In vitro feeding of Hyalomma excavatum and Hyalomma marginatum tick species.

The rearing of ticks is an important technique for studies aiming to elucidate the course and pathogenesis of tick-borne diseases (TBDs). TBDs caused by protozoans (Theileria, Babesia) and bacteria (Anaplasma/Ehrlichia) impose a serious constraint upon livestock health and production in tropical and sub-tropical regions where the distributions of host, pathogen, and vector overlap. This study focuses on Hyalomma marginatum, one of the most important Hyalomma species in the Mediterranean region, being a vector of the virus that causes Crimean-Congo haemorrhagic fever in humans, together with H. excavatum, a vector of Theileria annulata, an important protozoan of cattle. The adaptation of ticks to feeding on artificial membranes allows the creation of model systems that can be put to use examining the underlying mechanisms of pathogen transmission by ticks. Silicone membranes, in particular, offer researchers the flexibility to adjust membrane thickness and content during artificial feeding. The aim of the present study was to develop an artificial feeding technique using silicone-based membranes for all developmental stages of H. excavatum and H. marginatum ticks. Attachment rates after feeding on silicone membranes for females H. marginatum and H. excavatum were 8.33% (8/96) and 7.95% (7/88), respectively. The use of cow hair as a stimulant increased the attachment rate of H. marginatum adults in comparison to other stimulants. The engorgement of H. marginatum and H. excavatum females took 20.5 and 23 days with average weights of 307.85 and 260.64 mg, respectively. Although both tick species could complete egg-laying, and this was followed by hatching of larvae; their larvae and nymphs could not be fed artificially. Taken together, the results of the present study clearly indicate that silicone membranes are suitable for feeding of H. excavatum and H. marginatum adult ticks, supporting engorgement, laying of eggs, and hatching of the larvae. They thus represent a versatile tool for studying transmission mechanisms of tick-borne pathogens. Further studies are warranted to examine attachment and feeding behaviours in order to increase the success of artificial feeding of larvae and nymphal stages.

Prevalence of Theileria/Babesia Species in Ruminants in Burdur Province of Turkey.

PURPOSE: Theileriosis and babesiosis, two tick-borne haemoparasitic diseases (TBHDs) of ruminants, are caused by protozoan parasites of the genus Theileria and Babesia, respectively. Among them, some species are considered to be highly pathogenic causing serious economic losses to livestock holders especially in tropic and subtropic regions. Local and/or general control measures are needed to be applied to reduce economic impact of TBHDs. Prevalence studies are essential for the implementation and/or design of effective prevention and control measures based on true epidemiological data. Therefore, this study aimed to investigate the presence, prevalence and possible cross infections of Theileria/Babesia species between sheep, goat and cattle herds in Burdur province in Turkey. METHODS: A total of 964 blood samples were collected from sheep (n = 330), goat (n = 300) and cattle (n = 334) from five different districts of Burdur province. The samples were investigated for ovine and bovine Theileria/Babesia species using reverse line blot (RLB) hybridization assay. RESULTS: In small ruminants, T. ovis was the most abundant Theileria species detected in sheep with a rate of 79.69%. Among Babesia species, B. ovis and B. crassa were detected only in blood of goats (0.66%) and sheep (1.12%) as single and mixed infections, respectively. In cattle, T. annulata, B. bovis, Babesia spp. were detected in rates of 0.59%, 3.29%, 3.59%, respectively. CONCLUSION: Obtained results clearly indicated that no cross infections with Theileria/Babesia species occurred in small ruminant and cattle herds that use the same grazing area.

Investigation of Phlebotominae (Diptera: Psychodidae) Fauna, Seasonal Dynamics, and Natural Leishmania spp. Infection in Mugla, Southwest of Turkey.

