Genomic characterization of a novel virus of the family tymoviridae isolated from mosquitoes

Background

The family Tymoviridae comprises three plant virus genera, including Tymovirus, Marafivirus, and Maculavirus, which are found in most parts of the world and cause severe agricultural losses. We describe a putatively novel member of the family Tymoviridae, which is isolated from mosquitoes (Culex spp.), referred to as CuTLV.

Methods and Results

The CuTLV was isolated by cell culture, which replicates and causes cytopathic effects in Aedes albopictus C6/36 cells, but not in mammalian BHK-21 or Vero cells. The complete 6471 nucleotide sequence of CuTLV was determined. The genome of CuTLV is predicted to contain three open reading frames (ORFs). The largest ORF1 is 5307 nucleotides (nt) in length and encodes a putative polypeptide of 1769 amino acids (aa), which contains the conserved motifs for the methyltransferase (MTR), Tymovirus endopeptidase (PRO), helicase (HEL), and RNA-dependent RNA polymerase (RdRp) of the replication-associated proteins (RPs) of positive-stranded RNA viruses. In contrast, the ORF1 sequence does not contain the so-called “tymobox” or “marafibox”, the conserved subgenomic RNA promoter present in all tymoviruses or marafiviruses, respectively. ORF2 and ORF3 putatively encode a 248-aa coat protein (CP) and a proline-rich 149-aa polypeptide. The whole genomic nucleotide identity of CuTLV with other members of family Tymoviridae ranged from 46.2% (ChiYMV) to 52.4% (GFkV). Phylogenetic analysis based on the putative RP and CP genes of CuTLV demonstrated that the virus is most closely related to viruses in the genus Maculavirus.

Conclusions

The CuTLV is a novel virus related to members of the family Tymoviridae, with molecular characters that are distinct from those of tymoviruses, marafiviruses, and other maculaviruses or macula-like viruses. This is the first report of the isolation of a Tymoviridae-like virus from mosquitoes. Further investigations are required to clarify the origin, replication strategy, and the public health or agricultural importance of the CuTLV.     

Claviradulomyces, a new genus of Odontotremataceae from West African rainforest

A new genus, Claviradulomyces, morphologically typical of Odontotremataceae, is proposed to accommodate the new species Claviradulomyces dabeicola. The genus is characterised by its ornamented periphysoids, and long-cylindric, sigmoid, acuminate ascospores. Genetically, C. dabeicola falls within the Ostropales, but the Odontotremataceae have been very poorly sampled genetically and their phylogenetic relationship within the order is unclear. C. dabeicola has been collected consistently and exclusively on the rainforest tree Erythroxylum mannii in the Côte d'Ivoire and Ghana, where it is associated with bark abnormalities on branches and stems of living trees.     

New species of Microsporidia in the genus Amblyospora from blood-sucking mosquitoes in the family Culicidae

Two microsporidians of the genus Amblyospora from mosquito larvae of Aedes and Culex were studied in Tomsk region (Siberia). Peculiarities of the sporogony and the meiospore ultrastructure were studied by light and electron microscopy. The meiospores of A. caspius sp. n. from Aedes caspius and Ae. communis measured on smears, were c. 4.8 x 3.6 microns in size. A polar filament was anisofilar with 5 wide (290 nm) and 9 narrow (190 nm) coils arranged in a single layer. A polaroplast of both species was composed of two parts: anterior lamellae and posterior fibrillae. Meiospores of A. burlaki sp. n. from Culex pipiens measured c. 4.8 x 3.5 microns. A polar filament was anisofilar with 3 wide (260 nm) and 6 narrow (110 nm) coils. The meiospore of this species had a 260 nm thick multilayered exospore.     

