Seasonal variation in structure and function of gut microbiota in Pomacea canaliculata.

Gut microbiota is associated with host health and its environmental adaption, influenced by seasonal variation. Pomacea canaliculata is one of the world's 100 worst invasive alien species. Here, we used high-throughput sequencing of the 16S rRNA gene to analyze the seasonal variation of gut microbiota of P. canaliculata. The results suggested that the predominant gut microbial phyla of P. canaliculata included Firmicutes and Proteobacteria, which helped digest plant food and accumulate energy. The gut microbiota of P. canaliculata in summer group showed the highest diversity, whereas the winter group possessed the lowest, probably due to the shortage of food resources of P. canaliculata in winter. Principal coordinate analysis analysis based on unweighted unifrac and weighted unifrac indicated that the composition of gut microbiota of P. canaliculata significantly varied across seasons. Bacteroidetes tended to be enriched in summer by linear discriminant analysis effect size analysis. Actinobacteria and Cyanobacteria were extremely abundant in autumn, while Fusobacteria and Cetobacterium enriched in winter. In conclusion, the structure of the gut microbiota of P. canaliculata was significantly different among seasons, which was beneficial to the environment adaptation and the digestion and metabolism of food during different periods.

Transcriptome sequencing reveals the differentially expressed lncRNAs and mRNAs in response to cold acclimation and cold stress in Pomacea canaliculata.

BACKGROUND: Tolerance of low temperature has a significant impact on survival and expansion of invasive snail Pomacea canalicuata. Cold acclimation can enhance cold tolerance of Pomacea canalicuata. To elucidate the molecular mechanism of P. canaliculata's responses to cold acclimation and cold stress, a high-throughput transcriptome analysis of P. canaliculata was performed, and gene expression following artificial cold acclimation and then cold stress at 0 °C for 24 h was compared using RNA sequencing. RESULTS: Using the Illumina platform, we obtained 151.59 G subreads. A total of 5,416 novel lncRNAs were identified, and 3166 differentially expressed mRNAs and 211 differentially expressed lncRNAs were screened with stringent thresholds. The potential antisense, cis and trans targets of lncRNAs were predicted. Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that many target genes were involved in proteasome, linoleic acid metabolism and retinol metabolism under cold acclimation. The lncRNA of P. canaliculata could participate in cold acclimation by regulating the expression of E3 ubiquitin protein ligase, 26S proteasome non-ATPase dependent regulation subunit, glutathione S-transferase, sodium/glucose cotransporter and cytochrome P450. CONCLUSIONS: These results broaden our understanding of cold acclimation and cold stress associated lncRNAs and mRNAs, and provide new insights into lncRNA mediated regulation of P. canaliculata cold acclimation and cold stress response.

Genome-wide identification and functional analysis of the glutathione S-transferase (GST) family in Pomacea canaliculata.

Cold causes oxidative stress in living organisms, mainly caused by the accumulation of reactive oxygen species (ROS). In the antioxidant defense systems, Glutathione S-transferases (GSTs) play a vital role in the regulation of detoxification and redox balance of ROS. In this study, the P. canaliculata GST gene family were characterized using a genome-wide search of the conserved domain. Phylogenetic tree and domain composition analysis revealed that 30 PcGSTs belong to seven classes, including five in MAPEG, two in Mu, nine in Omega, 11 in Sigma, one in Rho, Theta and Alpha class, respectively. RNA-seq analysis revealed that most PcGSTs localized in mitochondria highly expressed in hepatopancreas, and most PcGSTs localized in cytoplasm highly expressed in kidney. A total of 12 PcGST genes were significantly up-regulated and PcGST12 was significantly down-regulated after cold acclimation. Ten PcGSTs were identified as DEGs under cold stress after cold acclimation. qRT-PCR revealed that the expression level of five PcGST genes were significantly varied during the cold acclimation. The present study investigated the characterization of the P. canaliculata GST gene family, extending our understanding of GST mediated cold acclimation and cold stress-response mechanisms in this invasive snail.

Characterization of the complete mitochondrial genome of the many-lined sun skink (Eutropis multifasciata) and comparison with other Scincomorpha species.

