Echinococcus granulosus Sensu Stricto and Echinococcus multilocularis in a Gray Wolf (Canis lupus) in Turkey: Further Evidence for Increased Risk of Alveolar Echinococcosis in Urban Areas.

OBJECTIVE: The aim of this study was to identify Echinococcus species by morphological and molecular means. METHODS: A dead gray wolf (Canis lupus) was found near Erzurum province and brought to the parasitology laboratory. Sedimentation and counting technique (SCT) and polymerase chain reaction (PCR) analysis were conducted. RESULTS: The SCT implications indicated that the wolf had a substantial worm burden (62,720 and 49,280 parasites) due to a co-infection of E. granulosus s.l. and E. multilocularis. Genus/species-specific PCR was used to analyze DNA extracted from adult worms and confirmed as E. granulosus s.s. and E. multilocularis, utilizing COI and 12S rRNA gene sequence analysis, respectively. CONCLUSION: This report presents the first co-detection of E. granulosus s.s. and E. multilocularis in a gray wolf found in an urban area in a highly endemic area for human echinococcosis in northeastern Turkey. The results emphasize that AE is not only a problem of rural areas, but also occurs in urban areas, which may pose a threat to public health. Therefore, surveillance in urban areas is crucial. The need to develop new control strategies for domestic and wildlife in the study area is also highlighted.

Molecular detection of Babesia and Theileria species/genotypes in sheep and ixodid ticks in Erzurum, Northeastern Turkey: First report of Babesia canis in sheep.

Piroplasmosis is a tick-borne protozoan disease caused by Babesia and Theileria species in ruminants. This study sought to determine the presence and prevalence of the agents causing piroplasmosis among sheep in Erzurum province, Turkey. It also sought to identify the tick species infesting the sheep and investigate the possible role of the ticks in the transmission of piroplasmosis. A total of 1621 blood samples and 1696 ixodid ticks from infested sheep were collected. Each blood sample and 115 tick pools were subjected to PCR assay. A total of 307 blood samples were found to be positive for Babesia spp. and Theileria spp. according to molecular analysis. The sequence analysis was revealed the presence of B. ovis (0.4%), B. crassa (0.4%), B. canis (0.4%), T. ovis (69.3%), Theileria sp. (26.6%), and Theileria sp. OT3 (2.9%) in 244 samples. The collected ticks were identified as D. marginatus (62.5%), Hae. parva (36.2%), Hae. punctata (1.1%), Rh. turanicus (0.1%), and H. marginatum (0.1%). The molecular analysis of the adult tick samples revealed T. ovis and T. annulata positivity in the D. marginatus pools, B. crassa and T. ovis positivity in the Hae. parva pools, and T. ovis positivity in the Hae. punctata pools. These results provide up-to-date data concerning tick-borne protozoan diseases of sheep and tick species infesting sheep in the region. The sheep breeding industry is an important livelihood for the region so it is essential to perform repeated studies on these pathogens in order to prevent disruptions to animal husbandry.

Cystic echinococcosis in slaughtered sheep in Erzurum province, Turkey.

Cystic echinococcosis (CE) is an important zoonotic infection caused by the larval stages of the genus Echinococcus. Turkey is a highly endemic region for CE and the disease is one of the major public health problems. The study was aimed to assess the situation of the CE in sheep in Turkey and also to provide data on circulating genotypes in the country. A total of 3319 sheep at slaughter were screened during the study. The prevalence of CE in the study area was 31.7% (1052/3319). The lungs were the most frequently CE infected organ (50%, 526/1052). Microscopic examination revealed that overall cyst fertility was 68.1%. Molecular analysis of partial fragments of 12S and COI gene regions were included for 351 selected cyst samples and all of them were identified as E. granulosus sensu stricto. Sequence analysis showed that the predominant genotype in the study areas was G1 (77.1%), and the rest were G3 (22.9%). The prevalence rate of CE in sheep in the study area is lower compared to previous years except for one province. Considering the high cyst fertility rate and the predominance of E. granulosus G1 which is particularly pathogenic to humans, calls for serious control measures like public awareness about the disease, sufficient dog deworming programs, continuity of monitoring the disease should be taken.

Zoonotic Babesia microti infection in wild rodents in Erzurum province, northeastern Turkey.

