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© 2022 John Wiley & Sons Ltd.We assessed genetic diversities among Ichthyophthirius multifiliis (Ich) field isolates collected from farmed rainbow trout (Oncorhynchus mykiss) in Turkey. The overall prevalence of Ich was 35.3% (634/1798). Five novel Ich genotypes (ImulTR1 and ImulTR3‒ImulTR6) were described based on mitochondrial cox-1 and nad1_b genes. The remaining genotype ImulTR2 was identical to the previously reported NY3 (or Ark9 and TW7) genotype from the United States and South Asia. Phylogenetic analysis indicated Turkish Ich isolates separated genetically into at least four distinct groups. Our study presents the first data on the genotypes of Ich in Turkey. We also provide evidence for the wide distribution of the NY3 genotype (or Ark9 and TW7) from the United States and South Asia to Turkey. Genetic diversities within the mitochondrial genes provided adequate resolution for describing novel genotypes and identifying the known genotype within Turkish Ich isolates. Description of the Ich genotypes allows for tracking of pathogen genotypes worldwide. Thus, we can better understand the connections between Ich outbreaks in the fisheries aquaculture.

© 2022 Elsevier B.V.Microsporidia are obligate intracellular fungus-like parasites that infect humans and animals worldwide. However, there is limited epidemiological data on the occurrence and molecular diversity of microsporidia in buffaloes worldwide. In the present study, fecal samples of 300 water buffaloes (Bubalus bubalis) in Kayseri, Sivas, and Samsun provinces of Turkey were investigated using two nested PCR assays targeting the rRNA of E. bieneusi and Encephalitozoon spp. All the fecal samples from water buffalo were found to be negative for Encephalitozoon spp. PCR positive isolates of E. bieneusi were bidirectionally sequenced for genotyping and phylogenetic analyses. Enterocytozoon bieneusi was the only microsporidian species identified in 8 water buffaloes with an overall molecular prevalence of 2.7%. Two known genotypes, YNDCEB-90 (n = 5) and J (n = 3) were identified by ITS sequence analysis. The YNDCEB-90 and J genotypes fall into zoonotic Group 1 and 2 of E. bieneusi in the phylogenetic tree, respectively. These findings suggested that water buffalo in Turkey are harbouring zoonotic genotypes of E. bieneusi and may have a significant risk for zoonotic transmission to humans. This is the first report of detecting E. bieneusi genotypes J and YNDCEB-90 in water buffaloes. Further insight into the epidemiology of E. bieneusi in water buffaloes in different geographical areas in Turkey will be highly important to have determined the public health significance of this pathogen.

Encephalitozoon spp. and Enterocytozoon bieneusi are well-known microsporidian pathogens, recently classified as fungi, infecting humans and reptiles, mammals, and birds. Budgerigars (Melopsittacus undulates) are the most preferred captive pet birds in the households. Prevalence and molecular data on microsporidian species in budgerigars are scarce worldwide. The aim of the present study was to investigate the occurrence and genotypes of Encephalitozoon spp. and E. bieneusi in budgerigars, and to reveal their zoonotic potential. A total of 143 fecal samples were collected from owned healthy budgerigars in Turkey. Encephalitozoon spp. and E. bieneusi were examined by nested PCR targeting the ribosomal internal transcribed spacer (ITS) region and sequenced for identifying Encephalitozoon spp. and E. bieneusi. The overall prevalence of E. hellem and E. bieneusi was 14.7% (21/143) and 3.5% (5/143), respectively. Two genotypes of E. hellem were identified, including one known 1A (n = 18) and a novel TURK1B (n = 3). In addition, we determined two E. bieneusi genotypes, including one known N (n = 2) and a novel TURKM1 (n = 3). E. hellem 1A and novel TURK1B clustered as a sister taxon, and genotype N and novel TURKM1 genotypes fall into group 2 of E. bieneusi in the phylogenetic tree. Novel genotypes of E. hellem and E. bieneusi were described for the first time in the avian host. Moreover, E. bieneusi genotype N was first detected in avian hosts in the present study. This study contributes to the current knowledge on the molecular epidemiology and transmission dynamics of E. hellem and E. bieneusi. Lay summary Spore producing microsporidia are ubiquitous, obligate, and intracellular fungus defined as emerging opportunistic pathogens of humans, livestock, companion animals, wild mammals, birds, and water worldwide. The occurrence of microsporidia in animals could be risky for human public health.

Vector-borne zoonotic diseases (VBZDs) are a major problem for public health and animal welfare all over the world. In recent years, there has been an alarming increase in VBZDs, mainly caused by new or re-emerging arboviruses, bacteria and parasites. The World Health Organization enumerated 10 threats to global health for 2019, notably emphasizing climate change and emerging pathogens as growing priorities. It is important to review potential threats and develop new control programs for rising threats against human health and safety. Changes in host and vector population diversity and density may affect pathogen transmission patterns and influence VBZD emergence processes. In addition to environmental and climate-related changes, human and animal migratory patterns pose future threats. The geographic location and habitat features of Turkey support the establishment of many arthropod species as vectors of various diseases. To date, a total of 107 zoonotic infections have been reported originating from Turkey. Arthropods transmit 19 of 107 such infections, including 2 mosquito-borne, 9 tick-borne, 1 sandfly-borne, 3 flea-borne, 1 simuliid-borne, 1 mite-borne and 2 fly-borne diseases. In this review, we focus on the present status of knowledge on VBZDs as a rising threat to public health in Turkey to provide a foundation for future control efforts. Keywords: Vectors, zoonoses, vector-borne diseases, Turkey.

The molecular prevalence and genotypes of Giardia duodenalis in cattle were investigated. A total of 450 fecal samples were collected from cattle in three provinces of Central Anatolia from August 2017 to July 2019. Genomic DNA was extracted from the fecal samples and used in molecular analysis carried out by nested PCR analyses of the β-giardin (bg) gene of G. duodenalis. Positive samples were further analyzed by nested PCR at two gene loci (triosephosphate isomerase (tpi) and glutamate dehydrogenase (gdh)) for genotyping of G. duodenalis isolates. PCR analyses of the bg gene indicated that the overall prevalence of G. duodenalis was 30.2%. However, lower rates were determined with PCR analyses for gdh and tpi loci. The sequence analyses of the bg, gdh, and tpi genes revealed the presence of zoonotic assemblage A and livestock-specific assemblage E. Combined-sequence analyses revealed that assemblage E was the most common in the study area. Our study provides the first data on the wide prevalence of livestock-specific assemblages E in cattle in Turkey. The prevalence of assemblage A in cattle also reveals the importance of cattle for zoonotic transmission of giardiasis in Turkey.

Dermanyssus gallinae, the poultry red mite is an obligatory blood-sucking ectoparasite and has been involved in the transmission of several pathogens between both animals and humans. In order to develop effective control strategies against D. gallinae, it is essential to determine their genetic compositions and investigate the potential vector roles of these mites. Therefore, we aimed to analyse the mt-COI gene regions of D. gallinae populations and investigate avian haemosporidian parasites in order to point out their risk potentials for the presence of possible transmission by these mites. In total, 1985 mites were collected from five laying hen breeding facilities in Central Anatolia Region in Turkey. The mt-COI sequence analyses of 100 individual mites identified 6 haplotypes (TD1 to TD6) in three haplogroups. Although the local lineages are common in the region, our findings also support the possibility of international transmission of D. gallinae. A total of 126 pools of D. gallinae were screened for investigation of three genera of avian haemosporidians. In total, five pools from three facilities were identified as positive for species of Plasmodium. These findings provide novel molecular data on the possible vector role of D. gallinae in the transmission of avian haemosporidians and draw attention to the risk potentials in poultry farms.