Characterizating the complete mitochondrial genome (mitogenome) of an organism allows detailed genomic studies in systematics and evolution. The present study decodes the mitogenome (17,062 bp) of the many-lined sun skink, Eutropis multifasciata, using next-generation sequencing. To compare the diversity of mitogenomic structure and investigate intraspecific evolutionary relationships among the Asian Scincomorpha, the mitogenomes of 46 other species were examined concurrently. Within the group, the size of mitogenomes varied predominantly in the length at their control regions. The Ka/Ks ratios of 12 protein codon genes (PCGs) were lower than 1.00, demonstrating that they were under relaxed or moderate purifying selection. However, the ND5 had a Ka/Ks ratio >1, and was considered to be under positive selection. Currently there are two superfamilies in Scincomorpha (i.e. Scincoidea and Lacertoidea), but phylogenetic analysis using Bayesian Inference and Maximum-Likelihood Estimations produced phylogenetic trees with three clades in Scincomorpha ((Scincoidea + Lacertoidea (part)) + Gymnophthalmidae)).

Molecular characterization of cathelicidin in tiger frog (Hoplobatrachus rugulosus): Antimicrobial activity and immunomodulatory activity.

Cathelicidins are an important antimicrobial peptide family and are expressed in many different vertebrates. They play an important role in the innate immune system of the host. However, amphibian cathelicidins are poorly understood. In this study, the cDNA of the cathelicidin gene was obtained from the skin transcriptome of tiger frog (Hoplobatrachus rugulosus). The predicted amino acid sequence of tiger frog cathelicidin (HR-CATH) comprises a signal peptide, a cathelin domain, and a mature peptide. The HR-CATH amino acid sequence alignment with other frog cathelicidins showed that the functional mature peptide is highly variable in amphibians, whereas the cathelin domain is conserved. A phylogenetic tree analysis showed that HR-CATH is most closely related to cathelicidin-NV from Nanorana ventripunctata. HR-CATH was chemically synthesized and its in vitro activity was determined. It had high antibacterial activity against Vibrio parahaemolyticus, Staphylococcus aureus, and the pathogenic bacterium Aeromonas hydrophila. HR-CATH damaged the cell membrane integrity of A. hydrophila according to a lactate dehydrogenase release assay and was able to hydrolyze the genomic DNA from A. hydrophila in a dose-dependent manner. Furthermore, in RAW264.7 cells (mouse leukemic monocyte/macrophage cell line), HR-CATH induced chemotaxis and enhanced respiratory burst. Our study shows that amphibian cathelicidin has antimicrobial activity and an immunomodulatory effect on immune cells.

Complete mitochondrial genome of Scincella modesta (Squamata: Scincidae).

The first complete mitochondrial genome sequence was determined for an oviparous lizard, Scincella modesta (Scincella, Scincidae). The total length of the complete mitochondrial genome was 17,511 bp, encodes 13 protein-coding genes, 22 tRNAs, 2 rRNA genes, and 2 non-coding regions. The overall base composition of S. modesta is A: 31.9%, T: 27.2%, G: 14.5%, and C: 26.5%. Most of the S. modesta mitochondrial genes are encoded on the H-strand except for the ND6 gene and eight tRNA genes, which are encoded on the L-strand. Mrbayes and ML tree based on 13 protein-coding genes indicated that S. vandenburghi is the sister group of the S. modesta within the genus Scincella. The complete mitogenome sequence of S. modesta provided fundamental data for resolving phylogenetic and genetic problems related to genus Scincella.

Acoustic divergence in advertisement calls among three sympatric Microhyla species from East China.

