversity in Kurdistan Province, West of Iran. Methods: A total number of 193 blood samples were randomly taken from jugular vein. Ixodid ticks were also collected from body surface of examined cattle in three sub-areas of the region, i.e. north, center and south. The genomic DNA was extracted and PCR was performed to amplify a 721-bp-long fragment of the 30 Kilo Dalton major merozoite surface antigen of T. annulata. Results: The overall prevalence was 50.2% (97/193) with lymphadenopathy (54.4%) and petechia in mucosal membrane (95%) of cross-breed cattle (24.9%) aged

Different N. caninum strains with distinct biological characteristics and pathogenicity have been introduced from various areas of the world. This study aimed to detect Neospora-infection in different hosts from Iran using the nested-Polymerase Chain Reaction (PCR) technique and to perform the phylogenetic analysis of N. caninum isolates using the Nc-5 gene. Different species of parasite hosts (cattle, dogs, rodents, cats, and birds) from the dairy farms with history of abortion with intensive breeding systems in Hamedan Province, Iran were included for sampling. The stillbirth fetuses of cattle, sheep, and goats were also collected from the slaughterhouse. The nested-PCR technique was used to identify the Neospora-DNA in the aborted fetal materials. The PCR products of the positive samples were sequenced and compared with other isolates in GenBank. Antibodies to N. caninum was found in 19.1% of dogs. Also, 11.4% of dogs were positive for Neospora-like oocysts using microscopy method. The molecular prevalence of N. caninum was 10.6% in dogs, 13.6% in cattle, 35.4% in the aborted fetuses of cows, 31.9% in rodents, 7.6% in pigeons, 1.9% in sparrows, 5.1% in cats, and 7% in the semen sample of bulls. A sampled coyote was positive while the stillbirth fetuses of sheep and goats were negative. The multiple sequence alignments indicated that 34 sequences of N. caninum bore a 94.9-99.3% similarity to each other. Hence, these sequences were classified into two major clusters (clusters A and B) and seven sub-groups (isolates 1-7). The sequence of coyote had a high similarity to the sequences derived from dogs and cows. Also, root diagram showed the isolates of current study and previous reported from Iran have a similar origin. This study provided a phylogenetic analysis of Neospora-infection in different hosts, and for the first time, explored Neospora-infection in coyotes from Iran. Different isolates were detected in the studied population in the region. Comparison of the sequences showed the farm dogs play an important role in the life cycle and distribution of the parasite; so the relationship of dogs with the herd and their access to aborted materials should be restricted. Also, regular control of rodents' population and culling of seropositive cows with a history of abortion is essential for cutting the chain of transmission.

Background: Neosporosis is considered as a ubiquitous disease in Iran and other countries. This research was expected to determine the prevalence and related risk factors of Neospora caninum in household dogs in Hamadan Municipality, Iran. Methods: A total of 184 whole blood was evaluated for the presence of antibodies to N. caninum by the enzyme-linked immunosorbent assay (ELISA). All seropositive animals were affirmed by molecular techniques. Results: Based on serology and molecular methods, N. caninum infection was detected in 4.9% (95% CI = 4.9 ± 3.1%) of animals. In addition, the highest infection rate was significantly recognized in female dogs (57.1%) with under 6 months old (54.4%). Additionally, the clinical signs of neosporosis were observed in 2 out of 4 positive dogs (P0.05) with breeding, food regime, housing, and direct contact with infected animals. Conclusions: In general, the serological and molecular outcomes were parallel together. It was concluded that this is a universal assessment of risk factors related to N. caninum in Iranian house dogs for the first time.

This study was conducted to analyse the serine protease of Fasciola hepatica eggs by specific substrates and inhibitors, and investigation of the effects of pH and temperature on proteases’ activity and stability. Adult worms were isolated from infected livers. After homogenisation, their protein levels were measured with the Bradford method. Total proteolytic activity of the Fasciola hepatica extract was evaluated with azocasein substrate at pH values from 2 to 12. N-benzoyl–arginine–p-nitroanilide (BApNA) trypsin and N-succinyl-alanine-alanine-prolin-phenylalanine-p-nitroanilide (SAAPFpNA) chymotrypsin substrates were used to measure specific protease activities. The effect of protease inhibitors phenylmethane sulfonyl fluoride (PMSF), pepsin, and ethylenediaminetetraacetic acid (EDTA) on these enzymes was evaluated. Estimation of optimum temperature and pH was performed in the temperature range of 10–90 °C and pH values from 2–12. The optimum pH activities for trypsin and chymotrypsin were at alkaline pH and for total proteolytic activity at acidic pH. The results using protease inhibitors showed that the eggs had serine protease activity. The optimum temperature activity of trypsin and chymotrypsin was 50 °C. These proteases were stable up to 40 °C. Due to the importance of pH and temperature in life cycle of Fasciola hepatica, these findings can be used for induction of some modifications in pH and preventing the activity of the enzyme for decrement of the efficacy of embryonic development and egg hatching of this zoonotic parasite.

