Merida virus, a putative novel rhabdovirus discovered in Culex and Ochlerotatus spp. mosquitoes in the Yucatan Peninsula of Mexico

Sequences corresponding to a putative, novel rhabdovirus [designated Merida virus (MERDV)] were initially detected in a pool of Culex quinquefasciatus collected in the Yucatan Peninsula of Mexico. The entire genome was sequenced, revealing 11 798 nt and five major ORFs, which encode the nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G) and RNA-dependent RNA polymerase (L). The deduced amino acid sequences of the N, G and L proteins have no more than 24, 38 and 43 % identity, respectively, to the corresponding sequences of all other known rhabdoviruses, whereas those of the P and M proteins have no significant identity with any sequences in GenBank and their identity is only suggested based on their genome position. Using specific reverse transcription-PCR assays established from the genome sequence, 27 571 C. quinquefasciatus which had been sorted in 728 pools were screened to assess the prevalence of MERDV in nature and 25 pools were found positive. The minimal infection rate (calculated as the number of positive mosquito pools per 1000 mosquitoes tested) was 0.9, and similar for both females and males. Screening another 140 pools of 5484 mosquitoes belonging to four other genera identified positive pools of Ochlerotatus spp. mosquitoes, indicating that the host range is not restricted to C. quinquefasciatus. Attempts to isolate MERDV in C6/36 and Vero cells were unsuccessful. In summary, we provide evidence that a previously undescribed rhabdovirus occurs in mosquitoes in Mexico.The authors thank Valeria Bussetti for expert technical assistance. This study was supported by the National Institutes of Health (awards 5R21AI067281, AI057158, 5R21AI067281 and AI088647), the United States Department of Defense and an intramural grant from Iowa State University. AEF is supported by a grant from the Wellcome Trust (award 106207).This is the final version of the article. It first appeared from the Microbiology Society via http://dx.doi.org/10.1099/jgv.0.00042

Merida virus, a putative novel rhabdovirus discovered in Culex and Ochlerotatus spp. mosquitoes in the Yucatan Peninsula of Mexico.

Sequences corresponding to a putative, novel rhabdovirus [designated Merida virus (MERDV)] were initially detected in a pool of Culex quinquefasciatus collected in the Yucatan Peninsula of Mexico. The entire genome was sequenced, revealing 11 798 nt and five major ORFs, which encode the nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G) and RNA-dependent RNA polymerase (L). The deduced amino acid sequences of the N, G and L proteins have no more than 24, 38 and 43 % identity, respectively, to the corresponding sequences of all other known rhabdoviruses, whereas those of the P and M proteins have no significant identity with any sequences in GenBank and their identity is only suggested based on their genome position. Using specific reverse transcription-PCR assays established from the genome sequence, 27 571 C. quinquefasciatus which had been sorted in 728 pools were screened to assess the prevalence of MERDV in nature and 25 pools were found positive. The minimal infection rate (calculated as the number of positive mosquito pools per 1000 mosquitoes tested) was 0.9, and similar for both females and males. Screening another 140 pools of 5484 mosquitoes belonging to four other genera identified positive pools of Ochlerotatus spp. mosquitoes, indicating that the host range is not restricted to C. quinquefasciatus. Attempts to isolate MERDV in C6/36 and Vero cells were unsuccessful. In summary, we provide evidence that a previously undescribed rhabdovirus occurs in mosquitoes in Mexico.The authors thank Valeria Bussetti for expert technical assistance. This study was supported by the National Institutes of Health (awards 5R21AI067281, AI057158, 5R21AI067281 and AI088647), the United States Department of Defense and an intramural grant from Iowa State University. AEF is supported by a grant from the Wellcome Trust (award 106207).This is the final version of the article. It first appeared from the Microbiology Society via http://dx.doi.org/10.1099/jgv.0.00042

Orthobunyavirus Antibodies in Humans, Yucatan Peninsula, Mexico

We performed a serologic investigation to determine whether orthobunyaviruses commonly infect humans in the Yucatan Peninsula of Mexico. Orthobunyavirus-specific antibodies were detected by plaque reduction neutralization test in 146 (18%) of 823 persons tested. Further studies are needed to determine health risks for humans from this potentially deadly group of viruses

