Molecular investigation of Anaplasma phagocytophilum and related strains among sheep flocks from different parts of Türkiye; with a note of phylogenetic analyses of Anaplasma phagocytophilum- like 1.

Anaplasma phagocytophilum is a vector-borne zoonotic pathogen and can infect various vertebrate hosts, especially cattle, sheep, goats, horses, and dogs. Molecular-based studies have revealed that the agent has a high genetic diversity and closely related strains circulate in hosts. In this study, 618 sheep blood samples obtained from different geographic regions of Türkiye were researched for A.phagocytophilum and related strains with PCR, RFLP, and DNA sequence analyses. The DNA of these pathogens was detected in 110 (17.79%) samples. RFLP assay showed that all positive samples were infected with A.phagocytophilum-like 1, whereas A.phagocytophilum-like 2 and A.phagocytophilum were not detected. Partial parts of 16 S rRNA gene of seven randomly selected positive samples were sequenced. The phylogenetic analyses of these isolates revealed that at least two A.phagocytophilum-like 1 isolates circulate among hosts in Türkiye and around the world. A.phagocytophilum-related strains have been reported in molecular-based studies over the last few years, but there is a lack of data on the vector competence, epidemiology, clinical symptoms, and genetic diversity of these pathogens. Therefore, large-scale molecular studies are still needed to obtain detailed data on the above-mentioned topics.

Anaplasma capra: a new emerging tick-borne zoonotic pathogen.

The genus Anaplasma includes A. marginale, A. centrale, A. bovis, A. ovis, A. platys, and A. phagocytophilum transmitted by ticks, some of which are zoonotic and cause anaplasmosis in humans and animals. In 2012, a new species was discovered in goats in China. In 2015, the same agent was detected in humans in China, and it was provisionally named Anaplasma capra, referring to 2012. The studies conducted to date have revealed the existence of A. capra in humans, domestic animals, wild animals, and ticks from three different continents (Asia, Europe, and Africa). Phylogenetic analyses based on gltA and groEL sequences show that A. capra clearly includes two different genotypes (A. capra genotype-1 and A. capra genotype-2). Although A. capra human isolates are in the genotype-2 group, goat, sheep, and cattle isolates are in both groups, making it difficult to establish a host genotype-relationship. According to current data, it can be thought that human isolates are genotype-2 and while only genotype-1 is found in Europe, both genotypes are found in Asia. Anaplasma capra causes clinical disease in humans, but the situation is not yet sufficient to understand the zoonotic importance and pathogenicity in animals. In the present review, the history, hosts (vertebrates and ticks), molecular prevalence, pathogenic properties, and genetic diversity of A. capra were evaluated from a broad perspective.

First Molecular Evidence of Babesia vogeli, Babesia vulpes, and Theileria ovis in Dogs from Kyrgyzstan.

Tick-borne parasitic diseases cause mild to severe infections among vertebrate hosts, including dogs. Species in the genus Babesia are important tick-borne pathogens and have worldwide distributions. Although there are data on the prevalence and distribution of Babesia species among dogs around the world, there is no information available in Kyrgyzstan, according to a literature review. In this study, 337 dogs were screened by nested PCR for the presence of the 18S small subunit ribosomal RNA (18S SSU rRNA) gene of piroplasm species. Overall prevalence was 6.23% (21/337) for Babesia/Theileria spp. DNA sequencing of positively tested samples revealed that eighteen samples were infected with Babesia vogeli (B. vogeli) (5.34%), two samples with B. vulpes (0.59%), and one sample with Theileria ovis (T. ovis) (0.29%). The phylogenetic analyses and nucleotide sequences in contrast with those present in GenBank revealed that two nucleotide substitutions (594th and 627th) were found between B. vogeli isolates, including ours, indicating that the mutation is relatively rare. The sequences of other pathogens obtained in this study confirmed 100% nucleotide identity with B. vulpes and T. ovis sequences in GenBank. To the best of our knowledge, B. vogeli, B. vulpes, and T. ovis were detected for the first time in dogs from Kyrgyzstan, and it is thought that results will contribute to the understanding of the epidemiology of canine tick-borne pathogens in the country.

Multi-sensor, multi-device smart building indoor environmental dataset.