Toxoplasmosis is a zoonotic food-borne disease with global distribution. The principal aim of this research was to detect the seroprevalence of T. gondii infection in cattle, dogs and cats in dairy farms with high incidence of abortion, as well as to identify associated risk factors in Hamedan, Iran. A number of four large dairy farms with high history of abortion were selected and evaluated for risk factors associated to T. gondii infection. Blood samples were taken from 400 of pregnant cattle, 85 of dogs and 35 of cats (all of the dogs and cats population from interring and around of farms were sampled). The samples were examined for presence of antibodies to T. gondii infection using ELISA technique. Seroprevalence of T. gondii infection were detected 13% in cattle, 47.1% in dogs and 40% in cats. With regard to cattle seropositivity and associated risk factors, significant differences were found regarding abortion history (P = 0.026), race (P = 0.035), rodents’ presence in farm (P = 0.006) and age groups (P = 0.011). Also, there were no significant differences between seroprevalence rate and stage of gestation (P = 0.998), type of breeding service (P = 0.561), stillbirth history (P = 0.302), dog contact with herd (P = 0.302) as well as poultry presence in the farm (P = 0.302). In both dogs and cats, the seroprevalence rate for females was higher than males (P 0.05). This is the comprehensive evaluation of risk factors associated to toxoplasmosis in Iranian dairy farms at the first time. Our finding indicated that toxoplasmosis may partly be responsible for transmission to humans and also abortion and economic losses in dairy farms. Improve the sanitary strategies for reducing the risk factors in animals’ husbandry and launching control programs are recommended.

Background: Dogs, especially stray dogs, act as the major carriers of different infectious and parasitic agents in the environment; hence, their possible transmission to humans is a public health concern. The principal aim of the present investigation was to evaluate the seroprevalence of Neospora caninum and Toxoplasma gondii infections in stray dogs of rural regions of Hamadan, West of Iran. Methods: During this cross-sectional survey in 2018, 180 blood samples were evaluated for the presence of antibodies to N. caninum and T. gondii using enzyme-linked immunosorbent assay (ELISA). Results: Seroprevalence of N. caninum and T. gondii infections was detected to be 5% (95% CI: 2.8-8.2%) and 35% (95% CI: 28.1-41.9%), respectively. In addition, coinfection was detected in 2.8% of animals. No significant differences were found between infection rate, sex, and age of animals regarding both parasites (P>0.05). Conclusions: This study provides the first insight into the infection of dogs in a region with the prevalence of N. caninum and T. gondii.

Placentally, transmission of Neospora caninum plays significant role on retaining of infection and economic losses in dairy farms. The objective of present study was to assess placental transmission of N. caninum in Iranian dairy cattle. A total of 476 blood samples of pregnant dairy cattle were randomly collected for serology test using ELISA technique. Genomic DNA was extracted and subjected for conventional PCR and Nested-PCR to amplify 330bp and 100bp fragment length of NC5 gene, respectively. Positive animals were follow-up during gestation till abortion and/or full term delivery occurred. Molecularly, 25.3% (118/476; 95% CI: 21%-28.6%) of examined animals were seropositive for N. caninum infection in Holstein cattle (15.1%) with2 years-old (75%) and history of abortion. The infection was detected. TT and abortion occurred in 13.6% and 3.57% of animals, respectively. Aborted cattle (94.1%, 16/17) were significantly infected to N. caninum and history of abortion was recorded in 42.4% of infected cattle. Placental transmission and abortion in naturally infected dairy cattle due to N. caninum was the first molecular evidence in all examined herds of cattle. It was also uncovered N. caninum had significant role on cattle abortion. Further studies for culling of seropositive cattle with history of abortion, associated risk factors related to neosporosis and lunch control program for reducing side effects of infection are essential in the region.

Iran is the second largest country in the Middle East with an area of about 1,650,000 km2 divided among 30 Provinces. It is bounded to the north by the Caspian Sea, to the east by Afghanistan and Pakistan, to the south by the Persian Gulf and Sea of Oman, and to the west by Iraq and Turkey. The center and east of the country are largely barren undulating deserts, while there are highlands of Zagros Mountains in the west, along the Turkish and Iraqi borders. In the north, the Alborz mountain ranges rise steeply from a fertile belt around the Caspian Sea (Rokni 2008) (Fig. 71.1).

Background Hepatozoon canis is a protozoan that is transmitted by the ixodid ticks. Ingesting the tick or a section of the tick organ which is infected by the mature oocysts containing infectious sporozoite is the main source of infection in dogs. Canine hepatozoonosis infects dogs in Iran, but the vector for Hepatozoon protozoa from Iran has never been demonstrated. The present study aims to detect H. canis in dogs and vector ticks in Iran. Methods During the period of 2018–2019, Blood samples and ixodid ticks were collected and examined using microscopical, molecular, and serological methods. A total of 246 blood samples were collected from the cephalic vein of pet, stray, and shelter dogs (103 stray, 99 shelter, and 44 pets) of both genders and varying ages in Northwest of Iran. Results Microscopically, infected neutrophils with Hepatozoon spp. were detected in 5 of 246 (2.03%) thin stained blood smears with low parasitemia. Indirect immunofluorescent antibody test (IFAT) was used to test the serum samples and antibodies against H. canis were detected in 31 (12.6%) of the serum samples. Molecularly, 23 out of 246 (9.34%) blood samples were found to be infected with H. canis. A comparison of the results of 3 diagnostic methods demonstrated a good agreement between IFAT and PCR and a poor agreement between microscopical examination with IFAT and PCR. There was no significant difference in different age groups and sex of sampled dogs. However, stray dogs had significant infection rate of than pets and shelter ones. In body inspection, 141 adult ticks (31 partially engorged females, 26 fully engorged female and 84 fed males) were collected from examined dogs and all ticks were belonging to species of Rhipicephalus sanguineus (sensu lato). Positive reaction to H. canis was observed in the genomic DNA of the 7 ticks (4.96%). A BLAST analysis of obtaining sequences isolated from both dogs and ticks indicated a 99–100% similarity with H. canis 18S rRNA gene sequences in GenBank. This is the first study in Iran to detect H. canis in R. sanguineus tick.

