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The establishment of good experimental models for Theileria sp. infection is important for theileriosis research. Routinely, infection of ticks is accomplished by feeding on parasite-infected animals (sheep, cows and horses), which raises practical and ethical problems, driving the search for alternative methods of tick infection. Artificial tick feeding systems are based mainly on rearing ticks on host-derived or hand-made artificial membranes. We developed a modified feeding assay for infecting nymphal stages of Hyalomma anatolicum ticks with Theileria lestoquardi, a highly pathogenic parasite of sheep. We compared two different membranes: an artificial silicone membrane and a natural alternative using mouse skin. We observed high attachment rates with mouse skin, whereas in vitro feeding of H. anatolicum nymphs on silicone membranes was unsuccessful. We could infect H. anatolicum nymphs with T. lestoquardi and the emerging adult ticks transmitted infective parasites to sheep. In contrast, similar infections with Rhipicephalus bursa, a representative tick with short mouth-parts that was proposed as a vector for T. lestoquardi, appeared not to be a competent vector tick species. This is the first report of an experimentally controlled infection of H. anatolicum with T. lestoquardi and opens avenues to explore tick-parasite dynamics in detail.

Background Cats as a definitive host have an important role in the epidemiology of toxoplasmosis in humans and animals. The aim of the study was to determine the frequency of Toxoplasma gondii infection and isolate and identify the genotypes of T. gondii in stray cats in the Mashhad suburb. Methods From April 2016 to August 2017, 175 fecal samples from stray cats and 31 brain samples from cats killed in driving accidents were collected. The fecal samples were examined by fecal flotation technique and T. gondii -specific PCR. The brain samples were investigated by T. gondii -specific PCR and consequently examined by mice bioassay. The DNA of T. gondii isolated was genotyped using SAG1, SAG2, SAG3, BTUB and GRA6 as PCR-restriction fragment length polymorphism (PCR-RFLP) markers. Results In the present study, Toxoplasma -like oocysts were microscopically observed in 2.2% (4/175) fecal samples. The presence of Toxoplasma oocysts was confirmed in one microscopy-positive sample by PCR. In addition, T . gondii DNA was detected in 4% (7/175) microscopy-negative samples using PCR. T. gondii was isolated from one brain PCR-positive sample by mice bioassay. The isolate was avirulent and many T. gondii cysts were observed in mice brain. The isolate was successfully genotyped by PCR-RLFP analysis. The isolated genotyped was type II. Besides, eight Toxoplasma- positive fecal samples contained insufficient DNA and only amplified at SAG-3 locus in PCR. These samples were also showed type II pattern at this locus. Conclusions Parasitological and molecular results showed low frequency of Toxoplasma infection in the stray cats, and identified the genotype of T. gondii isolate as type II, for the first time in Mashhad area, Khorasan Razavi Province.

Background Neospora caninum is an obligate intracellular protozoan that is well established as a causative agent of abortion in dairy cattle worldwide. Objectives The objective of this study was to determine the role of N. caninum infection in the abortion in dairy cattle in the Khorasan Razavi Province, Iran. Methods From 2022 to 2024, 105 aborted bovine foetuses were collected from dairy cattle in Khorasan Razavi province. Brain samples of aborted foetuses were tested using nested PCR and histopathological examination. In addition, blood samples were collected from dairy cattle that had aborted PCR‐positive foetuses and were analysed using enzyme‐linked immunosorbent assay (ELISA). Results and Conclusions In the present study, N. caninum infection was detected in 24.76% (26 out of 105) of aborted bovine foetuses by nested PCR analysis. The brain tissues of 20 bovine‐aborted foetuses were only suitable for histopathological examination. Lesions of the central nervous system were severe hyperaemia, perivascular cuffing, astrogliosis, mild encephalitis and focal necrosis. One foetus exhibited a 32‐µm N. caninum cyst within the brain tissue. IgG antibodies against N. caninum were identified in all dairy cattle that aborted infected foetuses through ELISA testing. Molecular, histopathological and serological findings strongly suggest that N. caninum plays a significant role in bovine abortion in dairy cattle in Khorasan Razavi Province, northeast Iran. Aborted bovine foetuses were collected from dairy cattle in Khorasan Razavi province. Brain samples of aborted foetuses were tested to detect N.caninum infection using nested PCR and histopathological examination. In addition, blood samples were collected from dairy cattle that had aborted PCR‐positive foetuses and were analysed using ELISA .

