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Anaplasmosis, caused by intracellular gram-negative bacteria Anaplasma marginale is one of the most frequently reported tick-borne disease (TBDs) in tropical and sub-tropical countries, including Pakistan. In the present study, a total of 428 cattle blood samples were collected to examine the prevalence and phylogenetic origin of A. marginale in two important livestock regions of Punjab Province in Pakistan, i.e. Lodhran and Dera Ghazi Khan Districts. In addition, association between occurrence of A. marginale in cattle blood and selected epidemiological factors has been also investigated. The presence of A. marginale genetic material was confirmed in 9% of the tested blood samples taken from cattle in Lodhran and in 17% from Dera Ghazi Khan. Prevalence of A. marginale was significantly higher in cattle from Dera Ghazi Khan. All the cattle breeds from both districts were equally susceptible to A. marginale infection. We reported higher prevalence of A. marginale in cattle living indoors or with other dairy animals in Dera Ghazi Khan district. However, no such relationship was observed in the Lodhran district. Sequencing of the msp1b gene shows 96-99% similarity of A. marginale in the study area to those reported from other parts of Pakistan, South Africa, and Israel. We recommend that large scale tick and tick-borne disease control strategies must be implemented in both districts.

Anaplasma centrale (A. centrale) is an obligate red blood cell residing tick transmitted rickettsiae that has not been studied extensively for its prevalence in cattle along with its epidemiology. Aim of this investigation was to report the seasonal prevalence, phylogeny and epidemiological parameters associated with the prevalence of A. centrale in cattle breeds enrolled from District Layyah in Punjab, Pakistan. A total of 844 blood samples [Cross breed = 300, Holstein Friesian = 244, Sahiwal breed = 300)] were collected from apparently healthy cattle along with epidemiological data during 2017–18. PCR amplified 426 base pair fragment from 16S rRNA gene of A. centrale in 14.4% (122/844) of cattle. Amplified 16S rRNA partial gene sequence of A. centrale were confirmed by DNA sequencing and deposited to GenBank. Highest A. centrale prevalence was observed in spring (24%) followed by autumn (12.4%) summer (10%) and winter (7.1%) seasons. Sahiwal breed (18.3%) was most susceptible to A. centrale infection followed by cross (12.3%) and Holstein Friesian breed (12.3%). 69/844 (8.2%) of Giemsa stained cattle blood smears were also found positive for Anaplasma spp. Farms where animal use to drink pool water and farms where dogs and other dairy animals were living with cattle had higher A. centrale prevalence. Female cattle and dogs having tick burden were found associated with A. centrale infection. Hematological profile was severely disturbed in A. centrale positive cattle. It is recommended that A. centrale should be screened in cattle, in addition to A. marginale, for the effective control of tick born diseases in Pakistan.

Anaplasma marginale, Theileria ovis and Theileria lestoquardi are intracellular pathogens that infect a wide variety of animals and cause enormous economic losses worldwide. The present investigation aims to report the occurrence and the phylogeny of these pathogens infecting sheep (N = 330) from Rajanpur District in Punjab (Pakistan) by using msp1b gene for A. marginale and 18 S rRNA for T. ovis and T. lestoquardi. Results revealed that prevalence rates of A. marginale, T. ovis and T. lestoquardi were 7.3%, 6.1% and 1.2%, respectively. Four (1.2%) and one (0.3%) sheep were found to be co-infected with two and three pathogens, respectively. Risk factor analysis revealed that rams were more infected with A. marginale compared to ewes (P = 0.015). In addition, it was observed that animals located in herds having dogs and those in small herds were found more infected, respectively, with T. ovis (P = 0.001) and T. lestoquardi (P = 0.009). Phylogenetic analysis revealed that Pakistani isolates of each detected pathogens clustered together and were closely related to other Pakistani isolates and those from worldwide countries such as Iran, Iraq, Turkey, Sudan, Tanzania, South Africa, and USA. This is the first molecular study reporting the natural infection of Pakistani sheep with these three pathogens. These data need to be taken into account in order to improve the productivity of the livestock sector which is one of main sources of income in the country.