Neospora caninum plays an important role in abortion and economic losses in dairy industry worldwide. The main target of this work was to detect the infection rate of N. caninum in various hosts in dairy farms for identifying the risk factors and applying appropriate control programs to reduce the number of abortions. The study was conducted in dairy farms with high incidence of abortion in Hamedan province, West of Iran. After the primary assessment, we conducted a controlling program for reducing the Neospora‑infection rate and associated abortions. Before implementing the control program, the seropositivity was 24.8% in cows (N=476 distributed in 10 farms) and 8.6% (N=185) in dogs. Abortion occurred in 3.57% of pregnant cows. 94.1% of aborted cows were positive for Neospora‑infection. Based on molecular technique, the infection rate was detected in 7.3% of cats (N=41), in 25% of pigeons (N=19)and in 11.8% of rodents (N=51). After the implementation of neosporosis control programs in the farms, the seropositivity of N. caninum decreased to 8.2% in cows and 2.9% in dogs. After the one‑year follow‑up, no cases of abortions were reported in the farms. This was the first parallel evaluation of Neospora‑infection and controlling programs in Iranian dairy farms. Regular control of rodents, retesting of seronegative animals and farm biosecurity measures are recommended for reducing the abortion incidence. The access of dogs to the herd and to aborted materials should be restricted.

Background: Fascioliasis is a worldwide zoonotic disease caused by the trematodes Fasciola hepatica in humans and animals. Proteases are essential for the survival of parasites and have important activities such as penetration, tissue migration, and egg hatching. This study was conducted to analyze cysteine protease of the miracidia and eggs of F. hepatica, and to assess the effects of pH and temperature on the proteases activity and stability. Methods: Adults F. hepatica were isolated from infected livers and were morphologically identified in 2018. Eggs collected from the adults and incubated in distilled water at 28 °C for 16 d to produce miracidia. The extract was collected from miracidia and eggs. A substrate for cathepsin B (Z-Arg-Arg-Pna) was used to assess the enzyme activity at different (2-12) pH levels. After homogenization, protein level was measured with Bradford method. Estimation of optimum temperature and pH was performed in the temperature range of 10-90 ° C and pH values from 2-12. Results: The highest activity of the miracidia and eggs enzyme extracts for Z-ArgArg-pNA was at pH 4. The miracidia extract was most stable at neutral pH and the eggs extract was most stable in acidic pH. The optimum temperature activity for both stages was 40 °C. These proteases were stable up to 40 °C. Conclusion: Upon the importance of pH and temperature in the life cycle of F. hepatica, the current findings can be used for induction of some modifications in pH and preventing the activity of the enzymes for decrement of the efficacy of miracidia penetration into the intermediate snails and egg hatching of this zoonotic parasite.

The bulk milk examination is a reliable screening tool for monitoring the quality of milk in the farms. The infection to Neospora caninum, Toxoplasma gondii and Brucella sp. Was evaluated in bulk milk samples of dairy farms in Hamedan province, West part of Iran. All the dairy farms (n = 149) were examined for N. caninum, T. gondii and Brucella infections using milk ring test (MRT), microbiology, serology (Enzyme-linked Immunosorbent Assay), and molecular techniques. Based on molecular methods, Brucella-infection was negative in all farms; while, 55 %, 5.4 % and 2.7 % of samples were positive for N. caninum, T. gondii and mix infection, respectively. The highest Neospora-infection was detected in the farms with history of abortion in fall and winter. There was significant association between Neospora-infection and the presence of dogs and rodents in the farms, herd size, and age of the animals. Also, a significant association was seen between Toxoplasma-infection and the presence of cats and rodents in the farms, as well as age of the animals. Average total bacterial count (TBC) was calculated 1.14 × 106±1.1 × 106. The highest TBC was in the farms from Central locations of studied area (5.7 × 106±2.24 × 106), farms with more than 120 animals (7.9 × 106±2.8 × 106), and farms with ≥50-months age (1.74 × 106±6.3 × 105) in spring and summer (6.9 × 106±3.7 × 106). The number of somatic cells was estimated between 1 × 104 and 2 × 106 (Average = 4.2 × 105±3.39 × 105). The current study was a comprehensive evaluation of Neospora, Toxoplasma and Brucella infections in milk samples of Iranian dairy farms for the first time. Neospora-infection is responsible for economic losses in the region. Health education and milk pasteurization are so helpful for inhibiting the milk borne diseases. To reduce the risk factors, predict and design the appropriate schemes like redundant of heterogeneous animals are recommended.

