A case of domestic goat parasitism by Neotrombicula heptneri (Acariformes: Trombiculidae) in Turkey

A chigger mite Neotrombicula heptneri Kudryashova, 1973, which was originally described from three gerbillid species in Iran, is for the first time recorded in Turkey and on the domestic goat Capra hircus L. It is the second chigger species found infesting livestock in Turkey.Russian Foundation for Basic ResearchRussian Foundation for Basic Research (RFBR) [13-04-00028-a]The authors wish to express their appreciation to Dr Naina I. Kudryashova (ZMMU), who gave them an opportunity to study the type material of N. heptneri. They are also grateful to two anonymous reviewers for useful comments. This research was supported by a grant from the Russian Foundation for Basic Research No. 13-04-00028-a (to A.A. Stekolnikov)

The study of exposure times and dose-escalation of tick saliva on mouse embryonic stem cell proliferation

The saliva of ticks contains numerous bioactive molecules with anti-hemostatic and immunomodulatory properties. Due to their abilities of self-renewal and pluripotency, stem cells hold considerable promise in the regenerative medicine and biomedical fields. The present study examines the viability and proliferation of mouse embryonic stem cells (mESCs) following the addition of tick salivary gland extracts obtained from three tick species (Dermacentor marginatus, Rhipicephalus bursa and Hyalomma marginatum) to the mESC medium in different quantities (0.2, 2, 20, 40, 80, and 160 µg/ml). On days 2, 5 and 7 of the treatment, the vitality and proliferation of the cells were determined with CellTiter-Glo and morphological tests. The results showed that the culture supplemented with D. marginatus salivary gland extract at a concentration of 80 µg/ml positively affected the proliferation rate of mESC. It was further shown that all concentrations of the salivary gland extracts derived from H. marginatum and R. bursa had a negative effect on the proliferation rate of mESC when compared to the controls

First evidence of predation of the ant species lasius alienus on the poultry red mite dermanyssus gallinae

The poultry red mite (PRM), Dermanyssus gallinae (De Geer, 1778) (Acari: Dermanyssi-dae), is a common and significant ectoparasite of the poultry industry worldwide. Although various biological, chemical, and physical methods have been attempted, an utterly successful control strategy has not been put forward yet. Our experimental investigations and observations revealed that the ant species Lasius alienus displays an effective predatory behavior on all biological stages of PRM. Our results also suggested that L. alienus is attracted by PRM­infested substrate at a distance. We concluded that predation by the ant on PRM is worth further investigation as it could possibly be an effective biological control strategy. © Kar S. et al

Predation of ant species Lasius alienus on tick eggs: impacts of egg wax coating and tick species

Several animal species, including ants, have been reported to be capable of predation on ticks. However, determining factors in most interactions between ticks and predators have not yet been fully deciphered. We hypothesized that the ant species Lasius alienus, which is unknown whether it has any impact on ticks, may exhibit predation on the eggs of tick species Hyalomma marginatum, H. excavatum, and Rhipicephalus bursa, and that the tick egg wax can be the main determinant in possible predation. In the study, 6300 tick eggs with the natural wax coating (waxed/untreated) and 2700 dewaxed tick eggs, the wax of which was removed in the laboratory, were repeatedly presented to the foraging workers belonging to three different ant nests in their natural habitat. Depending on the tick species and trials, the rate of the eggs carried by the ants ranged from 12.8 to 52.1% in the waxed and from 59.8 to 78.4% in the dewaxed eggs. It was observed that the dewaxing process both increased the interest of the ants in the eggs and resulted in a reduction in the variation associated with tick species. This study showed that L. alienus has a predatory effect on tick eggs, the severity of this impact is closely associated with the tick species, the tick-associated difference is caused by the species-specific property of the egg wax, and the variety in the protective effects of the wax seems to be an evolutional result of the biological and ecological adaptation process of the species

Black Soldier Fly Hermetia illucens (Linnaeus, 1758): Biology, Farming and Using for Animal Nutrition