A dataset of sensor measurements is presented. Our dataset contains discrete measurements of 8 IoT devices located in various places in a research lab at the University of Bristol. Nordic nRF52840 DK IoT devices periodically collects environmental data, such as temperature, humidity, pressure, gas, room light intensity, accelerometer; including also a measurement quality indicator. The measurements were taken every 10 seconds over a six-month period between February and September 2022. In addition, we provide Received Signal Strength Indicator (RSSI) of the IoT devices. The data files are formatted as CSV files. There are various software libraries available to access and read this file format. We provide "README.txt" file which explains the repository and how to use dataset. Each data file is named according to its creation date and, once it reaches a size of 1MB, it is compressed and archived. A new folder is created every week to store all the data files from that week automatically. The dataset can be used for drift detection such as malicious or anomaly detection algorithms. It can also be used for smart building applications like occupation detection. The dataset can be found at https://data.bris.ac.uk/data/dataset/fwlmb11wni392kodtyljkw4n2.

Molecular detection and phylogenetic analysis of Toxocara vitulorum in feces and milk samples from naturally infected water buffaloes.

Toxocara vitulorum infects cattle and water buffalo, leading to mild to severe infection in calves and has wide geographic distributions, especially in tropical and subtropical countries. This work aimed to assess the prevalence, distributions, and phylogeny of T.vitulorum in water buffaloes in different parts of Sivas, one of the essential buffalo-breeding areas in Türkiye. T.vitulorum was found in 42 (8.23%) and 54 (10.58%) fecal (n:510) samples using microscopic and molecular techniques, respectively. T.vitulorum was higher in animals aged 0-6 months compared to other groups. Furthermore, when animals aged 0-6 months were grouped within themselves, the prevalence of T.vitulorum in 1-3 month-old-animals was higher than in both younger than one month and older than three months. T.vitulorum was detected in fecal samples obtained from animals older than six months. In colostrum/milk samples (n:100), T.vitulorum-larvae were found in 4% and 10% with microscopic and molecular techniques, respectively. The larvae were detected in colostrum/milk samples in the mother between the 2nd and 28th days postpartum-period. The ITS-1-gene of 11 PCR-positive samples was sequenced. The 98.99-100% nucleotide identity was determined between our T.vitulorum isolates and those present in GenBank. In conclusion, this is the first molecular survey and phylogenetic analysis of T.vitulorum in fecal and colostrum/milk samples from naturally infected water buffaloes. Data obtained in this study will help to understand the life cycle and epidemiology of the nematode. Data also revealed that veterinarians should consider older animals as well as young animals in their control program of nematode infections in farms.

Molecular survey of Anaplasma phagocytophilum and related variants in water buffaloes: The first detection of Anaplasma phagocytophilum-like 1.

Anaplasma phagocytophilum infects various hosts and lead to mild to severe infection. Currently, two A.phagocytophilum-related variants have been documented in different countries. Although limited, there are studies revealing the presence of A.phagocytophilum in water buffaloes, but no study investigating A.phagocytophilum-like 1 and -like 2. A.phagocytophilum and related variants were investigated using PCR, PCR-RFLP, and DNA sequence analysis in water buffaloes in Türkiye. 364 buffalo blood samples were examined for A.phagocytophilum and related strains. Seven buffaloes were determined to be positive with PCR and PCR-RFLP revealed that all samples were A.phagocytophilum-like 1. According to the partial sequence of 16 S rRNA gene, A.phagocytophilum like-1 may split into two different variants. This work supplies the first molecular report of A.phagocytophilum-like 1 in water buffaloes. However, a lack of information is present on the pathogen's clinical manifestations and vector species. There is still a need to investigate vectors and clinical signs of the pathogen.

Molecular prevalence of bovine hemoplasmosis in Turkey with first detection of Mycoplasma wenyonii and Candidatus Mycoplasma haemobos in cattle and water buffalo.