Wild rodents are natural reservoir hosts of various pathogens, including Babesia microti. This study investigated the presence of B. microti in rodents from Erzurum province in Turkey. A total of 498 rodents and 21 rodent-fed ticks were analysed using the polymerase chain reaction (PCR) technique to test for the presence of B. microti. Babesia spp. were detected in three (0.6%) of the 498 rodent spleen samples. The Babesia-positive rodent species were identified as Microtus socialis by means of molecular analysis. The rodent-fed ticks comprised 15 Ixodes laguri and 6 Rhipicephalus sanguineus, none of which tested positive for Babesia spp. A sequence analysis of the 18S PCR amplicons confirmed the three Babesia-positive samples to be B. microti. The Erzurum isolates were 100% identical to the zoonotic Jena strain. The results of this study indicate the existence of zoonotic B. microti strains that may constitute a potential public health risk in Erzurum province. Future studies should determine the tick vector and other reservoir rodent species of B. microti in Erzurum.

A Strange Gingival Orifice in an Infant: A Case of Oral Myiasis.

Myiasis is a parasitic infection of humans and animals caused by dipterous larvae and is often seen in rural areas. For the first time in the literature as a case of gingival myiasis in an 18-month-old healthy child with good oral hygiene is reported. This case presents the clinical and histopathologic features of oral myiasis. The patient had a swelling in the upper lip for three days and a perforation in the gingiva. After mechanical removal of larvae intraoral symptoms resolved within six days. The larva was identified as Wohlfahrtia magnifica according to the histopathologic evaluation. Dentists working in rural areas should be aware of this very rare condition and its treatment.

The situation of echinococcosis in stray dogs in Turkey: the first finding of Echinococcus multilocularis and Echinococcus ortleppi.

Echinococcosis, caused by larval stage of the genus Echinococcus, is one of the most important zoonotic diseases worldwide. The purpose of this study was to determine the presence and prevalence of Echinococcus species in stray dogs of Erzurum, a highly endemic region for cystic echinococcosis (CE) and alveolar echinococcosis (AE) in Turkey. The study samples consisted of 446 stray dog faecal specimens collected from an animal shelter in Erzurum, Turkey, between October 2015 and February 2016. The faecal samples were collected from individual dogs for the isolation of taeniid eggs using the sequential sieving and flotation method (SSFM). Molecular analyses and sequencing revealed the prevalence of Echinococcus spp. as 14.13% (63/446) in faecal samples. The stray dogs harboured five different Echinococcus spp.: E. granulosus s.s. (G1/G3) (n = 41), E. equinus (G4) (n = 3), E. ortleppi (G5) (n = 1), E. canadensis (G6/G7) (n = 3) and E. multilocularis (n = 16). E. granulosus s.s. was the most abundant species. Surprisingly, the occurrence of E. multilocularis in dogs was revealed for the first time in Turkey. E. ortleppi was also reported for the first time in Turkey. These findings highlight a significant public health risk for human AE and CE, presenting useful baseline data on Echinococcus spp. infection in dogs for designing control strategies.

Echinococcus multilocularis in Red Foxes in Turkey: Increasing risk in urban.

This study was conducted to determine the occurrence of E. multilocularis in foxes and environmental fecal contamination by E. multilocularis in Erzurum, the most highly endemic region for AE in Turkey. The study materials consisted of 50 red fox carcasses collected from 20 counties of Erzurum, Turkey, between October 2015 and February 2016. After the application of the sedimentation and counting technique (SCT), E. multilocularis was identified through the identification of typical morphological structures. Fox fecal samples (n = 600) were also collected from these counties for the isolation of taeniid eggs using the sequential sieving and flotation method (SSFM). Then, the collected adult worms and taeniid eggs were subjected to molecular and sequence analyses. Mature E. multilocularis parasites were found in 42% (21/50) of the fox intestines, with a mean number of 7,806 (150-31,644). The severity of infection was higher in carcasses obtained from the central district (48.6%, 17/35) than in those obtained from the peripheral district (26.7%, 4/15). The prevalence of environmental fecal contamination with E. multilocularis was 10.5% (63/600) in fecal samples collected from all counties of Erzurum. This infection rate was higher in the central district (32.1%, 36/112) than in the peripheral district (5.5%, 27/488; P < 0.0001). In conclusion, contrary to expectation, the prevalence of E. multilocularis positivity was high in urban areas. This could have been due to alterations in the dietary habitats of definitive and intermediate hosts. Therefore, new control strategies are essential to eliminate human AE cases in the future as urbanization advances.

