Fuqiang Zhang

Virome Profiling of Bats from Myanmar by Metagenomic Analysis of Tissue Samples Reveals More Novel Mammalian Viruses

Bats are reservoir animals harboring many important pathogenic viruses and with the capability of transmitting these to humans and other animals. To establish an effective surveillance to monitor transboundary spread of bat viruses between Myanmar and China, complete organs from the thorax and abdomen from 853 bats of six species from two Myanmar counties close to Yunnan province, China, were collected and tested for their virome through metagenomics by Solexa sequencing and bioinformatic analysis. In total, 3,742,314 reads of 114 bases were generated, and over 86% were assembled into 1,649,512 contigs with an average length of 114 bp, of which 26,698 (2%) contigs were recognizable viral sequences belonging to 24 viral families. Of the viral contigs 45% (12,086/26,698) were related to vertebrate viruses, 28% (7,443/26,698) to insect viruses, 27% (7,074/26,698) to phages and 95 contigs to plant viruses. The metagenomic results were confirmed by PCR of selected viruses in all bat samples followed by phylogenetic analysis, which has led to the discovery of some novel bat viruses of the genera Mamastrovirus, Bocavirus, Circovirus, Iflavirus and Orthohepadnavirus and to their prevalence rates in two bat species. In conclusion, the present study aims to present the bat virome in Myanmar, and the results obtained further expand the spectrum of viruses harbored by bats.

Canine distemper outbreak in rhesus monkeys, China.

Since 2006, canine distemper outbreaks have occurred in rhesus monkeys at a breeding farm in Guangxi, People’s Republic of China. Approximately 10,000 animals were infected (25%–60% disease incidence); 5%–30% of infected animals died. The epidemic was controlled by vaccination. Amino acid sequence analysis of the virus indicated a unique strain.

Hepatitis virus in long-fingered bats, Myanmar.

During an analysis of the virome of bats from Myanmar, a large number of reads were annotated to orthohepadnaviruses. We present the full genome sequence and a morphological analysis of an orthohepadnavirus circulating in bats. This virus is substantially different from currently known members of the genus Orthohepadnavirus and represents a new species.

Characterization of a novel orthoreovirus isolated from fruit bat, China

BackgroundIn recent years novel human respiratory disease agents have been described for Southeast Asia and Australia. The causative pathogens were classified as pteropine orthoreoviruses with a strong phylogenetic relationship to orthoreoviruses of bat origin.ResultsIn this report, we isolated a novel Melaka-like reovirus (named “Cangyuan virus”) from intestinal content samples of one fruit bat residing in China’s Yunnan province. Phylogenetic analysis of the whole Cangyuan virus genome sequences of segments L, M and S demonstrated the genetic diversity of the Cangyuan virus. In contrast to the L and M segments, the phylogenetic trees for the S segments of Cangyuan virus demonstrated a greater degree of heterogeneity.ConclusionsPhylogenetic analysis indicated that the Cangyuan virus was a novel orthoreovirus and substantially different from currently known members of Pteropine orthoreovirus (PRV) species group.

Prevalence and genetic characterization of Toxoplasma gondii in bats in Myanmar.

ABSTRACT We detected Toxoplasma gondii in 29.3% (95% confidence interval [CI], 25.5% to 33.1%) of 550 insectivorous bats collected in Myanmar. The genotyping of these positive samples revealed they were closely related to or belong to clonal type I, which is highly virulent in mice, showing that these bats are potential reservoirs for T. gondii transmission.

Emergence of novel clade 2.3.4 influenza A (H5N1) virus subgroups in Yunnan Province, China.

From December 2013 to March 2014, a major wave of highly pathogenic avian influenza outbreak occurred in poultry in Yunnan Province, China. We isolated and characterized eight highly pathogenic avian influenza A (H5N1) viruses from poultry. Full genome influenza sequences and analyses have been performed. Sequence analyses revealed that they belonged to clade 2.3.4 but did not fit within the three defined subclades. The isolated viruses were provisional subclade 2.3.4.4e. The provisional subclade 2.3.4.4e viruses with six internal genes from avian influenza A (H5N2) viruses in 2013 were the novel reassortant influenza A (H5N1) viruses which were associated with the outbreak of H5N1 occurred in egg chicken farms in Yunnan Province. The HA genes were similar to subtype H5 viruses isolated from January to March of 2014 in Asia including H5N6 and H5N8. The NA genes were most closely related to A/chicken/Vietnam/NCVD-KA423/2013 (H5N1) from the subclade 2.3.2. The HI assay demonstrated a lack of antigenic relatedness between clades 2.3.4.4e and 2.3.4.1 (RE-5 vaccine strain) or 2.3.2.2 (RE-6 vaccine strain).

