Archives of Razi Institute
Volume:75 Issue: 2, Spring 2020

Coronaviruses (AvCoV) which include infectious bronchitis virus (IBV) and other bird coronaviruses belong to the genus gammacoronavirus, subfamily Coronavirinae. One of the most prominent representatives of gammacoronavirus genus is infectious bronchitis virus (IBV) which is a highly contagious viral pathogen of chickens causing considerable economic losses to the poultry industry. IBVs mostly affect the respiratory, urinary, and reproductive tracts leading to a substantial drop in production. Backyard poultry in the villages usually share their food and water with free flight birds which puts them at serious risk of disease transmission. Furthermore, the poor hygienic measurements which are often used in backyard flocks make them a potential reservoir for diseases that can be transferred to commercial poultry flocks. Live bird markets (LBMs) which receive live poultry to be resold or slaughtered and sold onsite play a significant role in spreading infectious diseases among the different bird species. In the present study, a number of 354 cloacal swab samples were collected from different bird species from LBMs of Gilan province. Subsequently, after RNA extraction, reverse transcription-polymerase chain reaction (RT-PCR) technique was carried out using specific primers of S1 gene to detect coronavirus infectious bronchitis virus. Two samples from backyard chickens were reported to be positive to coronavirus which were named Iran/Backyardchicken96/2017 and Iran/Backyardchicken94/2017. The results of the phylogenetic analysis demonstrated that these two isolates are placed in QX and IS-1494 strains, respectively. On a final note, the obtained results highlighted the role of live birds offered in LBMs in the epidemiology of IBV and the transmission of the virus to the industrial flock.

Infectious bronchitis is one of the most common diseases in the poultry industry in many countries, especially in the regions with a dense poultry farming industry. Gammacoronavirus is the etiologic agent of this disease, with the chickens and poultries as the natural reservoirs of the virus. Various strains of infectious bronchitis virus have been reported in poultry around the world. In terms of pathogenicity, this virus can induce a spectrum of diseases ranging from the moderate respiratory tract to kidney and reproductive diseases. The serotypes of this virus do not cause cross-immunity against each other. This issue makes it difficult to control the disease. Based on the analysis of the highly variable region of the glycoprotein S1 gene, the isolated strains in Iran were classified into seven different phylogenetic groups, including Massachusetts, QX, IS-720, IS-1494, 793/B, IR-1, and IR-2. The D1466 genotype has not been reported in the country; however, the killed vaccine is used in broiler breeder farms. In this study, tissue specimens were collected from 700 farms (i.e., broiler, egg-laying, and broiler breeder farms) suspected of infectious bronchitis within 2013-2017. The samples were examined using real-time reverse transcription-polymerase chain reaction. The D1466 genotype was not detected in any of the studied specimens. Due to the lack of immunity of the D1466 serotype against the dominant types in the country, one has to be careful in choosing the right vaccine. It is necessary to perform continuous monitoring of the circulation status of the various serotypes of viruses in the country to identify the dominant and possible new serotypes for the utilization of the appropriate vaccine.

Bovine subclinical mastitis is regarded as a devastating disease due to the economic costs imposed on dairy husbandry. Moreover, it is a hazard in the public sector in the cases of zoonotic bacteria because of the potential role of unpasteurized milk and dairy products to propagate the infectious agent to the human food chain. The present study aimed to evaluate the frequency, virulence content, and antimicrobial resistance profile of Shiga toxin-producing Escherichia coli (STEC) strains isolated from bovine subclinical mastitis in Kurdistan Province, West of Iran. A total of 400 bovine subclinical mastitis milk samples recognized in the California Mastitis Test were collected aseptically and analyzed for the presence of E. coli phenotypically and molecularly. The isolates were genotypically screened for stx1, stx2, and eae genes. Furthermore, O157:H7 STEC strain was searched among the isolates in a duplex polymerase chain reaction. The antimicrobial resistance scheme of the isolates was determined using the agar disk diffusion method. In general, 173 (43.25%) E. coli isolates were detected among which 39 (22.54%) isolates were STEC. The frequency of STEC virulence genotypes was stx2 (25 isolates, 64.10%), stx2+eae (6 isolates, 15.38%), stx1+stx2 (6 isolates, 15.38%), and stx1+stx2+eae (2 isolates, 5.12%). In addition, three O157: H7 strains were identified with the genetic content of stx1+stx2+eae (2 isolates) and stx1+stx2 (1 isolate). The most prevalent antimicrobial resistance was observed against streptomycin, tetracycline, and ampicillin. Gentamycin, amoxicillin-clavulanic acid, and trimethoprim-sulfadiazine were the most effective antibiotics against O157 strains, whereas gentamycin, ciprofloxacin, and nitrofurantoin were effective against non-O157 strains. The results revealed the significant role of STEC in bovine subclinical mastitis in the studied region. In addition, the distribution of O157:H7 strain and high prevalence of multidrug resistance among the isolates is a matter of concern. Therefore, there is a potential threat of human infection following the consumption of contaminated milk with STEC in Kurdistan Province, Iran.