Objective The identification and molecular characterization of the bot fly larvae from an infected human with naso-pharyngeal myiasis in Turkey were aimed in this study. Methods A total of 8 bot fly larvae from a 49-year-old woman with naso-pharyngeal infection in Adana province constituted the materials of this study. Morphological identification was performed on the larvae according to described keys. The barcode region of the CO1 gene from the genomic DNA extracts of the larvae was amplified and sequence analyses were utilized. Haplotype and genetic distance analyses were performed in CO1 sequences and a phylogenetic tree was built revealing phylogenetic relationships. Results All bot fly larvae were identified as second stage larvae of Oestrus ovis in terms of morphologic characteristics. There was no polymorphism among the CO1 sequences of all isolates leading to detection of a single novel haplotype. The newly characterized haplotype in this study clustered with the O. ovis haplotypes from Bosnia and Herzegovina, Croatia, Brazil, and Iran in a monophyletic clade with an overall identity of 99.5%. Interspecific genetic differences among the subfamilies of Oestridae were in the range of 19.8% to 30.8%. Conclusion This study has provided the first molecular characterization data on O. ovis larvae from an accidental human host in Turkey based on CO1 barcode sequences.

The protozoan Dientamoeba fragilis is one of the most common parasites in the diges-tive system of humans worldwide. The host range and transmission routes of D. fragi-lis, including the role of animals, are still ambiguous with few reports from non- human primates, sheep, rodents, pigs, a cat and a dog. In this study, we used microscopic and TaqMan qPCR analyses to investigate D. fragilisin 150 faecal samples from pet budger-igars (Melopsittacus undulatus) in the Central Anatolia Region of Turkey. Dientamoeba fragilis DNA was detected in 32 samples, resulting in a mean prevalence of 21.3%. In microscopic examination, trophozoites/cysts of D. fragilis were detected in 13 of 32 qPCR- positive samples. SSU rRNA sequence analyses of the qPCR- positive iso-lates identified genotype 1 of D. fragilis as predominant in budgerigars. Phylogenetic analyses of the SSU rRNA gene region clustered D. fragilis genotypes, as well as other trichomonads, in separate monophyletic clusters with bootstrap values ≥79.0. Our study provides the first evidence for the natural host status of pet budgerigars for D. fragilisand contributes to the knowledge of the epidemiology of this parasite. The high prevalence of genotype 1 of D. fragilis suggests that pet budgerigars are suitable reservoirs for zoonotic transmission. Our findings contribute to an increased aware-ness and knowledge of D. fragilis infections in the context of a one- health approach.

© 2022, The Author(s), under exclusive licence to Springer Nature B.V.Infections of avian haemosporidian parasites are regularly identified by molecular methods including multiplex PCR, which allows researchers to distinguish mixed infections of parasites from multiple genera. Here we extend the utility of a previously designed multiplex PCR by designing a primer set specific to parasites of the subgenus Haemoproteus (genus: Haemoproteus). The updated one-step multiplex PCR protocol we describe here allows for the detection of the genera Plasmodium and Leucocytozoon and the two subgenera (Haemoproteus and Parahaemoproteus) of the genus Haemoproteus. A sensitivity analysis showed that the multiplex PCR could amplify DNA of parasites in the subgenus Haemoproteus at very low levels of infection. We used this multiplex PCR to identify haemosporidian infections in 250 adult domestic pigeons (Columba livia) in Turkey. All samples were also screened by microscopy and a widely used nested PCR to compare with the results of multiplex PCR, to detect low levels of parasitemia, and to identify possible abortive infections. In total, 71 pigeons (28.4%) were found to be infected by all three methods. The multiplex PCR protocol successfully detected and discriminated both subgenera Haemoproteus and Parahaemoproteus infections. We compared our results with previous host species records to assess the host specificity of the parasite lineages we found. Our findings provide novel data on the prevalence of avian haemosporidians in domestic pigeons and demonstrate the utility of the new one-step multiplex PCR protocol for the determination of mixed avian haemosporidian infections. We expect that this protocol will contribute to a better understanding of the distribution, epizootiology, and ecology of avian haemosporidians.

A total of 100 fecal samples were collected from cats between June and October 2020 in Kayseri and Samsun provinces. Fecal samples were examined by nested polymerase chain reaction (PCR), targeting theOf the samples examined, Giardia intestinalis was determined in 8 samples (8.0%), whereas none of the samples were found positive forThis study adds to the current data on the molecular epidemiology of cryptosporidiosis and giardiasis in cats. The findings also highlight the potential risk of cats for public health concerning the zoonotic transmission dynamics ofKayseri ve Samsun illerinde, Haziran-Ekim 2020 tarihleri arasında toplam 100 kediden dışkı örneği toplanmıştır. Dışkı örnekleri,İncelemesi yapılan örneklerin 8’inde (%8,0)Bu çalışma, kedilerde Cryptosporidiosis ve Giardiasis’in moleküler epidemiyolojisi üzerine mevcut bilgi birikimine katkı sağlamıştır. Ayrıca, elde edilen çıktılar

Background Sequencing parasite genomes in the presence of host DNA is challenging. Sequence capture can overcome this problem by using RNA probes that hybridize with the parasite DNA and then are removed from solution, thus isolating the parasite DNA for efficient sequencing. Methods Here we describe a set of sequence capture probes designed to target 1035 genes (c. 2.5 Mbp) of the globally distributed avian haemosporidian parasite, Plasmodium relictum. Previous sequence capture studies of avian haemosporidians from the genus Haemoproteus have shown that sequencing success depends on parasitemia, with low-intensity, chronic infections (typical of most infected birds in the wild) often being difficult to sequence. We evaluate the relationship between parasitemia and sequencing success using birds experimentally infected with P. relictum and kept under laboratory conditions. Results We confirm the dependence of sequencing success on parasitemia. Sequencing success was low for birds with low levels of parasitemia (Plasmodium relictum is composed of multiple lineages defined by their mitochondrial DNA haplotype including three that are widespread (SGS1, GRW11, and GRW4); the probes successfully isolated DNA from all three. Furthermore, we used data from 25 genes to describe both among- and within-lineage genetic variation. For example, two samples of SGS1 isolated from different host species differed by 11 substitutions across those 25 genes. Conclusions The sequence capture approach we describe will allow for the generation of genomic data that will contribute to our understanding of the population genetic structure and evolutionary history of P. relictum, an extreme host generalist and widespread parasite. Graphical Abstract

A total of 250 droppings of tumbler pigeons (Columba livia domestica, Columbidae) were collected individually from different breeders in Turkey, to investigate the presence and genotyping of microsporidian species by nested PCR and to reveal their zoonotic potential. In the present study, Enterocytozoon bieneusi was the only microsporidian species identified in 35 pigeons with an overall molecular prevalence of 14.0%. Only one known genotype zoonotic Peru6 was identified in all positive samples according to the sequence analyses of the internal transcribed spacer region of ribosomal DNA of E. bieneusi. This study represents the first report of E. bieneusi in pigeons in Turkey. Our study also confirms the competence of breeding pigeons as hosts for the zoonotic Peru6 genotype, corroborating its potential role as a source of human infection and environmental contamination. Lay summary Microsporidia are spore-producing fungi defined as emerging opportunistic pathogens of humans. The occurrence of microsporidia in animals could be risky for human public health. Home kept breeding pigeons pose a high risk for transmission of the microsporidians to humans.