Cystic echinoccoccosis is a zoonotic and prevalent disease which causes health problems and economic looses worldwide. The biochemical components of hydatid cyst fluid (HCF) have important role in metabolism and growth of unilocular hydatid cyst. The aim of the current study was to determine HCF level of enzymes and electrolytes profiles in naturally infected Iranian domestic ruminants. The livers and lungs infected with hydatid cysts were collected and HCF was aspirated from a total of 100 cysts obtained from livers (50 cysts) and lungs (50 cysts) of slaughtered domestic ruminants. Biochemical measurement of HCF was undertaken using colorimetric technique, refractometer, flame photometer, and biuret method. The enzyme levels of HCF were also measured by using appropriate kits. The average HCF level of calcium (Ca) was found to be significant higher in cattle (7.49 ± 1.28 mg/dl) and goats (13.98 ± 5.11 mg/dl) lungs. The average HCF level of phosphorous (P) was significantly higher in cattle livers (0.82 ± 0.02 mg/dl) than other examined animals. Measurement of the average HCF level of magnesium (Mg) was significantly lower in camel lungs (11.8 ± 1.05 mg/dl) than that in cattle livers (10.56 ± 1.73 mg/dl). The highest average HCF levels of Natrium (Na, 122.8 ± 11.91 mEq/l) and Kalium (Ka, 7.18 ± 1.37 mEq/l) were measured in sheep livers. The average HCF level of albumin (Alb) in infected lungs was significantly lower in cattle (0.48 ± 0.01 mg/ml) than camels (0.95 ± 0.05 mg/ml). The average HCF level of total protein (TP) in infected lungs was also lower in sheep (0.51 ± 0.06 mg/dl) than goats (3.21 ± 0.51 mg/dl). The highest average HCF level of creatine phosphokinase (CPK, 1229.25 ± 13.21 U/ml) and lactate dehydrogenase (LDH, 363.62 ± 10.44 U/ml) were measured in infected lungs of camels. It was concluded that HCF level of enzymes and electrolytes had differences in examined Iranian domestic ruminants which may be used to screen and help in characterization and identification of strains of Echinococcus granulosus.

Trypanosoma evansi is a cosmopolitan protozoan which affects camelids and may cause illness and economic losses. The present study was aimed to determine the prevalence and geographic distribution of T. evansi in Iranian one-humped camels of South East Iran. A total of 369 camels were randomly examined from three parts of southeastern Iran from March to September 2015. Blood samples were taken from jugular vein and examined by using thin blood smear and mercuric chloride test. Ninety-five out of 369 examined camels (25.75%) with clinical signs (15.8%, 58/369) were found to be infected with T. evansi. The prevalence was significantly higher in camels more than 4 years-old (23.3%) with clinical signs (8.8%) than the other ones. There was no significant difference between the prevalence and sex (6.23% male and 19.51% female). Trypanosoma evansi infection was geographically found in all investigated regions with the highest prevalence in North (17.61%). The results indicated that T. evansi infection was prevalent in Iranian one-humped camels which could be useful finding to lunch control programs in the region.

Neospora caninum is a major cause of abortion and economic losses among dairy farms in Iran and other countries. The main goal of current investigation was to evaluate the presence of antibodies against N. caninum and associated risk factors in dairy herds with history of abortion in Hamedan province of West Iran. A total numbers of 476 and 185 blood samples of pregnant cattle and farm dogs from 10 dairy farms with history of abortion were randomly collected. Bulk milk sample was taken from each farm. All samples were subjected for detection of IgG antibody against N. caninum using ELISA technique. Of all examined animals, 24.8% of cattle and 8.65% of dogs were seropositive to N. caninum. The seroprevalence had significant differences with abortion, stillbirth, metritis, breed, close contact to dogs, wild carnivores, rodents, poultry, and pregnancy using artificial insemination method. There were no significant differences among seroprevalence and different age groups, number and stage of gestation, and herd population. In all investigated farms, bulk milk examination was positive. In examined dogs, there was significant difference between seroprevalence and sex. It was concluded that N. caninum infection may be responsible for abortion and economic losses in dairy farms of the region. This was also the first comprehensive report on associated risk factors to N. caninum infection in dairy farms in the region. Keywords: Neospora caninum, Dairy cattle, Risk factor, Serology, Abortion, Dogs

Toxoplasmosis is a zoonotic food-borne disease with global distribution. The principal aim of this research was to detect the seroprevalence of T. gondii infection in cattle, dogs and cats in dairy farms with high incidence of abortion, as well as to identify associated risk factors in Hamedan, Iran. A number of four large dairy farms with high history of abortion were selected and evaluated for risk factors associated to T. gondii infection. Blood samples were taken from 400 of pregnant cattle, 85 of dogs and 35 of cats (all of the dogs and cats population from interring and around of farms were sampled). The samples were examined for presence of antibodies to T. gondii infection using ELISA technique. Seroprevalence of T. gondii infection were detected 13% in cattle, 47.1% in dogs and 40% in cats. With regard to cattle seropositivity and associated risk factors, significant differences were found regarding abortion history (P = 0.026), race (P = 0.035), rodents’ presence in farm (P = 0.006) and age groups (P = 0.011). Also, there were no significant differences between seroprevalence rate and stage of gestation (P = 0.998), type of breeding service (P = 0.561), stillbirth history (P = 0.302), dog contact with herd (P = 0.302) as well as poultry presence in the farm (P = 0.302). In both dogs and cats, the seroprevalence rate for females was higher than males (P 0.05). This is the comprehensive evaluation of risk factors associated to toxoplasmosis in Iranian dairy farms at the first time. Our finding indicated that toxoplasmosis may partly be responsible for transmission to humans and also abortion and economic losses in dairy farms. Improve the sanitary strategies for reducing the risk factors in animals’ husbandry and launching control programs are recommended.

