Mitochondrial genomes of four slug moths (Lepidoptera, Limacodidae): Genome description and phylogenetic implications.

The family Limacodidae belongs to the superfamily Zygaenoidea, which includes 1672 species commonly referred to as slug moths. Limacodidae larvae are major pests for many economically important plant species and can cause human dermatitis. At present, the structure of the mitochondrial genome (mitogenome), phylogenetic position, and adaptive evolution of slug moths are poorly understood. Herein, the mitogenomes of Parasa lepida, Phlossa conjuncta, Thosea sinensis, and Setora sinensis were sequenced and compared with other available mitogenome sequences to better characterize the mitogenomic diversity and evolution of this moth family. The mitogenomes of P. lepida, P. conjuncta, T. sinensis, and S. sinensis were confirmed to be circular in structure with lengths of 15,575 bp, 15,553 bp, 15,535 bp, and 15,529 bp, respectively. The Limacodidae mitogenomes exhibited similar nucleotide composition, codon usage, RNA structure, and control region patterns, indicating the conservation of the mitogenome in the family Limacodidae. A sliding window, Ka/Ks, and genetic distance analyses revealed that the atp8 and nad6 genes exhibited the highest levels of variability and the most rapid evolutionary rates among the 13 protein-coding genes (PCGs) encoded in these Limacodidae mitogenomes, suggesting that they may offer value as candidate DNA markers. The phylogenetic analysis recovered the overall relationship as Tortricoidea + (Sesiidae + (Zygaenoidea + (Cossoidea/+Choreutoidea + (others)))). Within Zygaenoidea, Limacodidae was recovered as monophyletic, and the phylogenetic relationships were recovered as (Phaudidae + Zyganidae) + Limacodidae in all six phylogenetic trees. The analysis indicated that P. lepida, P. conjuncta, T. sinensis, and S. sinensis are members of the Limacodidae.

A rapid identification method for common astigmatid species based on multiplex polymerase chain reaction.

Astigmatid mites are economically significant pests of stored products and sources of inhalant allergens causing allergic rhinitis and asthma worldwide. The morphological identification of astigmatid mites at the species level is often a difficult task due to their small size, phenotypic similarity and lack of diagnostic characters. We used multiplex polymerase chain reaction (PCR) to identify astigmatid mite species, which could complement the morphological data for the species-specific identification of mites. Internal ribosomal transcribed spacer (ITS) sequences (i.e., partial 18S, the full length of ITS1-5.8S-ITS2 and partial 28S) from eight astigmatid species (Acarus siro, Tyrophagus putrescentiae, Suidasia nesbitti, Dermatophagoides pteronyssinus, Dermatophagoides farinae, Lepidoglyphus destructor, Chortoglyphus arcuatus and Gohieria fuscus) were obtained by DNA extraction and then sequenced after PCR amplification. Specific primers were designed in the ITS2 region manually. Results revealed that an identification method for eight common astigmatid species was established based on multiplex PCR, which should be effective for the identification of other species of mites by redesigning species-specific primers in future experiments.

Mitochondrial analysis of oribatid mites provides insights into their atypical tRNA annotation, genome rearrangement and evolution.

BACKGROUND: The mitochondrial (mt) genomes of Sarcoptiformes mites typically contain 37 genes. Although the loss of genes is rare in Sarcoptiformes mite mitogenomes, two of the six previously reported oribatid mites (Acariforms: Sarcoptiformes) are reported to have lost parts of their tRNA genes. To confirm whether the tRNA genes were indeed lost and whether the loss is universal, we re-annotated the available oribatid mite sequences and sequenced the mitogenome of Oribatula sakamorii. METHODS: The mitogenome of O. sakamorii was sequenced using an Illumina HiSeq sequencer. The mt tRNA gene was annotated using multi-software combined with a manual annotation approach. Phylogenetic analyses were performed using the maximum likelihood and Bayesian inference methods with concatenated nucleotide and amino acid sequences. RESULTS: The mitogenomes of O. sakamorii contained 37 genes, including 22 tRNA genes. We identified all mt tRNA genes that were reported as "lost" in Steganacarus magnus and Paraleius leontonychus and revealed certain atypical tRNA annotation errors in oribatid mite sequences. Oribatid mite mitogenomes are characterized by low rates of genetic rearrangement, with six or seven gene blocks conserved between the mitogenome of all species and that of ancestral arthropods. Considering the relative order of the major genes (protein-coding genes and rRNAs), only one or two genes were rearranged with respect to their positions in the ancestral genome. We explored the phylogenetic relationships among the available oribatid mites, and the results confirmed the systematic position of Hermannia in the Crotonioidea superfamily. This was also supported by the synapomorphic gene-derived boundaries. CONCLUSIONS: The tRNA "lost" phenomenon is not universal in oribatid mites. Rather, highly atypical secondary structure of the inferred mt tRNA genes made them unidentifiable using a single type of tRNA search program. The use of multi-software combined with a manual annotation approach can improve the accuracy of tRNA gene annotation. In addition, we identified the precise systematic position of Hermannia and validated that Astigmata is nested in Oribatida.