The health, growth and fertility of avian species can be negatively affected by parasite infection. This survey assesses the presence, variety and distribution of internal and external parasites among parrots and perching birds in Mashhad, Iran. This study examined 751 caged pet birds from different species and regions in Mashhad for faecal samples and 132 oral swabs for digestive tract parasites. Furthermore, skin scrapings were conducted on 14 canaries displaying dishevelled feathers. During the study, mortalities and moribund birds that had been referred underwent necropsies to examine internal parasites. Following the formol ether faecal examination, only one Malango parrot tested positive for Heterakoidea eggs among 751 faecal samples (0.13%). Further, one cockatiel showed evidence of parasitic infection with Eimeria spp. (0.13%). However, neither Cryptosporidium nor Giardia protozoa were detected in the samples (0%). Oral swabs revealed no evidence of Trichomonas (0%). On the other hand, skin scraping revealed the presence of the mite Dermanyssus in 7 out of 14 canaries with dishevelled feathers (50%). Of 25 moribund and weak budgerigars, 2 were infected with Acuaria in their proventriculus (8%). In addition, 3 out of 14 deceased myna birds carried the nematode Diplotriana in their coelomic cavities (21.42%). In conclusion, the rate of internal parasites has been relatively low in ornamental birds of Mashhad, whereas the prevalence of external parasites has been higher. This survey in Mashhad, Iran, explored the presence of gastrointestinal and external parasites in 751 ornamental birds. The results indicated that although gastrointestinal parasites were relatively uncommon, the prevalence of external parasites, such as mites in canaries and nematodes in some other species, was notably higher.

Malignant ovine theileriosis is caused by Theileria lestoquardi, which is highly pathogenic in sheep. Theileriosis involves different organs in ruminants. Little is known about the role of proinflammatory cytokines in the pathogenesis of T. lestoquardi infection. The aim of this study was to measure concentration changes of proinflammatory cytokines and immunoglobulin G (IgG) during an ovine experimental theileriosis and correlate it with clinical and haematological parameters. During an experimental study, seven healthy Baluchi sheep (four females and three males) about 6–8 months old were infected with T. lestoquardi by feeding of infected unfed ticks on the sheep’s ears. The infected sheep were clinically examined during the study and blood samples were collected on days 0, 2, 5, 7, 10, 12, 14, 17 and 21. The haematological parameters were analysed by an automatic veterinary haematology cell counter and the inflammatory cytokines interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and IgG were measured by enzyme-linked immunosorbent assay. All infected sheep had temperatures above 40 °C on days 3–4 post infection (PI). The maximum temperature was noted on day 7, and it remained high until day 21. The parasitaemia of T. lestoquardi infection increased from 0.01% (day 7 PI) to 3.3% (day 21 PI). The mean white blood cell (WBC), red blood cell (RBC), lymphocyte, neutrophil and platelet values slightly increased on day 2 PI and decreased by day 17 and day 21 PI. The percentage parasitaemia and fever had a negative correlation with the numbers of WBCs, RBCs, lymphocytes, neutrophils and platelets. The serum concentration of IL-6, TNF-α and IFN-γ cytokines increased and peaked on day 12 and thereafter decreased to levels lower than 0. Out of all tested cytokines, the concentration of IL-6 was significantly higher, as early as day 2 PI. No significant changes were observed for the IgG levels during the course of disease. A significant and strong correlation was observed between IL-6, TNF-α and IFN-γ values and a moderate correlation between IL-6 and the numbers of lymphocytes in the present study. A strong correlation was determined between the percentage parasitaemia and haematological parameters in T. lestoquardi-infected sheep. In addition, preliminary results indicate that the measurement of the serum concentrations of IL-6 in combination with haematological parameters could be considered a good marker to estimate the pathogenicity of T. lestoquardi strain.

A molecular study was undertaken to detect Theileria ovis, Theileria lestoquardi and Theileria annulatain sheep and tick vectors. Investigation was conducted from 2010 to 2011 in the south of Khorasan Razavi Province, Iran. A total of 150 blood samples were collected from 30 different sheep flocks. In addition, ixodid ticks were sampled from the same flocks. The stained blood smears were microscopically examined for the presence of piroplasms and a semi-nested polymerase chain reaction-restriction (PCR) was used for subsequent molecular speciation. Salivary glands were isolated from the ticks and subsequently analysed by semi-nested PCR. polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to differentiate between T. lestoquardi and T. annulata from PCR-positive samples. Theileria species infection was microscopically detected in 18.6% of blood smears. The presence of T. ovis and T. lestoquardi or T. annulata was detected by semi-nested PCR in 58.6% and 6.6% of blood samples respectively. In total, 169 ixodid ticks were collected from different areas of the province. The most prevalent ticks were Rhipicephalus turanicus (n = 155; 91.7% of the total), followed by Hyalomma anatolicum anatolicum (n = 8; 4.7%) and Hyalomma marginatum turanicum (n = 6; 3.5%). From an organ pooling of 33 ticks, three pools of salivary glands from R. turanicus were positive for Theileria species by semi-nested PCR. Of the three R. turanicus samples testing positive for Theileria species, two (6.1%) were positive for T. ovis and one (3.0%) for T. lestoquardi or T. annulata. Amongst the 11 PCR-positive samples for T. lestoquardi or T. annulata, 10 were positive for T. lestoquardi and one sample was positive for both T. lestoquardi and T. annulata using PCR-RFLP. The results also demonstrated that PCR-RFLP could be used for the detection of T. ovis. Based on the results, it can be concluded that T. ovis has a higher prevalence than T. lestoquardi, and that R. turanicus could be a possible vector for T. ovis and T. lestoquardi. Finally, the PCR-RFLP based on Msp1 restriction enzyme is a simple method for differentiation of Theileria species in sheep and ixodid ticks.

