Screening TLR4 Binding Peptide from Naja atra Venom Glands Based on Phage Display.

Toll-like receptor 4 (TLR4) is a crucial inflammatory signaling pathway that can serve as a potential treatment target for various disorders. A number of inhibitors have been developed for the TLR4 pathway, and although no inhibitors have been approved for clinical use, most have been screened against the TLR4-MD2 conformation. The venom gland is the organ of venomous snakes that secretes substances that are toxic to other animals. The level of gene transcription in venom glands is different from that in other tissues, includes a large number of biologically active ingredients, and is an important natural resource for the development of new drugs. We constructed a T7 phage display library using the cobra (Naja atra) venom gland from the Guangdong Snake Breeding Plant and performed three rounds of screening with TLR4 as the target, randomly selecting monoclonal phage spots for PCR followed by Sanger sequencing. The obtained sequences were subjected to length analysis, molecular docking, solubility prediction, and stability prediction, and a peptide containing 39 amino acids (NA39) was finally screened out. The BLAST results indicated that NA39 was a sequence in RPL19 (Ribosomal Protein L19). After peptide synthesis, the binding ability of NA39 to TLR4 was verified by the surface plasmon resonance (SPR) technique. In this study, a new peptide that can specifically bind TLR4 was successfully screened from the cobra venom gland cDNA library, further demonstrating the effectiveness of phage display technology in the field of drug discovery.

Chemical Dissolution-Assisted Ultrafine Grinding for Preparation of Quasi-Spherical Colloids of Zinc Oxide.

Submicron-sized quasi-spherical zinc oxide (ZnO) particles were prepared by wet ultrafine grinding in a stirred media mill under various conditions. The effects of parameters (i.e., solution type, acid or alkali concentration, solid content and grinding time) on the particle median size (d50), particle size distribution (PSD) and sphericity of ZnO particles was investigated. The results show that submicron-sized quasi-spherical particles (i.e., d50: 370 nm, uniformity coefficient (n) of 2.28 and sphericity of 0.91) can be obtained when the micron-sized ZnO particles are ground for 30 min in a CH3COOH solution at a concentration of 0.010 mol/L with 20 wt.% of solid content. The chemical dissolution of ZnO particles ground in the presence and absence of acetic acid is discussed. It is indicated that chemical dissolution accelerated due to the mechanochemical effects could reduce the particle size, obtain a narrower PSD and enhance the sphericity. In addition, the functions of selection and breakage were used to analyze the grinding mechanism of ZnO particles.

Description of a New Cobra (Naja Laurenti, 1768; Squamata, Elapidae) from China with Designation of a Neotype for Naja atra.

Taxonomic frameworks for medically important species such as cobras (genus Naja Laurenti, 1768; Squamata, Elapidae) are essential for the medical treatment of snake bites and accurate antivenin development. In this paper, we described the former N. kaouthia populations recorded from China as a new species and designated a neotype for N. atra-based morphological and mitochondrial phylogenetic analysis. The new species N. fuxisp. nov. was morphologically diagnosed from N. kaouthia by (1) regular single narrow crossband present on the middle and posterior parts of the dorsum (3-15, 7.9 ± 2.7, n = 32) and the dorsal surface of the tail (1-6, 4.2 ± 1.1, n = 32) of both adults and juveniles, buff-colored with dark fringes on both edges, vs. South Asian populations (n = 39) and Southeast Asian populations (n = 35) without cross bands, with irregular cross bands or multiple light-colored crossbands pairs, or densely woven lines; (2) small scales between the posterior chin shields, usually three (40%) or two (37%), rarely four (13%), or one (10%) (n = 30) vs. mostly one (81%) and rarely two (19%) (n = 28); (3) ventrals 179-205 (195.4 ± 6.7, n = 33) vs. South Asian populations 179-199 (188.7 ± 5.9, n = 12); Southeast Asian populations 168-186 (177.8 ± 4.9, n = 18). Phylogenetically, the new species forms an independent sister clade to the clade including N. atra, N. kaouthia, N. oxiana and N. sagittifera. Furthermore, the subspecies N. naja polyocellata should be resurrected and recognized as a full species, N. polyocellatacomb. nov., and the subspecies N. sumatrana miolepis should be resurrected.

Potential impact of land-use change on habitat quality in the distribution range of crocodile lizards in China.