The Mediterranean tick, Hyalomma marginatum, is the most important vector of Crimean-Congo haemorrhagic fever virus and several pathogens that cause animal and human diseases and economic losses to livestock production. Given the medical and veterinary importance of this tick species, we sequenced and characterized its mitochondrial genome (mitogenome) for the first time. We designed two new primer sets and combined long-range PCR with next generation sequencing to generate complete mitogenomes with deep coverage from 10 H. marginatum adults. The mitogenomes contained 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), two ribosomal subunits, two control regions, and three tick-box motifs. The nucleotide composition of the H. marginatum mitogenomes were A+T biased (79.76%) and exhibited negative AT- and GC- skews across most PCGs. All PCGs were initiated by ATK codons and two truncated termination codons were seen in the COX2 and COX3 genes. All tRNAs exhibited typical cloverleaf structures, except for tRNACys and tRNASer1. A total of 62 polymorphic sites defined ten unique haplotypes. Phylogenetic analyses based on the 13 PCGs of 56 tick species revealed that four Hyalomma species (H. marginatum, H. asiaticum, H. rufipes, and H. truncatum) formed a monophyletic clade with strong support. The results of this study provide a comprehensive resource for further studies on the systematics, population genetics, molecular epidemiology, and evolution of ticks.

The purpose of the present study was to establish experimental model of toxoplasmosis in pigeons, to investigate pathogenesis and compare tissue lesions by clinical, histopathological, serological and bioassay techniques. Total of 60 unknown aged pigeons (Columba livia), 21 males and 39 females were used. They were divided into groups as oral (Group I and II) and parenteral (Group III, IV, V and VI) infection groups (Table 1). While some pigeons in Group IV showed acute infection signs such as anorexia, weight loss, pale cockscomb, bend of head and neck and partial paralyze; chronic infection signs such as anorexia, weakness, weight loss, diarrhea, difficulties in breathing, and conjunctivitis were seen in Group IV, V, and VI. In necropsy, the pigeons in Group IV had hyperemia and focal hemorrhages in the meninges and brain; the pigeons in Groups V and VI had yellowish color of the liver, the pigeons in Group V had the pale chest muscles, pericardial thickening and opaqueness. There were no macroscopic findings in pigeons in Group I and III. Histopathological examination revealed nonsuppurative meningoencephalitis and tachyzoites and bradyzoite cysts formation of T. gondii in brain tissue, lymphoid cell infiltration and necrotic focal hepatitis and nephritis in Group IV. While pigeons in Group V had nonsuppurative focal myositis, myocarditis, hepatitis, gastritis, enteritis, pneumonitis, and necrotic pancreatitis, one of them had toxoplasma bradyzoite cyst in the sinusoid in the liver. In group VI, nonsuppurative focal hepatitis, myocarditis, nephritis and necrotic pancreatitis were detected in pigeons. Bioassay tests were performed with tissue samples taken from seropositive pigeons and parasitic tachyzoites were isolated from the peritoneal fluid of the mice. Seropositivity in the oral and parenteral groups was determined by Sabin-Feldman Dye Test (SFDT) and Indirect Hemagglutination Assay (IHA). As a result; in similar studies that will be performed investigating pathogenesis of Toxoplasmosis and subclinical cases that may be overlooked, serologic tests and bioassay applications should be used together for the diagnosis of toxoplasmosis.

A total of 1340 fresh fecal samples from farm and pet animals in Central Anatolia and the Middle Black Sea Region of Turkey were investigated using a PCR assay targeting the SSU rRNA of Blastocystis sp. An overall Blastocystis sp. prevalence of 19.4% (183/940) was found in farm animals, including cattle, sheep, water buffaloes, and chickens. Fecal samples of dogs, cats, and horses were negative. The highest prevalence of Blastocystis sp. was found in sheep (38.2%) among the farm animals. The SSU rRNA sequence analysis revealed two animal-specific subtypes, including ST10 in cattle and sheep and ST14 in water buffaloes. The zoonotic subtype ST7 was identified in chickens. Our results indicated a high prevalence of animal-specific subtypes in livestock and zoonotic subtype ST7 in chickens, highlighting the potential risk of chickens for zoonotic transmission of Blastocystis in the research area. This study is the first large-scale evaluation of Blastocystis in animal hosts in Turkey, and contributes to the molecular epidemiology and genetics of Blastocystis. Our results should be considered by authorities as an indication of the zoonotic importance of Blastocystis sp. and the need for surveillance in public health intervention programs.

Sürsal, Neslihan ( Aksaray, Yazar ), Globalization opens new market areas and affects food consumption habits, resulting in rapid and remarkable cultural change. Food habits such as consumption of raw fish meat have become popular, resulting in increased risk of emerging infectious diseases. Anisakis simplex sensu stricto (s.s) and A. pegreffii are the most common and important fish-borne zoonotic nematodes responsible for human anisakiasis, which occurs through the consumption of raw or undercooked fish as well as cooked fish due to their heat-stable allergens. Here, we investigated the prevalence, intensity, and abundance of Anisakis larvae in imported fish and ready-to-eat local fish products in Turkey. A total of 205 ready-to-eat fish products, 100 imported frozen Atlantic salmon (Salmo salar) fillets, and 100 imported frozen whole Atlantic mackerel (Scomber scombrus) were sampled from supermarkets, sushi restaurants, and fish markets. All samples were individually examined using a pepsin digestion technique. In total, 602 Anisakis type I larvae were recovered from 98/100 mackerel. No larvae were found in ready-to-eat products or frozen Atlantic salmon fillets. Overall, 8.8% of the larvae were found in the muscle tissue. The overall mean intensity and abundance of infection in mackerel were 6.14 and 6.02, respectively. The larvae were molecularly identified and their phylogenetic relationships with the relevant Anisakis sequences in GenBank were investigated. For this purpose, a subsample of randomly selected 100 Anisakis larvae were analyzed with PCR-RFLP of the ITS region. The larvae were identified as A. simplex (s.s.) (n = 87) and hybrids (n = 13). ITS and cox2 gene regions of all hybrids and randomly selected 50 A. simplex (s.s.) larvae were sequenced for species confirmation and phylogenetic analyses. No intraspecific nucleotide variation was found among the ITS sequences of either species. Seven and three haplotypes, respectively, were identified for A. simplex (s.s.) and hybrid species according to DNA polymorphism of the cox2 gene. Hybrids in our study clustered within the common A. simplex (s.s.) clade in the cox2 phylogenetic tree indicating the dominance of A. simplex (s.s) in the catching area of Atlantic mackerel. Consequently, our study indicates high occurrence of A. simplex (s.s.) larvae with an overall 98.0% prevalence in imported Atlantic mackerel, and highlights the importance of these fish as potential reservoirs for human allergic anisakiasis in Turkey and possibly in other countries.

Parasite species evolve by switching to new hosts, cospeciating with their current hosts, or speciating on their current hosts. Vector transmitted parasites are expected to speciate by host switching, but confirming this hypothesis has proved challenging. Parasite DNA can be difficult to sequence, thus well resolved parasite phylogenies that are needed to distinguish modes of parasite speciation are often lacking. Here, we studied speciation in vector transmitted avian haemosporidian parasites in the genus Haemoproteus and their warbler hosts (family Acrocephalidae). We overcome the difficulty of generating parasite genetic data by combining nested long-range PCR with next generation sequencing to sequence whole mitochondrial genomes from 19 parasite haplotypes confined to Acrocephalidae warblers, resulting in a well-supported parasite phylogeny. We also generated a well-supported host phylogeny using five genes from published sources. Our phylogenetic analyses confirm that these parasites have speciated by host switching. We also found that closely related host species shared parasites which themselves were not closely related. Sharing of parasites by closely related host species is not due to host geographic range overlap, but may be the result of phylogenetically conserved host immune systems.