Bordetellosis or turkey coryza, caused by Bordetella avium, has been an issue for turkey industry since its first description in 1967 when it was reported for the first time. Bordetella avium causes a highly contagious upper respiratory disease in turkeys. Therefore, this study aimed to isolate and characterize this species from commercial and backyard turkeys in Tehran, Isfahan, and Northern provinces of Iran. For the purpose of the study, 625 tracheal swabs were taken from 425 commercial poults and 200 backyard poults aged 2-6 weeks from September 2016 to September 2018. The swabs were immediately plated on MacConkey and blood agar plates and then pooled (5 swabs/pool) in tubes, containing 2 mL distilled water, to perform direct polymerase chain reaction (PCR) for the identification of B. avium. A total of 17 swab pools were found to be positive for B. avium. A subset of seven positive samples were sequenced for the flanking region of piuA gene. The analysis of the sequences indicated that the sequences were 98%, 96%, and 98% similar to B. avium 197N (AM167904.1), 4142 (AY925058.1), and 4156 (AY925068.1) sequences, respectively. To the best of our knowledge, the current study is the first attempt toward the molecular detection and characterization of B. avium in Iran. It is highly recommended to perform further studies to isolate, characterize, and differentiate the regional isolates in order to help the developing turkey industry of Iran meet the increasing demands for protein in the diet of the citizenry.

Human papillomavirus (HPV) has been associated with specific types of papillomas, lesions at particular anatomic sites, and malignancies. The HPV16 and HPV18 have been shown to play a role in a variety of carcinomas. The most documented HPV-associated cancer is cervical carcinoma. Suitable antigens are needed to be identified for the diagnostic tests and vaccines and the expression of L1 recombinant protein should be accelerated in papillomaviruses. Therefore, in this study, the expression of the L1 protein of HPV16 was evaluated and compared in insect cells using a plasmid and a baculovirus system. The expression of the L1 protein of HPV16 in insect cells was investigated using a plasmid (InsectDirect) and a baculovirus system (BacMagic). The expressed recombinant proteins were purified from the Sf9 lysate using Ni-NTA resin columns. The characterization of recombinant L1 protein expressed in both systems (BacMagic and InsectDirect) was performed using immunofluorescence, SDS-PAGE, western blot, and dot blot. The yields of the purified proteins from the plasmid- and baculovirus-based systems (10 ml culture; 107 cells) had the ranges of 455-495 µg/ml and 1.44-1.6 mg/ml as analyzed by spectrophotometer, respectively. The SDS-PAGE analysis of purified proteins revealed that the recombinant proteins with the expected size of 58 KDa were produced in both InsectDirect and baculovirus systems. A high degree (95%) of purification was achieved using this system as observed in SDS-PAGE. The purified L1 protein in the baculovirus system was clearly more efficient than the InsectDirect system. The results of this study indicate that the BacMagic system is an appropriate tool for large scale protein production and provides an alternative to the traditional baculovirus system. In addition, the InsectDirect system might provide a rapid and dependable pointer of whether a protein can be successfully produced in a baculovirus system. Both InsectDirect and BacMagic systems present remarkable savings in cost and time.

