Prevalence of Theileria spp. infection in sheep in South Khorasan province, Iran

The prevalence of Theileria spp. infection was studied in sheep in the South Khorasan province in Iran from 2003 to 2004. A total of 840 sheep from 34 flocks were clinically examined and investigated for the presence of Theileria spp. in the appropriate blood smears and any tick species on the body of the animals. In this study, 11.9% of sheep were infected with Theileria spp., with a parasitemia of 0.02–0.1%. Differences in the infection rates were statistically significant among different areas of the South Khorasan province. The highest prevalence was found in the Ferdows area (31.4%) and the lowest rate in the Nehbandan area (0.7%). The prevalence of Theileria spp. infection in males and females and between different age groups of sheep were not statistically significant. Seasonally, the prevalence of Theileria spp. infection in sheep reached its highest level in June (26.3%), whereas it decreased in July and August. It was found that 50.5% of the animals harbored Rhipicephalus sanguineus, 48.5% harboured Hyalomma anatolicum and 0.89% harboured Hyalomma dromedari.     

Molecular detection of Theileria sp. in ticks and naturally infected sheep

Seven healthy sheep and 10 sheep diagnosed with piroplasmosis based on clinical signs were tested for the presence of babesiae and theileriae. Using the molecular techniques, two species of theileriae were detected and characterized. Theileria ovis was present mostly in healthy sheep and in Rhipicephalus ticks collected from infected sheep. Theileria sp. OT3 parasite was detected mostly in ill animals which represent additional evidence to the possible pathogenic nature of Theileria sp. OT3. The presence of babesiae in sheep or in ticks was not determined. The results of this study showed that ovine piroplasmosis due to Theileria is present in Southern Croatia. It was concluded that clinical diagnosis of ovine piroplasmosis should be confirmed by molecular analysis in order to identify the species of piroplasm, to select the appropriate treatment and to exclude the threat for public health.     

Molecular detection of piroplasms in ixodid ticks infesting cattle and sheep in western Oromia, Ethiopia

In Ethiopia, ticks and tick-borne diseases are widely distributed and contribute to important economic losses. Several studies investigated the prevalence and species composition of ticks infesting ruminants; however, data on tick-borne pathogens are still scarce. During the study period from October 2010 to April 2011, a total of 1,246 adult ticks and 264 nymphs were collected from 267 cattle and 45 sheep in Bako District, western Oromia, Ethiopia. The study showed infestation of 228/267 (85.4 %) cattle and 35/45 (77.8 %) sheep with adult ticks. Overall, eight tick species, belonging to three genera (Amblyomma, Rhipicephalus, Hyalomma), were identified and Amblyomma cohaerens (n?=?577), Rhipicephalus evertsi evertsi (n?=?290), Rhipicephalus (Boophilus) decoloratus (n?=?287), and Amblyomma variegatum (n?=?85) were the more prevalent species. A statistically significant host preference in A. cohaerens for cattle and R. evertsi evertsi for sheep was noticed. Molecular detection of piroplasms, performed only for adult ticks of two species of the genus Rhipicephalus (R. evertsi evertsi and R. decoloratus), revealed an overall prevalence of 4 % (8/202) Theileria buffeli/sergenti/orientalis, 0.5 % (1/202) Theileria velifera, and 2 % (4/202) Theileria ovis. The study showed that tick infestation prevalence is considerably high in both cattle and sheep of the area, but with a low intensity of tick burden and a moderate circulation of mildly pathogenic piroplasm species.     

PCR-based detection of Theileria ovis in Rhipicephalus bursa adult ticks

Tick-borne diseases in ruminants are common in tropical and subtropical regions and lead to meat and milk production losses. In this study, polymerase chain reaction (PCR) was used to assess the presence of Theileria ovis in Rhipicephalus bursa ticks. We have demonstrated that the PCR enabled detection of T. ovis in field isolates of R. bursa collected from naturally infested sheep and goats in eastern Turkey. The sampling was done in spring season (between May and June 2004). A total of 420 R. bursa were collected and randomly selected 192 number of them (97 female and 95 male) were dissected. Primers specific for 520 bp fragments small subunit ribosomal RNA (ssu rRNA) gene of T. ovis amplified products from 37 of the 192 (19.27%) samples. The parasite was detected in 17 (17.52%) female and in 20 (21.05%) male ticks. Two T. ovis amplicons from the tick samples were purified and sequenced. The resulting sequences were identical to the nucleotide sequence of the Turkish sheep strain of T. ovis. These results showed that R. bursa might play an important role in the field as a natural vector of T. ovis.     