Meeting the food needs of growing world population is one of the most important global problems of our time. It is widely emphasized that the need will be much more serious in the coming decades and that radical measures should be taken at this point. The ongoing search for alternatives at this point has shown that insects can be one of the ideal sources for feeding of human and especially livestock. Many insect species have been focused in the related investigations, and the results showed that particularly black soldier fly is the most ideal species in many aspects. The species, which has advantages such as ease of production, feeding on organic wastes, potential for utilization of organic wastes as an organic fertilizer, being nature friendly and largely nutritional content of larvae, has begun to be widely produced in many parts of the world in recent decade. Related data and reports reveal the significance of this fly species in the worldwide food shortage issue with a very assertive certainty. In this review, the issues of biology, morphology, farming in Turkey conditions, nutritional content and use for animal nutrition of the fly have been elaborated in detail, and almost all the current data that both possible producers and related researchers can benefit from are presented in an integrated manner

A survey on anthelmintic resistance in strongyles to ivermectin and pyrantel and macrocyclic lactone-resistance in parascaris equorum

Bu çalışma, Makrosiklik Lakton (ML) grubu antelmentiklere dirençli Parascaris equorum'ların teşhis edildiği bir at çiftliğinde, atları enfekte eden dirençli parazitlerle ilgili son durumu tespit etmek ve Strongylidae enfeksiyonlarına karşı ivermektin ve pirantelin etkilerini belirleyerek, bunlarla ilgili bir direnç probleminin olup olmadığını araştırmak amacıyla yapılmıştır. Araştırmanın birinci döneminde, ML bileşiği olan abamektinin P. equorum'a karşı etkinliği araştırılmış ve 12 attan 11'inde etki %0 olarak saptanmıştır. Abamektin tedavisi sonrası Parascaris pozitif atlara uygulanan pirantel pamoat 8 atta %96-100, 3 atta ise %0-80 arası etki göstermiştir. Parascaris'le enfekte olan atlara tekrar uygulanan pirantel (iki kat doz) tüm atlarda %100 etkili bulunmuştur. Araştırmanın ikinci döneminde, iki grup atta ivermektin ve pirantelin Strongylidae enfeksiyonlarında 14. ve 28. gün etkilerine bakılmış ve her iki muayene gününde de etki %100 olarak saptanmıştır. Sonuç olarak, ML grubu antelmentiklerin çiftlik atlarındaki Parascaris'lere karşı artık tamamen etkisiz kaldıkları görülmüş, dirençli parazitlere ise pirantelin özellikle iki misli dozunun daha etkili olduğu bulunmuştur. Strongylidae enfeksiyonlarında ise ivermektin veya pirantel ile ilgili herhangi bir dirençlilik kaydedilmemiştir.This survey was carried out on a horse farm in order to acquire recent data on macrocyclic lactone-resistant Parascaris equorum which had been previously detected on this farm and to determine efficacies of ivermectin and pyrantel against strongyles in order to determine whether a resistance problem also exists in these parasites. In the first part of the study, abamectin was given to horses infected with P. equorum. In 11 out of 12 horses, zero efficacy (0%) was seen against P. equorum. Subsequently, horses which were Parascaris-positive after abamectin treatment received pyrantel pamoate and its efficacy was 0-80% in 3 animals and 96-100% 8. The remained Parascaris-infected horses were treated again with a double dose of pyrantel and it was fully effective in all horses. In the second part of the study, efficacy of ivermectin and pyrantel against strongyles infections was assessed 14 and 28 days after treatment and an efficacy of 100% was detected on both days. In conclusion, macrocyclic lactone anthelmintics were found to be non-efficacious against P. equorum whereas pyrantel at the double dose rate was effective against these resistant worms. Resistance to ivermectin or pyrantel was not detected in strongyles

Utility of a Sequence-Independent, Single-Primer-Amplification (SISPA) and Nanopore Sequencing Approach for Detection and Characterization of Tick-Borne Viral Pathogens

Currently, next generation sequencing (NGS) is the mainly used approach for identification and monitorization of viruses with a potential public health threat in clinical and environmental samples. To facilitate detection in NGS, the sequence-independent, single-primer-amplification (SISPA) is an effective tool for enriching virus sequences. We performed a preliminary assessment of SISPA-nanopore sequencing as a potential approach for screening tick-borne viruses in six specimens with detectable Crimean-Congo hemorrhagic fever virus (CCHFV) and Jingmen tick virus (JMTV) sequences. A comparison of unbiased NGS and SISPA followed by nanopore sequencing was carried out in 4 specimens with single and pooled ticks. The approach was further used for genome sequencing in culture-grown viruses. Overall, total/virus-specific read counts were significantly elevated in cell culture supernatants in comparison to single or pooled ticks. Virus genomes could be successfully characterized by SISPA with identities over 99%. Genome coverage varied according to the segment and total read count. Base calling errors were mainly observed in tick specimens and more frequent in lower viral loads. Culture-grown viruses were phylogenetically-related to previously-reported local viruses. In conclusion, the SISPA + nanopore sequencing was successful in generating data comparable to NGS and will provide an effective tool for broad-range virus detection in ticks