The malignant ovine theileriosis due to Theileria lestoquardi is highly pathogenic in sheep. During an experimental study, clinical and pathological findings were investigated in sheep infected with T. lestoquardi . The common clinical signs were high fever, emaciation, enlarged lymph nodes, anemia, icterus, nasal discharge, cough, severe pneumonia, circling, opisthotonus, recumbency, and death. Schizonts of Theileria were detected in impression smears of the lymph nodes, spleen, and liver. Gross lesions include pale mucous membranes, enlarged lymph nodes, splenomegaly, and hepatomegaly with gall bladder distention. Petechial and ecchymotic hemorrhages were observed on most of the surface of the serous membranes of the small and large intestines. In some cases, small ulcers were also present in the mucosal membrane of the abomasum. The lungs were congested and edematous, and petechial and ecchymotic hemorrhages were presented. The main microscopic lesions were characterized by lymphocytic infiltration with edema, congestion, and hemorrhage in many tissue sections. It was concluded that T. lestoquardi is highly pathogenic in sheep and the clinical signs and pathological lesions could be associated with a marked lymphoproliferative reaction in the different organs.

Dogs serve as definitive or intermediate hosts for some protozoan and helminth parasites, some of which are of zoonotic, and others cause severe infectious diseases in other animals. The aim of this study was to determine the prevalence of intestinal parasites in shelter dogs in Khorasan Razavi province, Iran. A total of 323 fecal samples were randomly collected from shelter dogs in Mashhad and Neyshabur areas. All samples were then transferred to the Parasitology Laboratory and examined detect intestinal protozoa and helminths using the formalin-ether technique. Out of the 323 fecal samples, 92 (28.48%) tested positive for intestinal parasites. The overall prevalence of intestinal parasites in shelter dogs in Mashhad and Neyshabur areas were 23.83 and 35.38%, respectively. Monospecific infection was found in 19.50% of dogs, while concurrent infection with two or more species was seen in 8.97% of dogs. The frequencies of the nine detected parasites were as follows: Sarcocystis sp. 12.38%, Giardia sp. 4.64%, Cystoisospora sp. 2.78%, Hammondia Neosporalike oocysts 2.16%, Taenia sp. 6.19%, Toxascaris leonina 7.43%, Dicrocoelium dendriticum 1.54%, Toxocara canis 0.92% and Capillaria sp. 0.30%. The results of this study showed the high frequency of intestinal parasites in shelter dogs. To control important zoonotic helminthic diseases such as echinococcosis and toxocariasis, it is recommended to eliminate stray dogs in cooperation with municipalities and veterinary departments in Iran. If stray dogs are transferred to shelters, they should receive regular treatment with effective anti-helminthic drugs.

[...]hydrogen peroxide and tetramethyl ben- zidine (TMB) were added and the plates were incubated for 15 min at room temperature. In addition to this, CCHF was first discovered in Crimea (located in the northwest of Iran)15. Because CCHF is a serious threat to Iran, im- ported animals, particularly camels, that carry a large number of ticks, should be screened and treated more carefully.

This comprehensive study was conducted on multi-purpose one-humped camel (Camelus dromedarius) sera and ticks to assess the epidemiological aspects of the Crimean-Congo hemorrhagic fever virus (CCHFV) in northeast Iran. From May 2012 to January 2013, eleven cities were randomly selected in the Khorasan Provinces of Iran as \"clusters,\" and at least 14 one-humped camels were sampled from each area. Reverse transcriptase polymerase chain reaction was used for the detection of the CCHFV genome in ticks. Sera were analyzed using specific enzyme-linked immunosorbent assay tests. Four hundred and eighty ixodid ticks were collected, and the genome of the CCHFV was detected in 49 (10.2%) out of 480 ticks. The CCHFV genome was detected in two out of four tick species, and in tick samples from three cities in Khorassan-e-Jonoobi. All three provinces, and six out of eleven cities, were CCHFV-specific IgG-positive. In total, nine (5.3%) out of 170 one-humped camels were IgG-positive. The highest rate of IgG-positive samples was found in Nehbandan (16.67%). Continued surveillance and strictly enforced importation and quarantine practices should be implemented to prevent human exposure and the on-going dispersal of infected ticks and livestock in these regions. It is recommended that acaricides be used to prevent CCHF transmission to humans, and to reduce the tick population. In addition, care should be taken by abattoirs workers and people who work with one-humped camels.