Raising small ruminants is the main source of income for farmers in Pakistan. Economic losses caused by Toxoplasma gondii to small ruminants have been reported worldwide, however reports on molecular detection of T. gondii are lacking in Pakistan despite a large goat population. The current study was carried out from March 2019 till February 2020 to report the seasonal and molecular prevalence of T. gondii in different breeds of goats located in Khanewal district of Punjab, Pakistan. A total of 898 blood goat samples were collected during the four seasons and screened for T. gondii DNA by using PCR based on the amplification of ITS-1 partial sequence. Out of 898 goats, 48 (5.3%) were found positive to T. gondii. The prevalence of T. gondii varied according to season (Chi square test,P = 0.016) and the highest prevalence was observed in goats tested during the summer (8.8%) followed by the spring (5.7%), the winter (4.4%) and the autumn season (2.2%). PCR products positive to T. gondii were confirmed by DNA sequencing and BLAST analysis. Phylogenetic study based on ITS 1 gene provided evidence that the amplified isolates of T. gondii were highly conserved in Pakistani goats. Buck (Fischer exact test, P = 0.002) and farms containing other dairy animals next to goats (Fischer exact test, P = 0.001) and farms with a water supply from pools (Fischer exact test, P = 0.001) were more infected with T. gondii. Infected goats had a reduction on red blood cell count (Two-sample t test, P = 0.01) and hemoglobin concentration (Two-sample t test, P = 0.03) and an increase in the number of monocytes (%) (Two-sample t test, P = 0.05), mean cell hemoglobin (Two-sample t test, P = 0.01) and serum creatinine (Two-sample t test, P = 0.01) as compared to T. gondii uninfected goats. In conclusion, we report a relatively low PCR based prevalence of T. gondii in goats from Khanewal district as previously the serum ELISA test based prevalence of T. gondii in Pakistani goats varied between 19-52%. Control measures should be taken to eradicate T. gondii infection in goats of the study area.

During early lactation, dairy cows may present a transient immunosuppressive state and develop anaplasmosis caused by Anaplasma marginale . In this study, clinical anaplasmosis in dairy cattle in the Thrace region of Turkey was investigated with respect to within-herd prevalence, vertical transmission, and genetic diversity. In March and September 2015, thirty lactating cows showed primary clinical signs of anaplasmosis, including fever, anaemia, decreased milk yield, anorexia, and laboured breathing. Symptoms disappeared in most cows after administration of long-acting oxytetracycline, but nine of them (30%) died. Following diagnosis based on clinical signs, microscopy and molecular findings, blood samples were collected from apparently healthy lactating cows (n = 184), pregnant heifers (n = 39) and newborn calves (n = 24). DNA was extracted from each sample and analyzed for the presence of major surface proteins (MSPs) of A. marginale , followed by sequencing to assess diversity of isolates. Microscopic examination of erythrocytes revealed A. marginale inclusion bodies in symptomatic cows. Examination of thin blood smears showed 3.8% of the lactating, clinically asymptomatic, cows to be infected with A. marginale , while nPCR detected 31.0% positive. A. marginale infection was not detected in pregnant heifers by either method. Congenital infection was found in one calf by nPCR. This is the first report of transplacental transmission of A. marginale in Turkey. The MSP4 sequence analyses showed high genetic diversity among the isolates, presenting 97.6-99.6% homology at the amino acid level. The sequences of MSP1a amplicons revealed genetic diversity providing three new tandem repeats.

Major surface protein 1 alpha (Msp1α) is a stable genetic marker for identifying Anaplasma marginale. Our aims were to investigate the genetic diversity of A. marginale based on microsatellites and tandem repeats of the msp1α gene and to determine the phylogenetic relationships among six isolates obtained from 63 dairy cows positive for the pathogen from Turkey and from strains worldwide. Msp1α microsatellite analysis revealed infection with three genotypes (B, C, G). Genotype B was found in cows held in an intensive management system, while genotypes C and G were found in grazing cows. Microsatellite sequences produced SD-ATG distances of 19 and 23 nucleotides, with 23 nucleotides predominating, showing high infection potential. Fourteen different tandem repeats of A. marginale were found in the samples, and four msp1α gene repeats designated Tr4, Tr5, Tr6 and Tr7 were newly described. The majority of A. marginale isolates exhibited more than three Msp1a tandem repeats, with a maximum of seven. The Msp1a tandem repeats α-73-73-73-73-73-73 were commonly observed in intensively managed cows. The phylogenetic analysis revealed A. marginale strains from Turkey clustering with those of the Americas, Africa, Asia and Europe, suggesting a history of cattle trade and transport.