Co-circulation of West Nile virus and distinct insect-specific flaviviruses in Turkey

Background: Active vector surveillance provides an efficient tool for monitoring the presence or spread of emerging or re-emerging vector-borne viruses. This study was undertaken to investigate the circulation of flaviviruses. Mosquitoes were collected from 58 locations in 10 provinces across the Aegean, Thrace and Mediterranean Anatolian regions of Turkey in 2014 and 2015. Following morphological identification, mosquitoes were pooled and screened by nested and real-time PCR assays. Detected viruses were further characterised by sequencing. Positive pools were inoculated onto cell lines for virus isolation. Next generation sequencing was employed for genomic characterisation of the isolates. Results: A total of 12,711 mosquito specimens representing 15 species were screened in 594 pools. Eleven pools (2%) were reactive in the virus screening assays. Sequencing revealed West Nile virus (WNV) in one Culex pipiens (s. l.) pool from Thrace. WNV sequence corresponded to lineage one clade 1a but clustered distinctly from the Turkish prototype isolate. In 10 pools, insect-specific flaviviruses were characterised as Culex theileri flavivirus in 5 pools of Culex theileri and one pool of Cx. pipiens (s. l.), Ochlerotatus caspius flavivirus in two pools of Aedes (Ochlerotatus) caspius, Flavivirus AV-2011 in one pool of Culiseta annulata, and an undetermined flavivirus in one pool of Uranotaenia unguiculata from the Aegean and Thrace regions. DNA forms or integration of the detected insect-specific flaviviruses were not observed. A virus strain, tentatively named as Ochlerotatus caspius flavivirus Turkey, was isolated from an Ae. caspius pool in C6/36 cells. The viral genome comprised 10,370 nucleotides with a putative polyprotein of 3,385 amino acids that follows the canonical flavivirus polyprotein organisation. Sequence comparisons and phylogenetic analyses revealed the close relationship of this strain with Ochlerotatus caspius flavivirus from Portugal and Hanko virus from Finland. Several conserved structural and amino acid motifs were identified. Conclusions: We identified WNV and several distinct insect-specific flaviviruses during an extensive biosurveillance study of mosquitoes in various regions of Turkey in 2014 and 2015. Ongoing circulation of WNV is revealed, with an unprecedented genetic diversity. A probable replicating form of an insect flavivirus identified only in DNA form was detected.U.S. Armed Forces Health Surveillance Board Global Emerging Infections Surveillance and Response System (AFHSB-GEIS) research; Walter Reed Army Institute of Research; Smithsonian InstitutionSmithsonian Institution; Georg Forster Research Fellowship (HERMES); Alexander von Humboldt Foundation, GermanyAlexander von Humboldt FoundationA U.S. Armed Forces Health Surveillance Board Global Emerging Infections Surveillance and Response System (AFHSB-GEIS) research award (to YML) supported this study. This research was performed in part under a Memorandum of Understanding between the Walter Reed Army Institute of Research and the Smithsonian Institution, with institutional support provided by both organisations. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The material to be published reflects the views of the authors and should not be construed to represent those of the United States Department of the Army or the United States Department of Defense. KE was a 2015 recipient of the Georg Forster Research Fellowship (HERMES) for Experienced Researchers, of the Alexander von Humboldt Foundation, Germany

Novel Tick Phlebovirus Genotypes Lacking Evidence for Vertebrate Infections in Anatolia and Thrace, Turkey