Horses might play an important role as reservoir hosts in the epidemiology of Enterocytozoon bieneusi, which is one of the most important zoonotic microsporidian pathogens, with a wide range of hosts. Nevertheless, limited information is available on the infection rates and genotypes of E. bieneusi in horses, and no data are available on the occurrence and molecular characteristics of E. bieneusi in horses in Turkey. We determined the prevalence of E. bieneusi among horses raised on farms from two provinces of Central Anatolia Region, by amplification of the partial small subunit ribosomal RNA gene using nested PCR. We identified the genotypes of E. bieneusi isolates by analyzing the ribosomal internal transcribed spacer (ITS) sequences. The overall prevalence of E. bieneusi was 18.7% (56/300), with no significant differences in infection rates among age groups or between genders of horses. Sequence analysis revealed eight genotypes: two known genotypes (ERUSS1, BEB6) and six novel genotypes (named ERUH2 to ERUH7). The genotype ERUSS1 was the most common and was found on all farms, age groups, and genders. Phylogenetic analysis clustered all the identified genotypes in ruminant-specific group 2. Our findings contribute to the molecular epidemiology of E. bieneusi.

Bu calisma, “Babesia bigemina Kayseri/Turkiye” susundan in vivo ve in vitro kultivasyon yapilarak ve pasajlanarak elde edilmis olan izolatlarin AMA-1 proteinlerinin molekuler karakterizasyonu ve ekspresyonu ile soz konusu proteinin attenuasyona bagli polimorfizm gosterip gostermedigini ortaya koymak amaciyla yapilmistir. Bu amacla, Babesia bigemina Kayseri/Turkiye susuna ait, in vivo ve in vitro izolatlardan genomik DNA ve total RNA izolasyonu sonrasinda hedef gen, PCR ile butun olarak amplifiye edilmis ve purifiye amplikonlar uygun vektorlere klonlanmistir. Klonlama sonrasinda rekombinant plazmidler, sekanslanarak izolatlara ait nukleotid dizilimleri GenBank’a kaydedilmistir . Nukleotid ve amino asit sekanslarin coklu ve ikili hizalama analizleri yapilmis ve izolatlarin filogenileri arastirilmistir. Elde edilen rekombinant plazmidler, E. coli BL21 (DE3) hucrelerine transforme edilerek IPTG ile induklenmis ve ekspresyonlari saglanmistir. Ekspresyon urunlerinde hedef proteinler SDS-PAGE ve Western Blot teknikleri ile analiz edilmistir. Kayseri/Turkey IV1 ve Kayseri/Turkey IT2 izolatlari nukleotid dizilimlerinin kendi aralarinda %99.7 identiklik gosterdigi ve 103. nukleotiddeki mutasyona bagli olarak amino asit diziliminde farklilik (%0.2) olustugu belirlenmistir. SDSPAGE ve Western Blot analizleri sonucu soz konusu iki izolata ait AMA-1 proteinlerinin 72 kDa molekuler agirliklarla farkli IPTG konsantrasyon duzeyleri ve zaman araliklarinda eksprese olduklari belirlenmistir. Sonuc olarak bu calisma ile Babesia bigemina Kayseri/Turkiye susunun in vivo ve in vitro pasajlarindan elde edilmis iki izolata ait AMA-1 nukleotid ve amino asit dizilimleri arasinda mutasyonel farkliliklari gosteren attenuasyona bagli polimorfik alanlar oldugu ortaya konulmustur

Kayseri Sultan Sazligi ekosisteminden toplanan 400 sivrisinek orneginin 296’si (%74.0) Culex pipiens, 63’u (%15.75) Aedes vexans ve 41’i (%10.25) Culiseta annulata olarak belirlenmistir. Ergin sineklerden bireysel DNA elde edilmis ve wsp gen bolgesi yonunden PZR analizleri gerceklestirilmistir. 119 (%40.2) Cx. pipiens disisinde Wolbachia pozitifligi saptanirken, erkek Cx. pipiens orneklerinde pozitiflik belirlenmemistir. Ae. vexans ve Cs. annulata orneklerinde de pozitiflik saptanmamistir. Wolbachia pozitif orneklerden secilen 10 izolatin filogenetik analizlerinde EruWolCpip1-10 izolatlari Wolbachia B super grubu ve wPip grubu icinde belirlenmistir. EruWolCpip1-9 izolatlarinin %100 identik olduklari ve EruWolCpip10 izolatiyla %0.5 genetik farklilik gosterdikleri saptanmistir. Daha once Kayseri’de Cx. pipiens’ten izole edilmis WolKys1 ve Cankiri’da Bovicola limbata’dan elde edilmis TrERUWolLice1 izolatlarinin, EruWolCpip1-9 izolatlariyla %100 benzerlik gosterdigi belirlenmistir. EruWolCpip1-9 izolatlarinin wPip grubundaki Cx. pipiens, Peribadotes rhomboidaria, Agriocnemis femina femina ve Eurema hecabe Wolbachia izolatlariyla %100 benzer olduklari gorulmustur. wPip grubundaki genetik farklilik %0.28±0.11 belirlenirken, wPip grubu ile ayni super gruptaki Dei, Con ve Ori gruplari arasindaki genetik farklilik sirasiyla %16.9±2.3, %12.5±1.9 ve %19.9±2.5 bulunmustur. A super grubundaki Haw, Riw, Mel, Aus, Mors, Uni, Pap ve Alba gruplari ile B super grubundaki wPip grubu arasinda genetik farklilik ise sirasiyla %23.5±3.0, %17.6±2.3, %19.1±2.5, %22.0±2.8, %21.0±2.7, %22.3±2.9, %22.0±2.8 ve %20.5±2.6 olarak saptanmistir. Farkli cografyalardaki degisik artropodlarda ayni veya farkli super gruplarda yer aldiklari belirlenmis Wolbachia izolatlari ile kendi izolatlarimiz arasindaki genetik farkliligin onemli duzeyde olabilecegi hipoteziyle yapilmis bu calisma sonucunda, ayni grubun izolatlari arasindaki farkliligin cok onemli olmadigi, ancak farkli super gruplardaki izolatlar arasindaki genetik farkliligin bir hayli onemli oldugu belirlenmistir. Bu sonuclar hipotezimizi dogrular nitelikte bulunmustur

