Serkan Bakirci

Geographical distribution of ticks in Turkey

Turkey’s tick fauna is composed of about 32 species in two families and ten genera in mammals, reptiles, and birds. The ticks of veterinary significance in the family Ixodidae comprise seven genera with 28 different species. Ixodes spp. are mostly seen in northern Turkey. It is likely that the high rainfall and the intensive forest in this area may contribute to this observation. To date, the following species have been found to be sporadically present: Amblyomma variegatum in Hatay province (border to Syria), Boophilus kohlsi in southeastern Turkey (border of Syria), Ornithodorus in Central and East Anatolia, and Otobius megnini in East Anatolia (Malatya Province). Ticks of the genera Haemaphysalis, Hyalomma, Boophilus, Dermacentor, Rhipicephalus, and Argas are widespread throughout Anatolia. Thus, their role in the epidemiology of important human and livestock diseases such as Crimean–Congo hemorrhagic fever, Lyme disease, and diseases caused by hemoparasites needs to be examined in greater detail.

In-vivo and in-vitro tick repellent properties of cotton fabric:

N,N-diethyl-m-toluamide (DEET) and eucalyptol were encapsulated by β-cyclodextrin (β-CD) using the kneading method at four different molar ratios. The products were applied to 100% cotton fabric through the impregnation method. Fourier transform infrared and differential scanning calorimetry analysis results confirmed that the inclusion complex occurred between β-CD and repellent agents at mole ratios of 1:3 and 1:2 for eucalyptol and DEET, respectively. The experiments carried out on Hyalomma marginatum showed that DEET and eucalyptol treated fabrics repel, inhibit and kill blood-feeding Hyalomma marginatum ticks in-vivo and in-vitro for a period of at least 48 and 72 hours under laboratory conditions. In-vitro essay showed good repellent activities for both DEET and eucalyptol. The results show that eucalyptol has more repellent effect than DEET. In the case of the in-vivo essay, a single washing cycle of eucalyptol: β-CD-treated fabric showed repellent activity.

Modelling the transmission dynamics of Theileria annulata: model structure and validation for the Turkish context.

SUMMARY A mathematical model that describes the transmission dynamics of Theileria annulata is proposed that consists of 2 host components: the Hyalomma tick population and a compartmental model of T. annulata infection in the cattle population. The model was parameterized using data describing tick infestation and the infection status of cattle in Turkey from 2006 to 2008. The tick attachment rates are highly seasonal and because of the temporal separation of infectious and susceptible ticks virtually all ticks are infected by carrier cattle, so that annual peaks of disease in cattle do not impact on infection in the Hyalomma tick population. The impact of intervention measures that target the tick population both on the host and in the environment and their impact on the transmission of T. annulata were investigated. Interventions that have a limited ‘one-off’ impact and interventions that have a more permanent impact were both considered. The results from the model show the importance of targeting ticks during the period when they have left their first host as nymphs but have yet to feed on their second host.

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

BackgroundTick-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.ResultsOverall, 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.ConclusionThe 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.

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.

Differential susceptibility of Hyalomma excavatum adults and nymphs to the entomopathogens Metarhizium anisopliae ARSEF 4556 and Steinernema carpocapsae

ABSTRACT This study investigated the virulence of the entomopathogenic fungus, Metarhizium anisopliae strain ARSEF 4556, and the entomopathogenic nematode Steinernema carpocapsae against adult and nymphal stages of the tick Hyalomma anatolicum excavatum. The fungus was tested at three different concentrations, 106, 107, and 108 conidia ml−1. Ticks were exposed to 4000 IJs of S. carpocapsae in 0.5 ml distilled water in Petri dishes lined with 2 layers of moist filter paper. Adult ticks were not susceptible to M. anisopliae, whereas the nymphs were highly susceptible. Nymph mortality was dose dependent with all ticks being killed at the highest dose 96 hours post-inoculation. In contrast, S. carpocapsae was much more effective against engorged adults than unfed adults or nymphs. Tick susceptibility was not influenced by gender. The mean mortalities of the unfed adult males, females, and nymphs were 30% ± 9.2%, 40% ± 7.5%, and 36% ± 4.7%, respectively. Nymphal mortality was 36% and there was a significant difference between nematode treatment and control group (P < 0.05).