Due to its geographical location, Mugla province is one of the most frequently used places by refugees. Although leishmaniasis have been previously reported in this region, there is a lack of information on the etiological agent and possible vectors. The main objectives of this study were; i) to investigate the sand fly fauna, ii) to reveal the natural Leishmania spp. infection in wild caught sand flies using molecular tools, and iii) to determine the annual seasonal dynamics of the sand flies in Mugla region. Totally, 2093 specimens belonging to 15 species [12 Phlebotomus, three Sergentomyia; 51 unidentified] were collected during the one-year (June 2016- June 2017) period. Of the collected sand flies, 1928 (92.12%) were caught by the Centers for Disease Control (CDC) light traps, while 165 (7.88%) of them were caught by sticky traps. Phlebotomus major sensu lato (s.l.), the potential vector of visceral leishmaniasis (VL) and canine leishmaniasis (CanL) in the Mediterranean and Aegean region, was detected in all sampling locations and found as the dominant taxon (n=1035; 49.45%) of the study area and followed by Phlebotomus tobbi (n=371; 17.72%). During the sampling period, sand fly activity was started in March and peaked in August. Sand fly population size reduced dramatically between mid-September and early October. The number of collected specimens was peaked in August, while there is only one sample collected both in November and March. The majority of the sand flies (78.66%) were collected at an altitude range of 200-400 m. Seventy-two monospecific pools were screened for the presence of Leishmania DNA by real time ITS1 PCR and 24 (nine P. major s.l., eight P. tobbi, two P. papatasi, two S. minuta, one P. alexandri, one P. similis, and one Phlebotomus (Transphlebotomus spp.) of them (33.8%) were found positive (L. infantum, L. tropica, and L. major). To the best of our knowledge, the presence of fifteen sand fly species and their distribution, seasonal dynamics, molecular detection of Leishmania parasites in Mugla province was reported for the first time. The presence of vector species in the study area, appropriate temperature and humidity conditions, long sand fly activity season, and presence of Leishmania parasite suggests that there is a serious risk in the transmission of leishmaniasis in Mugla.

Tick Bites on Humans in Southwestern Region of Turkey: Species Diversity

Objective: The aim of the present study was to determine tick species found on humans who suffered from tick bite in the Southwestern Anatolia Region, Turkey. Methods: Between January and October 2007, ticks were collected from people admitted to the city and/or town hospitals with complaints of tick bites in nine different provinces of Turkey. Genus and/or species of the ticks in adult, larva and nymph stages were identified microscopically. Identification was done using related taxonomic keys. Results: A total of 2.610 ticks were collected from humans who were admitted to the hospitals with complaints of tick bites in the Southwestern Anatolia Region in the present study. Of these, 1.858 samples were collected from the Aegean Region and the remaining 752 from the Mediterranean Region of the country. The ticks were identified as Hyalomma spp. (78.58%), Rhipicehalus spp. (18.89%), Ixodes spp. (0.88%), Dermacentor spp. (0.77%), Haemaphysalis spp. (0.61%), Argas spp. (0.23%), and Ornithodoros spp. (0.04%). Results indicated that the majority of the ticks were nymphs of Hyalomma spp. (n=1.582). Nymphal stage was most commonly encountered from the Aegean Region and the Mediterranean Region with a prevalence of 46.13% (n=1.204) and 14.48% (n=378) respectively. Within the collected adult ticks (n=913), the majority of the samples were identified as H. marginatum (n=233, 26.09%). Conclusion: The results indicate the high diversity of tick species infesting humans in the Southwestern Anatolia Region, Turkey. So, it is crucial to publish information on tick bite prevention, which would play an important role in reducing the incidence of tick-borne diseases.

The secreted Theileria annulata Ta9 protein contributes to activation of the AP-1 transcription factor.

Theileria annulata is an obligate intracellular protozoan parasite of the phylum Apicomplexa. Theileria sporozoites invade bovine leukocytes and develop into a multinucleate syncytial macroschizont that causes uncontrolled proliferation and dissemination of infected and transformed leukocytes. Activator protein 1 (AP-1) is a transcription factor driving expression of genes involved in proliferation and dissemination and is therefore a key player in Theileria-induced leukocytes transformation. Ta9 possesses a signal peptide allowing it to be secreted into the infected leukocyte cytosol and be presented to CD8 T cells in the context of MHC class I. First, we confirmed that Ta9 is secreted into the infected leukocyte cytosol, and then we generated truncated versions of GFP-tagged Ta9 and tested their ability to activate AP-1 in non-infected HEK293T human kidney embryo cells. The ability to activate AP-1-driven transcription was found to reside in the C-terminal 100 amino acids of Ta9 distant to the N-terminally located epitopes recognised by CD8+ T cells. Secreted Ta9 has therefore, not only the ability to stimulate CD8+ T cells, but also the potential to activate AP-1-driven transcription and contribute to T. annulata-induced leukocyte transformation.