Avian influenza (AI) virus (H9N2 and H5 subtypes) infections in birds cause major concerns around the world. The majority of the avian species, such as domestic, pet, and wild birds, are natural and experimental hosts of avian influenza viruses. There are global concerns about members of the Columbidae family, namely pigeons or doves, for their role as the potential interspecies bridge in influenza A viruses ecology. The acquired scientific data in this regard is still not clear since there are doubts about whether or not they transmit viruses between susceptible populations, and spread viruses among farms during outbreaks. To monitor H5 and H9 influenza virus infection status in the rural, backyard, and domestic birds, an annual active surveillance program was performed from September to October 2016. In December 2016, an outbreak of highly pathogenic avian influenza (HPAI) virus subtype H5N8 was detected in a layer farm in Tehran province, Iran. The present research was conducted to study H9N2 or H5 infections in pigeons within HPAI H5N8 2016 outbreaks and annual national AI surveillance in Iran. For this purpose, cloacal swabs and tissue samples (trachea, lung, brain, liver, heart, pancreas, and cecal tonsil) were collected and examined by real-time reverse transcription-polymerase chain reaction (RT-PCR) method and virus isolation. Results of the tests performed on the swab and tissue samples were negative for H5 nor H9N2 viruses. The samples in real-time RT-PCR that after three passages still showed negative results in HA and molecular tests were considered negative. Moreover, the Newcastle disease virus was isolated in most of the samples taken from dead pigeons, after inoculation in embryonated chicken eggs.

Infectious bursal disease (IBD) is a highly contagious disease in young chickens worldwide. The major strategy for the prevention and control of IBD virus (IBDV) is vaccination. Therefore, the present study aimed to compare the immunogenicity of four commercially available IBD vaccines on broilers (Ross 308) that were raised in areas with very virulent IBDV infection history. Two commercial broiler farms with four standard poultry houses were selected in Alborz (n=6,250 birds per house) and Khorasan Razavi (n=8,000 birds per house) provinces of Iran. In each farm, the houses were randomly assigned to one of the four IBD intermediate vaccine brands including Dn, Vc, Ch, and Razi. The birds in Alborz were vaccinated against IBDV via drinking water at 18 and 22; and 15 and 21 days of age in Alborz and Khorasan Razavi flocks, respectively. The enzyme-linked immunosorbent assay antibody titers against IBDV were measured in 20 birds per group at 1, 28, 35, and 42 days of age. In addition, production attributes including body weight, feed conversion ratio, mortality, and production index were measured during the research period. According to the findings, the IBD antibody titers were not affected by the vaccine brands at 28, 35, and 42 days of age (P>0.05). Following the second IBD vaccination, an increasing trend in IBD antibody titers was noted in the Razi vaccine as well as other brands at days 35 and 42 compared to the previously recorded titers (P<0.05). Moreover, the production attributes of the flocks receiving various IBDV vaccine brands were not different (P>0.05). Regarding the productivity indices and high immunogenicity levels, the results indicated that the potential of the IBD Razi vaccine was comparable to the other investigated brands of commercial IBD vaccines, and nominated it as an immunogenic candidate vaccine for use in commercial broilers.

Multifactorial neonatal calf diarrhea (NCD) is one of the main causes of mortality in calves under 1 month of age. Studies have shown that pro-inflammatory cytokines (i.e., interleukin (IL)-1β, IL-6, and tumor necrosis factor-alpha [TNF-α]) can increase in many pathophysiological conditions. In addition, IL-10 as one of the most important anti-inflammatory cytokines contributes to the inhibition of pro-inflammatory cytokines. Few studies have addressed cytokine levels in calves with NCD. Therefore, the goal of this study was to evaluate the plasma concentrations of pro-inflammatory cytokines, as well as IL-10, in the calves naturally infected with NCD syndrome. For this purpose, 87 neonatal calves were monitored for 1 month. Out of this population, 10 cases were diagnosed with NCD and studied as the case group. In addition, a control group was considered that consisted of 10 age- and gender-matched neonatal calves without any diseases. From each calf, 5 ml blood sample was collected into EDTA tubes by jugular venipuncture. The samples were then centrifuged, and the extracted plasmas were aliquoted and stored at -20 °C. Finally, the plasma concentrations of pro-inflammatory cytokines (i.e., IL-1β, IL-6, and TNF-α) and anti-inflammatory cytokine IL-10 were measured using the ELISA kits based on the manufacturer's protocols. The results showed that the plasma concentrations of pro-inflammatory cytokines were significantly higher in the case group than in the control group. However, no significant difference was found between the two groups in terms of the plasma concentrations of IL-10. This study indicates that pro-inflammatory cytokines could be used to recognize the immune system response to NCD.

