Spotted Fever Group Rickettsia Infecting Ticks (Acari: Ixodidae), Yak (Bos grunniens), and Tibetan Sheep (Ovis aries) in the Qinghai-Tibetan Plateau Area, China.

The Qinghai-Tibet Plateau Area (QTPA) has a complex natural ecosystem, causing a greatly increased risk of spreading various tick-borne diseases including rickettsial infections, which are regarded as one of the oldest known vector-borne zoonoses. However, the information of one of its pathogen, spotted fever group Rickettsia (SFG Rickettsia), is limited in tick vectors and animals in this area. Therefore, this study focused on the investigation of SFG Rickettsia in tick vectors, yaks (Bos grunniens), and Tibetan sheep (Ovis aries) in the QTPA. A total of 1,000 samples were collected from nine sampling sites, including 425 of yaks, 309 of Tibetan sheep, 266 of ticks. By morphological examination, PCR, and sequencing, we confirmed the species of all collected ticks. All tick samples, all yak and Tibetan sheep blood samples were detected based on SFG Rickettsia ompA and sca4 gene. The results showed that all tick samples were identified to be Haemaphysalis qinghaiensis, and the positive rates of SFG Rickettsia were 5.9% (25/425), 0.3% (1/309), and 54.1% (144/266) in yaks, Tibetan sheep, and ticks, respectively. All positive samples were sequenced, and BLASTn analysis of the ompA gene sequences of SFG Rickettsia showed that all positive samples from animals and ticks had 99.04-100% identity with yak and horse isolates from Qinghai Province, China. BLASTn analysis of the sca4 gene sequences of SFG Rickettsia showed that all positive samples had 97.60-98.72% identity with tick isolates from Ukraine. In addition, the phylogenetic analysis showed that all the SFG Rickettsia ompA and sca4 sequences obtained from this study belong to the same clade as Rickettsia raoultii isolated from livestock and ticks from China and other countries. Molecularly, this study detected and characterized SFG Rickettsia both in the tick vectors and animals, suggesting that the relationship between SFG Rickettsia, tick species and animal hosts should be explored to understand their interrelationships, which provide a theoretical basis for preventing control of this pathogen.

The mitochondrial transfer RNAAsp A7551G mutation may contribute to the clinical expression of deafness associated with the A1555G mutation in a pedigree with hearing impairment.

The role of mitochondrial (mt)DNA variations in hearing loss have been studied extensively; in particular, the well­known pathogenic A1555G mutation in the human mitochondrial 12S ribosomal RNA gene is associated with aminoglycoside­induced and non­syndromic hearing loss. The present paper described a Chinese pedigree with hearing impairments. We first performed polymerase chain reaction and direct sequence analysis for the mtDNA genes. Additionally, the GJB2 gene mutations were also genotyped. Notably, this family had a very high penetrance of deafness (66.7 and 33.3%; including and excluding aminoglycoside use, respectively). Sequence analysis of the mtDNA genes from the matrilineal relatives identified the occurrence of A1555G mutation, as well as the tRNAAsp A7551G mutation. The A7551G mutation occurred at position 37 in the anticodon stem of tRNAAsp, which is extremely conserved among various species. The nucleotide at this position is often chemically modified and thus contributes to the maintenance of functional tRNAAsp, therefore, this mutation may cause an imbalance in the level of tRNAAsp and lead to mitochondrial dysfunction which is involved in the pathogenesis of hearing loss. Taken together, the findings of the present study demonstrated that the A7551G mutation may have contributed to the deafness phenotype caused by the A1555G mutation.

Effect of microwave chlorine depleted pyrolyzate on the combustion characteristics of refuse derived fuel derived from package waste.