Complete genome sequence of T’Ho virus, a novel putative flavivirus from the Yucatan Peninsula of Mexico

Abstract Background We previously reported the discovery of a novel, putative flavivirus designated T’Ho virus in Culex quinquefasciatus mosquitoes in the Yucatan Peninsula of Mexico. A 1358-nt region of the NS5 gene was amplified and sequenced but an isolate was not recovered. Results The complete genome of T’Ho virus was sequenced using a combination of unbiased high-throughput sequencing, 5′ and 3′ rapid amplification of cDNA ends, reverse transcription-polymerase chain reaction and Sanger sequencing. The genome contains a single open reading frame of 10,284 nt which is flanked by 5′ and 3′ untranslated regions of 97 and 556-nt, respectively. Genome sequence alignments revealed that T’Ho virus is most closely related to Rocio virus (67.4% nucleotide identity) and Ilheus virus (65.9%), both of which belong to the Ntaya group, followed by other Ntaya group viruses (58.8–63.3%) and Japanese encephalitis group viruses (62.0–63.7%). Phylogenetic inference is in agreement with these findings. Conclusions This study furthers our understanding of flavivirus genetics, phylogeny and diagnostics. Because the two closest known relatives of T’Ho virus are human pathogens, T’Ho virus could be an unrecognized cause of human disease. It is therefore important that future studies investigate the public health significance of this virus

Gonotrophic cycle estimate for Culex quinquefasciatus in Mérida, Yucatán, México

West Nile virus (WNV) has been present in the Yucatán State, México, since 2002. Culex quinquefasciatus, one of the main vectors of WNV transmission in the United States, is also common in Mexico and may be a key vector of WNV transmission to humans in the Yucatán. The aim of this study was to determine the length of the gonotrophic cycle and the survival rates of Cx. quinquefasciatus from Mérida, Yucatán, during the rainy versus the dry season. Mosquitoes were collected during 25-day periods in October (rainy season) and in April (dry season), and captured females were classified by abdominal appearance (freshly fed, late-stage fed, half gravid, and subgravid). To determine the age structure as nulliparous and parous females and to calculate the gonotrophic cycle through a time series and the mosquito survival, we used Davidson formulae. Also, vitellogenesis analysis to monitor egg maturity was conducted during both seasons. Cross-correlation data suggested a similar length of the gonotrophic cycle (4 days) in both seasons. Oogenic development required a minimum of 72 h in each season. However, survival of the mosquito population collected in the rainy season was significantly higher (0.91) with a mean temperature of 28 ± 1.57°C than was survival in the dry season (0.78) with a mean temperature of 29 ± 1.10°C. Survival, although higher during the rainy season, did not influence the length of the gonotrophic cycle of Cx. quinquefasciatus in Yucatán

Detection of Flaviviruses and Orthobunyaviruses in Mosquitoes in the Yucatan Peninsula of Mexico in 2008

A total of 191,244 mosquitoes from 24 species were collected in the Yucatan Peninsula of Mexico from January to December 2008, and tested for the presence of cytopathic virus by virus isolation in Vero cells. Eighteen virus isolates were obtained, all of which were orthobunyaviruses. These were identified by reverse transcription-polymerase chain reaction (RT-PCR) and nucleotide sequencing as Cache Valley virus (n = 17) and South River virus (n = 1). A subset (n = 20,124) of Culex quinquefasciatus collected throughout the year was further tested by RT-PCR using flavivirus-specific primers. Flavivirus RNA was present in this mosquito species year-round. The overall flavivirus minimal infection rate, expressed as the number of positive mosquito pools per 1000 mosquitoes tested, was 7.7 and the monthly flavivirus minimal infection rates ranged from 4.3 to 16.6. Approximately one-third of the RT-PCR products were sequenced and all corresponded to Culex flavivirus, a recently discovered insect-specific flavivirus