Clostridium septicum, the anaerobic toxigenic bacterium is the agent that causes dangerous disease in man and animals. There is a lethal toxin of the bacterium namely alpha toxin. The ɑ-toxin has hemolytic, necrotic and lethal activities. Today, Razi Vaccine and Serum Research Institute of Iran produced the C. septicum vaccine in the form of bacterin/toxoid. Because of some problems, the vaccine needs to improve on an industrial scale. The study is going to find an appropriate supplement to improve growth and ɑ-toxin production. Three strains of C. septicum (vaccine, NH1 and NH8 strains) were cultured in the basic vaccine media. Magnesium sulfate, Copper, Ferrous, yeast extract, and trace elements plus vitamins' solution were added to the basic vaccine media in different cultures. The effect of the ingredients on the growth was measured by a spectrophotometer and the α-toxin secretion was assayed by hemolysin test. Growth of the bacterium and α-toxin secretion were increased by Magnesium (80 mg/l) in NH8 and vaccine strains significantly. The black precipitate was difficult to dissolve in magnesium media that must be solved. Trace elements plus vitamins solution mildly influence on NH1strain growth and toxin secretion. Other supplements (Cu, Fe, yeast extract) were not showen any significant changes in the growth and α-toxin production of C. septicum. Overflowing peptone (4%) in the vaccine media, fixes essentials of proteolysis activity, allows the sufficient growth and toxin production without Cu, Fe, and yeast extract. Due to essentially of Mg for growth, extra magnesium was added for improvement of media culture. The study suggests for Magnesium addition in the C. septicum vaccine media during production procedure after precipitation solving problem.

Leishmaniasisis an important tropical cutaneous disease that is endemic in the Middle East, including Iran. There is no consensus on the appropriate therapeutic method, dose, and duration for this disease. The pentavalent antimonial compounds are the first-line treatments of leishmaniasis. With regard to the resistance of this disease against drugs and its treatment failure in some patients, the present study was conducted to investigate the role of alternating magnetic field (AMF) in the treatment of cutaneous leishmaniasis lesionsin mice. To this end, 45 male Balb/c mice with the age of 3-4 weeks and weight of 18-20 g were purchased from the Pasteur Institute, Tehran, Iran, to be investigated. The mice were infected with Leishmania promastigote (2×106) injected in the upper end of the tail.After 3 weeks, the animals were screened for developing lesions. Finally, 15 mice were selected and randomly divided into three equal groups of positive control (treated with the standard drug), negative control (without treatment), and experimental (subjected to AMF at a frequency of 50 kHz for 30 min on a daily basis for 4 weeks). The subjects were followed up for 4 weeks, and the size of their lesions was measured weekly. The data were analyzed by repeated measures test in SPSS software (version 20) at a p-value of < 0.05. There was no significant difference between the experimental and positive control groups (P>0.05). However, the negative control group showed a significant difference with the positive control and experimental groups (P=0.0001). As the findings indicated, AMF was seemingly able to decrease the size of lesions to the same extent as the standard drug. Consequently, AMF could be suggested as a noninvasive and complementary tool against cutaneous leishmaniasis. However, it is required to perform more studies to address different aspects of this domain.

Goat warble-fly infestation is one of the parasitic diseases caused by the larvae of przhevalskiana spp. It is known to be a major challenge for health and welfare in infested goats and causes high economic losses in livestock worldwide. The detection of goat grub was previously conducted by direct palpation of second and third stage larvae in the back and flank site of the live animals or visual evaluation of the carcasses in slaughterhouses. However, due to the small size of the first instar larvae of przhevalskiana (less than 1 mm during emerging from the egg), some of the infected cases were ignored and recorded as negative samples. Immunodiagnostic procedures as easy and cost-effective diagnostic methods provide early detection of myiasis in living animals (even when the larvae are still migrating or are undetectable in the animal body).This study was conducted to evaluate the competitive enzyme-linked immunosorbent assay (ELISA) system in order to detect the antibodies of przhevalskiana larvae in the goat sera. In order to prepare the larval antigen, 200 first instar larvae of przhevalskiana were collected from the subdermal region of the back and flanks of the infested goats in Khoramabad slaughterhouse, Khoramabad, Iran, from September 2017 to March 2018. Totally, 37 and 46 sera samples were taken from the infected and uninfected goats. The sensitivity and specificity at cut-off 3SD were obtained at 89.18% and 84.78%. Moreover, the measures of inter-and intra-assay coefficients of variability to express the precision or replicability of ELISA kit results were 5.33% and 2.81%, respectively.