Prevalence of ixodid ticks on cattle and sheep southeast of Iran

A survey was carried out to investigate the prevalence of hard tick species (Acari: Ixodidae) on cattle and sheep southeast of Iran. A total of 972 ticks were collected from 280 infested cattle and 1,207 ticks were collected from 632 infested sheep during activating seasons of ticks in 2008–2009. The species collected from cattle were Hyalomma marginatum marginatum (50.92%), Hyalomma anatolicum excavatum (25.61%), Hyalomma anatolicum anatolicum (8.12%), Hyalomma asiaticum asiaticum (1.85%), and Rhipicephalus sanguineus (13.47%) while the species collected from sheep were R. sanguineus (36.37%), H. marginatum marginatum (30.65%), H. anatolicum excavatum (19.05%), H. asiaticum asiaticum (10.52%), Hyalomma detritum (3.14%), and Dermacentor marginatus (0.24%). The results show that, H. marginatum marginatum, H. anatolicum excavatum, and R. sanguineus are dominant tick species in the surveyed area.     

Occurrence and antimicrobial resistance of emergent Arcobacter spp. isolated from cattle and sheep in Iran

This study is conducted to determine the occurrence and antimicrobial resistance of Arcobacter spp. isolated from clinically healthy food animals. A total of 308 samples from cattle (200) and sheep (108) were collected from Shiraz slaughterhouse, southern Iran to investigate the presence of the important Arcobacter spp. using cultivation and Polymerase Chain Reaction (PCR) methods. Antimicrobial susceptibility of Arcobacter isolates was determined for 18 antibiotics using disk diffusion method. Among 308 samples, 27 (8.7%) and 44 (14.28%) were positive for the presence of Arcobacter species with cultivation and PCR procedures, respectively. The predominant species was A. butzleri in both cattle (58.33%) and sheep (55%). In addition, concurrent incidence of the species was observed in 25% of the positive samples. All Arcobacter isolates were resistant to rifampicin, vancomycin, ceftriaxone, trimethoprim and cephalothin. The isolates showed high susceptibility to tetracycline, oxytetracycline, erythromycin, ciprofloxacin, kanamycin, amikacin, gentamicin and enrofloxacin. No significant difference among cattle and sheep isolates in resistance pattern was observed. The results indicate that cattle and sheep are significant intestinal carriers for Arcobacter spp. Moreover, tetracycline and aminoglycosides showed great effects on Arcobacter species in antibiogram test and can be used for treatment of human Arcobacter infections.     

Prevalence of subclinical coccidiosis in broiler-chicken farms in the municipality of Mashhad, Khorasan, Iran

A survey of the prevalence of subclinical coccidiosis in broiler-chickens was conducted in the municipality of Mashhad, Khorasan, Iran. Eighty-four chicken farms were randomly selected; from each farm, five birds per 10 000 were sampled (as was litter). Serial scraping of the intestinal lining was done in chicks at 3rd and >6th week of age.

The farm-level prevalence of subclinical coccidiosis was 38% (95% CI: 28, 48.47%). Uni- and multi-variable associations were tested between each variable. An increased risk of infection in the broiler was associated with the larger farm, with older chickens, and if the chicken farm were sampled in the winter or spring, using coccidiostat in the food was not associated. The peak oocyst score in the litter occurred at >6th week of age. Most farms (97%) had E. acervulina; (41%) had E. maxima and (12%) had E. tenella.     

牛羊梨形虫病及媒介蜱综合防治药物筛选

天祝县莫客片一带牛梨形虫病发病率达24.6%,病死率28.6%;羊发病率34.2%,病死率27.5%,与往年相比变化不大,笔者从蜱控制、梨形虫病预防加治疗相结合综合防控入手,作控制、预防药物试验对比,筛选药效长、低毒、安全、广谱药物,结果显示用敌杀死环境灭蜱,同时注施可诺注射液控制蜱感染,用蒿甲醚注射液或盐酸咪唑苯脲,预防和治疗梨形虫病,能有效降低当地牛羊梨形虫病的发病率和病死率。     

Molecular epidemiological survey of Theileria orientalis in Thua Thien Hue Province, Vietnam

Theileria orientalis is a benign bovine protozoan parasite that occasionally causes serious economic loss in the livestock industry. We report the findings of a molecular epidemiological survey of T. orientalis in 94 Vietnamese yellow cattle, 43 water buffaloes, 21 sheep, 21 goats and 85 blood-sucking ticks of cattle in the Thua Thien Hue province of Vietnam. The major piroplasm surface protein (MPSP) gene of T. orientalis was detected using polymerase chain reaction from 13 cattle (13.8%), 11 water buffaloes (25.6%), 1 sheep (4.8%) and 9 ticks (10.6%). Phylogenetic analysis using MPSP gene sequences showed the presence of seven genotypes, four previously categorized genotypes (Types 1, 3, 5 and 7) and three new genotypes (Types N-1, N-2 and N-3).     

