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

Infection dynamics of Theileria annulata over a disease season following cell line vaccination

Tropical theileriosis is a tick-borne haemoparasitic disease of cattle caused by the protozoan parasite Theileria annulata. Globally, the economic impact of the disease is immense and enhanced control measures would improve livestock production in endemic regions. Immunisation with a live attenuated vaccine is an effective and widely used control method, however, the repeated use of live vaccines may have an impact on the field parasite population at a genetic level. Additionally, there has been an increasing number of reports of vaccine breakthrough cases in recent years. Thus, the present study was designed to evaluate the genetic composition of a parasite population over a disease season in a locality where live cell line vaccination is practised. A diverse range of parasite genotypes was identified and every T. annulata positive cattle blood sample harboured multiple parasite genotypes. An alteration in the major genotype and an increasing multiplicity of infection in individual animals was observed over the course of the disease season. Vaccination status was found not to effect within-host multiplicity of infection, while a significantly higher number of genotypes was detected in grazed cattle compared to non-grazed ones. A degree of genetic isolation was evident between parasite populations on a micro-geographic scale, which has not been reported previously for T. annulata. Analysis of parasite genotypes in vaccinated animals suggested only a transient effect of the vaccine genotype on the genetic diversity of the T. annulata population. The vaccine genotype was not detected among clones of two vaccine ‘breakthrough’ isolates and there is no suggestion that it was responsible for disease. The obtained data indicated that in the system studied there is no apparent risk of introducing the vaccine genotype into the population with only a transient effect on the genetic diversity of the parasite population during the disease season

Selection of genotypes harbouring mutations in the cytochrome b gene of Theileria annulata is associated with resistance to buparvaquone

Buparvaquone remains the only effective therapeutic agent for the treatment of tropical theileriosis caused by Theileria annulata. However, an increase in the rate of buparvaquone treatment failures has been observed in recent years, raising the possibility that resistance to this drug is associated with the selection of T. annulata genotypes bearing mutation(s) in the cytochrome b gene (Cyto b). The aim of the present study was: (1) to demonstrate whether there is an association between mutations in the T. annulata Cyto b gene and selection of parasite-infected cells resistant to buparvaquone and (2) to determine the frequency of these mutations in parasites derived from infected cattle in the Aydın region of Türkiye. Susceptibility to buparvaquone was assessed by comparing the proliferative index of schizont-infected cells obtained from cattle with theileriosis before and/or after treatment with various doses of buparvaquone, using the 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colourimetric assay. The DNA sequence of the parasite Cyto b gene from cell lines identified as resistant or susceptible was determined. A total of six nonsynonymous and six synonymous mutations were identified. Two of the nonsynonymous mutations resulted in the substitutions V135A and P253S which are located at the putative buparvaquone binding regions of cytochrome b. Allele-specific PCR (AS-PCR) analyses detected the V135A and P253S mutations at a frequency of 3.90% and 3.57% respectively in a regional study population and revealed an increase in the frequency of both mutations over the years. The A53P mutation of TaPIN1 of T. annulata, previously suggested as being involved in buparvaquone resistance, was not detected in any of the clonal cell lines examined in the present study. The observed data strongly suggested that the genetic mutations resulting in V135A and P253S detected at the putative binding sites of buparvaquone in cytochrome b play a significant role in conferring, and promoting selection of, T. annulata genotypes resistant to buparvaquone, whereas the role of mutations in TaPIN1 is more equivocal

In vitro feeding of all life stages of two-host Hyalomma excavatum and Hyalomma scupense and three-host Hyalomma dromedarii ticks