Trypanosoma evansi is a pathogenic African animal protozoan, affecting livestock and wild animals worldwide and Iran. The present study was carried out to detect the infection of T. evansi in one-humped camels (Camelus dromedarius) of southeastern Iran. Over six months, a total numbers of 370 dromedary camels were randomly selected from three sub-areas located in southeastern Iran in 2015. Blood samples were taken from jugular vein and examined by using micro-hematocrit centrifugation (MHCT) and PCR techniques. Genomic DNA was extracted and PCR was performed to amplify a fragment of the mini-chromosome satellite DNA TBR1/2 of T. evansi. The overall prevalence was 31.35% (116/370). The highest T. evansi infection was significantly in adult camels (24.05%, 89/370) with chronic clinical signs (11.89%, 44/370). There was significant difference between prevalence and sex (9.46% male and 21.89% female). Only 19.19% (71 out of 370) of the infected camels were from the plain areas. The highest T. evansi infection rate was significantly recorded in the camels of north (19.19%, 71/370) part of the region. The molecular analysis was uncovered high level of infection with T. evansi in camel herds which can help to establish effective control programs in the region.

Fasciolosis is a zoonotic parasitic disease caused by the trematode Fasciola hepatica. The proteases are essential for the survival of parasites. The present study was aimed to determine serine proteases activities in miracidia of F. hepatica and evaluate the effects of pH and different inhibitors on the serine proteases activities. Adult F. hepatica helminths were removed from naturally infected livers of the slaughtered cattle and crushed. The eggs were incubated at 28.00 ˚C for 16 days. The released miracidia were homogenized and total proteolytic activity of the extract of miracidia at different pH values were evaluated. Serine proteases activities were determined using specific substrates. The inhibitory effects of chemical and herbal inhibitors on the enzymes were also assessed. The extract of miracidia hydrolyzed azocasein with optimum activity at pH 8.00. The optimum pH effect on serine proteases activities was found at alkaline pH. Phenylmethylsulfonyl fluoride and Bowman-Birk inhibitors inhibited and decreased the proteases activities in the miracidia extract. It was concluded that there were proteases activities in miracidia of F. hepatica which were inhibited by chemical and herbal inhibitors.

Background: The stray cats are considered as the sources of emerging humans and domestic livestock pathogens and the zoonoses of public health importance. The present study was aimed to elucidate intestinal helminth infections and infestation with ectoparasites of the stray cats of Ahar City, northwestern Iran. Methods: Totally, 51 stray cats were randomly trapped from different parts of the city between Mar and Nov 2013. The cats were assessed for ectoparasites by hair brushing, skin scraping, acetate tape preparation and othic swabs. They were euthanized and inspected for helminths infection. Results: Overall prevalence of helminths and flea were 44/51 (86.3%) and 31/51 (60.78%), respectively. The infection rates were significantly different among different age groups (P

The protozoan Neospora caninum (Apicomplexa: Sarcocystidae), respectively infects canids and ruminants as the most definitive and intermediate hosts. A diagnosis of neosporosis is generally made on the basis of clinical signs together with the detection of high levels of antibodies in serum. The present study compares the serum electrolyte profile (Ca, K, Mg, Na, P) and serum enzyme level (ALP, AST, CPK) of non-infected dogs with those of stray dogs naturally infected with N. caninum. The indirect fluorescent antibody test (IFAT) revealed that 17 of the 137 analysed serum samples (12.41%) of the stray dogs were seropositive (Sp) to N. caninum. Serum levels of the electrolytes and the enzymes were evaluated in the 17 Sp and 28 seronegative (Sn, 20.44%) dogs using common enzyme kits, spectrophotometry and flame photometry techniques. The average serum level of ALP, AST, and CPK were found to be significantly higher in the Sp stray dogs. Measurements of the average serum levels of Ca, K, Mg, and P were higher in Sp than Sn stray dogs, with the average K level being significantly higher in seropositive stray dogs. It was concluded that evaluation of serum enzyme and electrolyte levels may be used to screen N. caninum infection in stray dogs.

Background: The present study was carried out to detect the infection of larval stages of Trichobilharzia species in the snail Lymnaea auricularia in northwestern Iran based on DNA analysis. Methods: A total number of 320 snails of L. auricularia were sampled from four water-bodies located in the suburb of Urmia City, North West Iran, during May to November 2011. The snails were first microscopically inspected for the infection with larval stages of trematodes. Genomic DNA was extracted from the snails and PCR was performed to amplify a fragment of the ribosomal DNA of Trichobilharzia species in the infected snails. Results: Microscopic examinations indicated that 11.25% (36 out of 320) of the snails were infected with larval stages of trematodes, while the PCR patterns showed a much higher infection rate (31.25%, 100/320). According to the PCR, the infections were caused by the larval stages of T. szidati (21.56%, 69/320) and T. franki (9.69%, 31/320) or both of them (8.44%, 27/320). The infected snails were observed in three out of the four studied sites. The highest infection rate in a single site was 50% (25/50). Only 7.81% (25 out of 320) of the infected snails were from the plain areas, while the remaining was from high altitudes. Conclusion: Results of this study contribute the utility of the employed technique for quick and accurate detection of the infection with trichobilharzian species in their intermediate host snails, which may have potential zoonotic role in the region.

Background: The present study was carried out to detect the infection of larval stages of Trichobilharzia species in the snail Lymnaea auricularia in northwestern Iran based on DNA analysis. Methods: A total number of 320 snails of L. auricularia were sampled from four water-bodies located in the suburb of Urmia City, North West Iran, during May to November 2011. The snails were first microscopically inspected for the infection with larval stages of trematodes. Genomic DNA was extracted from the snails and PCR was performed to amplify a fragment of the ribosomal DNA of Trichobilharzia species in the infected snails. Results: Microscopic examinations indicated that 11.25% (36 out of 320) of the snails were infected with larval stages of trematodes, while the PCR patterns showed a much higher infection rate (31.25%, 100/320). According to the PCR, the infections were caused by the larval stages of T. szidati (21.56%, 69/320) and T. franki (9.69%, 31/320) or both of them (8.44%, 27/320). The infected snails were observed in three out of the four studied sites. The highest infection rate in a single site was 50% (25/50). Only 7.81% (25 out of 320) of the infected snails were from the plain areas, while the remaining was from high altitudes. Conclusion: Results of this study contribute the utility of the employed technique for quick and accurate detection of the infection with trichobilharzian species in their intermediate host snails, which may have potential zoonotic role in the region.