Eustigmaeus johnstoni Zhang - Gerson (Acari: Stigmaeidae) a Parasitic Mite Species Detected in Phlebotomus papatasi (Diptera: Psychodidae) Specimens Collected from Aydin, Turkey.

OBJECTIVE: We aimed to report the presence of the parasitic mite Eustigmaeus johnstoni Zhang - Gerson (E. johnstoni) on the sand fly species Phlebotomus papatasi (P. papatasi) collected in Aydin province, Turkey. METHODS: Sand flies were collected from nature by CDC light traps in Nazilli town in Aydin province and were separated according to collection localities. Head and genital areas were cut and mounted for species identification, and midgut contents were checked for the possible presence of Leishmania parasites under a light microscope. Mites detected in sand flies were diagnosed on the species level based on the literature. RESULTS: A total of 360 female and 378 male sand flies were caught. During dissection and preparation, two female E. johnstoni parasitic mite specimens belonging to the genus Eustigmaeus were detected on the abdominal pleura of P. papatasi specimens. CONCLUSION: To our knowledge, this is the first study to report the presence E. johnstoni on P. papatasi in Turkey. Previous studies showing the relationship between parasitic mites and sand flies have indicated that this relationship is not accidental. More studies are needed to understand this relationship for obtaining more detailed information.

Selection of Genetic Markers to Determine Diversity in Theileria annulata Populations after Recombination.

OBJECTIVE: Selecting polymorphic mini- and microsatellite markers to determine genetic diversity and chromosomal regions exhibiting elevated rates of recombination in Theileria annulata populations after recombination. METHODS: The Unipro UGENE software was used to select markers. A score at which 10 times more tandem repeats (TRs) were identified in the real DNA sequence than those in the scrambled sequences of T. annulata was used as the cutoff. TRs containing minimum three nucleotides in length for microsatellite and six nucleotides for minisatellite regions and having a repeat motif identity between 96%-100% with the unit size repeated minimum three times were screened through the whole genome using the suffix array algorithm. RESULTS: A total of 359 minisatellites and 8973 microsatellites were identified. TRs were screened one by one through the whole genome; mini- and microsatellites representing a single region and having suitable regions for primer design were selected based on their localization on T. annulata chromosomes, their repeat motif identity (>96%), and their repeat length (<1500 bp). The primers used to amplify selected candidates were designed, and each primer was used to check 27 different isolates of T. annulata. CONCLUSION: In the present study, a total of 13 polymorphic mini- and microsatellite markers located on the different chromosomes were selected to determine the population diversity of T. annulata.

Prevalence of tick-borne haemoparasites in small ruminants in Turkey and diagnostic sensitivity of single-PCR and RLB.

BACKGROUND: Tick-borne haemoparasitic diseases (TBHDs), caused by Theileria, Babesia, Anaplasma and Ehrlichia, are common in regions of the world where the distributions of host, pathogen and vector overlap. Many of these diseases threaten livestock production and some also represent a concern to human public health. The primary aim of this study was to determine the prevalence of the above-mentioned pathogens in a large number of blood samples (n = 1979) collected from sheep (n = 1727) and goats (n = 252) in Turkey. A secondary aim was to assess the diagnostic sensitivity of a number of species-specific polymerase chain reaction (PCR) tests and the reverse line blotting (RLB) assay. DNA samples were screened using species-specific PCR for the presence of Theileria ovis, Theileria sp. MK, T. lestoquardi, T. uilenbergi, T. luwenshuni, Babesia ovis, Anaplasma ovis and A. phagocytophilum while RLB was undertaken to test for the presence of all known Theileria, Babesia, Anaplasma and Ehrlichia species. The diagnostic sensitivity of these two approaches was then compared in terms of their ability to detect single species and mixed infections. RESULTS: Overall, 84 and 74.43% of the small ruminants sampled were identified as hosting one or more pathogen(s) by species-specific PCR and RLB respectively. The presence of Theileria sp. OT1, T. luwenshuni and T. uilenbergi in Turkey was revealed for the first time while the presence of Babesia motasi, B. crassa and T. separata in Turkish small ruminants was confirmed using molecular methods. A high prevalence of mixed infection was evident, with PCR and RLB approaches indicating that 52.24 and 35.42% of animals were co-infected with multiple species, respectively. More than 80% of the mixed infections contained T. ovis and/or A. ovis. The RLB approach was found to be capable of detecting mixed infections with species such as Theileria sp. OT1, Theileria sp. OT3, T. separata, B. crassa and Babesia spp. CONCLUSION: The results indicated that pathogens causing TBHDs are highly prevalent in sheep and goats in Turkey. The diagnostic sensitivity of species-specific single PCR was generally higher than that of RLB. However, the latter approach was still capable of identifying a high proportion of individuals containing mixed-species infections. The use of species-specific single PCR is recommended to accurately estimate pathogen prevalence and to identify co-infected hosts.