Molecular and phylogenetic characterization of Theileria spp. parasites in autochthonous bovines (Mirandesa breed) in Portugal

A survey was conducted during the months of April-June 2003 in the northeast Portugal (Bragan?a district) in order to characterize the hemoparasite population of an autochthonous Mirandesa breed of Bos taurus. The polymerase chain reaction (PCR) analysis of the bovine blood revealed that 3 out of 116 animals were infected with Theileria and/or Babesia parasites, while reverse line blot hybridisation (RLB) analysis showed that these animals were infected with Theileria buffeli/orientalis. Cloning and sequencing confirmed the RLB results. Database sequence searches combined with phylogenetic analysis of the partial 18S ribosomal RNA gene sequences obtained enabled us to place the parasites in question as members of the T. buffeli/orientalis group, confirming the PCR/RLB diagnosis.     

Molecular detection of Hepatozoon canis in dogs and ticks in Shaanxi province,China

Hepatozoon canis, transmitted by Rhipicephalus sanguineus, is a tick-borne pathogen and causes canine hepatozoonosis. Until now, only limited previous studies were conducted on the molecular detection and characterization of Hepatozoon sp. in dogs in China. Blood samples were collected from 93 sick dogs that were clinically diagnosed as babesiosis but tested negative for Babesia, and 103 apparently healthy dogs, as well as their infesting ticks in Xi’an and Hanzhong cities, Shaanxi province of China. PCR amplifying partial 18S rRNA gene was used to detect the DNA of Hepatozoon sp. Genetic and phylogenetic analysis were performed to determine the Hepatozoon species. Our results demonstrated that H. canis was identified from the sick dogs and the infested ticks in Hanzhong, with no significant differences of prevalence between both genders and ages. No positive blood or tick samples were found in Xi’an. Moreover, all the 18S rRNA gene sequences recovered from both dogs and the infested ticks showed a high genetic similarity with each other, and also presented a close relationship with other known sequences in and outside China. In conclusion, H. canis was identified in babesiosis-suspected dogs and ticks infesting them in Shaanxi, China, although the association between clinical signs and H. canis need further study.     

The value of dosing sheep with tetramisole,in the Marand area of North Western Iran

Summary The value of using three doses of an anthelmintic over a period of six months has been demonstrated in a commercial sheep flock in Marand area of North Western Iran. Reduction in worm egg counts and resultant increases in body weight were shown, and the economic benefits calculated.

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Sumario Los resultados de este trabajo en un hato comercial ovino demuestran el valor de usar tres dosis de un anti-helmíntico sobre un periodo de seis meses. Se demostraron reducciones en la cuenta de huevos de gusanos y la consiguiente ganacia de peso. Se hace un analisis del beneficio económico de este último punto.

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Résumé Les résultats de ce travail effectué dans un troupeau de moutons du commerce démontrent la valeur de l’emploi de trois doses anthelminthiques sur une période de 6 mois. On y montre la réduction du nombre des oeufs émis, l’augmentation résultante du poids des animaux, en même temps qu’on analyse le bénéfice économique qu’on en tire. El valor de dosificar ovinos con tetramizole, en el area de marand de Iran noroeste.

     

Molecular detection of Anaplasma phagocytophilum in cattle and Ixodes persulcatus ticks

The tick-borne pathogen, Anaplasma phagocytophilum (A. phagocytophilum), the causative agent of human granulocytic anaplasmosis (HGA), is increasingly becoming a public health concern as an aetiological agent for emerging infectious disease. We found A. phagocytophilum infection in a pooled sample of field-collected Ixodes persulcatus (I. persulcatus) ticks from one district in Hokkaido, Japan. Thus, to further investigate the prevalence in field-collected ticks, we used PCR assays targeting the A. phagocytophilum gene encoding 44 kDa major outer membrane protein (p44) for screening of I. persulcatus ticks and samples from cattle from pastures. Out of the 281 I. persulcatus ticks, 20 (7.1%) were found to harbor A. phagocytophilum DNA. The infection rate for A. phagocytophilum in cattle was 3.4% (42/1251). In future studies, it will be necessary to investigate effects of the infection in order to understand its pathogenesis of A. phagocytophilum in domestic animals.     