Hemoplasma species can cause infection varying from mild to severe in a wide range of hosts, including cattle and water buffalo. Two hemoplasma species, Mycoplasma wenyonii and Candidatus Mycoplasma haemobos, have been reported in cattle and water buffalo from different parts of the world to date. There was a lack of information on the presence and distribution of these pathogens in Turkey despite the negative economic impact on livestock production. This study aimed to develop a duplex PCR assay amplifying the 16S rRNA gene, in order to analyze DNA samples obtained from 297 cattle and 360 water buffaloes, and to determine the molecular prevalence of bovine hemoplasma species in Sivas province. Bovine hemoplasma species were found in 94 of 297 (31.64%) cattle and in 17 of 360 (4.72%) water buffaloes in this study. Randomly selected six positives PCR products (three samples each species) obtained from cattle and water buffaloes were sequenced, and the consensus sequences were uploaded to GenBank. Nucleotide similarity of 96.97-100% was determined between M. wenyonii isolates obtained in this study and those of M. wenyonii isolates present in the GenBank database, whereas C. Mycoplasma haemobos isolates from this study shared 99.04-100% homology with the C. Mycoplasma haemobos isolates uploaded to the GenBank. With the current study, the molecular presence of M. wenyonii and C. Mycoplasma haemobos were documented for the first time in cattle and water buffaloes in Turkey. Considering the rate of prevalence, veterinarians should take precautions against bovine hemoplasma species to protect animal health.

Molecular Survey of Anaplasma phagocytophilum and Related Strains in Sheep and Goats from Sivas; with a High Prevalence of Anaplasma phagocytophilum-like 1

Objective: This study aimed to investigate Anaplasma phagocytophilum and related strains (A. phagocytophilum-like 1 and like 2) in sheep and goats for the first time in Sivas province with molecular techniques. Methods: The study material was composed of 247 animal (159 sheep and 88 goats) blood samples from four districts of Sivas province (Sivas City Center, Kangal, Koyulhisar, and Yildizeli). A. phagocytophilum and related strains were screened with polymerase chain reaction (PCR), PCR-RFLP, and DNA sequence analysis. Results: A. phagocytophilum related strains were found in 44.93% (111/247) of the small ruminants using PCR. The infection rate was 45.91% (73/159) in sheep and 43.18% (38/88) in goats. In this study, 110 samples were positive for only A. phagocytophilum-like 1, while A. phagocytophilum-like 1 and like 2 were mix-infection in one sample. A. phagocytophilum was not detected in sheep or goats. Two randomly selected PCR products were sequenced in both directions, and the consensus sequences were deposited on the GenBank under accession numbers: ON598644 and ON598645. Nucleotide similarity of 99.34-100% was determined between A. phagocytophilum-like 1 isolates obtained in this study and those of A. phagocytophilum-like 1 isolates present in the GenBank database. Conclusion: This study provides the first molecular data on A. phagocytophilum-like 1 and like 2 in Sivas province. Considering the high positive rate of the A. phagocytophilum-like 1 in sheep and goats, there is a paucity of data on clinical symptoms and vector species of the pathogen. Therefore, further studies are needed to investigate the vector tick species and clinical symptoms of the pathogen in the host.

The detection and phylogenetic analysis of Anaplasma phagocytophilum-like 1, A. ovis and A. capra in sheep: A. capra divides into two genogroups.

In this study, the presence, prevalence, and genotypes of Anaplasma phagocytophilum, A. ovis, and A. capra in sheep were investigated based on 16 S SSU rRNA, groEL, and gtlA gene-specific polymerase chain reaction (PCR), respectively. The sequences of the genes were used for detection of the phylogenetic position of the species. Additionally, a restriction fragment length polymorphism (RFLP) were carried out for discrimination of A. phagocytophilum and related variants (A. phagocytophilum-like 1 and 2). The prevalence of Anaplasma spp. was found as 25.8% (101/391), while it was found that A. ovis, A. phagocytophilum-like 1, and A. capra are circulating in the sheep herds in Kyrgyzstan, according to the PCRs, RFLP and the partial DNA sequencing results. The positivity rates of A. phagocytophilum-like 1, A. ovis, and A. capra genotype-1 were 6.9, 22.5, and 5.3%, respectively. A total of 32 (8.2%) sheep were found to be mix infected. Moreover, phylogenetic analyses and sequence comparison with those available in the GenBank showed that A. capra formed two distinct genetic groups (A. capra genotype-1 and A. capra genotype-2). Considering the zoonotic potential of these species, it may be necessary to make changes in the interpretation of anaplasmosis cases in animals and there is a need for further studies to determine the pathogenicity of the species/genotypes circulating in animals.