We screened ticks and human clinical specimens to detect and characterize tick phleboviruses and pathogenicity in vertebrates. Ticks were collected at locations in Istanbul (Northwest Anatolia, Thrace), Edirne, Kirklareli, and Tekirdag (Thrace), Mersin (Mediterranean Anatolia), Adiyaman and Sanliurfa (Southeastern Anatolia) provinces from 2013-2018 and were analyzed following morphological identification and pooling. Specimens from individuals with febrile disease or meningoencephalitic symptoms of an unknown etiology were also evaluated. The pools were screened via generic tick phlebovirus amplification assays and products were sequenced. Selected pools were used for cell culture and suckling mice inoculations and next generation sequencing (NGS). A total of 7492 ticks were screened in 609 pools where 4.2% were positive. A phylogenetic sequence clustering according to tick species was observed. No human samples were positive. NGS provided near-complete viral replicase coding sequences in three pools. A comprehensive analysis revealed three distinct, monophyletic virus genotypes, comprised of previously-described viruses from Anatolia and the Balkans, with unique fingerprints in conserved amino acid motifs in viral replicase. A novel tick phlebovirus group was discovered circulating in the Balkans and Turkey, with at least three genotypes or species. No evidence for replication in vertebrates or infections in clinical cases could be demonstrated.Armed Forces Health Surveillance Board, Global Emerging Infections Surveillance and Response System (AFHSB-GEIS), United States of America [P0034_18_WR]; US ArmyUnited States Department of Defense [W911QY-16-C-0160]The study was supported in part by the Armed Forces Health Surveillance Board, Global Emerging Infections Surveillance and Response System (AFHSB-GEIS), United States of America (FY18 award P0034_18_WR (PI: Yvonne-Marie Linton) under US Army subcontract W911QY-16-C-0160)

Isolation and genomic characterization of Culex theileri flaviviruses in field-collected mosquitoes from Turkey

Vector surveillance for the arthropod-borne infections has resulted in the isolation of a growing number of novel viruses, including several flavivirus strains that exclusively replicate in insects. This report describes the isolation and genomic characterization of four insect-specific flaviviruses frommosquitoes, previously collected from various locations in Turkey. C6/36 Aedes albopictus and Vero cell lines were inoculated with mosquito pools. On C6/36 cells, mild cytopathic effects, characterized as rounding and detachment, were observed in four pools that comprised female Culex theileri mosquitoes. Complete (3 isolates, 10,697 nucleotides) or near-complete (1 isolate, 10,452 nucleotides) genomic characterization was performed in these culture supernatants via next generation sequencing. All strains demonstrated high genetic similarities, with over 99% identity match on nucleotide and amino acid alignments, revealing them to be different isolates of the same virus. Sequence comparisons identified the closest relative to be the Culex theileri flavivirus (CTFV) strains, originally characterized in Portugal. Phylogenetic analyses demonstrated that the isolates remained distinct as a cluster but formed amonophyletic group with CTFV strains, and shared a common ancestor with Quang Binh or related Culex flaviviruses. The organization of the viral genome was consistent with the universal flavivirus structure and stem-loops; conserved motifs and imperfect tandem repeats were identified in the non-coding ends of the viral genomes. A potential ribosomal shifting site, resulting in the translation of an additional reading frame, was detected. The deduced viral polyprotein comprised 3357 amino acids and was highly-conserved. Amino acid variations, presumably associated with adaptive environmental pressures, were identified. These isolates comprise the first fully characterized insect-specific flaviviruses in Turkey. Their impact on West Nile virus circulation, which is also endemic in the study region, remains to be explored. (C) 2016 Elsevier B.V. All rights reserved.Armed Forces Health Surveillance Center, Global Emerging Infections Surveillance and Response System (AFHSC-GEIS), United States [W81XWH-11-2-0174]; Georg Forster Research Fellowship (HERMES) for Experienced Researchers by Alexander von Humboldt Foundation; National Research Council (NRC) Research Associateship Award at the Walter Reed Army Institute of ResearchThis study was partially supported by The Armed Forces Health Surveillance Center, Global Emerging Infections Surveillance and Response System (AFHSC-GEIS), United States (W81XWH-11-2-0174) (with Yvonne-Marie Linton as the principal investigator). KE is a recipient of the Georg Forster Research Fellowship (HERMES) for Experienced Researchers by the Alexander von Humboldt Foundation, 2015. This manuscript was prepared whilst YML held a National Research Council (NRC) Research Associateship Award at the Walter Reed Army Institute of Research. This research was performed in part under a Memorandum of Understanding between the Walter Reed Army Institute of Research and the Smithsonian Institution, with institutional support provided by both organizations. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript. The material to be published reflects the views of the authors and should not be construed to represent those of the US Department of the Army or the US Department of Defense