An epidemiological study on ovine babesiosis in the Mashhad suburb area,province of Khorasan,Iran

The prevalence of Babesia spp. infection was studied in sheep of the Mashhad area in Iran from 1998 to 2000. A total of 677 sheep originating from 115 flocks were clinically examined and investigated for the presence of Babesia spp. in appropriate blood smears and any tick species on the body of the animals. The study revealed that the infection rate for Babesia ovis and Babesia motasi were 167 (24.6%) and 4 (0.5%), respectively. Double (mixed) infections occurred in 21 (3%) sheep. Differences in infection rates were statistically non-significant between male and female sheep and between different age groups. Seasonally, the prevalence of Babesia spp. infection started to increase in April and reached highest values in August (56%), while a decrease was observed in September, reaching the lowest levels In February and March. The study demonstrated that 1.7% of sheep infected with B. ovis and 50% of sheep infected with B. motasi exhibited clinical signs. Sheep infected with B. motasi showed the highest levels of parasitemia. We found that 550 (73%) of the animals harbored Rhipicephalus sanguineus; 166 (21%) Hyalomma marginatum; 19 (2.5%) Dermacentor daghestanicus; 14 (1.8%) Hyalomma anatolicum; 6 (0.66%) Hyalomma asiaticum; and one (0.13%) Haemaphysalis punctata. The examination of 727 tick haemolymph samples and 52 tick egg smears showed that one sample (0.2%) of haemolymph of R. sanguineus, two (1.2%) haemolymphs of H. marginatum and two (2%) eggs of R. sanguineus harbored kinetes morphologically matching the criteria described for B. ovis.     

Detection of Theileria and Babesia species in ticks collected from cattle

The present study was carried out to detect tick species that infest cattle, and Theileria and Babesia species transmitted by these ticks in Kayseri province (Turkey). A total of 300 cattle were examined for tick infestations. Of the 300 cattle, 117 (39%) were infested with ticks. A total of 1160 ticks belonging to 11 Ixodid genera were collected from the infested animals and their shelters. The most prevalent tick species was Boophilus annulatus 26.37% (306/1160) followed by Hyalomma marginatum marginatum 21.12% (245/1160) and Rhipicephalus turanicus 18.7% (217/1160). The collected ticks were separated into 43 tick pools, according to their species. These pools were examined for bovine Theileria and Babesia species (Theileria sp., Babesia sp., Theileria annulata, T. buffeli/orientalis, Babesia bigemina, B. bovis and B. divergens) by using the reverse line blotting method (RLB). Of the 43 tick pools examined, 6 (14%) were infected with B. bigemina, 4 (9.3%) with T. annulata, and 1 (2.3%) with Babesia sp., whereas 1 (2.3%) displayed mixed infection with T. annulata + B. bigemina. The sequence and phylogenetic analyses of Babesia sp., which could not be identified to the species level by RLB, were performed. In the phylogenetic tree, Babesia sp. (Kayseri 1) grouped with Babesia sp. (Kashi 2), Babesia sp. (Kashi 1), Babesia sp. (Xinjiang) and B. orientalis with 96.8-100% identity.     

Infectivity and cross-immunity studies of Theileria lestoquardi and Theileria annulata in sheep and cattle: I. In vivo responses.

In a series of experiments, sporozoite stabilates of a Theileria lestoquardi (Lahr) and a T. annulata (Ankara) stock prepared from Hyalomma anatolicum anatolicum ticks, were used to examine the infectivity of both parasite species for sheep and cattle and to study the development of cross-immunity between these parasite species. In the first experiment sheep and cattle were inoculated with T. lestoquardi sporozoites. Surviving animals and naive sheep and cattle were, in the second experiment, inoculated with T. annulata. In the third experiment, naive sheep and sheep previously infected with T. annulata, were inoculated with T. lestoquardi. The following responses to inoculations were monitored: clinical and haematological signs of infection, appearance of parasitic stages of the parasites in lymph node biopsies and in peripheral blood and serological response to T. lestoquardi and T. annulata schizont antigens. While T. lestoquardi readily infected sheep and caused severe disease, it did not infect cattle. On the other hand, T. annulata infected both cattle and sheep. However, whereas cattle became severely affected, infected sheep showed mild clinical symptoms only and piroplasms did not develop. Despite their different behaviour in the host species examined, cross-immunity studies suggested that the parasite species are very closely related. Experiments in sheep indicated that T. lestoquardi infection protected against subsequent T. annulata infection. On the other hand, recovery from T. annulata infection did not prevent infection by sporozoites of T. lestoquardi, resulting in the establishment of schizonts and their subsequent development into piroplasms, although it protected against the major clinical effects of T. lestoquardi infection.     