Molecular Characterization of Cryptosporidium in Bats from Yunnan Province, Southwestern China

Abstract: The present study was conducted to investigate the prevalence and genotypes of Cryptosporidium in bats. A total of 247 bats, belonging to Rhinolophus sinicus, Rousettus leschenaultia, Aselliscus stoliczkanus, and Hipposideros fulvus, were collected in Yunnan Province, Southwestern China, and the intestinal tissues were examined for Cryptosporidium infection by PCR amplification of the small subunit ribosomal RNA (SSU rRNA). The overall infection rate was 7.7% (95% CI, 4.5 to 11.0%), with R. sinicus having the highest level at 9.5% (95% CI, 2.8 to 16.1%) followed by A. stoliczkanus at 7.8% (95% CI, 2.2 to 18.9%), H. fulvus at 7.2% (95% CI, 1.1 to 13.4%), and R. leschenaultia at 5.7% (95% CI, 1.2 to 15.7%). DNA sequence and phylogenetic analyses based on SSU rRNA revealed the presence of 2 novel genotypes, designated as Cryptosporidium bat genotype I in A. stoliczkanus and R. sinicus and Cryptosporidium bat genotype II in R. leschenaultia, R. sinicus, and H. fulvus. This is the first report of Cryptosporidium genotypes in bats. Further biological and genetic characterization is needed to determine the relationship of the 2 novel genotypes to established Cryptosporidium species–genotypes.

Identification of a novel orthohepadnavirus in pomona roundleaf bats in China

Bats in Myanmar, Gabon, and Panama have been found to harbor diverse hepadnaviruses. Here, we report a novel hepadnavirus in 4 of 20 pomona roundleaf bats from Yunnan province, China. This virus contains 3,278 nucleotides (nt) in the full circularized genome, with four predicted open frames (ORFs) reading in the same direction. Full genomic sequence comparisons and evolutionary analysis indicate that this virus is a member of a new species within the genus Orthohepadnavirus

Epidemiological and molecular characteristics of emergent dengue virus in Yunnan Province near the China-Myanmar-Laos border, 2013-2015.

BackgroundYunnan Province is located in southwestern China and neighbors the Southeast Asian countries, all of which are dengue-endemic areas. In 2000–2013, sporadic imported cases of dengue fever (DF) were reported almost annually in Yunnan Province. During 2013–2015, we confirmed that a large-scale indigenous DF outbreak emerged in cities of Yunnan Province near the China-Myanmar-Laos border.MethodsEpidemiological characteristics of DF in Yunnan Province during 2013–2015 were evaluated by retrospective analysis. A total of 232 dengue virus (DENV)-positive sera were randomly collected for sequence analysis of the capsid/premembrane region of DENV from patients with DF in Yunnan Province. The envelope gene of DENV isolates was also amplified and sequenced. Phylogenetic analyses were performed using the neighbor-joining method with the Tajima-Nei model.ResultsPhylogenetically, all DENV-positive samples could be classified into DENV-1 genotype I and DENV-2 Asian I genotype during 2013–2015 and DENV-4 genotype I in 2015 from Ruili City; and DENV-3 genotype II in 2013 and DENV-2 Cosmopolitan genotype in 2015 from Xishuangbanna Prefecture.ConclusionsOur results indicated that imported DF from patients from Laos and Myanmar was the primary cause of the DF epidemic in Yunnan Province. Additionally, DENV strains of all four serotypes were identified in indigenous cases in Yunnan Province during the same time period, while the dengue epidemic pattern observed in southwestern Yunnan showed characteristics of a hypoendemic nature: circulation of DENV-1 and DENV-2 over consecutive years.

Larvicidal Activity of Essential Extract of Rosmarinus officinalis Against Culex quinquefasciatus

Constituents in rosemary (Rosmarinus officinalis) have been shown to have larvicidal activity against invertebrates. In order to explore the properties of crude extract of rosemary further, we studied the chemical composition and its activity against dichlorodiphenyltrichloroethane (DDT)-susceptible, DDT-resistant, and field strains of Culex quinquefasciatus larvae. The major components of R. officinalis were found to be eucalyptol and camphor, with relative percentages of 10.93% and 5.51%, respectively. Minor constituents included limonene, (+)-4-carene, isoborneol, 1-methyl-4-(1-methylethylidene)-cyclohexene, and pinene. The median lethal concentration (LC50) values of the essential oil of R. officinalis against DDT-susceptible, DDT-resistant, and field strains of larvae of Cx. quinquefasciatus were 30.6, 26.4, and 38.3 mg/liter, respectively. The single median lethal dose (LD50) in Kunming mice was 4752 mg/kg. Essential oils from R. officinalis may, therefore, provide an effective natural plant product for use in mosquito prevention and control.