Theileriosis is a widespread and economically important disease of small ruminants in Pakistan. Ruminants are the intermediate hosts in the lifecycle of Theileria spp., with ticks of the family Ixodidae being the definitive hosts. To better understand the distribution and prevalence of theileriosis in Pakistan, a molecular survey was performed in small ruminants from the Lower Dir district of the Khyber Pakhtunkhwa province. A total of 200 healthy sheep and goats were screened from Maidan, Samar Bagh and Munda districts of district Dir Lower, Pakistan during December (2017) to April (2018). DNA samples were screened through nested PCR using universal primers. The amplified 492–498 bp amplicon was subjected to RLB analysis which was based on the hypervariable of the 18S rRNA gene to test for the presence of genotypes of Theileria in blood samples. A phylogeny was constructed to determine the species of Theileria genotypes. Nested PCR results indicated 53.5% prevalence of one or more Theileria genotypes in the blood of the host animal. From RLB assay, 27 animals (13.5%) showed infection with only a single species of Theileria while 80 animals (40%) showed coinfection by multiple Theileria spp. Based on the 18S rRNA phylogeny, the unknown genotype is of the species Theileria luwenshuni and is closely related to Chinese isolates. The present finding is the first report on molecular diagnosis of Theileria luwenshuni in small ruminants in Pakistan.

Anaplasma species are obligate intracellular rickettsial pathogens that affect the health of humans and other animals. Clinical cases of anaplasmosis caused by Anaplasma phagocytophilum were evaluated, and the frequency of bovine Anaplasma species was determined in cattle. Blood samples and thin blood smears were collected from 10 cattle exhibiting clinical signs of tick-borne fever. In addition, blood samples were collected from 123 apparently healthy cattle from the same area. DNA was screened by reverse line blot assay for the presence of the hypervariable V1 region of the 16S rRNA gene of Anaplasma/Ehrlichia species. Intracytoplasmic inclusion bodies of A. phagocytophilum were observed in neutrophils of 6 sick animals. Parasitemia ranged from 0.2 to 1.6% in individual slides. Reverse line blot showed 45.1% (60/133) of the sampled cattle to be positive for one or more of five Anaplasma species. The frequency of single infections was 20.3% (27/133), while mixed infections were found in 24.8% (33/133) of samples with six different combinations of species and a maximum of four pathogens detected. A. phagocytophilum was the most prevalent (41/133, 30.8%) followed by Anaplasma marginale (25/133, 18.8%), Anaplasma centrale (24/133, 18%), Ehrlichia sp. strain Omatjenne (18/133, 13.5%) and Anaplasma bovis (1/133, 0.7%). This is the first report of A. bovis in a cow from Turkey. This is also the first report of clinical cases caused by A. phagocytophilum in cattle from the country. Therefore, A. phagocytophilum should be taken into account as differential diagnosis in cases of high fever and anorexia in pastured animals.

This study was undertaken in two different climatic areas of Turkey to determine the presence of tick-borne pathogens of medical and veterinary importance. The ticks were removed from humans, pooled according to species and developmental stages, and analyzed by PCR, reverse line blot (RLB) and sequencing. Of the 2333 removed ticks from 10 species, 1238 (53.06%) were obtained from the arid cold zone, and the remaining 1095 (46.93%) were obtained from the humid zone. The removed ticks were identified as Hyalomma marginatum, Hyalomma detritum, Hyalomma excavatum, Rhipicephalus bursa, Rhipicephalus turanicus, Rhipicephalus sanguineus, Dermacentor marginatus, Haemaphysalis punctata, Haemaphysalis sulcata, Ixodes ricinus, Haemaphysalis and Ixodes spp. nymphs. The dominant species was L ricinus (61.27%) in the humid zone, whereas the Haemaphysalis spp. nymph dominated (30.29%) in the arid zone. Infection rates were calculated as the maximum likelihood estimation (MLE) with 95% confidence intervals (CI). Of the 169 pools tested, 49 (28.99%) were found to be infected with the pathogens, and the overall MLE of the infection rate was calculated as 2.44% (CI 1.88-3.17). The MLE of the infection varied among tick species, ranging from 0.85% (CI 0.23-2.34) in Haemaphysalis spp. nymph to 17.93% (CI 6.94-37.91) in D. marginatus. Pathogens identified in ticks included Theileria annulata, Babesia ovis, Babesia crassa, Anaplasma/Ehrlichia spp., Anaplasma ovis, Ehrlichia canis, Anaplasma phagocytophilum, Hepatozoon canis and Hepatozoon fells. Most tick pools were infected with a single pathogen. However, four pools infected with H. canis displayed infections with B. crassa, A. phagocytophilum and E. canis. The sequencing indicated that Anaplasma/Ehrlichia spp. was 100% identical to the sequence of Ehrlichia sp. Firat 2 and 3 previously identified from Hyalomma anatolicum. (C) 2013 Elsevier B.V. All rights reserved.