Infectivity and cross-immunity studies of Theileria lestoquardi and Theileria annulata in sheep and cattle: II. In vitro studies.

In the studies previously reported, the tick-borne protozoan parasites Theileria lestoquardi and Theileria annulata were shown to differ in their capacity to infect sheep and cattle. In the studies presented here, these findings were further supported. In vitro infectivity of T. lestoquardi and T. annulata sporozoites for peripheral blood mononuclear cells of sheep and cattle were determined by analysis of cell cultures for cell proliferation, the detection of parasites in Giemsa-stained cytospin smears and the establishment of continuously growing schizont-infected cell lines. In the same way, the development of schizont-infected cells into continuously growing cell lines was studied with material isolated ex vivo from the sheep and cattle undergoing primary infections described elsewhere. Comparisons were also made between development of ex vivo cell lines from animals undergoing primary infections with those of the animals undergoing challenge infection with the other parasite species. Theileria species specific primers were used in a PCR to determine the identity of the parasites in the cell lines. These in vitro studies confirmed earlier observations that T. lestoquardi was unable to infect cattle, whereas infection of all sheep with T. annulata was proven. Moreover, earlier indications of the development of partial cross-immunity in sheep of T. annulata to T. lestoquardi and vice versa were strengthened. These findings may thus have consequences for the understanding of the epidemiology of T. lestoquardi infections of sheep. On the other hand. since piroplasms were not demonstrated in sheep infected with T. annulata, such sheep will not be infective to ticks and will consequently be unlikely to play a role in the maintenance and transmission of T. annulata to cattle.     

Molecular detection of Culicoides spp. and Culicoides imicola, the principal vector of bluetongue (BT) and African horse sickness (AHS) in Africa and Europe

Bluetongue (BT) and African Horse Sickness (AHS) are infectious arthropod-borne viral diseases affecting ruminants and horses, respectively. Culicoides imicola Kieffer, 1913, a biting midge, is the principal vector of these livestock diseases in Africa and Europe. Recently bluetongue disease has re-emerged in the Mediterranean Basin and has had a devastating effect on the sheep industry in Italy and on the islands of Sicily, Sardinia, Corsica and the Balearics, but fortunately, has not penetrated onto mainland France and Spain. To survey for the presence of C. imicola, an extensive light-trap network for the collection of Culicoides, was implemented in 2002 in southern mainland France. The morphological identification of Culicoides can be both tedious and time-consuming because its size ranges from 1.5 to 3 mm. Therefore, an ITS1 rDNA polymerase chain reaction (PCR)-based diagnostic assay was developed to rapidly and reliably identify Culicoides spp. and C. imicola. The aim of this work was to set up a rapid test for the detection of C. imicola amongst a pool of insects collected in areas at risk for BT. The sequence similarity of the rDNA (nuclear ribosomal DNA), which is greater within species than between species, is the foundation of its utilisation in species-diagnostic assays. The alignment of the 11 ITS1 sequences of Culicoides obtained from Genbank and EMBL databases helped us to identify one region in the 5' end and one in the 3' end that appear highly conserved. PCR primers were designed within these regions to amplify genus-specific fragments. In order to set up a C. imicola-specific PCR, another forward primer was designed and used in combination with the previously designed reverse primer. These primers proved to be highly specific and sensitive and permitted a rapid diagnostic separation of C. imicola from Culicoides spp.     

Naturally occurring infections of cattle with Theileria lestoquardi and sheep with Theileria annulata in the Sudan

Theileria annulata is endemic in northern Sudan, hindering all efforts at upgrading cattle for milk production. T. lestoquardi clinical cases occur throughout the year and causes annual outbreaks that result in substantial losses in sheep. In the northern Sudan both cattle and small ruminants are frequently raised together and/or share common grazing grounds at river banks. In an attempt to evaluate field cross infectivity of Theileria lestoquardi and T. annulata in cattle and sheep respectively, a PCR analysis was carried out on samples collected from closely reared sheep and cattle using both T. annulata and T. lestoquardi specific primers. A total of 19 sheep out of 51 (37.3%) were positive for T. lestoquardi while four sheep (7.8%) showed T. annulata specific amplicons. A total of 38 out of 52 (73.1%) surveyed cattle were PCR positive for T. annulata and only two (3.8%) showed T. lestoquardi specific bands. These findings indicate complex epidemiology of both infections in areas where both parasites are transmitted by the same vector and call for further investigations of this phenomenon.