ZIKA virus isolated from mosquitoes: a field and laboratory investigation in China, 2016

A field investigation of arboviruses was conducted in Dejiang, Guizhou Province in the summer of 2016. A total of 8,795 mosquitoes, belonging to four species of three genera, and 1,300 midges were collected. The mosquito samples were identified on site according to their morphology, and the pooled samples were ground and centrifuged in the laboratory. The supernatant was incubated with mosquito tissue culture cells (C6/36) and mammalian cells (BHK-21) for virus isolation. The results indicated that 40% (3,540/8,795) were Anopheles sinensis, 30% (2,700/8,795) were Culex pipiens quinquefasciatus, and 29% (2,530/8,795) were Armigeres subbalbeatus. Furthermore, a total of eight virus isolates were obtained, and genome sequencing revealed two Zika viruses (ZIKVs) isolated from Culex pipiens quinquefasciatus and Armigeres subbalbeatus, respectively; three Japanese encephalitis viruses (JEVs) isolated from Culex pipiens quinquefasciatus; two Banna viruses (BAVs) isolated from Culex pipiens quinquefasciatus and Anopheles sinensis, respectively; and one densovirus (DNV) isolated from Culex pipiens quinquefasciatus. The ZIKVs isolated from the Culex pipiens quinquefasciatus and Armigeres subbalbeatus mosquitoes represent the first ZIKV isolates in mainland China. This discovery presents new challenges for the prevention and control of ZIKV in China, and prompts international cooperation on this global issue.     

Genome sequence of a novel member of the genus Psychrobacter isolated from Antarctic soil

Psychrobacter spp. have shown characteristics indicating remarkable capabilities at subzero temperatures that identify them as potential model organisms for the study of low-temperature adaptations. Here we present the draft genome sequence of Psychrobacter sp. PAMC 21119, which was isolated from permafrost soil of Antarctica; this information could provide insight into adaptation and evolution strategies under extreme environmental conditions.     

Manitoba virus, a new rhabdovirus isolated from Culex tarsalis mosquitoes collected in Manitoba, Canada

A rhabdovirus, Mn 936-77, was isolated from a pool of two Culex tarsalis collected on August 16, 1977, from Morris, Manitoba. Isolate Mn 936-77 was not pathogenic for suckling Swiss white mice inoculated by the intracerebral route. The virus propagated in three vertebrate cell lines (Vero, primary chick embryo, mouse neuroblastoma), but apparently not in Aedes albopictus C6/36 cells. Isolate Mn 936-77 did not react by amplified enzyme-linked immunosorbant assay with 230 viruses of proven or possible arbovirus etiology or by immunofluorescence with 88 members of the family Rhabdoviridae. Isolate Mn 936-77 appears to be a newly discovered virus for which the name Manitoba virus is proposed.     

Genome sequencing and phylogenetic analysis of Banna virus(genus Seadornavirus,family Reoviridae) isolated from Culicoides

In an investigation of blood-sucking insects and arboviruses, a virus(YN12243) was isolated from Culicoides samples collected in the Sino-Burmese border region of Yunnan Province, China. The virus caused cytopathic effect(CPE) in C6/36 cells and passaged stably. Polyacrylamide gel analysis showed that the genome of YN12243 was composed of 12 segments of double-stranded RNA(dsRNA), with a distribution pattern of 6-6. The nucleotide and amino acid sequences of the coding region(1.12 segments) were17,803 bp and 5,925 amino acids in length, respectively. The phylogenetic analysis of VP1 protein(RdRp) revealed that YN12243 belonged to genus Seadornavirus of family Reoviridae, and further analysis indicated that YN12243 belongs to the Banna virus(BAV) genotype A2. Additionally, YN12243 was located in the same evolutionary cluster as BAV strains isolated from different mosquito species, suggesting that the BAV isolated from Culicoides does not have species barriers. These results indicate that Culicoides can also be a vector for BAV. In view of the hematophagous habits of Culicoides on cattle, horses, deer, and other large animals, as well as the possibility of spreading and causing a variety of animal arboviral diseases, it is important to improve infection detection and monitor the BAV in large livestock.     