Many species of animals play the role of intermediate host for Neospora caninum. Although the neosporosis has a global distribution, however there is no information on goats' infection in Iran. The main goal of present study was to determine the seroprevalence of N. caninum in goats of western Iran. A total of 450 sera were taken and analyzed for antibodies to N. caninum using ELISA technique. Of those, 28 (6.2%) examined goats were found to be seropositive (95%, CI=6.2±2.2%). In which male goats (11.1%) had the highest IgG against N. caninum with more than 2 years-old (54.4%). There was significant difference between prevalence and age. Positive association were in animals with history of abortion (18/30, 60%) and IgG against N. caninum. No correlations were found between male and female. This was the first report on N. caninum infection in Iranian goats and may be responsible for abortion and economic losses in goat husbandry in the region.goat, Neospora caninum, ELISA, Iran.

OBJECTIVE Ectoparasitism is an important factor associated with the poor production of indigenous chickens. The present study was undertaken to estimate the prevalence and ectoparasite diversity in indigenous chickens of the Dalahu region in the western part of Kermanshah province, Iran. METHODS A total of 600 indigenous chickens (250 roosters and 350 hens) were randomly examined for the presence of different ectoparasites over the period April to September 2011. Ectoparasites were collected from different parts of chicken body using a hand lens, magnifying glass, and flashlights. The samples were preserved in 70% alcohol and cleared in lactophenol. RESULTS The overall prevalence of ectoparasites was 52.8% (66% hens and 34.4% roosters) (p

The Family Lymnaeidae is comprised of large aquatic snails that are intermediate hosts of parasitic flukes. In the present study, radular morphology, teeth length, and tooth formula of field-collected lymnaeid snails including Galba truncatula (Muller, 1774), Lymnaea stagnalis (Linnaeus, 1758), Radix auricularia (Linnaeus, 1758), and Radix gedrosiana (Annandale and Prashad, 1919) were investigated. The conical and dextral freshwater snails were collected from their natural habitats in West Azerbaijan province, northwestern Iran, in fall 2010. The snails were rinsed, measured, and taxonomically identified. The radula of all specimens was extracted and stained using Mallory II. Besides the morphological variations, the mean length of radular teeth significantly increased with the body size of the examined lymnaeid snails. Morphometric investigation indicated that the mean length of lateral (L) (427.86 ± 6.9 µm) and marginal (M) (427.82 ± 6.86 µm) radular teeth of Lymnaea stagnalis differed signifi...

Ticks have been reported as the most abundant ectoparasites infesting domestic ruminants in Iran. This study reports tick diversity, fauna and seasonal distribution on the domestic ruminants and close surrounding of them in the natural habitat of Sanandaj suburb, Iran, from March 2009 to February 2010. A total number of 1140 ruminants (398 cattle, 508 sheep and 234 goats) in four seasons were randomly selected. Argasid ticks were also collected randomly from 100 distinct sites. Of all animals examined, 37.19% (424/1140) were infested with a total of 1144 ixodid ticks. Of 1360 soft ticks, four species of argasid ticks of two genera (Argas and Ornithodoros) were identified. The ixodid tick infestation in the north was significantly higher than in other parts of the region. The highest numbers of hard ticks were collected from sheep in the region. The ixodid tick indices were 2.92, 2.2 and 3.34 for cattle, sheep and goats, respectively. There was no significant difference between an animal's sex and tick pre...

Cryptosporidium is a ubiquitous enteropathogen protozoan infection affecting livestock worldwide. The present study was carried out to determine the prevalence of Cryptosporidium infection in different age groups of dromedary camels in northwestern Iran from November 2009 to July 2010. A total number of 170 fecal samples were collected and examined using modified Ziehl-Neelsen (MZN) staining under light microscope. Examination of stained fecal smears revealed that 17 camels (10%) were positive for Cryptosporidium-like. The prevalence of Cryptosporidium-like was significantly higher in camel calves (< 1 years old) (20%) than other age groups, in which the diarrhoeic calves had the prevalence of 16%. In adult camels the prevalence was 6.5%. There was no significant difference in the prevalence of Cryptosporidium-like between male and female camels. It is concluded that Cryptosporidium infection is a problem in camel husbandry and could be of public health concern in the region.

The pond snail Lymnaea auricularia serves as an intermediate host for many digenian species. In West Azerbaijan province, northwestern Iran, the cercariae infection investigation was undertaken in L. auricularia from May to November 2010. Of 6759 collected Lymnaeid snails, 370(5.5%) L. auricularia snails were identified. Cercariae infection was found in a number of 276 (74.56 %) snails. The results showed that removed cercariae from L. auricularia belonged to Echinostomcercariae (96.38 %) and Furcocercariae (3.62 %) which were found in two out of 28 sites during the course of study. In summer and fall, the highest prevalence of cercariae infection was recorded for both identified cercariae. It is concluded that L. auricularia could be an important intermediate host of large group digenian trematodes in the region, which is necessary to take consideration in the control program of trematode infection.