Thermogravimetric-Fourier Transform Infrared Spectroscopy was conducted to evaluate the combustion characteristics of refuse derived fuel (RDF) adding of microwave chlorine depleted pyrolyzate in the mass proportion of 5-15%. It studied the catalyze effect of chlorine depleted pyrolyzate on RDF combustion performance. The combustion process of RDF could be divided into four stages. The temperature range of the most significant combustion stage of 10-RDF was much more extensive than another three ones. According to the FTIR analysis, the addition of chlorine depleted pyrolyzate might promote the combustion of CH4 and carbonyls to CO2 and H2O earlier. Based on the distributed activation energy model (DAEM), the E value of RDF with chlorine depleted pyrolyzate added was much lower than that with no chlorine depleted pyrolyzate added. The chlorine depleted pyrolyzate enhanced the combustion performance of RDF with the lower ignition, lower burnout temperature, better combustion ability, better flammability and more outstanding combustion performance. The best combustion characteristic was obtained when the dosage of chlorine depleted pyrolyzate was 10%.

[Ecological compensation standard in Dongting Lake region of returning cropland to lake based on emergy analysis].

The annual emergy and currency value of the main ecological service value of returning cropland to lake in Dongting Lake region from 1999 to 2010 was calculated based on emergy analysis. The calculation method of ecological compensation standard was established by calculating annual total emergy of ecological service function increment since the starting year of returning cropland to lake, and the annual ecological compensation standard and compensation area were analyzed from 1999 to 2010. The results indicated that ecological compensation standard from 1999 to 2010 was 40.31-86.48 yuan x m(-2) with the mean of 57.33 yuan x m(-2). The ecological compensation standard presented an increase trend year by year due to the effect of eco-recovery of returning cropland to lake. The ecological compensation standard in the research area presented a swift and steady growth trend after 2005 mainly due to the intensive economy development of Hunan Province, suggesting the value of natural ecological resources would increase along with the development of society and economy. Appling the emergy analysis to research the ecological compensation standard could reveal the dynamics of annual ecological compensation standard, solve the abutment problem of matter flow, energy flow and economic flow, and overcome the subjective and arbitrary of environment economic methods. The empirical research of ecological compensation standard in Dongting Lake region showed that the emergy analysis was feasible and advanced.

Molecular cloning and expression analysis of inhibitor of growth protein 3 (ING3) in the Manila clam, Ruditapes philippinarum.

Inhibitor of growth protein 3 (ING3), a new member of ING family, is involved in the regulation of various processes. In this study, a full-length cDNA of ING3 (named as RpING3) was cloned from the gill of Ruditapes philippinarum by rapid amplification of cDNA ends method for the first time. The cDNA obtained was 1442 bp exclusive of poly (A) residues with a 1248 bp open reading frame encoding 415 amino acids. The RpING3 protein has a calculated molecular weight of 46.59 kDa and isoelectric point of 6.62. Two conserved motif and some functional sites were found. Tissue distribution analysis of the RpING3 mRNA revealed that the RpING3 expression level was much higher in gill and digestive gland while lower in mantle, foot and adductor muscle. The temporal expression of RpING3 in digestive gland after lead exposure was recorded by quantitative real-time PCR. The result showed that RpING3 was rapidly up-regulated at 6 h post-exposure and reached tenfold of the control group. These results suggest that RpING3 dependent signaling pathway is present in Manila clam and RpING3 may play important roles in protecting cells from heavy metal damage in R. philippinarum.

Isolation and genetic characterization of hantaviruses carried by Microtus voles in China.

To gain more insights into hantavirus distribution in China, Microtus fortis were caught in Jilin province and M. maximowiczii in the Inner Mongolia Autonomous Region. Hantavirus specific RNA was detected by RT-PCR in 3 out of 26 M. fortis and 5 out of 64 M. maximowiczii. Two hantaviruses (Fusong-Mf-682 and Yakeshi-Mm-59) were isolated successfully in cell culture and their S and M segment nucleotide sequences were determined. Phylogenetic analysis of the S and M segment sequences revealed that the Mf-originated strains from Fusong were closely related to Vladivostok hantavirus (VLAV) with 99% nucleotide identity, but differed from the Yakeshi-Mm strains, with an amino acid divergence of more than 8.8% for the N protein and 11.8% for the GnGc proteins. Yakeshi-Mm strains were closely related to the Khabarovsk hantavirus (KHAV) isolated earlier from M. fortis in Khabarovsk, with an amino acid sequence identity of more than 98.4% for the S segment and 95.6% for the M segment. On phylogenetic trees, Yakeshi-Mm strains clustered together with KHAV and Topografov virus (TOPV) carried by Lemmus sibiricus. The results suggest that the hantavirus carried by M. fortis in China belongs to VLAV type and should be considered as a distinct hantavirus species. They also suggest that M. fortis is the natural host of VLAV (including Fusong-Mf strains), whereas M. maximowiczii is the natural host of KHAV including Yakeshi-Mm strains. Thus, in addition to Hantaan, Seoul, Dabieshan and Puumala-like Hokkaido viruses, at least two other hantaviruses, namely KHAV and VLAV, are circulating in China.