Ticks are blood-sucking ectoparasites and can transmit various pathogens of medical and veterinary relevance. The life cycle of ticks can be completed under laboratory conditions on experimental animals, but the artificial feeding of ticks has attracted increased interest as an alternative method. This study represents the first report on the successful in vitro feeding of all life stages of two-host tick species, Hyalomma scupense and Hyalomma excavatum, and the three-host tick Hyalomma dromedarii. The attachment and engorgement rates of adults were 84% (21/25) and 76% (19/25) for H. scupense females. For adult H. excavatum and H. dromedarii, 70% (21/30) and 34.4% (11/32) of the females attached and all attached females successfully fed to repletion. The oviposition rates of the artificially fed females were 36.4%, 57.1% and 63.1% for H. dromedarii, H. excavatum and H. scupense, respectively, with a reproductive efficiency index varying between 44.3 and 60.7%. For the larvae, the attachment and engorgement rates were 44.2% (313/708) and 42.8% (303/708) for H. dromedarii, 70.5% (129/183) and 56.8% (104/183) for H. excavatum and 92.6% (113/122) and 55.7% (68/122) for H. scupense. The attachment and engorgement rates for the nymphs were 90.2% (129/143) and 47.6% (68/143) for H. dromedarii, 66.7% (34/51) and 41.2% (21/51) for H. excavatum, and 44.1% (30/68) and 36.8% (25/68) for H. scupense. Molting rates of the immature stages varied between 71.3% (216/303) and 100% (68/68) for the larvae and between 61.9% (13/21) and 96% (24/25) for the nymphs. The successful in vitro feeding of all stages of the three Hyalomma species makes this method a valuable tool for tick research, with potential applications in studies on the pathogens transmitted by these tick species such as Theileria annulata

Multifunctional 3D printing of heterogeneous hydrogel structures

Multimaterial additive manufacturing or three-dimensional (3D) printing of hydrogel structures provides the opportunity to engineer geometrically dependent functionalities. However, current fabrication methods are mostly limited to one type of material or only provide one type of functionality. In this paper, we report a novel method of multimaterial deposition of hydrogel structures based on an aspiration-on-demand protocol, in which the constitutive multimaterial segments of extruded filaments were first assembled in liquid state by sequential aspiration of inks into a glass capillary, followed by in situ gel formation. We printed different patterned objects with varying chemical, electrical, mechanical, and biological properties by tuning process and material related parameters, to demonstrate the abilities of this method in producing heterogeneous and multi-functional hydrogel structures. Our results show the potential of proposed method in producing heterogeneous objects with spatially controlled functionalities while preserving structural integrity at the switching interface between different segments. We anticipate that this method would introduce new opportunities in multimaterial additive manufacturing of hydrogels for diverse applications such as biosensors, flexible electronics, tissue engineering and organ printing

Geographical distribution of ticks in Turkey

Bu proje, Avrupa Birliği tarafından desteklenmiştir. Integrated Consortium Ticks Tick Borne Dis 3 (ICTTD-3, IC18-CT95-0009)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.Avrupa Birliğ