Zika virus: a new threat from mosquitoes

正In the winter of 2015,the first dengue vaccine(Dengvaxia)was approved for clinical use.Dengue virus is the most prevalent mosquito-transmitted viral pathogen in humans(Qin and Shi,2014).Even though the approved Dengvaxia has its weaknesses(e.g.,low protection against serotype-2virus),it has been recently licensed to Mexico,Philippines,     

Molecular evidence for trypanosomatids in Culex mosquitoes collected during a West Nile virus survey

Adult mosquitoes were previously collected and tested for West Nile virus during an intense WNV outbreak in 2003-2004 along the Cache la Poudre River in Colorado, USA. A subset of these mosquitoes was also tested for infection with trypanosomatids using nested PCR to amplify 18S rRNA. Of the 69 pools of Culex pipiens that were screened for both pathogens, 4.3% were positive for WNV and 11.6% tested positive for trypanosomes; no pools were found to be co-infected with both pathogens. One hundred and forty-three pools of Culex tarsalis, considered to be the principal WNV vector in this area, were tested in the same manner. 7.7% were positive for WNV and 20.3% of these pools tested positive for trypanosomes. Five pools of C. tarsalis were found to be co-infected with both pathogens, which was approximately 2.2 times more frequent than would be expected if these pathogens are independent of each other. Sequencing and maximum parsimony analysis of 18S rRNA revealed that four of the isolates arise in or near clades of described avian trypanosomes, likely indicating that these are vectored pathogens between birds and mosquitoes. Unexpectedly, the majority (24/28, 86%) of our positive samples form their own separate clade within the order Trypanosomatida with 100% bootstrap support. We have identified a potential new clade of trypanosomatids that exist within important mosquito vectors and discuss the potential ecological connections between these trypanosomes, arboviruses and mosquitoes.     

Density, survival and dispersal of Anopheles gambiae complex mosquitoes in a West African Sudan savanna village

Abstract. To obtain information on adult populations of Afrotropical malaria vector mosquitoes, mark-release-recapture experiments were performed with Anopheles females collected from indoor resting-sites in a savanna area near Ouagadougou, Burkina Faso, during September 1991 and 1992. Results were used to estimate the absolute population densities, daily survival rates, and dispersal parameters of malaria vectors in that area.
In 1991 a total of 7260 female Anopheles were marked and released, of which 106 were recaptured in the release village and 6 in the neighbouring villages, a total recapture rate of 1.5%. The following year 13, 854 female Anopheles were released and 116 recaptured in Goundri and 8 in the neighbouring villages, a total recapture rate of 0.9%. Recaptures were found in three of eight villages near Goundri. Nearly all of the recaptured mosquitoes were An.gambiae s.l. Of these, molecular determination revealed that An.gambiae s.s. and An.arabiensis were present in a ratio of -2:3.
Two simple random models of dispersal were simulated and the parameters of the models determined by searching for the least-squared fit between simulated and observed distributions. The mean distance moved by individual mosquitoes, estimated in this way, ranged 350–650 m day^-1, depending on die model and the year considered. Population densities were estimated using the Lincoln Index, Fisher-Ford and Jolly's methods. The estimates of population size had high standard errors and were not particularly consistent. A 'consensus' value of 150,000–350,000 mosquitoes is believed to apply for ht An.gambiae s.l. female population. Survival was estimated to be 80–88% per day.     

Microsporidia of the genus Parahtelohania (Microspora: Amblyosporidae) from blood-sucking mosquitoes of the genus Anopeles (Diptera: Culicidae) from the South of the West Siberia

Five new microsporidian species of the genus Parathelohania have been found in the fat body of blood-sucking mosquitoes collected in various water basins in the South of West Siberia: Parathelohania divulgata sp. n., P. formosa sp. n., P. sibirika sp. n., P. teguldeti sp. n., and P. tomski sp. n. Processes of merogony, sporogony, and ultrastructure of spores was were investigated by means light and electronic microscopy.     