Buffaloes as new hosts for Anaplasma capra: Molecular prevalence and phylogeny based on gtlA, groEL, and 16S rRNA genes.

Anaplasma capra is a tick-borne pathogen that was discovered for the first time in goats in China in 2012. The studies carried out from the first detection in China to the present have revealed the presence of this species in eight countries including Angola, France, Iranian, South Korea, Kyrgyzstan, Malaysia, Spain, and Türkiye in three continents (Africa, Asia, and Europe). It has also been determined that humans, sheep, cattle, dog, and wild animals are the hosts of A. capra. It was investigated whether water buffaloes were the host of A. capra using nested-PCR and DNA sequencing in this study. The prevalence of A. capra in Turkish water buffalo herds was investigated and phylogenetic analyzes were performed on the basis of gltA, groEL, and 16S rRNA genes. A total of 364 water buffalo blood samples were examined in terms of A. capra using gltA gene species-specific nested-PCR. A. capra were detected in 52 of 364 (14.28%) blood samples. There was no statistically significant difference between the prevalence, gender, and age parameters. The gltA, groEL, and 16S rRNA genes in randomly selected three positive samples were sequenced. A. capra isolates obtained from water buffaloes in this study shared 85.20-100%(gltA), 89.84-100%(groEL), and 99.82-100%(16S rRNA) nucleotide similarity with A.capra isolates present in GeneBank. Phylogenetic analyses of gtlA and groEL genes revealed that A. capra divided in two different genogroups. In conclusion, this study revealed that water buffalo is a new host of A. capra. However, comprehensive studies are needed to determine the pathogenicity, vectors, and biological properties of A. capra in this new host.

The first molecular detection of Anaplasma capra in domestic ruminants in the central part of Turkey, with genetic diversity and genotyping of Anaplasma capra.

Tick-borne diseases have been an increasing threat to human and animal health all over the world. Anaplasmosis is one of the emerging tick-borne diseases and has zoonotic potential. A new novel species, which was detected in China in 2010-2012 and provisionally named Anaplasma capra in 2015, causes zoonotic infections and infects many different animal species. In this study, we investigated the presence of A. capra in domestic ruminants from Turkey. A total of 468 blood samples (cattle, sheep, and goat) were examined by the gltA gene-specific nested polymerase chain reaction, revealing the presence of A. capra in six samples (1.28%): one of them from cattle (0.41%) and the other five from sheep (3.22%). According to DNA sequences results of the gltA gene, A. capra isolates identified in the present study were shown high nucleotide similarity with A. capra isolates detected from different hosts. However, the nucleotide differences were detected in the same nucleotide positions between A. capra isolates. For this reason, we thought that at least two different A. capra genotypes could be circulating in the world. As a result, it is seen that A. capra, which was determined to be a new species with zoonotic potential, was revealed in European and Asian countries and in different hosts. In order to raise awareness about human anaplasmosis infections, it is important to reveal the prevalence of the species in the world. The emergence of A. capra in Turkey reveals the need for a re-evaluation of the human and animal health risk analysis in terms of anaplasmosis.

First molecular detection of Anaplasma species in cattle from Kyrgyzstan; molecular identification of human pathogenic novel genotype Anaplasma capra and Anaplasma phagocytophilum related strain.

Anaplasmosis is a rickettsial infection with significant effects on human and animal health, and the discovery of new species or genotypes with zoonotic potential in recent years has increased this importance. The aim of this study was to provide the first assessment of the molecular etiology and prevalence of bovine anaplasmosis in Kyrgyzstan (specifically in the Chuy, Talas, Djalal-Abad, Naryn, and Issyk-Kul regions). The prevalence of bovine anaplasmosis was determined as 1.7% (6/358). PCR and partial DNA sequencing results of the 16S ribosomal RNA (rRNA) gene revealed that Anaplasma centrale, A. phagocytophilum like-1, and the human pathogenic novel genotype A. capra are circulating in cattle herds in Kyrgyzstan. Six DNA nucleotide sequences obtained in this study were deposited in GenBank under the following accession numbers: A. centrale (MW672117, MW672118, MW672119, MW672120), A. phagocytophilum (MW672121), and A. capra (MW672115).