Identification of candidate transmission-blocking antigen genes in Theileria annulata and related vector-borne apicomplexan parasites

BackgroundVector-borne apicomplexan parasites are a major cause of mortality and morbidity to humans and livestock globally. The most important disease syndromes caused by these parasites are malaria, babesiosis and theileriosis. Strategies for control often target parasite stages in the mammalian host that cause disease, but this can result in reservoir infections that promote pathogen transmission and generate economic loss. Optimal control strategies should protect against clinical disease, block transmission and be applicable across related genera of parasites. We have used bioinformatics and transcriptomics to screen for transmission-blocking candidate antigens in the tick-borne apicomplexan parasite, Theileria annulata.ResultsA number of candidate antigen genes were identified which encoded amino acid domains that are conserved across vector-borne Apicomplexa (Babesia, Plasmodium and Theileria), including the Pfs48/45 6-cys domain and a novel cysteine-rich domain. Expression profiling confirmed that selected candidate genes are expressed by life cycle stages within infected ticks. Additionally, putative B cell epitopes were identified in the T. annulata gene sequences encoding the 6-cys and cysteine rich domains, in a gene encoding a putative papain-family cysteine peptidase, with similarity to the Plasmodium SERA family, and the gene encoding the T. annulata major merozoite/piroplasm surface antigen, Tams1.ConclusionsCandidate genes were identified that encode proteins with similarity to known transmission blocking candidates in related parasites, while one is a novel candidate conserved across vector-borne apicomplexans and has a potential role in the sexual phase of the life cycle. The results indicate that a ‘One Health’ approach could be utilised to develop a transmission-blocking strategy effective against vector-borne apicomplexan parasites of animals and humans.

Investigation of the cases presenting to Dokuz Eylül University Hospital with tick bites

OBJECTIVE Determination of the properties of ticks, which are the vectors of many infectious diseases and the patients with tick bites are important for the prevention of these diseases. For tist reason, the purpose of this study is to determine the important properties of the cases presenting with tick bites to Dokuz Eylul University Hospital and of the removed ticks from the cases. METHODS Two hundred seventy three of 294 patients, who presented with tick bites to Dokuz Eylül University Hospital, Izmir, were included in the study. Demographic parameters, symptoms related to tick borne diseases of the patients and the species and other characteristics of ticks removed from humans, also the tick population in the related habitat were investigated. RESULTS Removed ticks were classified into five genera. The overwhelmingly dominant genera were Hyalomma and it comprised 52.4% of the collection. The majority of these ticks were nymphs. The majority (11.4%) of removed adult ticks were Rhipicephalus sanguineus (R. sanguineus). In most cases (60.7%) the ticks were removed from the patients by medical staff. CONCLUSION There was no significant difference in removing ticks without damage between the health personnel and the patient or relatives (p=0.133).

[Detection of Anaplasma / Ehrlichia Species of Cattle and Ticks in Aydın Region].

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 Aydın. 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.

[Gerbils, As Experimental Animals (Meriones unguiculatus): Is A Good Role Model for Leishmania major?].

OBJECTIVE This study aimed to observation the possible visceralization tendency and dissemination of L. major amastigotes in gerbils (Meriones unguiculatus) using a classic smear technique, inoculated into enriched Novy-MacNeal-Nicolle (NNN) culture and polymerase chain reaction (PCR) assay for diagnosis of infection. METHODS In this study, L. major isolated from a man who 18 years old, living in Bitlis province of Turkey. This strain also was utilized to infect gerbils. A total of 1 × 10(8)/mL promastigotes were inoculated to 10 gerbils. Necropsy was performed on infected gerbils for monitoring the visceralization tendency of the parasites. Tissue samples were prepared from each animal and stained by Giemsa and inoculated into NNN culture. However, a real-time PCR assay was performed to confirm the infection the clinical material. RESULTS Examination of Giemsa-stained tissue smears showed that infected animals with L.major were positive for Leishmania amastigotes in all tissues at the first month post infection and Leishmania promastigotes were cultured at 26°C in culture flasks containing NNN. Melting curve analyses of ribozomal internal transcribed spacer 1 (ITS-1) PCR showed the peak concordant with L. major. CONCLUSION As a result, the present study confirmed by both Giemsa-stained smears and PCR, visceralization and dissemination of L. major amastigotes, the principal cause of zoonotic cutaneous leishmaniasis in gerbils.