A total of 1064 adult ticks identified as Hyalomma marginatum ( n = 965), Rhipicephalus turanicus ( n = 86), Haemaphysalis inermis ( n = 9) and Rhipicephalus bursa ( n = 4) along with 20 egg masses derived from H. marginatum ( n = 18) and R. bursa were screened for Babesia occultans . The ticks were combined into 328 pools and infection rates were calculated as the maximum likelihood estimation (MLE) with 95% confidence intervals (CI). Of these pools, 23 (7%) were positive for B. occultans , and the overall MLE was calculated as 2.27% (CI 1.48–3.35). Infection prevalence in tick species was 7.09% for H. marginatum (22/310, MLE 2.41, CI 1.55–3.59) and 7.14% for R. turanicus (1/14, MLE 1.11, CI 0.07–5.18). None of H. inermis and R. bursa was infected with the parasite. Host-seeking H. marginatum females displayed an infection prevalence of 1.68% (2/119, MLE 0.63, CI 0.11–2.06), while no parasite DNA was detected in the males. The prevalence of infection in feeding H. marginatum varied, ranging from 27.27% (9/33, MLE 4.61, CI 2.26–8.52) in males to 25.92% (7/27, MLE 8.22, CI 3.72–15.75) in females. Of the egg masses examined, 4 (22.22%) of H. marginatum and 1 (50%) of R. turanicus were infected with B. occultans . To the best of our knowledge, this is the first time that B. occultans has been detected in R. turanicus.

Crimean-Congo hemorrhagic fever virus (CCHFV) is the causative agent of a tick-borne disease with high mortality Fates in humans. The distribution of CCHFV includes over 30 countries in Asia, the Middle East, southeastern Europe, and Africa. It was first recognized in Turkey in 2002, with an increasing number of cases reported between 2002 and 2009. Recent analysis of complete genome sequences of CCHFV isolates has revealed that the genomic plasticity of the virus is surprisingly high for an arthropod-borne virus. We have determined the complete nucleotide and deduced amino acid sequences of strain CCHFV Turkey-Kelkit06 isolated from the blood of a patient in an endemic region of Turkey in 2006. The complete sequence length of the CCHFV Turkey-Kelkit06 strain is 19,186 nt, consisting of a 1673 nt S segment, a 5364 nt M segment, and a 12,149 nt L segment. Based on the analysis of S, M, and L segments, CCHFV Turkey-Kelkit06 clustered in Group V, which represents the Europe/Turkey geographic lineage. Although glycoproteins encoded by the M gene are the most variable part of the CCHFV Turkey-Kelkit06 strain, some functional domains of the glycoproteins are well conserved. Here, we report the complete sequence and genome organization of the CCHFV Turkey-Kelkit06 strain and its phylogenetic relationship to other strains of CCHFV. Collecting data on viral sequences among isolates from CCHF epidemics may provide valuable information regarding the molecular basis of the epidemic potential of the virus. (C) 2009 Elsevier B.V. All rights reserved.

Theileria spp. infect wild and domestic ruminants in the tropical and subtropical regions of the world. Two species, T. lestoquardi and T. ovis, are suspected to cause ovine theileriosis in Iran. The epidemiological aspects of ovine theileriosis in Iran are poorly understood, and further investigations by sensitive and precise techniques are required. In this study, the use of a nested PCR for amplification of a fragment of the 18S ribosomal DNA from virtually all species of Theileria is described. For differentiation of various Theileria spp. a RFLP assay was developed as a diagnostic tool enabling direct, concurrent, highly specific and sensitive identification of Theileria spp. The sensitivity of the nested PCR for Theileria species was 10−5% parasitemia. Restriction fragment length polymorphism (RFLP) of the PCR products allowed differentiation between three different Theileria species (T. annulata, T. lestoquardi and T. ovis) and seems to be useful for differentiation of other species such as T. separata and Theileria spp. china. From 100 field blood samples obtained from sheep in East and South-East Iran, 56% were positive for Theileria spp. by nested-PCR compared with 21% by microscopic examination. Out of 56 positive samples, 12.5% (7/56) were positive for T. ovis and 87/5% (49/56) were positive for T. lestoquardi. This is the first report in which T. ovis has been detected in Iran using molecular identification techniques.

