Abstract
Purpose: In order to screen for invasive mosquito species and associated viruses, we performed a field survey for mosquitoes in the Black Sea region of Anatolia where Aedes aegypti and Aedes albopictus were previously recorded. Methods & Materials: Mosquitoes were collected from 31 sites in Artvin, Trabzon and Rize provinces during 2016-2017. The specimens were identified morphologically and pooled according to collection site and species. Selected specimens were processed for DNA barcoding via cytochrome oxidase I amplification and sequencing, for confirmation of the morphological species identification. Virus screening was carried out using polymerase chain reaction (PCR) assays targeting alpha and flaviviruses, as well as recently-described novel rhabdovirus Merida-like virus Turkey (MERDLVT), followed by sequencing for characterization. Results: A total of 756 mosquitoes that comprise Ae. albopictus (675, 89,2%), Ae. aegypti (61, 8,1%) and Culex pipiens sensu lato (20, 2,6%) were collected and grouped in 65 pools. No amplification was observed in any pool via generic alphavirus PCR. Generic flavivirus PCR was reactive in 7 and 8 pools collected in 2016 and 2017, respectively. Cell fusing agent virus (CFAV) sequences were characterized in 4 pools (6,1%) of Ae. albopictus (n = 2) and Ae. aegypti (n = 2), collected in 2016. Aedes flavivirus (AFEV) sequences were characterized in 6 pools (9,2%) of Ae. albopictus (n = 5) and and Ae. aegypti (n = 1), collected in 2016 (n = 3) and 2017 (n = 3). Sequences of West Nile virus (WNV) was detected in 5 pools (7,6%) of Ae. albopictus (n = 4) and and Cx. pipiens s.l. (n = 1), collected in 2017. In phylogenetic analyses, the WNV sequences clustered with local and global lineage 1 clade 1a strains. Moreover, partial L and N gene sequences of MERDLVT were identified in the Cx. pipiens s.l. pool, coinfected with WNV. Conclusion: This is the initial detection of WNV and MERDLVT in field-collected mosquitoes from the Black Sea region. Although no alphavirus sequence could be demonstrated, presence of Ae. albopictus and Ae. aegypti indicates ongoing risk for potential spread.
Highlights
No amplification was observed in any pool via generic alphavirus polymerase chain reaction (PCR)
Cell fusing agent virus (CFAV) sequences were characterized in 4 pools (6,1%) of Ae. albopictus (n = 2) and Ae. aegypti (n = 2), collected in 2016
Partial L and N gene sequences of Merida-like virus Turkey (MERDLVT) were identified in the Cx. pipiens s.l. pool, coinfected with West Nile virus (WNV)
Summary
A Hospital based Cross sectional descriptive study conducted in JSS tertiary care hospital, Mysuru. Study included 55 stool samples of children aged between 1 to 60 months who presented to Department of Pediatrics, JSS hospital, both as in and out patients with complaints of diarrhea and dysentery. All stool samples were inoculated on to selective and non-selective media with filtration using a 0.45 m membrane filters and incubated in microaerophilic conditions using the candle jar at temperatures 37 ◦C and 42 ◦C. The culture isolates were confirmed by standard phenotypic tests. A simplex polymerase chain reaction (PCR) with universal Campylobacter primers and primers specific for C.jejuni and C.coli, was performed on the DNA extracted from the stool filtrates
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