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.

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.

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.

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.

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.