Prevalence of Larval-Stage Dicrocoeliidae (Digenea) Trematodes in Helix lucorum (Mollusca: Pulmonata) in Van Province.

OBJECTIVE: The aim of this study was to investigate the prevalence of larval-stage Dicrocoeliidae trematodes in Helix lucorum, a land snail found in Van Province. METHODS: Helix lucorum snails were collected in April, May, and June 2017 from Edremit and Gevas, the central districts of Van Province, especially from natural areas where ruminants predominate. The snails were anesthetized with magnesium chloride, were removed from their shells, and their digestive glands were disrupted. The disrupted parts were examined under a microscope. RESULTS: In Van Province, H. lucorum snails were found to be intermediate hosts for Dicrocoelium trematodes with a prevalence of 22%. The larval stages detected in the microscope are photographed and shown in detail. The number of infection with larval stages of the parasite was found to be highest in May. CONCLUSION: Helix lucorum the land snail, serves as an intermediate host for some developmental stages of the Dicrocoeliid trematodes, is also consumed as nutrients by humans in some countries. Based on the obtained results in this study, it can be concluded that this snail would have important effects on animal health in the Van region which has a hard climate and a border with Iran.

Identification and Analysis of Immunodominant Antigens for ELISA-Based Detection of Theileria annulata.

Tropical or Mediterranean theileriosis, caused by the protozoan parasite Theileria annulata, remains an economically important bovine disease in North Africa, Southern Europe, India, the Middle East and Asia. The disease affects mainly exotic cattle and imposes serious constraints upon livestock production and breed improvement programmes. While microscopic and molecular methods exist which are capable of detecting T. annulata during acute infection, the identification of animals in the carrier state is more challenging. Serological tests, which detect antibodies that react against parasite-encoded antigens, should ideally have the potential to identify carrier animals with very high levels of sensitivity and specificity. However, assays developed to date have suffered from a lack of sensitivity and/or specificity and it is, therefore, necessary to identify novel parasite antigens, which can be developed for this purpose. In the present study, genes encoding predicted antigens were bioinformatically identified in the T. annulata genome. These proteins, together with a panel of previously described antigens, were assessed by western blot analysis for immunoreactivity, and this revealed that four novel candidates and five previously described antigens were recognised by immune bovine serum. Using a combination of immunoprecipitation and mass spectrophotometric analysis, an immunodominant protein (encoded by TA15705) was identified as Ta9, a previously defined T cell antigen. Western blotting revealed another of the five proteins in the Ta9 family, TA15710, also to be an immunodominant protein. However, validation by Enzyme-Linked Immunosorbent Assay indicated that due to either allelic polymorphism or differential immune responses of individual hosts, none of the novel candidates can be considered ideal for routine detection of T. annulata-infected/carrier animals.

[Detection of Anaplasma / Ehrlichia Species of Cattle and Ticks in Aydin Region]. / Aydin Yöresinde Sigirlarda ve Kenelerde Anaplasma / Ehrlichia Türlerinin Belirlenmesi.

OBJECTIVE: The aim of this study is to detect the Anaplasma/Ehrlichia species of cattle and ticks and to provide knowledge on the prevalence of these species during sampling periods. METHODS: A total of 679 blood and 186 tick samples were collected from the Osmanbükü, Akçaova, Dalama, and Söke districts of Aydin. The samples were screened with genus polymerase chain reaction (PCR) for Anaplasma/Ehrlichia spp., species-specific polymerase chain reaction for Anaplasma marginale and A. centrale, and nested PCR for A. bovis and A. phagocytophilum. RESULTS: A. centrale was detected in Söke during September and in Dalama and Akçaova during March, June, September, and December. A. marginale was detected in Osmanbükü during June; in Söke during March and December; in Akçaova during June, September, and March; and in Dalama during the entire sampling period. A. phagocytophilum was detected in all regions during the entire sampling period. None of the samples were positive for A. bovis. Mixed infections were detected in 50 blood samples. A. marginale and A. phagocytophilum were detected in the tick samples. CONCLUSION: In this study, A. phagocytophilum was abundantly detected compared with A. marginale and A. centrale. A. phagocytophilum and A. centrale were extensively found in Akçaova and A. marginale was mostly seen in Dalama. Parasites were extensively detected in September and March. The analysis indicated that collected ticks were infected with different Anaplasma/Ehrlichia species.

