RNA viromes from terrestrial sites across China expand environmental viral diversity.

Environmental RNA viruses are ubiquitous and diverse, and probably have important ecological and biogeochemical impacts. Understanding the global diversity of RNA viruses is limited by sampling biases, dependence on cell culture and PCR for virus discovery, and a focus on viruses pathogenic to humans or economically important animals and plants. To address this knowledge gap, we generated metatranscriptomic sequence data from 32 diverse environments in 16 provinces and regions of China. We identified 6,624 putatively novel virus operational taxonomic units from soil, sediment and faecal samples, greatly expanding known diversity of the RNA virosphere. These newly identified viruses included positive-sense, negative-sense and double-strand RNA viruses from at least 62 families. Sediments and animal faeces were rich sources of viruses. Virome compositions were affected by local environmental factors, including organic content and eukaryote species abundance. Notably, environmental factors had a greater impact on the abundance and diversity of plant, fungal and bacterial viruses than of animal viromes. Our data confirm that RNA viruses are an integral part of both terrestrial and aquatic ecosystems.

Diverse and abundant resistome in terrestrial and aquatic vertebrates revealed by transcriptional analysis.

Despite increasing evidence that antibiotic resistant pathogens are shared among humans and animals, the diversity, abundance and patterns of spread of antibiotic resistance genes (ARGs) in wildlife remains unclear. We identified 194 ARGs associated with phenotypic resistance to 13 types of antibiotic in meta-transcriptomic data generated from a broad range of lower vertebrates residing in both terrestrial and aquatic habitats. These ARGs, confirmed by PCR, included those that shared high sequence similarity to clinical isolates of public health concern. Notably, the lower vertebrate resistome varied by ecological niche of the host sampled. The resistomes in marine fish shared high similarity and were characterized by very high abundance, distinct from that observed in other habitats. An assessment of ARG mobility found that ARGs in marine fish were frequently co-localized with mobile elements, indicating that they were likely spread by horizontal gene transfer. Together, these data reveal the remarkable diversity and transcriptional levels of ARGs in lower vertebrates, and suggest that these wildlife species might play an important role in the global spread of ARGs.

Newly identified viral genomes in pangolins with fatal disease.

Epizootic pathogens pose a major threat to many wildlife species, particularly in the context of rapidly changing environments. Pangolins (order Pholidota) are highly threatened mammals, in large part due to the trade in illegal wildlife. During July to August 2018 four sick wild pangolins (three Manis javanica and one Manis pentadactyla) exhibiting a variety of clinical symptoms were rescued by the Jinhua Wildlife Protection Station in Zhejiang province, China. Although three of these animals died, fortunately one recovered after 2 weeks of symptomatic treatment. Using meta-transcriptomics combined with reverse transcription polymerase chain reaction (RT-PCR), we identified two novel RNA viruses in two of the dead pangolins. Genomic analysis revealed that these viruses were most closely related to pestiviruses and coltiviruses, although still highly genetically distinct, with more than 48 and 25 per cent sequence divergence at the amino acid level, respectively. We named these Dongyang pangolin virus (DYPV) and Lishui pangolin virus (LSPV) based on the sampling site and hosts. Although coltiviruses (LSPV) are known to be transmitted by ticks, we found no evidence of LSPV in ticks sampled close to where the pangolins were collected. In addition, although DYPV was present in nymph ticks (Amblyomma javanense) collected from a diseased pangolin, they were not found in the local tick population. Epidemiological investigation revealed that both novel viruses might have been imported following the illegal international trade of pangolins. Hence, these data indicate that illegal wildlife trafficking not only threatens the status of pangolin populations, but may also spread epizootic pathogens.

[Distribution patterns and pollution assessments of heavy metals in the Spartina alterniflora salt-marsh wetland of Rudong, Jiangsu province].

To understand the ecological impact of Spartina alterniflora on the coastal wetland environment, field survey was carried out in July, 2010, over the intertidal areas of Rudong coast, Jiangsu province; sediment samples were collected from a series of stations with different conditions of vegetation cover and S. alterniflora growth. The contents of eight heavy metals, together with sediment composition and total organic carbon were analyzed to reveal the distribution patterns of the heavy metals. Environmental quality status was evaluated using both the index of geoaccumulation (I(geo)) and the index of the Håkanson ecological risk. The analytical results showed that the average contents of Pb, Cd, As, Hg, Cr, Cu, Ni and Zn were below the standard for the Category I sediment quality, among which Cd, Hg, Ni and Zn exceeded the sediment background value of the region. On the whole, the contents of eight heavy metals in vegetation areas were higher than those associated with the adjacent bare flat areas. These data sets indicate a non-polluted condition in term of I(geo) estimation; however, a critical state of low to moderate degrees of pollution and a low level of risk were deduced according to the index of the Håkanson potential ecological risk. Both indices suggested that the pollution level of Hg and Cd were the highest among the eight metals measured. Along the transection from seaward to landward, the contents of As, Cu and Hg, their indices of I(geo) and Håkanson ecological risk all showed an increasing tread, in accordance with the condition of vegetation cover. Along the coastline with S. alterniflora being distributed in patchiness, all metal contents and their ecological risk level values for the marshes were higher than those for the unvegetated sediments nearby; moreover, except for Hg, other seven metals exhibited relatively low values than those in the shore-normal section with a better S. alterniflora growth. These findings indicate that S. alterniflora is one of important factors to enrich the heavy metal in tidal flat sediment. Thus, ecological risk of the heavy metal is reduced or blocked, due to the filtering effect of salt-marsh, which prevents metals from entering the open sea directly. The distribution of heavy metal is influenced by a combination of colonization time of vegetation, chemical form of metals and their origins.