Morphological and molecular data of Hepatozoon ursi in two brown bears (Ursus arctos) in Turkey.

Species of Hepatozoon Miller, 1908 are vector-borne parasites that infect domestic and wild animals worldwide. Hepatozoon ursi Kubo, Uni, Agatsuma, Nagataki, Panciera et al., 2008 was reported from bears (Ursidae) in Japan and India. The present study represents the first report of infection with H. ursi in Turkish brown bears (Ursus arctos Linnaeus) by microscopic and molecular analysis. Two dead brown bears were found in Uzundere and Pasinler districts of Erzurum. Blood and visceral organ (spleen and liver) samples were delivered to laboratory by the Nature Conservation and National Parks officers. Detected gamonts were evaluated based on morphological features and confirmed as gamonts of H. ursi. The size of gamonts and parasitemia were 8.2 × 3.5 µm (6.9-8.7 × 3.0-3.9 µm; n = 12) and 0.6% (6/1000 leukocytes), respectively. The blood and visceral organ samples were positive for species of Hepatozoon by PCR targeting partial sequence of 18S rDNA. Sequence analysis of newly obtained sequences of H. ursi showed 98.8-100% identity with previously sequenced isolates of H. ursi. Sequences of H. ursi from Erzurum were identical to each other and showed 100% identity with isolates of H. ursi from ticks Ixodes ricinus (Linnaeus), Rhipicephalus turanicus Pomerantzev and Hyalomma marginatum Koch collected from two brown bears in Turkey (GenBank accession numbers MN463021, MN463022, MN905023). Analysis of partial sequences of the 18S rRNA gene of H. ursi showed that Turkish isolates differ in NT substitutions found at three different positions [72 (A→G), 537 (A→G) and 570 (A→T)]. This study provides morphological and molecular data of H. ursi infection in brown bears from two districts of Erzurum, Turkey. Further studies are needed to elucidate whether brown bears have any eco-epidemiologic importance in the life cycle of H. ursi in wildlife.

Molecular Characterization of Echinococcus multilocularis and Echinococcus granulosus from Cysts and Formalin-Fixed Paraffin-Embedded Tissue Samples of Human Isolates in Northeastern Turkey.

Erzurum province of Turkey is known to be highly endemic for alveolar echinococcosis (AE) and cystic echinococcosis (CE). In this study, we confirmed Echinococcus multilocularis cases, searched genetic variations of the isolates, and-for the first time-determined the genotypes of Echinococcus granulosus s.l. infecting humans in the province. A total of 5 alveolar and 106 hydatid cysts as well as 23 formalin-fixed paraffin-embedded (FFPE) samples that were diagnosed as AE were collected from hospitals between 2015 and 2017. Partial sequences of two mitochondrial genes were amplified to detect E. multilocularis and E. granulosus sensu lato with conventional polymerase chain reactions (PCRs) and genotypes confirmed by sequencing. PCR amplification of a partial 12S rRNA gene on an alveolar cyst and FFPE tissue samples yielded the expected bp in 5 cysts and 19 of 23 FFPE samples; all Erzurum E. multilocularis isolates were confirmed by sequencing. Phylogenetic analysis of the isolates indicated that some of them were identical to European isolates, whereas some of them were identical to Asian isolates. Off all hydatid cyst samples, 101 (95.2%) yielded the expected bp (94 with 12S rRNA-PCR and 7 with COI-PCR). Sequence analysis showed that 98 (97%) of them corresponded to the G1 genotype, whereas 3 (3%) corresponded to the G3 genotype. Results of the study emphasize that E. multilocularis isolates of Erzurum, based on short sequencing, are similar to both European and Asian isolates, and the G1 genotype of E. granulosus is the main causative agent of human CE in Erzurum.