This study was carried out on seven flocks of ewes suffered from late abortion and neonatal mortality with the prevalence rate of infection reported as 13.95%. The blood and tissue samples were collected from the aborted ewes in several flocks of Duhok province, Kurdistan Region, Iraq. Serological analysis indicated that all the aborted ewes were confirmed positive for agglutination to Toxoplasma gondii (T. gondii)antibody. The investigation of the aborted fetuses showed the blood-stained fluid in the thoracic and abdominal cavity. Most of the aborted fetuses had also enlarged, congested, and friable livers and lungs. The placenta was swollen, reddish, and friable, and its cotyledons also spotted with whitish foci. T. gondii tachyzoites were also demonstrated in the placental sections of some aborted ewes. Severe congestion, necrosis, and infiltration of multinucleated cells were the most predominant histopathological changes of the aborted fetuses, as well as presented tissue cysts, tachyzoites, and bradyzoites in the liver, brain, heart, and lung. There were also several clusters of dark purple banana-shaped T. gondii tachyzoites within the brain and heart tissues in most of the examined aborted fetuses in different flocks. T. gondii tachyzoites were also detected from the peritoneal ascites of mice inoculated experimentally 12 days following the infection. Moreover, T. gondii tissue cysts were detected from the impression smears of the mice brains 32 days after the infection. Accordingly, the demonstration of T. gondii in Giemsa-stained impression smears associated with characteristic histopathological changes of different organs is a great fundamental method for the diagnosis of T. gondii in aborted cases.

Peripheral nerve disorders are the most common neurological problems; therefore, it is important to intervene to treat or stop the resulting side effects. This study aimed to investigate the effect of oat extract on experimental sciatic nerve injury in rats. Totally, 50 adult male rats were divided into five groups (n=10). Group 1 was exposed to sham condition, and group 2 was regarded as the control group (nerve injury without treatment). Moreover, groups 3-5 were subjected to sciatic nerve injury, and they received oral gavages of the oat extract (100, 200, and 400 mg/kg), respectively. Subsequently, 2 and 4 weeks later, the rats were euthanized for pathological evaluation of nerve repair. The results showed an increase in the formation of the perineurium and epineurium dose in the oat-treated groups (100, 200, and 400 mg/kg), compared to the control group after 2 weeks (P<0.05). Furthermore, the presence of inflammatory cells in the oat extract-treated groups (100, 200, and 400 mg/kg) decreased, compared to that in the control group after 2 weeks (P<0.05). In addition, the swelling of the axon significantly decreased in the oat extract-treated groups (200 and 400 mg/kg), compared to the control group (P<0.05). However, the axon dose-dependently increased in oat-treated groups (100, 200, and 400 mg/kg), compared to that in the control group after 4 weeks (P<0.05). These results suggest that oat extract has positive effects on sciatic nerve repair in rats.

This study investigated a person with an AB0 discrepancy. Her blood group initially typed at the birth as AB Rh+ (positive); however, it was B Rh+ (positive) or Rh- (negative) when she was in her teens. At room temperature, her erythrocytes were agglutinated by anti-B, and the agglutination was significantly weaker at 37 ºC. As a result, her erythrocytes did not absorb anti-B but anti-A. Furthermore, her erythrocytes were agglutinated by anti-A at 37 ºC with signs of hemolysis in the presence of complement. The unwashed erythrocytes were also agglutinated in an antiglobulin test by polyclonal anti-A at 37 ºC and by heated polyclonal anti-A and anti-A MAB 2-8 at room temperature. Moreover, her serum agglutinated A erythrocytes at room temperature with less activity at 37 ºC; however, it agglutinated B erythrocytes at 37 ºC. The ability of the erythrocytes of this person to absorb anti-A came along with the agglutination of her erythrocytes at 37 ºC by polyclonal serum and decreased activity of the serum to agglutinate A erythrocytes at 37 ºC, compared to room temperature. The absence of anti-B absorbance by the person’s erythrocytes was accompanied by the presence of anti-B in the serum, which was active at 37 ºC. The incubation of the person’s serum with 0 erythrocytes induced the ability of erythrocytes to absorb anti-A and to be hemolyzed by anti-A in the presence of complement in accordance with the person’s characteristics of erythrocytes. The reaction of absorption and agglutination at room temperature and 37 ºC by heated serum with the use of complement may help to reveal both weak A and B antigens and anti-A and anti-B antibodies while AB0 blood typing.