Gastrointestinal parasites are among the most common parasitic infections found in stray cats, which might act potential helminthic parasites to domestic cats. The objective of this study was to determine the prevalence of gastrointestinal parasites in stray cats in the city of Azarshahr, which is located in East Azerbaijan province, Iran. A cross—sectional study was conducted on 50 necropsied stray cats, trapped and collected from different geographic regions of Azarshahr. From a total 50 stray cats examined, 15 (30 %) were female and 35 (70 %) were male. Overall 47 cats (94 %) were identified as infected with at least one of the endoparasites. The prevalence of parasites found were: Taenia taeniaeformis (60 %), Dipylidium caninum (58 %), Taenia hydatigera (24 %), Mesocestoides lineatus (78 %), Ancylostoma tubaeforme (14 %), Toxascaris leonina (30 %), Toxocara cati (78 %), Physaloptera praeputialis (10 %), and Syphacia obvelata (10 %). Contamination rate for zoonotic parasites of cat was greater than expected in AzarShahr region. Therefore, appropriate control measures should be taken and preventive methods should be applied.

A total number of 457 Makui sheep were examined because of lameness in one or more limbs. Clinical examination of the affected flocks revealed heavy tick infestation on their limbs. Hosts usually had only one aggregate of ticks on each limb. Attachment of two or more ticks on the pastern caused non-weight bearing lameness in the lambs. Lameness was more prevalent (17%) in lambs than adults (3.5%). Hyalomma anatolicum anatolicum and H. asiaticum asiaticum were the most prevalent species detected on all affected animals. Heavy infestation of the hosts with these ticks could be the main cause of lameness. One aggregate of the ticks caused lameness in lambs. It is likely that considerable tissue damage caused by long mouthparts of the ticks and inflammatory reaction could lead to lameness.

Background: Infection with Ornithobilharzia turkestanicum has been reported in a wide range of animals worldwide. This study was undertaken to assess the util-ity of polymerase chain reaction (PCR), for detecting the infection with O. turke-stanicum larvae stages in Lymnaea gedrosiana. Methods: A total of 6,759 Lymnaeidae snails were collected from six aquatic habitats in West Azarbaijan, northwest Iran. Of these, the snails of L. gedrosiana were identified. To detect infected L. gedrosiana with the larval stages of O. turke-stanicum, they were subjected for cercarial shedding and molecular examinations. The genomic DNA was extracted and PCR was performed to specifically ampli-fy a fragment of the nuclear 28SrRNA gene of O. turkestanicum. Results: Of all collected snails, 5.4% (365/6,759) were the snails of L. gedrosiana. The cercarial shedding method revealed that 23.56% (86/365) of the snails were infected. The PCR patterns confirmed that 28.77% (105/365) snails of L. gedrosiana were infected with larval stages of O. turkestanicum. The infected snails were observed in five studied sites. The highest infection rate (66.66%, 20/30) was recorded in the snails of Ghargologh in the northern part. Only 35.24% (37/105) of the infected snails were from the plain areas, whereas the remaining existed in high altitudes. Conclusion: It was concluded PCR method could be an efficient and fast method for uncovering the actual rate of infection with larval stages of O. turke-stanicum in the snails of L. gedrosiana. This method can be also useful for the do-mestic animals and public health management programs in the country.

Linguatula serrata (Fröhlich, 1789) is a ubiquitous parasite species belonging to pentastomids, which has been reported frequently from domestic herbivores of Iran. This study was carried out to elucidate the pathological changes caused by the nymphal stage of L. serrata in the mesenteric lymph nodes (MLNs) of goats. Jejunal and ileal MLNs of slaughtered goats infected with L. serrata were collected and fixed in 10% neutral formal saline. Histopathological changes included follicular necrosis accompanied by a loss of lymphocytes and oedematous cystlike spaces. Granulomatous lesions were formed by mononuclear cell infiltration and necrotic tissue. Some lymphocytes were apoptotic with chromatolysis. A more frequent vascular abnormality was vasculitis. These findings show that larval migration is the main factor to initiate trauma and cause irritation in the MLNs of goats. Since goats are habitual intermediate hosts in Iran, rigorous inspection of the MLNs of slaughtered goats is recommended in order to decrease the chance of human infection through the consumption of raw visceral organs of goats.

The protozoan intestinal parasite Cryptosporidium commonly infects cattle throughout the world and Iran. The present study was undertaken to determine the abundance and associated risk factors of Cryptosporidium infection in cattle herds of northwestern Iran. A total number of 246 fecal samples from 138 (56.1%) diarrheic (D) and 108 (43.9%) non-diarrheic (ND) cattle were randomly collected and examined by fecal smears stained with Ziehl-Neelsen. For molecular specification, DNA was extracted from collected Cryptosporidium oocysts and a fragment of 1325 bp in size from 18S rRNA gene was amplified. The overall prevalence of Cryptosporidium infection was 22.3% (55/246). The prevalence of Cryptosporidium infection in examined calves less than 6 month-old was significantly higher than adult cattle. C. parvum and C. andersoni were identified in 20.3% (50/246) and 2.03% (5/246) of examined cattle, respectively. The highest prevalence of C. parvum infection was found in D calves < 6 month-old (13.4%, 33/246), while C. andersoni was only detected in ND cattle (8.9%, 22/246). There was significant difference in the prevalence between male than female cattle. There was no significant difference between prevalence and seasons of investigation. It was concluded that C. parvum was the prevalent species in younger animals compared to older ones as a potentially zoonotic agent in the region.