Distribution of tick species on cattle in the Western Anatolia

Bu çalışma Batı Anadolu Bölgesi sığırlarında bulunan kene türlerini, mevsimsel aktivitelerini ve yaygınlıklarını belirlemek amacıyla planlanmış ve Haziran 2006-Mayıs 2008 tarihleri arasında Manisa, İzmir ve Aydın illerine bağlı dokuz köyde yürütülmüştür. Belirlenen çalışma merkezlerine yirmi dört ay boyunca ayda bir düzenli olarak gidilmiş, bu merkezlerdeki 75 hayvan barınağı ve bu barınaklardaki sığırlar çalışma boyunca 8018 kez kene varlığı açısından incelenmiştir.Araştırma süresince sığırların %23,01'inin kene ile enfeste olduğu saptanmış ve sığırlardan toplam 19679 erişkin kene toplanmıştır. Toplanan bu kenelerden 3318 (%16,86)'i Manisa'da, 8980 (%45,63)'i İzmir'de ve 7381 (%37,51)'i Aydın ilinde bulunmuştur. Çalışma merkezlerindeki sığırlarda erişkin olarak 12 kene türü (Ixodes ricinus %2,88, Rhipicephalus (Boophilus) annulatus %0,14, Dermacentor marginatus %4,49, Hyalomma anatolicum %0,86, Hyalomma excavatum %18,89, Hyalomma detritum %13,68, Hyalomma marginatum %37,39, Hyalomma rufipes %0,07, Haemaphysalis parva %0,04, Rhipicephalus bursa %4,22, Rhipicephalus sanguineus %0,04, Rhipicephalus turanicus %17,24) saptanmıştır. Tür tayini yapılamayan kene oranı ise %0,06 olarak belirlenmiştir. Teşhis edilen kene türlerinin mevsimsel aktivitelerine bakıldığında, Ixodes ricinus'un erişkinlerinin, Kasım ayının başlarında başlayıp, Mayıs ayı başlarına kadar görüldüğü, Rhipicephalus (Boophilus) annulatus'un, Temmuz, Ağustos ile Ekim, Kasım aylarında gözlendiği, Dermacentor marginatus'un Eylül ? Haziran (Mayıs ayı hariç) ayları arasında görüldüğü tespit edilmiştir. Çalışma yürütülen bölgede Hyalomma türlerinin erişkinlerinin tüm yıl boyunca görüldüğü, ancak kış aylarında sınırlı sayılarda kaldığı belirlenmiştir. Haemaphysalis parva türünün sığırlar üzerinde sadece Ekim, Kasım aylarında az sayıda olduğu saptanmıştır. Rhipicephalus türleri ise daha çok ilkbahar ve yaz aylarında tespit edilmiştir.Hyalomma marginatum (%37,39) ve Hyalomma excavatum (%18,89) bölgede en yaygın kene türleri olarak tespit edilmiş olup, Ege Bölgesinde daha önceleri bildirilmeyen Ixodes ricinus (%2,88) , Dermacentor marginatus (%4,49) ve Haemaphysalis parva (%0,04) sığırlarda saptanmıştır. Bunun dışında daha önce Türkiye'de bildirilmeyen Hyalomma rufipes (%0,07) türü de Ege Bölgesindeki sığırlarda bulunmuş olup, bu kene türünün varlığı Türkiye'de ilk kez belirlenmiştir.The aims of the present study were to determine the identity, seasonal activity and distribution of tick species of cattle in the Western Anatolia of Turkey between June2006- May 2008. Nine villages within three cities, viz. Manisa, Izmir and Aydin, were included in the study and a total of 75 animal cernals were visited monthly for a period of 24 months and cattle were examined 8018 times for the presence of ticks.In this study, 23,01 % of cattle were determined to be infested with ticks. A total of 19679 adult ticks were collected. 3318 (16,86%), 8980 (45,63%), 7381 (37,51%) out of 19679 adult ticks were collected from Manisa, Izmir and Aydin, respectively. Twelve tick species (Ixodes ricinus 2,88%, Rhipicephalus (Boophilus) annulatus 0,14%, Dermacentor marginatus 4,49%, Hyalomma anatolicum 0,86%, Hyalomma excavatum 18,89%, Hyalomma detritum 13,68%, Hyalomma marginatum 37,39%, Hyalomma rufipes 0,07%, Haemaphysalis parva 0,04%, Rhipicephalus bursa 4,22%, Rhipicephalus sanguineus 0,04%, Rhipicephalus turanicus 17,24%) were identified. The percantage of unidentified ticks was 0,06%. Among the identified tick species, adult Ixodes ricinus occurred between 1st of November and begining of May. Occurrence of Rhipicephalus (Boophilus) annulatus, between July and August and October and November, on cattle. Dermacentor marginatus was observed between September and June (excluding May). Adults of Hyalomma species were observed during the whole year, although they were more abundant in summer time than that observed in winter. Rhiphicephalus species were found to be more abundant in the spring and summer.Hyalomma marginatum (37,39%) and Hyalomma excavatum (18,89%) were determined to be the most abundant tickspecies. Species that have not been reported previously, namely, Ixodes ricinus (2,88%), Dermacentor marginatus (4,49%) and Haemaphysalis parva (0,04%), were detected on cattle in the Aegean Region of Turkey. In addition, the present study demonstrates that Hyalomma rufipes (0,07%), a species that is not known to exist in Turkey, is present in cattle in the Aegean Region of Turkey.ADÜ Araştırma Fon