Single dose of secnidazole treatment against naturally occuring Giardia duodenalis infection in Sakiz lambs / Tratamiento con dosis única con secnidazol en corderos naturalmente infectados con Giardia duodenalis

Objetive. The purposes of this study were to determine whether secnidazole administered at a single dose of 10 mg/kg, orally, lessens or eliminates Giardia cyst shedding, and to validate the benefit of secnidazole treatment on clinical signs and faecal consistency in lambs naturally infected with Giardia duodenalis. Materials and methods. To this extent weaned 12 weeks of age lambs were selected and randomly assigned into two groups based on placebo (group C, n=7 untreated control group) or treatment (group S, n=10 lambs treated with a single dose of secnidazole at 10 mg/kg). Results. On days 0 and 10, before and after treatment, respectively, reduction in cyst excretion was determined. The faecal consistency and general health were recorded at each sampling day. Conclusions. During the study there was a high (99.98%) reduction in cyst excretion in the secnidazol treatment group compared to the positive control group on day 10, resulting in a significant (p<0.001) reduction, making secnidazole highly effective treatment option.

Objetivo. El objetivo de este estudio fue determinar si la administración oral de una dosis única de secnidazol de 10mh/kg reduce o elimina la presencia de quistes de Giardia y validar el beneficio del tratamiento con secnidazol en los signos clínicos y la consistencia fecal de corderos infectados naturalmente con Giardia duodenalis. Materiales y métodos. Se seleccionaron corderos destetados de 12 semanas de edad que fueron asignados al azar a dos grupos con base a placebo (grupo C, n=7, grupo de control sin tratar) o tratamiento (grupo S, n=10 corderos con tratamiento de dosis única de secnidazol de 10 mg/kg). Resultados. En los días 0 y 10, antes y después del tratamiento, respectivamente, se determinó una merma en la excreción de quistes. La consistencia de la materia fecal y el estado general de salud fue registrado en cada muestreo diario. Conclusiones. Durante el estudio hubo una alta reducción en la excreción de quistes (99.98%) en el grupo tratado con secnidazol comparado con el grupo control positivo en el día 10, resultando en una disminución significativa (p<0.001), haciendo del tratamiento con secnizadol una opción altamente efectiva.

Development of a multiplex PCR assay for simultaneous detection of Theileria annulata, Babesia bovis and Anaplasma marginale in cattle.

Tropical theileriosis, bovine babesiosis and anaplasmosis are tick-borne protozoan diseases that impose serious constraints on the health and productivity of domestic cattle in tropical and sub-tropical regions of the world. A common feature of these diseases is that, following recovery from primary infection, animals become persistent carriers of the pathogen and continue to play a critical role in disease epidemiology, acting as reservoirs of infection. This study describes development and evaluation of multiplex and single PCR assays for simultaneous detection of Theileria annulata, Babesia bovis and Anaplasma marginale in cattle. Following in silico screening for candidate target genes representing each of the pathogens, an optimised multiplex PCR assay was established using three primer sets, cytob1, MAR1bB2 and bovar2A, for amplification of genomic DNA of T. annulata, A. marginale and B. bovis respectively. The designed primer sets were found to be species-specific, generating amplicons of 312, 265 and 166 base pairs, respectively and were deemed suitable for the development of a multiplex assay. The sensitivity of each primer pair was evaluated using serial dilutions of parasite DNA, while specificity was confirmed by testing for amplification from DNA of different stocks of each pathogen and other Theileria, Babesia and Anaplasma species. Additionally, DNA preparations derived from field samples were used to evaluate the utility of the single and multiplex PCRs for determination of infection status. The multiplex PCR was found to detect each pathogen species with the same level of sensitivity, irrespective of whether its DNA was amplified in isolation or together with DNA representing the other pathogens. Moreover, single and multiplex PCRs were able to detect each species with equal sensitivity in serially diluted DNA representing mixtures of T. annulata, B. bovis and A. marginale, and no evidence of non-specific amplification from non-target species was observed. Validation that the multiplex PCR efficiently detects single and mixed infections from field samples was demonstrated. The developed assay represents a simple and efficient diagnostic for co-detection of tropical theileriosis, bovine babesiosis and anaplasmosis, and may be a valuable tool for epidemiological studies aimed at assessing the burden of multiple infection with tick-borne pathogens and improving control of the associated diseases in endemic regions.