Bu çalışmada, Haziran-Ağustos 2014 tarihleri arasında Kayseri yöresinde çeşitli bölgelerden toplanmış sivrisinek örneklerinde Culex pipiens kompleks biyotipleri ve Cx. torrentium moleküler olarak araştırılmıştır. Saha çalışmaları boyunca toplam 1052 dişi sivrisinek örneklemesi yapılmış ve bunların 315’i (%29.9) morfolojik teşhis analizleriyle Cx. pipiens kompleks ve/veya Cx. torrentium olarak teşhis edilerek ayrılmıştır. Ayrılan örneklere ait genomik DNA izolatlarının ilk basamak Real time PCR analizlerinde tamamının Cx. pipiens kompleks nesillerine ait oldukları saptanmış, Cx. torrentium pozitifliği belirlenmemiştir. İkinci basamak Real time PCR analizlerinde Cx. pipiens komplekste belirlenen izolatların, 311’inin Cx. pipiens form pipiens nesillerine ait oldukları tespit edilmiş, Cx. pipiens form molestus pozitifliği görülmemiştir. Kalan 4 örnek, analizlerde her iki biyotip yönünden de amplifikasyon göstermemiştir. Real time PCR sonuçlarının konfirmasyonu ve hibrit nesillerin araştırılması amacıyla tüm örneklere ait izolatların ACE-2 ve CQ11 microsatellite DNA analizleri gerçekleştirilmiştir. Analizlerde Cx. pipiens form pipiens olarak belirlenen tüm izolatların ilgili DNA markerları ile pipiens biyotipine özgü bant profilleri sergilediği görülmüş ve izolatların konfirmasyonu sağlanmıştır. Real time PCR analizlerinde biyoformlar yönünden amplifikasyon göstermeyen 4 izolatın microsatellite DNA analizleriyle Cx. pipiens form pipiens ve Cx. pipiens form molestus hibritleri oldukları tespit edilmiştir. Cx. pipiens form pipiens ve hibrit nesillere ait birer izolatın mitokondrial cytochrome oxidase I (mt-COI) gen bölgesine göre barkodlaması yapılmış ve filogenetik analizleri gerçekleştirilmiştir. Her iki izolatın sekans analiz sonuçlarına göre yeni bir haplotip oldukları belirlenmiş ve haplotip bazında Cx. pipiens komplekse ait GenBank’ta kayıtlı homolog nesillerle filogenetik ilişkileri ortaya konmuştur, The biotypes of Culex pipiens complex and Cx. torrentium were molecularly investigated in the mosquito specimens collected from different regions of Kayseri between June and August 2015 in this study. A total of 1052 female mosquito specimens were sampled during the field surveys and 315 (29.9%) out of these were morphologically identified and reserved as Cx. pipiens complex and/or Cx. torrentium. The first step Real time PCR analyses of genomic DNA isolates from these reserved specimens revealed that all the specimens were belong to Culex pipiens complex and there was no Cx. torrentium positivity. Among the isolates determined in Culex pipiens complex, 311 were determined as belong to lineages of Cx. pipiens form pipiens according to second step Real time PCR analyses and there was no positivity for Cx. pipiens form molestus. The remaining 4 isolates showed no amplification for both two biotypes. ACE-2 and CQ11 microsatellite DNA analyses were carried out on the isolates from all specimens in order to confirm the real time PCR results and investigate the hybrid lineages. All the isolates determined as Cx. pipiens form pipiens showed specific banding pattern for pipiens biotype with the related DNA markers in the analyses result in the confirmation of the isolates. The 4 isolates that did not show amplification for both biotypes in Real time PCR assays were designated as the hybrids of Cx. pipiens form pipiens and Cx. pipiens form molestus with the microsatellite DNA analyses. DNA barcoding and phylogenetic analyses were performed on one isolate belong to each Cx. pipiens form pipiens pipiens and hybrids based on mitochondrial cytochrome oxidase I (mt-COI) gene region. Both isolates were determined as new haplotypes according to the sequence analyses and their phylogenetic relations based on haplotypes with the available homolog linages belong to Cx. pipiens complex in GenBank were revealed

Objective In this study, forming of experimental toxoplasmosis in quails; clinical, pathological, and serological determination of tissue lesions and bioassay techniques, which were aimed to compare them and determine pathogenesis. Methods A total of 120 one-year-old female quails (Coturnix coturnix japonica) were divided into oral infection, parenteral infection, and control groups. The oral group was infected with 0.5 ml inoculum suspension containing 106 tachyzoites of Toxoplasma gondii, whereas the control group was administered 0.5 ml of saline. The parenteral group was further divided into the following four subgroups: intraperitoneal, intramuscular, intravenous, and cloacal. The quails of the parenteral group were also divided into two groups and one by control group within itself for the 105 and 104 doses of the tachyzoite inoculums. Results Because of acute toxoplasmosis, death occurred in a quail that as intramuscularly infected with 105 tachyzoites; the quail exhibited neurological clinical symptoms such as torticollis, ataxia, and tremor. In histopathologic examination, T. gondii tissue cysts were detected in infected quails that were intramuscularly infected with 105 tachyzoites. Mouse trials were conducted using tissues of seropositive quails and isolated from peritoneal fluids infected mice. By Sabin-Feldman dye test and indirect hemagglutination test, seropositivity was observed in quails infected with 105 and 104 tachyzoites. Conclusion Similar studies and subclinical cases, which may overlooked was concluded for diagnosis of toxoplasmosis with useful bioassay applications and serological tests.

Avian haemosporidians (Haemosporida) represent a globally distributed, species-rich multiparasite-multihost host-parasite system. Each year, many of these parasite lineages are carried between temperate and tropical regions by migratory birds. While several factors can limit the transmission of avian haemosporidians to new areas, recent studies have shown that some abundant parasites can sometimes disperse and be transmitted in new areas to become emerging infectious diseases. In this study, we investigated the prevalence and diversity of avian haemosporidian parasites in Sultan Marshes National Park (SMNP), a major stopover site in the eastern Mediterranean flyway, and we evaluated the potential for avian haemosporidians in SMNP to be transmitted to areas outside of their known distributions. We sampled a total of 565 migratory and resident birds belonging to 39 species and 23 families. We applied both molecular and microscopic methods to detect and identify avian haemosporidian infections and also quantified the frequency of potential abortive infections. We identified a total of 52 different mitochondrial cytochrome b (cyt b) parasite lineages belonging to the genera Plasmodium (N = 12), Haemoproteus (N = 31), and Leucocytozoon (N = 9) in 193 (34.2%) infected birds. Ten of the lineages were reported for the first time. Our findings show that numerous parasite lineages are actively transmitted among resident bird species of SMNP. Our findings also revealed new parasite-host interactions while considering the role of possible abortive infections. The relatively high frequency of presumed abortive infections suggests that analyses of datasets generated only by PCR-based methods should be interpreted with caution. We also compared the prevalence and distribution of avian haemosporidian infections in both resident and migratory bird species and showed that haemosporidian prevalence was related to bird migratory behavior. The results of this study contribute to a better understanding of the ecological and genetic adaptations associated with changes in transmission areas of avian haemosporidian parasites.

Bovine babesiosis is a global tick-borne disease that causes important cattle losses and has potential zoonotic implications. The impact of bovine babesiosis in Turkey remains poorly characterized, but several Babesia spp., including B. bovis, B. bigemina, and B. divergens, among others and competent tick vectors, except Rhipicephalus microplus, have been recently identified in the country. Bovine babesiosis has been reported in all provinces but is more prevalent in central and highly humid areas in low and medium altitude regions of the country housing approximately 70% of the cattle population. Current control measures include acaricides and babesicidal drugs, but not live vaccines. Despite the perceived relevant impact of bovine babesiosis in Turkey, basic research programs focused on developing in vitro cultures of parasites, point-of-care diagnostic methods, vaccine development, “omics” analysis, and gene manipulation techniques of local Babesia strains are scarce. Additionally, no effective and coordinated control efforts managed by a central animal health authority have been established to date. Development of state-of-the-art research programs in bovine babesiosis to address current gaps in knowledge and implementation of long-term plans to control the disease will surely result in important economic, nutritional, and public health benefits for the country and the region.