This study was carried out to determine the presence and distribution of tick-borne haemoprotozoan parasites (Theileria and Babesia) in apparently healthy cattle in the East Black Sea Region of Turkey. A total of 389 blood samples were collected from the animals of various ages in six provinces in the region. Prevalence of infection was determined by reverse line blot (RLB) assay. The hypervariable V4 region of the 18S ribosomal RNA (rRNA) gene was amplified with a set of primers for members of the genera Theileria and Babesia. Amplified PCR products were hybridized onto a membrane to which generic- and species-specific oligonucleotide probes were covalently linked. RLB hybridization identified infection in 16.19% of the samples. Blood smears were also examined microscopically for Theileria and/or Babesia spp. and 5.14% were positive. All samples shown to be positive by microscopy also tested positive with RLB assay. Two Theileria (T annulata and 7: buffelilorientatis) and three Babesia (B. bigemina, B. major and Babesia sp.) species or genotypes were identified in the region. Babesia sp. genotype shared 99% similarity with the previously reported sequences of Babesia sp. Kashi 1, Babesia sp. Kashi 2 and Babesia sp. Kayseri 1. The most frequently found species was 7: buffeli/orientalis, present in 11.56% of the samples. T. annulata was identified in five samples (1.28%). Babesia infections were less frequently detected: B. bigemina was found in three samples (0.77%), B. major in two samples (0.51%) and Babesia sp. in five samples (1.28%). A single animal infected with T. buffeli/orientalis was also infected with B. bigemina. (C) 2008 Elsevier B.V. All rights reserved.

Polymerase chain reaction (PCR) was used to assess the presence and the frequency of Babesia ovis infection in the adult Rhipicephalus bursa and their hosts in Elazig province located in eastern Turkey. Tick and blood samples were collected from 32 sheep and 28 goats of four selected herds. A total of 226 R. bursa were randomly selected from the collected ticks and their salivary glands were dissected out in 0.85% saline under stereo microscope. DNA amplification method revealed that the frequency of B. ovis infections in the ticks and the small ruminants were 16.37% (37/226) and 6.66% (4/60), respectively. Three positive products, two of which were from the salivary glands of R. bursa and the other from sheep blood were purified from agarose gel and sequenced. The results showed that nucleotide sequences were identical to the previously reported nucleotide sequences of B. ovis. It is concluded that R. bursa might play an important role in the field as a natural vector of the parasite.

Detection and identification of Theileria and Babesia species in 920 apparently healthy small ruminants in eastern Turkey, as well as parasite genetic diversity, was investigated using a specifically designed reverse line blot (RLB) assay. The hypervariable V4 region of the 18S ribosomal RNA (rRNA) gene was amplified and hybridized to a membrane onto which catchall and species-specific oligonucleotide probes were covalently linked. Three Theileria and one Babesia genotype were identified. Comparison of the Theileria genotypes revealed 93.6-96.2% similarity among their 18S rRNA genes. Two Theileria shared 100% and 99.7% similarity with the previously described sequences of T. ovis and Theileria sp. OT3, respectively. A third Theileria genotype was found to be clearly different from previously described Theileria species. The genotype was provisionally designated as Theileria sp. MK. The Babesia genotype shared 100% similarity with Babesia ovis. The survey indicated a high prevalence of piroplasm infections in small ruminants (38.36%). Theileria spp. prevalence was 36.08%. Prevalence of B. ovis was 5.43%. The most abundant Theileria species identified was T. ovis (34.56%) followed by Theileia sp. MK (1.30%) and Theileria sp. OT3 (0.43%).