Distribution and seasonal activity of tick species on cattle in the West Aegean region of Turkey.

The aim of the present study was to determine the identity, seasonal activity and distribution of tick species of cattle in the West Aegean region of Turkey between June 2006 and May 2008. Nine villages within three provinces, viz. Manisa, Izmir and Aydin, were included in the study and a total of 75 animal barns were visited monthly for a period of 24 months and 443 cattle were examined for the presence of ticks. It was determined that 23% of cattle were infested with ticks. A total of 19,679 adult ticks were collected. The most abundant tick species was Hyalomma marginatum (33.5%) and H. excavatum (16.9%) in the study area. Seasonal appearance of the adult ticks varied among species. Adult ticks of the Hyalomma genus were present throughout the year, although in smaller numbers during the winter. Species of Rhipicephalus were detected in all seasons except autumn. Rhipicephalus (Boophilus) annulatus was identified in July and August, Haemaphysalis parva was detected during the autumn. Ixodes ricinus and Dermacentor marginatus were identified during spring, autumn and winter. The study demonstrated the presence of I. ricinus, D. marginatus, Hyalomma rufipes and Hae. parva for the first time in the West Aegean region of Turkey.

[Some biological features of Hyalomma marginatum in the laboratory]. / Laboratuvarda Hyalomma marginatum'un Bazi Biyolojik Özellikleri.

OBJECTIVE: This study has been planned to investigate some biological features of Hyalomma marginatum, which is one of the important vectors in the world and in our country. METHODS: The study started with the adult H. marginatum which were collected from cattle in the field and attempts were made for these to be adapted to the laboratory by breeding in the laboratory throughout a generation. Unfed adult ticks and unfed larvae were bred on the rabbit ear while unfed nymphs were bred on the calf ear. The biologic development stages of this tick kind, except for sucking blood stage, passed in an incubator which was at a temperature of 28°C and 85% relative humidity, while the unfed active stages passed in an incubator which was at 18°C and 85% relative humidity to conserve for a long time. RESULTS: Unfed females on rabbits were full and fell in approximately 15 days, began to ovulate 20.5 days later and ovulated for 16 days. The larvae which hatched out approximately 29 days later became active in 8.5 days. These larvae sucked blood from rabbits for 14.5 days and abandoned these animals as full nymphs. Unfed adult ticks, which existed as a result of these nymphs casting off their skin in approximately 26 days, became active by completing their cutinization in approximately 10 days. CONCLUSION: At the end of the study, it was determined that H. marginatum showed 2 different behaviors in rabbits. According to this, the total life cycle in the laboratory environment changed by between 97 and 182 days and lasted approximately 138.5 days.

Evaluation of cytochrome b as a sensitive target for PCR based detection of T. annulata carrier animals.

Bovine tropical theileriosis, caused by the tick-borne protozoan Theileria annulata, imposes a serious constraint upon breed improvement programmes and livestock production in tropical and sub-tropical regions of the world. Animals that recover from primary infection serve as carriers and play a critical role in the epidemiology of the disease, acting as reservoirs of infection. However, conclusive identification of carrier animals can be problematic. This study describes assessment of candidate target genes for PCR assay-based detection of T. annulata infected carrier animals. Following in silico screening and rejection of three major multi-copy gene families, an assay based on PCR amplification of a 312 bp segment of the T. annulata gene for cytochrome b (Cytob1 assay) was established. Sensitivity was evaluated using serial dilutions of blood obtained from experimentally infected calves, while specificity was confirmed by testing DNA representing twelve different T. annulata stocks and other Theileria and Babesia species. Direct comparison with other target genes and published data indicated that Cytob1 PCR-based assays provide the greatest level of sensitivity, combined with a high level of specificity and the ability to detect different T. annulata genotypes. It can be concluded that the cytochrome b gene is the optimal target for PCR amplification and its incorporation in a Reverse Line Blot Assay offers the most sensitive method yet devised to detect the parasite in carrier animals. The use of this assay will increase the accuracy of epidemiological studies aimed at improving disease control in endemically unstable regions.

