Ecotoxicity assessment of triphenyl phosphate (TPhP) exposure in Hoplobatrachus rugulosus tadpoles.

Triphenyl phosphate (TPhP), a widely used aromatic organophosphate flame retardant, is known to accumulate in organisms through water, air, and soil, consequently, causing toxicity. This study is the first to evaluate the acute and sub-chronic toxicities of TPhP to amphibians. In the acute toxicity analysis, the 96-h median lethal concentration (LC50) for GS35 Hoplobatrachus rugulosus tadpoles was 2.893 mg/L, and the 10% effect concentration (EC10) was 289 µg/L. After two weeks of exposure to low TPhP concentrations, the survival and metamorphosis rates of H. rugulosus tadpoles decreased, and the metamorphosis time was prolonged as the TPhP concentration increased. The threshold concentration that affected tadpole survival and metamorphosis time was 50 µg/L and 100 µg/L, respectively. No significant differences were observed in the condition factor and hepatic somatic index of the tadpole after metamorphosis; however, tadpole body mass and TPhP concentration were negatively correlated. Further, TPhP inhibited the expressions of Cu-Zn sod and cat, thereby reducing the activities of superoxide dismutase and catalase in the tadpole liver. The threshold for affecting gene expression and enzymatic activity was 100 µg/L. These findings provide significant insights on the stress ecology of aquatic organisms.

Microbial Diversity of the Chinese Tiger Frog (Hoplobatrachus rugulosus) on Healthy versus Ulcerated Skin.

The Chinese tiger frog (Hoplobatrachus rugulosus) is extensively farmed in southern China. Due to cramped living conditions, skin diseases are prevalent among unhealthy tiger frogs which thereby affects their welfare. In this study, the differences in microbiota present on healthy versus ulcerated H. rugulosus skin were examined using 16S rRNA sequences. Proteobacteria were the dominant phylum on H. rugulosus skin, but their abundance was greater on the healthy skin than on the ulcerated skin. Rhodocyclaceae and Comamonadaceae were the most dominant families on the healthy skin, whereas Moraxellaceae was the most dominant family on the ulcerated skin. The abundance of these three families was different between the groups. Acidovorax was the most dominant genus on the healthy skin, whereas Acinetobacter was the most dominant genus on the ulcerated skin, and its abundance was greater on the ulcerated skin than on the healthy skin. Moreover, the genes related to the Kyoto Encyclopedia of Genes and Genomes pathways of levels 2-3, especially those genes that are involved in cell motility, flagellar assembly, and bacterial chemotaxis in the skin microbiota, were found to be greater on the healthy skin than on the ulcerated skin, indicating that the function of skin microbiota was affected by ulceration. Overall, the composition, abundance, and function of skin microbial communities differed between the healthy and ulcerated H. rugulosus skin. Our results may assist in developing measures to combat diseases in H. rugulosus.

[Effect of Down-Regulation of ANRIL on Proliferation and Apoptosis of Kasumi-1 Cells and Its Potential Mechanism].

OBJECTIVE: To investigate the down-regulation of ANRIL (Antisense non-coding RNA in the INK4 Locus) effects on proliferation and apoptosis of Kasumi-1 cells and its related molecular mechanism. METHODS: Recombinant lentivirus was used to construct ANRIL down-regulation Kasumi-1 cells (sh-ANRIL group) and its control cells (sh-NC group). A fluorescence microscope was used to observe the transfection efficiency, RT-qPCR was used to detect knockdown efficiency and ANRIL expression in PBMCs and MBMCs of patients with acute myeloid leukemia (AML). Proliferation and apoptosis of Kasumi-1 cells were assayed by CCK-8 method and flow cytometry. Western blot was employed to detect the expression of PI3K, AKT, p-AKT, and relevant protein after down-regulation of ANRIL in Kasumi-1 cells. RESULTS: ANRIL overexpressed significantly in PBMCs and MBMCs of patients with AML, the transfection efficiency of recombinant lentivirus carrying sh-ANRIL and sh-NC on Kasumi-1 cells exceeded 90%, and the knockdown efficiency was 70%. When DNR was administrated for 24, 48, and 72 hours, the cell inhibition rate of the sh-ANRIL group was (47.40±1.49)%, (69.11±0.51)% and (91.82±1.10)%, which were significantly higher than those of the sh-NC group, respectively (P<0.05). The apoptotic rate in the sh-ANRIL group was (10.29±0.58)%, which was significantly higher than (5.42±0.67)% of the sh-NC group (P<0.01). After DNR treatment for 24 hours, the apoptotic rate of the sh-ANRIL group was (54.41±1.69)%, which was significantly higher than (38.28±1.42)% of sh-NC group (P<0.001). Western blot revealed that the protein levels of PI3K, p-AKT, PCNA, and BCL-2 in the sh-ANRIL group were reduced significantly than those in the sh-NC group, while the BAX protein expression increased. CONCLUSION: ANRIL may affect the proliferation and apoptosis of Kasumi-1 cells through PI3K/AKT signaling pathway. ANRIL is a potential therapeutic target for AML.

Advertisement calls of the lesser spiny frog Quasipaa exilispinosa (Liu and Hu, 1975) (Anura: Dicroglossidae).

There are currently only eleven species of Quasipaa (Anura: Dicroglossidae) distributed in southern and southwestern China to central Vietnam, southeastern Thailand, and southwestern Cambodia and presumably also in Laos (Frost 2020). Eight species of Quasipaa are currently known in China: Q. boulengeri (Günther 1889), Q. courtoisi (Angel 1922), Q. exilispinosa (Liu Hu 1975), Q. jiulongensis (Huang Liu 1985), Q. shini (Ahl 1930), Q. spinosa (David 1875), Q. yei (Chen, Qu, and Jiang 2002), and Q. verrucospinosa (Bourret 1937). The first seven species are endemic to China (AmphibiaChina 2020). These species have similar morphological traits and are capable of introgressive hybridization between the closely related species of this genus(Zhang et al. 2018). Despite the detailed acoustic analysis that is available to identify a variety of species and is beneficial to the study of anuran taxonomy (e.g., Microhyla species; Chen et al. 2020), advertisement calls have only been reported in detail for Q. spinosa (Yu Zheng 2009; Chen et al. 2012; Shen et al. 2015) and Q. shini (Kong et al. 2016), whereas the call for Q. exilispinosa was briefly described from observations in captivity (Voitel 2000).

Two complete mitochondrial genomes of the black-spotted stout newt (Pachytriton brevipes) and their relative phylogenetics relationship with other Salamandridae.

The black-spotted stout newt (Pachytriton brevipes) is widely distributed in the mountains of southeastern China. In this study, we sequenced and determined the complete mitochondrial genome (mitogenomes) of two P. brevipes samples collected in Mount Wuyi. The assembled mitogenomes were 16,298 bp and 16,301 bp in length, and contained 13 protein-coding genes (PCGs), two ribosomal RNA genes, 22 transfer RNA genes, one non-coding region, and one control region. The phylogenetic analysis indicated the two samples of black-spotted stout newt grouped together and are the sister group of P. feii.