BACKGROUND: Species-specific advertisement calls are the main mechanism of transmitting information between individuals in anuran amphibians and are therefore indispensable for anuran survival and reproduction. Survey methods that monitor these calls can be used for rapid species recognition, behavioral experiments, and conservation monitoring. In this study, we described in detail 10 call parameters from three sympatric species in the genus Microhyla and analyzed the differences in call parameter among these species to provide a basis for systematic monitoring, acoustic analysis and taxonomic study of this genus. METHODS: The quantitative analyses of temporal and spectral call parameters were used in our study for the advertisement calls of three sympatric Microhyla species (M. beilunensis, M. fissipes and M. heymonsi) in Zhejiang Province, East China. RESULTS: Our results showed the following: (1) Significant differences existed among the three sympatric Microhyla species in call duration (CD), call interval (CI), number of pulses (NP), pulse rate, call intensity (CIT), dominant frequency (DF) and frequency of the first to fourth formants (F1, F2, F3 and F4). (2) Some spectral parameters (DF, F1 and F3) were negatively correlated with the body size of the vocalizing individuals in each species. (3) The coefficients of variation within individuals (CVw) for CIT, DF and F1-F4 were smaller than 5%, whereas the CVW for CI was larger than 10% in each species. (4) The principal component analysis and discriminant function analysis showed that call parameters could distinguish the three Microhyla species. (5) The phylogenetic generalized least squares analysis showed that phylogenetic relationships affected CD and NP against snout-vent length (SVL), DF and NP against CD, and NP against DF, but not of DF against SVL; based on the phylogenetic analysis, CD and NP were not related to SVL, but DF was negatively related to SVL.

The combined effects of temperature and aromatase inhibitor on metamorphosis, growth, locomotion, and sex ratio of tiger frog (Hoplobatrachus rugulosus) tadpoles.

BACKGROUND: The tiger frog (Hoplobatrachus rugulosus) is widely raised by many farms in southern region of China as an economically edible frog. The growth, development, and sexual differentiation of amphibians are influenced by temperature and steroid hormone level. However, the problem of hormone residues is caused by the addition of exogenous hormones in frog breeding, it is worth considering whether non-sterol aromatase inhibitors can be used instead of hormones. METHODS: In our study, H. rugulosus tadpoles were subjected to two water temperatures (29 °C and 34 °C) and three letrozole concentrations in the feed (0, 0.1 and 1 mg/g) to examine the effects of temperature, aromatase inhibitor and their interaction on metamorphosis, locomotion, and sex ratios. A G-test and contingency table were used to analyze the metamorphosis rate of tadpoles and the survival rate of froglets after feeding for 90 days. A G-test was also used to analyze sex ratios in different treatment groups. RESULTS: Metamorphosis time and body size (snout-vent length, body mass and condition factor) were significantly different between the two temperature treatments. Metamorphosis time was longer and body size was increased at 29 °C compared to those at 34 °C. Letrozole concentration and the temperature × letrozole interaction did not affect these variables. The jumping distance of froglets following metamorphosis was positively associated with the condition factor; when controlling for condition factor, jumping distance was not affected by temperature, letrozole concentration and their interaction. Temperature and letrozole concentration also did not affect metamorphosis and survival rate. Sex ratio of the control group (0 mg/g letrozole) was 1:1 at 29 °C, but there were more males at 34 °C. The sex ratios of H. rugulosus treated with letrozole at 29 °C and 34 °C were significantly biased toward males, and male ratio increased as letrozole concentration increased. Furthermore, more males were produced at 34 °C than at 29 °C at each letrozole concentration.

Partial mitochondrial genome of the Sanchiang Tree Toad Hyla sanchiangensis (Anura: Hylidae).

We reported the partial mitochondrial genome (mitogenome) for Hyla sanchiangensis (Anura: Hylidae), a arboreal frog and endemic in China. The length of partial mitogenome of H. sanchiangensis was 15,664 bp, and contained PCGs (COX1-3, ND1-6, ND4L, ATP6, ATP8 and CYTB), 2 ribosomal RNA genes, 22 transfer RNA genes, and 2 non-coding regions of a L-strand replication origin and a partial loop region. The overall base composition of the sequence is 29.91% A, 29.86% T, 14.58% G, and 25.65% C, with a total A + T content of 59.77%. The result of phylogenetic analysis showed that H. sanchiangensis formed a clade with other species belonging to the genus of Hyla. This mitogenome data could help in evolutionary biology and population genetics of the Hylid species.