A case of naso-ophthalmic myiasis.

Myiasis is defined as an occupational disease and it is mainly seen in people who are in contact with animals, in particular sheep. We detected nasal and external ophthalmomyiasis caused by Oestrus ovis in a 33-year-old male who worked as a research assistant in the faculty of agriculture. The presenting complaints were severe foreign body sensation and discharge in the nose, cough, and foreign body sensation in the right eye. Ophthalmologic examination revealed many moving larvae in the right eye of the patient. Larvae were removed and nasolacrimal duct irrigation was performed through the inferior canaliculus. Endoscopic nasal examination also showed larvae colonies. The larvae were removed under local anesthesia and were taken to the parasitology laboratory, wherein Oestrus ovis was identified. Minimal endoscopic sinus surgery showed nonspecific chronic inflammation.

[Prevalence of tropical theileriosis in cattle in the Aydin Region and determination of efficacy of attenuated Theileria annulata vaccine]. / Aydin Ili Sigirlarinda Tropikal Theileriosisin Yayginligi ve Theileria annulata Sizont Asisinin Sahada Etkinliginin Degerlendirilmesi.

The present study was carried out to find out the prevalence of tropical theileriosis in the Aydin region and to determine immune status of cattle vaccinated with Theileria annulata schizont vaccine using the indirect fluorescence antibody test (IFAT) during and after the disease season. A total of 236 out of 466 cattle found to be seronegative with IFAT were vaccinated in the Aydin region (Cine, Incirliova, Nazilli, and Centrum) before the disease season (March). The remaining cattle (230/466) served as controls. Blood samples were collected from all cattle following the vaccination once in each month of April, June, September, and December (a total of 4 times) to determine the incidence of the disease using microscopic and serologic evaluations. Results indicated that the incidence of the disease varied among the regions. Clinical theileriosis was found in a total of 22 cattle (10 vaccinated, 12 unvaccinated) during the disease season.

[Gastro-intestinal helminths detected by fecal examination in stray dogs in the Aydin province]. / Aydin yöresi sokak köpeklerinde disk bakisina göre saptanan mide bagirsak helmintleri.

Fecal specimens of a total of 200 dogs were examined by native, Fulleborn's floatation and Benedek's sedimentation methods to determine the prevalence of gastro-intestinal helminth infections in stray dogs in the Aydin Municipality Animal Shelter and the Kusadasi Municipality Animal Shelter. Helminth infections were encountered in 82 (41%) of the fecal samples examined. One cestode egg and four nematode eggs were found in the infected fecal samples. The helminth eggs found were identified as follows: Taenia spp. (7.5%), Toxacara spp. (20%), Toxascaris leonina (1%), Uncinaria stenocephala (21%) and Trichuris vulpis (1.5%). No trematode eggs and nematode larvae were found in this study.

A parasitological, molecular and serological survey of Hepatozoon canis infection in dogs around the Aegean coast of Turkey.

Canine hepatozoonosis is caused by the tick-borne protozoon Hepatozoon spp. The prevalence of the infection in the Aegean coast of Turkey was investigated by examination of blood smear parasitology and polymerase chain reaction (PCR) using blood samples from 349 dogs collected from Central Aydin, Kusadasi, Selcuk, Central Manisa, Bodrum and Marmaris within the Aegean coast of Turkey. The indirect fluorescent antibody test (IFAT) for the detection of Hepatozoon canis antibodies was also used to detect the exposure rate to H. canis. PCR amplifying a 666bp fragment of 18S rRNA gene of Hepatozoon spp. was used in the epidemiological survey. The prevalence of Hepatozoon spp. infection was 10.6% by blood smear parasitology and 25.8% by PCR. IFAT revealed that 36.8% of serum samples were positive for antibodies reactive with Hepatozoon spp. The PCR products of 18S rRNA gene of Hepatozoon spp. isolated from six infected dogs, one isolate originating from each of the six different locations, were sequenced. The results of sequence analysis indicate that they are closely related to Indian and Japanese isolates of H. canis. This is the first epidemiological study on the prevalence of H. canis infection in the dog, in Turkey.