Endoparasites Determined by Fecal Examination in Sheep in Erzurum Province

Objective: The aim of the current study was to determine the presence and prevalence of Eimeria and helminth species in sheep raised in Erzurum province by using fecal examination. Methods: Faecal samples were collected from a total of 784 sheep raised in Aziziye, Yakutiye and Palandöken districts between February-March 2019. The samples were examined by Fulleborn's flotation, Benedect sedimentation, and Baermann-Wetzel methods. Results: Eimeria spp. and helminths were found in 49.36% (387/784) and 74.11% (581/784) of the samples, respectively. Identified Eimeria species were as follows: E. parva (59.68%), E. ovina (51.67%), E. faurei (47.80%), E. ahsata (39.27%), E. granulosa (36.62%), E. punctata (28.42%), E. pallida (26.09%), E. ovinoidalis (18.34%), E. crandallis (16.79%), E. intricata (15.76%), E. weybridgensis (11.36%) and E. marsica (6.20%). Helminth species identified at genus/species level were Dicrocoelium spp. (33.91%), Fasciola spp. (5.68%), Paramphistomum spp. (2.58%), Moniezia spp. (5.85%), Trichostrongylid type egg (49.05%), Marshallagia spp. (38.73%), Nematodirus spp. (20.98%), Trichuris spp. (14.46%), Protostrongylus spp. (18.42%), Dictyocaulus filaria (2.41%) and Muellerius capillaris (1.38%). Conclusion: Parasitic diseases cause important economic losses in livestock industry. In following years, it is aimed to plan prevention and control strategies for the parasites detected in this area in line with the data of this study and to share this data with the animal breeders.

Echinococcus multilocularis in a Eurasian lynx (Lynx lynx) in Turkey.

Echinococcus multilocularis is the causative agent of alveolar echinococcosis (AE), one of the most threatening zoonoses in Eurasia. Human AE is widespread in the Erzurum region of Turkey, but the situation of the disease in intermediate and definitive hosts is unknown. A Eurasian lynx (Lynx lynx) was killed in a traffic accident in the north of Erzurum, and was taken to our laboratory. Sedimentation and counting technique (SCT), DNA isolation and polymerase chain reaction (PCR) analysis were performed. The SCT results showed that the lynx was infected with E. multilocularis with a medium (745 worms) worm burden. The DNA of adult worms obtained from the lynx was analyzed with a species-specific PCR, and the worms were confirmed to be E. multilocularis by 12S rRNA gene sequence analysis. This is the first report of E. multilocularis from Eurasian lynx in Turkey.

First detection of Echinococcus multilocularis in rodent intermediate hosts in Turkey.

Echinococcus multilocularis is the causative agent of alveolar echinococcosis (AE), a potentially fatal zoonotic disease. Large parts of Turkey are considered as endemic for E. multilocularis. The aim of this study was to determine the occurrence of metacestode of E. multilocularis in wild rodents in Erzurum, an endemic region for human AE in Turkey. During the sampling period, a total of 498 rodents were trapped in twenty counties of Erzurum Province. Suspected lesions were observed on the livers of 48 rodents, and then partial fragment of mitochondrial 12S rRNA gene was PCR-amplified. Five liver samples exhibited E. multilocularis infection. The prevalence of E. multilocularis for Microtus spp. was 1·3%. All of the infected rodents had fertile metacestodes. Infected rodents were morphologically and molecularly analysed and were confirmed to be Microtus irani by the mitochondrial cytochrome b gene sequence analysis. This is the first report of the presence of E. multilocularis in rodent intermediate hosts in Turkey. Our findings of infected M. irani with protoscoleces show that this rodent can act as suitable intermediate host for E. multilocularis' life cycle in Turkey. However, there was a complete lack of data on the infection of carnivores from the country. An extensive survey is recommended to determine the prevalence of E. multilocularis in definitive hosts in this endemic region.

First Molecular Characterization of Echinococcus multilocularis in Turkey.

This study aimed to find out the occurrence of Echinococcus multilocularis in foxes in Erzurum province, the highest endemic region for human alveolar echinococcosis in Turkey. The sedimentation and counting technique was used to reveal adult Echinococcus spp. in the intestines of foxes. One out of the 10 foxes was infected with E. multilocularis. The adult worms were analyzed morphologically and molecularly and were confirmed to be E. multilocularis by species-specific PCR. Pairwise comparisons between the 12S rRNA sequences of the E. multilocularis isolate from Erzurum and other E. multilocularis isolates showed 100% similarity of the Erzurum isolate with European isolates. With this study, the presence of E. multilocularis in a fox in Erzurum was confirmed by PCR, and molecular identification of E. multilocularis is reported for the first time in Turkey.