Neospora caninum, a protozoan parasite, causes abortions in cattle, as well as neurological disorders and reproductive problems in dogs. This study aimed to investigate the serological and the molecular prevalence of N.caninum among foxes and dogs using indirect fluorescent antibody test (IFAT) and nested-polymerase chain reaction (PCR). For this purpose, 288 and 95 both fecal and serum samples of dogs and foxes were collected, respectively, from around industrial and traditional dairy flocks in different parts of Sanandaj, Kurdistan Province, Iran, from 2013 to 2015. The sera were examined using IFAT, and fecal samples were microscopically assessed for detecting Neospora oocyst and by nested-PCR. The findings revealed that N.caninum seroprevalence were 4.86% and 4.21% in dogs and foxes, respectively. In addition, no Neospora oocysts were found microscopically and by PCR. Since this study is the first serological and molecular investigation of N.caninum among both dogs and foxes in Sanandaj,  the findings of indicated that stray dogs is a main source of N.caninum infection in dairy farms in Sanandaj, Iran.

Some of the medicinal plants have antibacterial contents and appear to be proper alternatives for antibiotics in the treatment of streptococcal disease, which causes plenty of mortalities in fish farms annually. Therefore, this study investigated the therapeutic effect of Aloe vera and Salvia officinalis hydroethanolic extracts against Streptococcus iniae in rainbow trout. Plant extracts components were analyzed by Gas chromatography-mass spectrometry method and tested in vitro against S. iniae by disk diffusion assay. In in vivo, 480 rainbow trout (10±0.1 g) were distributed in 9 groups (with 3 replication), and all groups (except for the first group as the negative control that was injected with 100 µl of physiologic serum) were injected by 100 µl of LD50 (3.66×108.5CFU/ml) of S. iniae suspension, intraperitoneally. The fish of groups were treated by A. vera and S. officinalis extracts in doses 0 (positive control group was fed by commercial diet without plant extract), 0.5, 1, and 1.5% (supplemented diet) and 80 mg/kg body weight erythromycin for the next 10 days. At the end of the study period, tissue samples of the gills and livers of all groups were taken for the histopathological lesion assay. The results showed that A. vera and S. officinalis had antibacterial components as Cineol, and S. iniae was sensitive to both A. vera (MBC=4.067 mg/ml) and S. officinalis (MBC=5.185 mg/ml) extracts. At the end of the treatment period, there were no significant differences among erythromycin, A. vera (1.5%), and A. vera (1%) in terms of the mortality of the infected fish (P˂0.05). Moreover, A. vera (1.5%) showed a significantly lower mortality rate, compared to the positive control (P˂0.05). A. vera (1.5%) was the best group to moderate all histopathological lesions, compared to other groups. Accordingly, A. vera (1.5 %) is useful to treat streptococcosis (caused by S. iniae) and alter gill and liver histopathological lesions in rainbow trout.

The purpose of the present study was to investigate the Theileria and Babesia infection in sheep using polymerase chain reaction (PCR) assay in Baneh, Iran. Theileria and Babesia are apicomplexan parasites that have both vertebral and invertebrate hosts. These protozoa, which are transmitted by tick vectors, are considered to be the most important causes of parasitic diseases in Iran.The detection methods of Babesia and Theileria spp. are morphological examination, serology tests, and more recently, molecular assays, such as PCR.  In this study, a total of 66 blood samples were collected and analyzed using specific primers for Theileria annulata, T. ovis, T. lestoquardi, and Babesia ovis. Two PCR methods were used, namely semi-nested PCR and competitive PCR. Based on the results of the PCR assay of 66 sheep blood samples, B. ovis, T. ovis, T. lestoquardi, and T. annulata were detected in 57 (86.4%), 28 (42.4%), 0, and 16 (24%) cases, respectively. Detection of low levels of protozoan infection with high morbidity in the tested animals shows their status as a carrier that keeps the infection in the region and extends the protozoan life cycle. Another important factor is the geographical situation of Baneh as a border city since the hemoprotozoan infection is present in this region. Moreover, piroplasmida infection was found in Iraq and other neighboring provinces. Therefore, animal husbandry in Baneh is at the risk of infection with Babesia and Theileria. The collected data in this study are useful for reaching a better understanding of the epizootiology of theileriosis and babesiosis, in order to control and prevent the diseases in this region.