Examining Effective Communication Levels Of Student Athletes: A Sample On Football As An Sports Branch

This study was conducted with the aim of examining the contribution of football to the communication levels of the student athletes who are at different grade levels and play in amateur sports clubs in terms of some variables. The sample of the study which was conducted by using general screening model is composed of 174 volunteer students selected through simple random method. In the research, “Scale of Effective Communication in Team Sports” was used as the data collection tool. As a result of the research, the athletes in the sample were given the answers to the scale questions; In the sub-dimension of acceptance and positive conflict, the participants of secondary education, aged 15 to 18, with family income of 5001 ? and over, playing in striker and midfielder position and playing football in the 1st Leagues, compared to the other participants, It was determined that the participants who were educated at university level, 23-26 age group, family income level of 5001 ? and above, playing in goalkeeper and striker position and playing football in school team and amateur leagues were more effective in communicating with teammates than the other participants. In addition, while the statistical value of education, age and family income level showed a significant difference in both sub-dimensions, it was found that they did not show a significant difference in the variables of play area and sporting level. Key Words: Communication, Athlete, Student, School Team, Amateur Sports.Bu araştırma futbol spor dalının; farklı okul düzeylerinde okuyan ve amatör spor kulüplerinde oynayan sporcu öğrenciler üzerindeki iletişim düzeylerine olan katkısının bazı değişkenlere göre inceleyebilmek amacıyla yapılmıştır. Genel tarama modeli kullanılarak yapılan araştırmanın örneklemini basit tesadüfü yöntemle seçilmiş gönüllü 174 sporcu öğrenci oluşturmaktadır. Araştırmada veri toplama aracı olarak “Takım Sporlarında Etkili İletişim Ölçeği” kullanılmıştır. Araştırma sonucunda örneklemdeki sporcu öğrencilerin ölçek sorularına vermiş oldukları cevaplardan hareketle; kabul etme ve pozitif çatışma alt boyutunda ortaöğretim düzeyinde eğitim alan, 15 ile 18 yaş aralığında olan, 5001 TL ve üzerinde aile gelirleri olan, forvet ve orta saha mevkiinde oynayan ve 1. Liglerde futbol oynayan katılımcıların diğer katılımcılara göre daha, ayırt etme ve negatif çatışma alt boyutunda ise üniversite düzeyinde eğitim alan, 23-26 yaş aralığında olan, 5001 TL ve üzeri aile gelir düzeyi olan, kaleci ve forvet mevkiinde oynayan ve okul takımı ve amatör liglerde futbol oynayan katılımcıların diğer katılımcılara göre daha fazla puan ile takım arkadaşlarıyla etkili iletişim içerisinde oldukları belirlenmiştir. Ayrıca araştırmada her iki alt boyutta da istatistiki değer olarak eğitim, yaş ile aile gelir düzeyi değişkenlerinde anlamlı bir fark gösterirken, oynadıkları oyun bölgesi ve spor yapma düzeyi değişkenlerinde anlamlı bir fark göstermediği saptanmıştır. Anahtar kelimeler: İletişim, Sporcu, Öğrenci, Okul Takımı, Amatör Spor

Investigation of Opinions of Seventh Grade Students about Socio-Scientific Issues by means of Science Teaching Based on Common Knowledge Construction Model

The aim of this study is to investigate the opinions of seventh-grade students on socio-scientific issues through science teaching based on a common knowledge construction model. The study was carried out with 25 grade seven students attending a secondary school in the spring term of the 2016-2017 academic year. Within case study research methodology, one of qualitative research methods, was applied. In the study, the Socio-Scientific Issues Evaluation Form and semi-structured interview questions were used for data collection. In analyzing the data content analysis were used. The findings indicated that the students found socio-scientific issues were complex, open-ended and mostly debatable issues which did not any have clear-cut answer. The CKCM lessons sequence was found to be influential on the development of the 7th grade students’ decision-making skills regarding socio-scientific issues, problem-solving skills in daily lives and the development of science literacy. To clarify the effect of the CKCM on socio-scientific issues, future studies should be conducted with a larger sample from different grades