De novo sequence of the mitochondrial genome of Tyrophagus putrescentiae (Acari: Sarcoptiformes) including 22 tRNA sequences and the largest non-coding region.

In this study, we de novo sequenced and analyzed the circular mitochondrial genome (mitogenome) of Tyrophagus putrescentiae. It was 14,156 bp long and contained a complete set of 37 genes, contrary to the initial published sequences; it included 22 tRNA sequences and the largest non-coding region. The mtDNA gene order of T. putrescentiae was found to be identical to that of Aleuroglyphus ovatus, Caloglyphus berlesei, and Rhizoglyphus robini (all Acaroidea). Most tRNAs of T. putrescentiae lack at least a D-arm or T-arm. Tyrophagus putrescentiae tRNAs also shared considerable structural and sequence similarity with the tRNAs of other reported Acaroidea species that have the full set of tRNAs. The largest non-coding region was located between trnF and trnS1, and it contained a microsatellite-like (AT)n sequence, short palindromic sequences, and several hairpin loops, as observed in other reported Acaroidea species (excepting Tyrophagus longior).

Association of Immune Factors with Drug-Resistant Tuberculosis: A Case-Control Study.

BACKGROUND Presently, studies of factors associated with drug-resistant tuberculosis (TB) focus on patients' socio-demographic characteristics and living habits, to the exclusion of biochemical indicators, especially immune factors. This study was carried out to determine whether immune factors are associated with drug-resistant TB. MATERIAL AND METHODS A total of 227 drug-resistant pulmonary TB patients and 225 drug-susceptible pulmonary TB patients were enrolled in this study. Information on socio-demographic characteristics and biochemical indicators were obtained through their clinical records. Non-conditional logistic regression was used to analyze the association of these indicators with drug-resistant TB. RESULTS There were significant differences in re-treatment, marital status, alanine aminotransferase (ALT), blood uric acid (BUA), carcino-embryonic antigen (CEA), T-spot, and CD3 and CD4 counts between the 2 groups. In multivariable analysis, re-treatment [Odds Ratio (OR)=5.290, 95% Confidence Interval [CI]=2.652-10.551); CD3 (OR=1.034, 95% CI=1.001-1.068); CD4 (OR=1.035, 95% CI =1.001-1.070) and IgM (OR=1.845, 95% CI=1.153-2.952) were associated with drug-resistant TB. CONCLUSIONS These results suggest the need for greater attention to re-treatment cases and immune function when treating drug-resistant TB.

[Composition and diversity of Acaroid mites' communities in indoor environments of kindergartens in Wuhu City].

OBJECTIVE: To understand the breeding situation of Acaroid mites in indoor environments of kindergartens in Wuhu City, so as to provide the evidence for its prevention and control. METHODS: From March to June and September to December in 2014, dust samples were collected from 15 kindergartens of 3 ranks every month. Acaroid mites in the samples were isolated, identified and counted. RESULTS: Totally 360 samples were selected, and 169 samples (46.94%) had mite infestation. Pyroglyphidae accounted for the most in the population, with the breeding rate of 45.00%. Totally 18 504 mites were found, and Dermatophagoides farina and Dermatophagoides pteronyssinus were the dominant species, with the composition ratios of 30.21% (5 590 mites) and 17.83% (3 300 mites), respectively. The third-ranked kindergartens were in the most serious situation concerning with 81.67% (98/120) of the breeding rate of Acaroid mites. The differences were statistically significant among the kindergartens with different ranks (F = 6.048, χ2 = 73.523, both P < 0.05). There were no significant differences about the Acaroid mite population among different grades in kindergartens (F = 0.132, χ2 = 2.377, both P > 0.05). CONCLUSION: Acaroid mites in the kindergartens in Wuhu City were in serious condition, and the population and species composition of Acaroid mite community is influenced by the human behavior and indoors environments.

[Investigation of Acaroid mites breeding in stored dry fruits].

OBJECTIVE: To study the species and density of Acaroid mites breeding in stored dry fruits. METHODS: The samples from the dried fruit stores and warehouses were collected, and the mites breeding in them were separated, then the slides with mites were prepared and observed by a light microscope for species identification and counting. The indexes such as the breeding density, species richness index, diversity index and evenness index were calculated. RESULTS: Totally 12 species of Acaroid mites belonging to 6 families and 10 genera were obtained from the total 49 samples. The dominant mite species were Carpoglyphus lactis, Tyrophagus putrescentiae, Acarus siro, and Caloglyphus berlesei. The breeding densities of mites in longans, filberts and plum candies were 79.78, 48.91, 35.73 mites/g, respectively, which were higher than those in other dry fruits. The seasonal variation experiment of mites found that the average breeding density of acaroid mites was higher in July and October, the richness index and diversity index reached the highest value in July, and the evenness index was higher in January and April. The observation of the growth and decline of Acaroid mites under the artificial condition found the number of Caloglyphus berlesei declined sharply and Tyrophagus putrescentiae first increased and then decreased. CONCLUSION: The pollution of Acaroid mites is serious in the stored dried fruits, for which the positive prevention and control measures to the mite breeding should be taken to reduce the harm.