Intestinal system helminths of red foxes and molecular characterization Taeniid cestodes.

Red foxes (Vulpes vulpes) are the most prevalent wild carnivores in the world and definitive hosts of many pathogenic parasites for humans and farm animals. These animals travel great distances in search of prey and nests, and cause contamination of large geographic areas with parasites. For this reason, monitoring the parasitic pathogens of red foxes is particularly important in terms of public and animal health. The goal of this study was to determine the intestinal helminths and molecular characterization of Taenia species of red foxes in Turkey. In this study, 103 red fox intestines obtained from 29 provinces of Turkey were examined with sedimentation and counting technique. Collected helminths were diagnosed according to their morphologic features. Additionally, further molecular analysis (PCR and DNA sequencing) was performed for the identification of Taeniid cestodes. At the end of the study, it was determined that 87.37% (90/103) of red foxes were infected with at least one helminth species. Detected helminths and their prevalence's were Mesocestoides sp. (56.31%), Joyeuxiella echinorhynchoides (33%), Taenia polyacantha (15.53%), Dipylidium caninum (0.97%), Pterygodermatites affinis (51.45%), Toxascaris leonina (45.63%), Uncinaria stenocephala (33%), Oxynema numicidum (20.38%), Toxocara canis (14.56%), Ancylostoma caninum (12.62%), and Trichuris vulpis (1.94%), respectively. Additionally, Pachysentis sp. (37.69%), Centrorhynchus sp. (0.97%) (Acantocephala), and nymphs of Linguatula serrata (20.38%) (Arthropoda) were also detected in the same intestinal samples. This is the most comprehensive study that has been conducted on the intestinal helminthes of red foxes in Turkey. To the best of our knowledge, molecular characterization of T. polyacantha and the detection of O. numicidum, A. caninum, Pachysentis sp., and Centrorhynchus sp. are the first reports in red foxes in Turkey. Our study revealed that red foxes are important hosts for many intestinal helminth species and are link between domestic and sylvatic cycles of these parasites.

Updated checklist of Culicoides Latreille (Diptera: Ceratopogonidae) of Turkey with ten new records.

We investigated the Culicoides fauna in Turkey during the years 2016-2019 in the process of entomological surveillance for arboviral diseases. The entomological survey was conducted at 104 sampling stations in 51 provinces in Turkey during four consecutive years. There were approximately 450,000 specimens and 59 identified species collected during the surveillance. Ten species were newly recorded for Turkey: C. chiopterus, C. grisescens, C. paradoxalis, C. santonicus, C. poperinghensis, C. sergenti, C. tbilisicus, C. comosioculatus, C. haranti, and C. univittatus. Identification of C. chiopterus and C. grisescens was confirmed using species-specific PCR and DNA sequencing. With our recent findings, previous data were critically reviewed and updated, and the number of Culicoides species has been increased to 71 for Turkey. The presence of C. chiopterus has particular importance due to its potential vector status for bluetongue virus (BTV) and Schmallenberg virus (SBV). This study presents result of the first large-scale integrated faunistic survey on Culicoides species in Turkey.

First Parasitological Data on a Wild Grey Wolf in Turkey with Morphological and Molecular Confirmation of the Parasites.

INTRODUCTION: The grey wolf (Canis lupus) is the natural host of many parasites. These animals travel quite long distances to search for prey and nests, causing parasites to spread over large areas; therefore, determination of the parasites carried by grey wolves is important. METHODS: In this study, we used both morphological and molecular methods for parasitological identification of helminth species. For this purpose, the material obtained after necropsy was examined by macroscopic, microscopic, and molecular (multiplex PCR and DNA sequencing) methods. RESULTS: No pathological lesions and parasites were detected in the macroscopic examination of the trachea, lungs, heart, liver, spleen, stomach, and kidneys. The parasites collected from the intestines and diaphragm muscles were identified as Taenia hydatigena, Mesocestoides litteratus and Trichinella britovi. CONCLUSION: The aim of this study was to determine the helminth species in a dead grey wolf from wildlife. To the best of our knowledge, with this study, Taenia hydatigena, Mesocestoides litteratus and Trichinella britovi were detected for the first time in a grey wolf in Turkey.