Posthodiplostomum Dubois, 1936 species have worldwide distribution. Metacercariae of P. cuticola that is common in Europe causes “black spot” disease in freshwater fishes. A limited number of data on mitochondrial and ribosomal sequences of P. cuticola are available in the GenBank. In addition, there is no sequencing data on trematodes belonging to genus of Posthodiplostomum in Turkey. The aim of this study was to investigate the presence of Posthodiplostomum metacercariae in freshwater chubs (Squalius cephalus) and to reveal the molecular characterization of mt-COI and ITS-2 gene regions of the identified isolates. Posthodiplostomum metacercariae were detected in the gills of two (1.6%) out of 123 fish. The ITS-2 gene sequences of the isolates (POSTC1-C2) (MN701652-3) found in this study showed 99.8-100% identity with P. cuticola sequences isolated from skin, muscle and fins of different fish species in Czech Republic and Hungary. In addition, the mt-COI gene sequences of the POSTC1-C2 isolates (MN700658-9) showed 99.1-98.9% identity with P. cuticola isolate (cercariae) (KX931424) from Planorbis planorbis in Lithuanian. This study provides first molecular and phylogenetic characterization of P. cuticola isolates found in chubs (Squalius cephalus) in Turkey., Posthodiplostomum Dubois, 1936 türleri dünya genelinde oldukça yaygınlık göstermektedirler. Avrupa’da yaygın olan P. cuticola türünün metaserkeri tatlı su balıklarında siyah nokta “black spot” hastalığına sebep olmaktadır. GenBank veri tabanında P. cuticola türüne ilişkin mitokondriyal ve ribozomal sekans kaydı sınırlıdır. Diğer yandan, Türkiye’de Posthodiplostomum cinsinde yer alan trematodların sekans karakterizasyonuna dair herhangi bir veri bulunmamaktadır. Bu çalışmada, tatlı su kefallerinde (Squalius cephalus) Posthodiplostomum metaserkerlerinin araştırılması ve tespit edilen izolatların mt-COI ve ITS-2 gen bölgelerinin moleküler karakterizasyonlarının yapılması amaçlanmıştır. İncelenen 123 balıktan ikisinin (%1,6) solungaçlarında Posthodiplostomum metaserkerleri saptanmıştır. Çalışmada tespit ettiğimiz izolatların (POSTC1-C2) (MN701652-3) ITS-2 gen bölgesine ait nükleotid dizilimleri, Çek Cumhuriyeti ve Macaristan’da farklı balık türlerinin deri, kas ve yüzgeçlerinden izole edilen P. cuticola izolatları ile %99,8-100 arasında identiklik göstermiştir. Yine POSTC1-C2 izolatlarının (MN700658-9) mt-COI gen bölgesine ait nükleotid dizilimleri, Litvanya’da Planorbis planorbis’te bulunan P. cuticola izolatı (serker) (KX931424) ile %99,1-98,9 oranında identik bulunmuştur. Bu çalışma ile Türkiye’de ilk kez tatlı su kefalinde (Squalius cephalus) bulunan P. cuticola izolatlarının moleküler ve filogenetik karakterizasyonları ortaya konulmuştur.

Microsporidia are opportunistic pathogens that infect a wide range of invertebrates and vertebrates. To assess the potential role of dogs in the transmission of these zoonotic pathogens, a total of 282 fecal samples from dogs in the Central Anatolia Region of Turkey were analyzed by utilizing species specific polymerase chain reaction for the four most frequent human microsporidia. Two microsporidia species were recognized in 41 samples (14.5%). Encephalitozoon intestinalis was detected in 35 samples (12.4%) and it was the most common microsporidium. The second microsporidium, E. cuniculi, was identified in six (2.1%) of the samples. Sequence analysis of the intergenic spacer of the ribosomal ribonucleic acid (RNA) internal transcribed spacer (ITS) gene revealed the presence of three E. intestinalis haplotypes closely associated with each other. No polymorphic region was found among the ITS sequences of E. cuniculi isolates and they were characterized as genotype III. This study provides the first data on the zoonotic microsporidia species from dogs in Turkey.

The goal of our study was to investigate the molecular prevalence of Giardia intestinalis in naturally infected horses in Kayseri, Central Anatolia Region in Turkey, to determine the molecular characterization of the obtained isolates and to exhibit the potential role of horses in zoonotic transmission of G. intestinalis. Fecal samples were randomly collected from totally 150 horses with clinically healthy between March and June of 2018. After the genomic DNA extractions, 25 (16.6%) of the 150 fecal samples, were found positive for G. intestinalis by nested PCR analyses of beta-giardin gene. Phylogenetic analysis of the beta-giardin gene sequences of G. intestinalis showed that the sequences detected in this study belonged to assemblage A that is regarded as zoonotic. Our study is the first report on the presence of G. intestinalis in horses in Turkey. The findings of the present study indicate that future research studies are required to determine molecular epidemiology and geographical distribution of G. intestinalis infections in horses nationwide. In addition, this study also may be helpful to assess the zoonotic potential for public health of G. intestinalis infections. (C) 2019 Elsevier Inc. All rights reserved.

Hypodermins A (HA), B (HB), and C (HC) of warble flies are modulatory antigens involved in host inflammation and immune responses during migration of the warble fly larvae through host connective tissues. In the current study, molecular characteristics of the genes encoding HA, HB, and HC were revealed from cDNA constructs of third-instar larvae of Hypoderma bovis. The open reading frame (ORF) of each hypodermin gene was amplified with modified gene-specific primers, and the resulting PCR products were cloned into pGEM-T Easy Vector to produce recombinant plasmids (rHA, rHB, and rHC). The ORF sequences of rHA, rHB, and rHC genes are 705 bp, 771 bp, and 783 bp long and encode proteins of 234, 256, and 263 amino acids with predicted sizes of 25.74 kDa, 27.79 kDa, and 28.51 kDa, respectively. The rHC gene was subcloned into the pET 100/D-TOPO Expression Vector, and the recombinant HC was purified using affinity chromatography. Western blotting indicated that rHC was recognized by the sera of cattle naturally infested with H. bovis. The rHC and a synthetic peptide (sHC) containing its linear B cell-specific epitope were evaluated as serological markers in indirect ELISA (iELISA) for the diagnosis of bovine hypodermosis. Both sHC and rHC iELISAs had sensitivity values equal to or higher than 90 % and specificity values of 100 %. A total of 200 serum samples from cattle in the Central Anatolia Region of Turkey were also analyzed by rHC and sHC-iELISAs to reveal the seroprevalence of bovine hypodermosis. The results of both iELISAs were consistent with one another and revealed a hypodermosis prevalence of 62 %. Our study provides the first data on molecular characterization of hypodermin genes of H. bovis and indicates the efficacy of recombinant antigen and peptide-based iELISA for serodiagnosis of bovine hypodermosis.

INCI, Abdullah/0000-0003-1614-0756; Duzlu, Onder/0000-0002-6951-0901; Pekmezci, Gokmen Zafer/0000-0002-7791-1959; Yildirim, Alparslan/0000-0001-9868-0363 WOS: 000463575500003 PubMed: 30645737 ObjectiveThis preliminary study was conducted to reveal that the molecular identity of Encephalitozoon spp. and Enterocytozoon bieneusi in indoor domestic cats' fecal samples from Turkey was screened using the PCR.Materials and MethodsNested PCR was performed using MSP and EBITS primers.All of the amplification products were sequenced to identify the microsporidia species.ResultsFour (5.5%) and three (4.1%) genomic DNA isolates of the fecal samples from 72 indoor domestic cats showed amplification of the ITS regions of E. bieneusi and Encephalitozoon spp., respectively. Two different genotypes, D and IV, of E. bieneusi were determined in two cats each based on the ITS sequence analyses. Moreover, Encephalitozoon spp. sequence analyses revealed three isolates belonging to E. intestinalis.ConclusionsThispreliminary study has provided the first molecular data on the zoonotic genotypes of E. bieneusi and E. intestinalis in cats in Turkey. Furthermore, E. bieneusi genotype IV (accession number MG727664) was submitted to GenBank for the first time in the Western Palearctic Region as hosted by a domestic cat. Additionally, E. intestinalis (accession number MG570080) was also submitted to GenBank as a valid ITS sequence for the first time as hosted by a domestic cat worldwide.