[Zoonotic trematode species identified in domestic animals from Huainan area].

OBJECTIVE: To understand the zoonotic trematode species identified in domestic animals from Huainan area, Anhui Province, so as to supply the evidence for the prevention of zoonoses. METHODS: The livestock/definitive hosts were commercially available on category basis, and sacrificed for obtaining the parasitic samples via dissecting the viscera. The specimens were microscopically identified, with reference to the descriptions in the previous literature. RESULTS: By identification, sorting and classification, 41 species of internal trematodes were found so far in the domestic animals fed in Huainan areas, in which 23 species were zoonotic trematodes and 18 were internally parasitic in livestock, belonging to 4 orders, 12 families and 21 genera, and occurred in 9 definitive hosts including chicken, ducks, geese, pigs, cattle, buffaloes, sheep, goats and dogs, respectively. CONCLUSION: The findings suggest that the zoonotic trematodes belong to many kinds and prevalent in the domestic animals in Huainan areas, and such prevalence should call for high attention since it is urgent risk factors for zoonoses in this geographic area.

The complete mitochondrial genome of the brown leg mite, Aleuroglyphus ovatus (Acari: Sarcoptiformes): evaluation of largest non-coding region and unique tRNAs.

The circular mitochondrial genome (mitogenome) of Aleuroglyphus ovatus was sequenced. It was 14,328 bp long, and consisted of 37 coding genes including 13 protein-coding genes, two ribosomal RNA genes, and 22 transfer RNA genes. This is the first description of the complete mitogenome of a species in the Acaridae (Acari: Sarcoptiformes). The mtDNA gene order for A. ovatus is identical to those of Dermatophagoides farinae and D. pteronyssinus, but distinctly different from the mtDNA of other Acari. Most inferred tRNA genes of A. ovatus are extremely truncated (48-62 bp), lack stem-loops on either the T- or D-arm (except the trnK), and are unable to fold into the canonical tRNA cloverleaf structure. The largest non-coding region (378 bp) contained several conserved sequences involved in the regulation of mitogenome replication, including one core sequence (ACAT) associated with termination of the J-strand replication and several hypothetical stem-loop structures. The microsatellite-like (AT)n sequence in the largest non-coding region was observed in two other Astigmata species, but it has not been found in other Acari.

[Investigation on species and community ecology of cheyletoid mites breeding in the stored traditional Chinese medicinal materials].

OBJECTIVE: To investigate the species of Cheyletoid mites breeding status in the stored traditional Chinese medicinal materials and the relationship between its community and habitats. METHODS: A total of 60 samples of the traditional Chinese medicinal materials were collected from Huainan, Wuhu and Xuancheng in China. The mites were isolated with shakesieve shock, directicopy, Tullgren funnel, waternacopy and redricopy, and identified and counted under the light microscope. RESULTS: Cheyletoid mites were represented in 48 of the 60 samples, and the breeding rate accounted for as high as 80.0% (48/60). Totally, 6 species of Cheyletoid mites were identified, which belonged to Cheyletus, Acaropsis, Cheletomorpha, Eucheyletic and Pseudocheyles genera under the family of Cheyletidae. In the three investigated areas, the average breeding density of Cheyletoid mites was 109.75 heads/g, and average richness index was 1.54. The index of species diversity was 2.55 and the number of evenness was 0.95. CONCLUSION: This result entails positive prevention and control of the mite breeding in storage and processing of herbal materials.

Application of beta-(2-chloroaroyl) thioacetanilides in synthesis: an unusual and highly efficient access to thiochromeno[2,3-b]pyridine derivatives.

A series of unusual fused tricyclic thiochromeno[2,3-b]pyridines were successfully synthesized by tandem [3 + 3] annulation and SNAr of beta-(2-chloroaroyl) thioacetanilides with activated 4-arylidene-2-phenyloxazol-5(4H)-ones or aromatic aldehydes and ethyl 2-cyanoacetate under microwave irradiation, respectively. Because of the existence of the o-chloro atom of beta-(2-chloroaroyl) thioacetanilides, these reactions were very mild, convenient, and ortho-selective to form new fused tricyclic target molecules. In the domino processes, at least seven reactive distinct chemical sites were involved and up to three new covalent bonds and one tricycle with only cis configuration were generated. A synthetic study and mechanistic proposal for these transformations are presented.

[Survey on natural nidus of Metorchis orientalis in Huaihe River Basin].

Parafossarulus striatulus, Pseudorasbora parva and brood ducks are involved in the lifecycle of Metorchis orientalis. Natural nidi of M. orientalis are confirmed in Huaihe River Basin.