Rubrobacter bracarensis sp. nov., a novel member of the genus Rubrobacter isolated from a biodeteriorated monument

Three actinobacteria strains isolated from a green biofilm covering the biodeteriorated interior walls of Vilar de Frades Church (Portugal) were studied using a polyphasic approach. The three strains were aerobic, non-spore forming and Gram-positive. Phylogenetically, the most closely related described species was Rubrobacter radiotolerans (94.2-94.3% and 81.9-82.5% similarities for the 16S rRNA and rpoB gene sequences, respectively). The fatty acid profile was dominated by anteiso-C(17:1) ω9c, and MK-8 was the only menaquinone present. These data clearly showed that the three strains could represent a new species, for which we propose the name Rubrobacter bracarensis sp. nov., with strain VF70612_S1(T) (=CECT 7924=DSMZ 24908) as the type strain.     

Complete genome sequence of a novel picorna-like virus isolated from Spodoptera exigua

The complete genome sequence and the gene organization of a novel insect picorna-like virus, Spodoptera exigua virus (SeV), were determined. The genomic RNA of the SeV was 9501 nt in length excluding the poly(A) tail and contained a single, large open reading frame (nt 392–9424) encoding a 3010 aa polyprotein. Sequence comparisons with other viral polyproteins revealed that the consensus sequences for picornavirus RNA helicase, cysteine protease, and RNA-dependent RNA polymerase (RdRp) proteins are found on the genome in that order from the 5′ to the 3′ end. In terms of sequence similarity, identity, and genome organization, SeV resembled insect picorna-like viruses belonging to the genus Iflavirus. A phylogenetic analysis based on the eight conserved domains in the RdRp sequence showed that SeV was most closely related to the Perina nuda virus and Ectropis obliqua picorna-like virus, suggesting that these three insect picorna-like viruses might share a common ancestor.     

Transmission dynamics of West Nile virus in mosquitoes and corvids and non-corvids

There are more than 300 avian species that can transmit West Nile virus (WNv). In general, the corvid and non-corvid families of birds have different responses to the virus, with corvids suffering a higher disease-induced mortality rate. By taking both corvids and non-corvids as the primary reservoir hosts and mosquitoes as vectors; we formulate and study a system of ordinary differential equations to model a single season of the transmission dynamics of WNv in the mosquito–bird cycle. We calculate the basic reproduction number and analyze the existence and stability of the equilibria. The existence of a backward bifurcation gives a further sub-threshold condition beyond the basic reproduction number for the spread of the virus. We also discuss the role of corvids and non-corvids in spreading the virus. We conclude that knowledge of the relative abundance of corvid bird species and other mammals assist us in accurate estimation of the epidemic of WNv.     

Dietzia natronolimnaios sp. nov., a new member of the genus Dietzia isolated from an East African soda lake

Two novel alkaliphilic aerobic organotrophic bacteria have been isolated from a moderately saline and alkaline East African soda lake. The new isolates grow at pH values between 6 and 10, with a pH optimum for growth of 9.0, and at a salt concentration between 0% and 10% (w/v). Phylogenetic analysis based on 16S rDNA sequence shows that these isolates are very closely related (99.6% similarity) and are members of the monospecific genus Dietzia (98.8% and 98.7% similarity). DNA/DNA hybridization revealed a relatedness of 83% between the two isolates, but only 8% between them and the type strain Dietzia maris. The G + C content as measured by thermal denaturation is 66.1 mol%. Phenotypic comparisons between D. maris and one isolate showed that they share very similar morphological and chemotaxonomic properties, but differ significantly in carbon source utilization profiles and halotolerance in alkaline medium. We propose a second species of this genus which we name Dietzia natronolimnaios (type strain 15LN1 = CBS 107.95). Received: October 14, 1997 / Accepted: February 26, 1998