The prevalence of Cryptosporidium species in calves and heifers with relation to diarrhea from several herds was investigated in this study. Fecal samples were collected from 135 and 120 pre-weaned calves and 79 and 130 heifers raised in the Central Anatolia (CAR) and Mediterranean Regions (MR) of Turkey, respectively. A total of 86 post-weaned calves in CAR were also included in the study. For diagnostic comparison, all samples were examined by microscopic examination, SSU rRNA nested PCR and TaqMan real-time PCR for the presence of oocyst and Cryptosporidium DNA. In total, 102 (34.0 %) and 93 (37.2 %) of the examined samples from CAR and MR were found positive for Cryptosporidium DNA with both nested PCR and real-time PCR analyses, respectively with an overall prevalence of 35.5 %. The diagnostic sensitivity and specificity of microscopic examination were determined as 68.7 % and 100.0 % compared to molecular tools, respectively. RFLP and sequence analyses of the SSU rRNA from the PCR products revealed that 138 (70.8 %) out of 195 positive isolates were C. parvum further confirming the species-specific real-time PCR results. Among the remaining 57 (29.2 %) positive isolates, 30 (15.4 %) and 27 (13.8 %) were characterized as C. ryanae and C. bovis, respectively. C. parvum was the dominant species in pre-weaned calves especially with diarrhea while C. bovis and C. ryanae were mostly found in post-weaned calves and heifers. The sequence analyses of the gp60 gene of C. parvum isolates revealed two subtypes (IIaA13G2R1, IIaA14G1R1) belonging to zoonotic family IIa, with IIaA13G2R1 being the most common in diarrheic calves.

WOS: 000457510400057, PubMed: 30553894, Identification of Culicoides (Diptera: Ceratopogonidae) biting midges to species has become important due to their potential role in the transmission of arboviruses such as bluctongue virus, bovine ephemeral fever virus, Akabane virus, African horse sickness virus, epizootic haemorrhagic disease virus and Schmallenberg virus. In several studies, molecular tools, used for the identification of biting midges, revealed the presence of cryptic and undescribed species especially within Pulicaris complex. The presence of cryptic species within species complexes raise questions about their role in viral disease transmission as there are apparent differences in the vectorial capacity between closely related species. In this study, we analyzed the mitochondria' DNA cytochrome oxidase I (COI) gene sequences of species within the Pulicaris complex present in Turkey and determined their phylogenetic relationships. Twenty-one haplotypes within the already described species C. pulicaris P1, C. lupicaris, C. lupicaris 12, C. newsteadi, C. newsteadi N1, C. punctatus, C. fagineus F2 and C. flavipulicaris were determined from the study areas. The molecular analysis revealed further two haplotypes belonging to new non described cryptic species named as C. lupicaris L3 and Culicoides WBS corresponding to C. lupicaris and Fagineus complex which diverged by 17.9% to 25.7% and 18.7% to 31.8%, respectively from other species in the subgenus Culicoides. Genetic divergence within species was < 2.0% and phylogenetic analyses of the COI dataset revealed 22 different monophyletic separate clades within two major cluster. The results of this study emphasize the applicability of COI sequences as a diagnostic marker for differentiating Culicoides species and revealing cryptic species., Erciyes University Research FundErciyes University [TSA-2015-5762], This research was funded by the Erciyes University Research Fund with the project code TSA-2015-5762.

OBJECTIVE The aim of this study is to examine the molecular characterization of moth flies (Diptera: Psychodidae) based on their mitochondrial DNA sequences and determine the vectorial potential and damage caused by moth flies in future researches. METHODS A total of 240 adult moth flies were collected from toilet, bathroom, and basement walls of houses from different locations of the Kayseri region between May 2016 and April 2017. The polymerase chain reaction (PCR) analyses were performed using primer pairs, specificaly targeting the mitochondrial cytochrome oxidase c subunit I (mt-COI) gene of adult flies. RESULTS In total, five isolates were gel purified and sequenced for molecular characterization and phylogenetic analyses. Two species, namely Telmatoscopus albipunctatus (ERU-Telmatos3 and ERU-Telmatos6) and Psychodidae sp. (ERU-Psycho1,4,5), were successfully identified with the sequence alignment of isolates. According to the phylogenetic analysis, it was determined that the ERU-Telmatos3 and ERU-Telmatos6 isolates are clustered in the haplogroup A, while the ERU-Psycho1,4,5 isolate was clustered within the haplogroup B. The ERU-Psycho1 isolate was characterized as a new haplotype within the haplogroup B. CONCLUSION This study represents the first molecular characterization and phylogenetic status of moth flies in Turkey. The obtained findings should be the first step in the future investigation based on detecting the transmission of bacterial pathogens by moth flies.

The aim of this study was to determine the presence and prevalence of Wolbachia bacteria in natural population of fleas (Insecta: Siphonaptera) in Turkey, and to exhibit the molecular characterization and the phylogenetic reconstruction at the positive isolates with other species in GenBank, based on 16S rDNA sequences. One hundred twenty-four flea samples belonging to the species Ctenocephalides canis, C. felis, and Pulex irritans were collected from animal shelters in Kayseri between January and August 2017. All flea species were individually screened for the presence of Wolbachia spp. by polymerase chain reaction (PCR) targeting the 16S ribosomal RNA gene. According to PCR analyses, Wolbachia spp. were found prevalent in C. canis and P. irritans fleas, while it was not detected in the C. felis species. Totally, 20 isolates were purified from agarose gel and sequenced with the same primers for molecular characterization and phylogenetic analyses. The sequence analyses revealed 17 polymorphic sites and 2 genetically different Wolbachia isolates, representing two different haplotypes in two flea species. The distribution patterns, molecular characterization, and phylogenetic status of Wolbachia spp. of fleas in Turkey are presented for the first time with this study. Understanding of the role of Wolbachia in vector biology may provide information for developing Wolbachia-based biological control tools.

Zoonotic infections are globally important diseases and lead to huge economic losses in both low- and middle-income and high-income countries. Global warming, environmental and ecological changes, illegal movement of animals and humans, regional civil wars, and poverty are predisposing factors for the emergence of zoonotic infections and their distribution worldwide; they are also a big threat for the future. In addition, environmental pollution and antimicrobial resistance are immense serious threats and dangers to prevent and control zoonotic infections. The natural location of Turkey allows many emerged or re-emerged infections with zoonotic characteristics by animal movements, such as bird immigrations, and by human movements due to civil wars as seen with regional refugees. Numerous zoonotic diseases, including 37 bacterial, 13 fungal, 29 viral, 28 parasitic (3 trematodes, 7 cestodes, 10 nematodes, and 8 protozoan), and totally 107 infections, have been reported from Turkey to date. Additionally, many ectoparasitic zoonoses within 15 different arthropod groups and one leech infestation have been reported from Turkey to date. The "One Health" initiative is particularly relevant for developing strategies to combat zoonotic diseases. In this article, we review the occurrence of zoonotic diseases in man and animals in Turkey in the light of the "One Health" perspective.