Detection of phylogenetically distinct Puumala-like viruses from red-grey vole Clethrionomys rufocanus in China.

In order to investigate whether Puumala virus (PUUV) or PUUV-like virus is present in China, Clethrionomys rufocanus and C. rutilus were captured in the Jilin province during the spring and autumn of 2002-2003 for detection of PUUV viral RNA by RT-PCR and confirmation of PUUV-positive antigens by an immunofluorescence assay. PUUV-positive RNA was identified in six out of 121 C. rufocanus but not in any of the 41 C. rutilus. Complete S and partial M sequences (nt 1,316-1,598 and 2,687-3,089) were amplified by RT-PCR directly from some of the antigen positive lung tissues and subjected to nucleic acid sequencing. It was found that the Chinese PUUV-like viruses were related most closely with the PUUV strains with 77.7-81.7% identity at the nucleotide level and 91.7-97% identity at the amino acid level for S segment, and with 77-78.8% identity at the nucleotide level and 91.5-92.6% identity at the amino acid level for the partial M segment (nt 1,316-1,598). Genetic analysis indicated that the Chinese PUUV-like viruses shared the highest level of identity with the viruses which circulate in C. rufocanus in the Far East region of Russia with 85.1-87.4% identity at the nucleotide level and 95.9% identity at the amino acid level for the partial M segment (nt 2,687-3,089), respectively. Phylogenetic analysis revealed that the Chinese PUUV-like viruses are distinct from those identified from Japan, South Korea, Europe or Russia. These results indicate that PUUV-like virus is present in China in addition to Hantaan, Seoul and Dabieshan viruses.

Isolation and characterization of hantavirus carried by Apodemus peninsulae in Jilin, China.

To provide a better understanding of hantavirus epidemiology in China, Korean field mice (Apodemus peninsulae) and striped field mice (Apodemus agrarius) were captured in Jilin province, China, where haemorrhagic fever with renal syndrome (HFRS) is endemic. Hantavirus antigens were detected in eight of the 130 A. peninsulae individuals and in four of the 193 A. agrarius individuals by using an immunofluorescence assay. Partial S and M segments were amplified from all of the antigen-positive samples. Furthermore, two hantaviruses (CJAp89 and CJAp93) were isolated successfully in cell culture and the entire S and M segments were amplified from one of them (CJAp93). Phylogenetic analysis of these sequences (partial or complete) showed that hantaviruses carried by A. peninsulae and A. agrarius form two distinct lineages, although viruses carried by A. peninsulae are similar to those isolated previously from A. agrarius in China and from HFRS patients in Russia. However, the viruses detected in A. peninsulae in China are genetically different from those detected in A. peninsulae in other countries. These data suggest that A. peninsulae is also a natural host for HTNV in north-eastern China.

[Identification of Puumala like viruses in China].

BACKGROUND: To confirm if Puumala like viruses exist in China. METHODS: RNA was extracted from lungs of bank voles captured in the Northeast China, partial S segments genome of Puumala viruses were amplified and sequenced. RESULTS: 926 bp cDNA of S segments of Puumala like virus was amplified and sequenced. The phylogenetic analysis revealed that the Puumala like viruses found in China were most close to that found in Far East region of Russia. CONCLUSIONS: Puumala like virus does exist in Northeast China, and the nucleotides sequence of the viruses have high homolog to Puumala viruses found in Russia.