Effects of HIV-1 tat on secretion of TNF-α and IL-1ß by U87 cells in AIDS patients with or without AIDS dementia complex.

OBJECTIVE: To explore the role of HIV-1 tat gene variations in AIDS dementia complex (ADC) pathogenesis. METHODS: HIV-1 tat genes derived from peripheral spleen and central basal ganglia of an AIDS patient with ADC and an AIDS patient without ADC were cloned for sequence analysis. HIV-1 tat gene sequence alignment was performed by using CLUSTAL W and the phylogentic analysis was conducted by using Neighbor-joining with MEGA4 software. All tat genes were used to construct recombinant retroviral expressing vector MSCV-IRES-GFP/tat. The MSCV-IRES-GFP/tat was cotransfected into 293T cells with pCMV-VSV-G and pUMVC vectors to assemble the recombinant retrovirus. After infection of gliomas U87 cells with equal amount of the recombinant retrovirus, TNF-α, and IL-1ß concentrations in the supernatant of U87 cells were determined with ELISA. RESULTS: HIV-1 tat genes derived from peripheral spleen and central basal ganglia of the AIDS patient with ADC and the other one without ADC exhibited genetic variations. Tat variations and amino acid mutation sites existed mainly at Tat protein core functional area (38-47aa). All Tat proteins could induce U87 cells to produce TNF-α and IL-1ß, but the level of IL-1ß production was different among Tat proteins derived from the ADC patient's spleen, basal ganglia, and the non-ADC patient's spleen. The level of Tat proteins derived from the ADC patient's spleen, basal ganglia, and the non-ADC patient's spleen were obviously higher than that from the non-ADC patient's basal ganglia. CONCLUSION: Tat protein core functional area (38-47aa) may serve as the key area of enhancing the secretion of IL-1ß. This may be related with the neurotoxicity of HIV-1 Tat.

[Distribution patterns of heavy metals in surficial sediment and their influence on the environment quality of the intertidal flat of Luoyuan Bay, Fujian coast].

Intertidal flats represent a typical environmentally fragile and sensitive zone. In order to investigate the environmental quality of the intertidal zone in Luoyuan Bay, field survey was carried out in 2009. Contents of heavy metals (Co, Cr, Cu, Ni, Pb, V and Zn) in the surficial sediment were measured using an inductively coupled plasma optical emission spectrometer (ICP-OES). Moreover, the impact on the environment quality was evaluated with the potential ecological risk method. The average contents of the heavy metals Co, Cr, Cu, Ni, Pb, V and Zn in the surficial sediment were 20.48, 77.82, 23.24, 40.67, 36.25, 134.75 and 111.21 mg x kg(-1), respectively. The heavy metal contents in the Spartina alterniflora salt-marsh were apparently higher than those in the bare flat. Further, the heavy metal concentrations found in the present study were generally higher than the background values of the coastal regions of Fujian Province, but lower than those associated with the Pearl River estuary. According to principal component and correlation analyses, industrial wastewater, mineral exploration and degradation of organic matter were the main sources of heavy metals in the area investigated. The results of potential ecological risk evaluation indicated that the intertidal zone as a whole can be ranked as "moderate potential ecological risk". Ni and Co were the major pollutants among the metals in consideration; the pollution related to Pb was less significant. The level of potential ecological risk of the Spartina alterniflora slat-marsh was higher than that of the bare flat. The sequence of potential ecological risk for the heavy metals was Ni > Co > Cu > Pb > Cr > V > Zn.

[Determination of trace chromium(VI) by catalytic spectrofluorimetry using pyronine Y-H2O2-Cr(VI) system].

A simple and highly sensitive new kinetic catalytic fluorimetric method was proposed for the determination of trace chromium (VI), based on the catalytic effect of trace amounts of chromium (VI) on the oxidation of Pyronine Y by hydrogen peroxide in acetic acid-sodium acetate buffer medium leading to a decrease in the fluorescence intensity. The optimum conditions and kinetic properties of the catalytic reaction were also studied. The apparent activation energy and the apparent rate constant are 159. 92 k * x mol(-1) and 5. 7X10-2 s-1 respectively. The linear range of the calibration curve is 0. 02-0. 24 microg x mL(-1) and the detection limit is 0. 012 microg x mL(-1). The present method was applied to the determination of chromium(VI) in river water, and industrial and electroplating waste water with good results.

[Headspace solid-phase microextraction-gas chromatography-mass spectrometry for analysis of volatile components from pollen pini].

Headspace solid-phase microextraction HS-SPME) technique was employed to extract the volatile compounds from Pollen Pini. The volatile components were isolated and identified successfully by gas chromatography-mass spectrometry (GC-MS). The volatiles were collected using several SPME parameters (including extraction time and temperature). The best response was obtained using a PA fiber by when the extraction temperature was 70 degrees C, extraction time was 30 minutes and desorption time was 10 time. Analysis was performed by GC-MS. Seventy components were identified by HS-SPME. The main components were fatty compounds. The results showed that the HS-SPME technique could be used to extract the volatile components from HS-SPME-GC-MS.