Pekmezci, Gokmen Zafer/0000-0002-7791-1959; Yildirim, Alparslan/0000-0001-9868-0363; Yetismis, Gamze/0000-0001-5260-3450; Duzlu, Onder/0000-0002-6951-0901; INCI, Abdullah/0000-0003-1614-0756; CILOGLU, ARIF/0000-0003-2695-7102 WOS: 000435595600013 PubMed: 29721658 The metacercariae of Clinostomum species which known as yellow grubs have zoonotic potential by infecting humans. In the present study, a total of 403 freshwater fish specimens belonging to different genera from Central Anatolia Region of Turkey were examined for yellow grub metacercariae infections. Only three specimens belonging to Squalius cephalus were found to be infected with metacercariae with a prevalence on this host species of 2.4% and an overall prevalence of 0.7%. All the metacercariae were morphologically identified as Clinostomum complanatum. Partial fragments of mitochondrial cytochrome oxidase I (mt-COI) gene and internal transcribed spacer 2 (ITS-2) were amplified for sequence and phylogenetic analyses. The sequence analyses of ITS-2 and mt-COI revealed three and nine polymorphic sites leading to detection of four and five haplotypes within the related gene regions, respectively. Moreover, the intraspecific genetic distances for C. complanatum isolates ranged from 0.0 to 0.7% for ITS-2 and 0.0 to 1.4% for mt-COI data sets. Consequently, the present study has provided first combined morphologic and molecular data on C. complanatum infecting Turkish freshwater fishes. Erciyes University Scientific and Technological Research Center, Kayseri, Turkey [TCD-2016-6795] This work was funded by the Erciyes University Scientific and Technological Research Center, Kayseri, Turkey (project number, TCD-2016-6795).

Bu calisma, Erzurum yoresi sigirlarinda Babesia bovis ve B. bigemina yayginliginin Real Time PCR ile arastirilmasi ve saptanan izolatlarin molekuler karakterizasyonlari amaciyla yapilmistir. Bu amacla 2011-2012 tarihleri arasinda farkli yas, cinsiyet ve irklarda, klinik olarak saglikli gorunumlu toplam 300 sigirdan EDTA’li tuplere tam kan ornekleri toplanmistir. DNA ekstraksiyonunu takiben B. bovis ve B.bigemina icin sirasiyla msa-2c ve rap-1 gen bolgelerine ozgu primerlerle Sybergreen ve TaqMan prob bazli Real Time PCR analizleri gerceklestirilmistir. Incelenen 300 kan orneginin 28’inde (%9,3) B. bovis, 17’sinde (%5,7) B. bigemina saptanmistir. Orneklerde miks enfeksiyona rastlanmamistir. B. bovis pozitiflerden 4’unun, B. bigemina pozitiflerden 3’unun msa-2c ve rap-1 gen bolgelerine gore sekans analizleri yapilmistir. B. bovis izolatlarinin pairwise kiyaslamalarinda kendi aralarinda, Turkiye’den diger bazi ve dunyadaki diger izolatlarla aralarinda sirasiyla %0,6, %2,3 ve %7,5 genetik farklilik belirlenmistir. B. bigemina izolatlarinin kendi aralarinda %0,3 ve dunyadaki diger izolatlarla aralarinda %0,5 genetik farklilik saptanmistir. B. bovis ve B. bigemina yonunden pozitif belirlenen orneklerin parazit turu temel alinarak sigirlarin yas, cinsiyet ve irk ozelliklerine gore istatistiksel analizlerinde; yas ve irk ozelliklerine gore istatistiksel bir farklilik belirlenmezken (p>0,05), cinsiyete bagli farklilik B. bovis icin onemsiz (p>0,05), B. bigemina icin ise onemli (p

Accurate species identifications are the essential first step in understanding the medical, economic, and ecological importance of black flies. The utility of DNA barcoding based on cytochrome c oxidase subunit 1 (COI) sequences was evaluated for identifying six common species of Palearctic black flies in the subgenus Wilhelmia, including several that are virulent pests. Chromosomally identified larvae from Turkey and Germany and COI sequences in GenBank were analyzed. Intraspecific genetic divergence was 0.7-3.5% (mean 1.6%), whereas interspecific genetic divergence was 2.7-16.9%. On the basis of COI barcodes, the six nominal species of Simulium (Wilhelmia) were clustered in three distinct clades with high levels of genetic divergence, using maximum likelihood and Bayesian analyses. All specimens of Simulium equinum (L.), Simulium pseudequinum Seguy, and Simulium paraequinum Puri were correctly identified. However, >75% of identifications were ambiguous for Simulium lineatum (Meigen) and Simulium turgaicum Rubtsov (Meigen) because of overlapping intra-and interspecific divergence of the two species and Simulium balcanicum (Enderlein), all three of which are chromosomally similar and nearly isomorphic. Phylogenetic evaluation showed that S. balcanicum, S. equinum, S. pseudequinum, and S. paraequinum were monophyletic, with high bootstrap and posterior probability values, but it also showed that S. lineatum and S. turgaicum were paraphyletic, each clustering in two distinct groups, suggesting the presence of cryptic taxa. Although DNA barcoding provided a partialmeans of identification and indications of additional biodiversity, other- molecularmarkers are needed to clarify the limits of all pest species of the subgenus Wilhelmia.

Haemonchus contortus is one of the most important gastrointestinal nematodes (GINs) infecting sheep, goats, and cattle worldwide. We developed a SYBR Green real-time PCR (qPCR) assay for detection and quantification of H. contortus by using specific primers based on a conserved region of the mitochondrial cytochrome oxidase subunit I (mt-COI) gene, and evaluated this technique in the detection of H. contortus infections in cattle in Central Anatolia Region of Turkey. The newly developed qPCR assay successfully discriminated H. contortus from other GIN species infecting cattle in the specificity evaluations, with a specific melt peak of 77.5 degrees C. Our results revealed the efficient amplification of the proposed target COI region within the range of plasmid copies, from 2 x 10 (6) to 2 x 10 (1) per mu l, with 96.9 % efficiency, R-2 value of 0.999, and a slope of -3.398. Among the 920 cattle fecal samples from the field, 58 samples (6.3 %) were positive with qPCR assay, whereas 45 samples (4.9 %) were positive, as determined by larval culture, suggesting the utility of SYBR Green qPCR. Phylogenetic characterization of the partial COI gene of H. contortus isolates was also evaluated for 100 eggs and third stage larvae recovered from positive cattle faecal samples, which were verified with the qPCR assay prior to analyses. COI sequences were classified into three haplotypes (THC1 to THC3) with intraspecific nucleotide differences of 0.50 to 0.76 %. Phylogenetic analyses revealed that the haplotypes grouped with H. contortus isolates from several countries in a monophyletic cluster, with evidence of at least two main haplogroups. Overall, the SYBR Green qPCR assay was highly specific and sensitive, suggesting that it can be used for screening of H. contortus infections in livestock populations in epidemiological studies and the control of this important parasite.

Organisms with vast distributions often represent geographical mosaics of cryptic species. The black fly Simulium (Wilhelmia) lineatum is among the most widely distributed members of the family Simuliidae, ranging from the British Isles to eastern China. Rather than viewing S. lineatum as a possible aggregate of multiple species, taxonomists have suggested a more inclusive taxon with additional synonyms. Accordingly, S. lineatum is an ideal candidate for testing the hypothesis that a wide geographical distribution signals the presence of more than one species. A cytogenetic approach was used to probe the macrogenome of S. lineatum and other taxa proposed by taxonomists as conspecific. The banding patterns in the polytene chromosomes of 480 larvae from 15 countries across the Palearctic Region revealed 128 rearrangements of the complement. All rearrangements were autosomal and 89% were inversions nonrandomly distributed among species and among chromosome arms. The analyses clarify long-standing confusion over previously proposed names and reveal a longitudinal succession of four species sequentially replacing one another from west to east: Simulium lineatum s.s., Simulium balcanicum, Simulium turgaicum, and Simulium takahasii. Thus, S. turgaicum is recalled from synonymy and the other three species are validated. Within the most-represented species, S. balcanicum, the frequency of inversions follows a longitudinal gradient with a north–south bias; as the distance between the sites increases along this north-west–south-east axis, the similarity of inversion frequencies between sites decreases. Validation of the concept that broadly distributed black flies are composites of structurally similar species provides a framework for guiding discovery of additional biodiversity. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 114, 163–183.