Fasciolosis is an important disease in veterinary medicine worldwide, and is a cause of great economic loss in livestock husbandry in Iran. This study was aimed to determine prevalence of Fasciola gigantica infection in field-collected snails of Radix gedrosiana in northwestern Iran. The snails were collected from 28 perennial and seasonal freshwater habitats from May to December 2010 and identified. A fragment of 618 bp of 28s rRNA gene was amplified by polymerase chain reaction (PCR). The PCR products were subjected to restriction fragment length polymorphism (RFLP) using DraII and AvaII enzymes. PCR-RFLP patterns revealed that 3.12% of the snails were infected with F. gigantica. It was also found that the infected snails had a limited distribution over the water bodies located in the central part of the region. It was concluded that PCR-RFLP was a reliable approach to detect Fasciola infection in pond snails and may be useful to establish control measures for livestock and humans' fasciolosis in the region.

To investigate anti-Neospora caninum antibodies in stray dogs living in Urmia city, 135 blood samples were collected. Serum samples were screened for detection of anti-N. caninum IgG antibodies using indirect fluorescent antibody test (IFAT; ≥50). Antibodies were seen in 36 (27%) of 135 dogs. The IFAT antibody titers were as follows: 1:50 in 16 dogs, 1:100 in ten dogs, 1:200 in six dogs, 1:400 in one dog, 1:800 in two dogs, and 1:1,600 in one dog. There were no significant differences in seroprevalence of Neospora infection between different genders (p > 0.05). The seropositive results were increased with age and the differences were statistically significant (p < 0.05). The results confirm the presence and exposure of stray dogs to N. caninum in Urmia city and the importance of this protozoan as a cause of disease in dogs of the region.

The prevalence of ovine Eimeria parasitism was monitored in 240 sheep in the western part of Iran, in the period from 2003-2004. The overall prevalence of coccidiosis in sheep was found to be 19.2%. The identified species included three pathogenic species, i.e. E. ahsata (10%), E. ovinoidalis (31%) and E. ovina (10%) and three non-pathogenic species, i.e. E. faurei (29%), E. parva (10%) and E. intricata (10%). Of 240 sheep, single infection was not observed and all 19.2% of infected sheep had mixed infections with at least three species. Consistency and intensity had a significant correlation with age. The sex and age of the sheep had a significant effect on prevalence, as well. The results of this study showed that Eimeria infection was prevalent in sheep in the western part of Iran., Promatrana je prevalencija invazije parazitima roda Eimeria u 240 ovaca u zapadnom dijelu Irana u razdoblju od 2003. do 2004. Prevalencija kokcidioze iznosila je 19,2%. Ustanovljene su tri patogene vrste, E. ahsata (10%), E. ovinoidalis (31%) and E. ovina (10%), i tri nepatogene vrste E. faurei (29%), E. Parva (10%) and E. intricata (10%). U svih 19,2% pozitivnih ovaca ustanovljena je mješovita invazija s najmanje tri vrste. Učestalost i jačina invazije bila je u korelaciji s dobi ovaca. Spol je također imao značajan učinak na prevalenciju. Rezultati istraživanja pokazuju da je invazija vrstama roda Eimeria česta u ovaca u zapadnom dijelu Irana.

The present study was conducted to investigate the prevalence of canine gastrointestinal helminths in guardian and herding dogs of Gilanegharb suburb during March 2012 to April 2013. A total number of 138 fecal samples were collected. The overall prevalence of gastrointestinal helminths in the studied animals was 44.92%. The mean of prevalence the gastrointestinal helminths was as follows; Toxocara canis (17.39%), the most prevalent helminths in dogs, Ancylostoma caninum (7.97%), Taenia spp. (10.14%) and mixture of these infections (9.42%). Our findings also indicated that young dogs, under 1 year of age, show a higher overall prevalence than that of older dogs, over 12 months of age (P

An investigation into ectoparasites of sheep and goats was carried out in the northwest region of Iran. One thousand two hundred goats and 1200 sheep in 14 flocks which were less than eight years old were subjected to examination in three sub-divided areas from April 2003 to March 2004. Ectoparasites (ticks, mites, lice, and fleas) were collected from 77 sheep (6.4%) and 119 goats (9.9%), with an overall prevalence of 8.2%. Ticks were the most frequent ectoparasites. Adult ticks (849) were collected and identified: the highest number belonged to the Rhipicephalus bursa (90.7% of sheep and 88.8% of goats), followed by R. sanguineus (6.9%), Boophilus annulatus (2.4%), plus Ornithodoros lahorensis (2.6%). All goats were infested with two species, including R. bursa (88.8%) and R. sanguineus (11.4%). Ixodid tick distributions per animal were 2.5 for sheep and 4.3 for goats. Fifty-two sheep (67.5%) and 85 goats (71.4%) were infested with three species of lice. These were Damalina ovis (58.8%) and D. caprae (71.4%), Haematopinus species (on sheep 76.6%, and on goats 62.2%), Linognathus stenopsis (36.1%), L. ovillus (29.4%). The seasons of highest infestation were fall and winter (50%), the least being spring and summer. Of these, two sheep (2.6%) were infested by Sarcoptes scabiei, maximum infestation occurring in winter. Ctenocephalides felis felis comprised of all the fleas collected (16.8% of goats, and 13% of sheep). Flea infestation was more widespread in fall and winter (10.2%) than in spring and summer (6.1%).