The heat shock protein 70 (hsp70) genes of Babesia gibsoni, B. canis canis, B. canis vogeli, and B. canis rossi isolated from infected dogs were cloned by polymerase chain reaction (PCR) and sequenced. In the nucleotide sequence and the predicted amino acid sequence of the gene, the parasites were very similar to each other. The nucleotide sequences of the hsp70 gene had more variety than those of 18S nuclear subunit ribosomal DNA (18S rDNA). A phylogenetic analysis of these sequences and comparisons with sequences from other Babesia and Theileria species revealed that all canine babesial isolates analyzed in the present study were closely related to each other and formed one cluster. Additionally, a phylogenetic analysis of Babesia and Theileria species showed that these parasites could be divided into three groups: group A including canine babesial isolates, B. divergens, B. odocoilei, B. bovis, B. caballi, and B. ovis; group B including Theileria annulata, T. orientalis, and T. cervi; and group C including B. microti and B. rodhaini. These results suggested that a phylogenetic analysis of the hsp70 gene sequence might be helpful in classifying Babesia and Theileria species, and that canine babesial isolates might be closely related to each other, indicating their evolution from the same ancestry.

This study provides the first molecular data on infection Theileria of sheep in Turkey. A total of 218 blood samples were collected from sheep in five distinct geographical locations of eastern Turkey. Theileria piroplasm DNAs, extracted from sheep blood, were subjected to the polymerase chain reaction (PCR) procedures, using specific primers for Theileria spp. and Theileria lestoquardi. Blood smears were examined for Theileria piroplasms by microscopic observation. In PCR analysis, the expected amplicons were obtained from 90 (41.2%) blood samples with a molecular size of 1098 bp for Theileria spp. whereas none were amplified by Theileria lestoquardi-specific primers. Piroplasms of Theileria spp. were detected in 34 (15.5%) of the samples.

This study was carried out to determine the prevalence and distribution of tropical theileriosis in cattle in eastern Turkey by microscopical, serological and molecular methods. A total of 1561 whole blood, 1505 serum and 1483 blood smear samples were collected from cattle of various breeds and ages in 11 towns of Eastern Turkey. Theileria annulata piroplasm DNA extracted from cattle blood was amplified by polymerase chain reaction (PCR) using species-specific primers. Serum antibodies against T. annulata were investigated by indirect fluorescence antibody test (IFAT). Blood smears were examined for Theileria piroplasms by microscopical examination (ME). In the examination of DNA extracted from 1561 blood samples, an amplicon with the size of 721bp was obtained in 37.8% (590/1561) of these samples. Serum antibodies against T. annulata and piroplasm of Theileria spp. were detected in 34.9% (526/1505) and 19.7% (293/1483) of the samples, respectively. The differences between ME and PCR results and between ME and IFAT results were statistically significant (P < 0.05). In contrast, there was no significant difference between the PCR and IFAT results. A total of 179 ticks (136 female; 43 male) belonging to Hyalomma spp. were collected from cattle from three towns. Ticks were identified to be Hyalomma anatolicum anatolicum on the basis of morphological features.

Aktas, Munir/0000-0002-3188-8757 WOS: 000188537100001 PubMed: 15036571 A total of 2388 cattle and 442 shelters, from two provinces (Elazig and Malatya) endemic for tropical theileriosis in the east of Turkey, were studied for Hyalomma tick populations from July 1993 to July 1995 in Elazig and from May 1998 to January 1999 in Malatya. Four thousand five hundred and eighty one of 7455 Hyalomma ticks were collected from cattle, the other ticks (2874) were collected from shelters. All of the ticks collected from shelters were Hyalomma anatolicum anatolicum. Two thousand eight hundred and ninety five (63.1%) of 4581 Hyalomma ticks collected from cattle were H.a. anatolicum. 23.8% (1047/4581), 11.7% (536/4581) and 0.6% (3/4581). of Hyalomma ticks were Hyalomma anatolicum excavatum, Hyalomma detritum and Hyalomma marginatum marginatum, respectively. A total of 5909 Hyalomma adult ticks collected from cattle (3362/5909) and shelters (2447/5909) were dissected and salivary glands were stained with Methyl-green/Pyronin method. Thousand one hundred and fifty (46.9%) of 2447 H.a. anatolicum collected from shelters and 412 (19.1%) of 2147 H.a. anatolicum collected from cattle were positive for Theileria infection. Twenty (2.4%) of 820 H.a. excavatum and 23 (4.6%) of 495. H. detritum collected from cattle were positive. The mean number of infected acini per infected male and female ticks collected from cattle were 11.3 and 22.4 in H.a. anatolicum, 4 and 6.8 in H.a. excavatum, 17.9 and 18.3 in H. detritum, respectively. In H.a. anatolicum collected from shelters, the above rates were 11.8 and 17.6 in male and female ticks, respectively. The prevalence and intensity of Theileria infection was greater in female ticks than in males. (C) 2003 Elsevier B.V. All rights reserved.