The over-exploitation of land resources poses a serious threat to biodiversity on a global scale. Changes in land-use and human exploitation have had a major impact on wild populations and their habitat in China. We assessed how habitat quality has changed over time (1995-2020). Specifically, we analyzed how the habitat quality of crocodile lizard has changed over time based on multi-temporal land-use data (1995, 2000, 2010, 2015 and 2020) using a land-use transfer matrix and habitat quality model. The results showed that the main landscape types in the study area were arable land (21.21% of the area) and woodland (69.59% of the area) during the period. Construction land (land used for development) had decreased by 991 km2, a decrease rate of 59.84% from 1995 to 2000, and increased to 2349 km2, an increase rate of 71.69% from 2000 to 2020. The proportion of grasslands and areas with water were negligible and overall, did not vary significantly in size over the study period. The main feature of land use change in the study area was the loss of grasslands and woodlands through development. The habitat quality model indicated that habitat quality was highest and degradation was lowest in Dayao mountain, Guxiu town, Qichong village and Beituo town. Habitat quality improved in Daguishan and Luokeng areas. Habitat quality was good in Daping mountain and Linzhouding, but they were highly fragmented with patches of low-quality habitat of varying sizes. Habitats were severely degraded in the Dateng Gorge area. The rate of habitat degradation has slowed over time in the study area, but gradually increased in degradation intensity, and low-quality habitats were widely distributed and overlapped with the crocodile lizards distribution area. We recommend that protected areas for the crocodile lizard be more closely monitored and managed to halt further decline in habitat quality.

Climate Change and Dispersal Ability Jointly Affects the Future Distribution of Crocodile Lizards.

Crocodile lizards (Shinisaurus crocodilurus) are an endangered, 'living fossil' reptile from a monophyletic family and therefore, a high priority for conservation. We constructed climatic models to evaluate the potential impact of climate change on the distribution of crocodile lizards for the period 2000 to 2100 and determined the key environmental factors that affect the dispersal of this endangered species. For the construction of climatic models, we used 985 presence-only data points and 6 predictor variables which showed excellent performance (AUC = 0.974). The three top-ranked factors predicting crocodile lizard distribution were precipitation of the wettest month (bio13, 37.1%), precipitation of the coldest quarter (bio19, 17.9%), and temperature seasonality (bio4, 14.3%). Crocodile lizards were, just as they are now, widely distributed in the north of Guangdong Province in China and Quang Ninh Province in Vietnam at the last glacial maximum (LGM). Since the LGM, there has been an increase in suitable habitats, particularly in east-central Guangxi Province, China. Under future global warming scenarios, the potential habitat for crocodile lizards is expected to decrease significantly in the next 100 years. Under the most optimistic scenario, only 7.35% to 6.54% of suitable habitat will remain, and under the worst climatic scenario, only 8.34% to 0.86% of suitable habitat will remain. Models for no dispersal and limited dispersal showed that all crocodile lizards would lose habitat as temperatures increase. Our work contributes to an increased understanding of the current and future spatial distribution of the species, supporting practical management and conservation plans.

Comparison of Gut Microbiota Diversity Between Captive and Wild Tokay Gecko (Gekko gecko).

Captive animals and wild animals may exhibit different characteristics due to the heterogeneity of their living environments. The gut microbiota play an important role in the digestion and absorption, energy metabolism, immune regulation, and physiological health of the host. However, information about the gut microbiota of captive and wild Gekko gecko is currently limited. To determine the difference in gut microbiota community composition, diversity, and structure between captive and wild geckos, we used the Illumina miseq platform to conduct high-throughput sequencing and bioinformatics analysis of the v3-v4 hypervariable region of 16S rRNA in 54 gecko samples. Our results showed that Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria were the dominant gut microbiota phyla of the gecko. The dominant genera comprised mainly Pseudomonas, Burkholderia-caballeronia-paraburkholderia, Ralstonia, Romboutsia, and Bacteroides. Captive geckos had significantly higher alpha diversity and potential pathogenic bacteria than wild populations. Moreover, significant differences in beta diversity of gut microbiota were observed between two populations. Functional prediction analysis showed that the relative abundance of functional pathways of wild geckos was more higher in metabolism, genetic information processing and organismal system function than those in captive geckos. Total length significantly affected gut microbial community (R2 = 0.4527, p = 0.001) and explained 10.45% of the total variation for gut microbial community variance between two groups. These results may be related to differences in diet and living environment between two populations, suggesting that the management of captive populations should mimic wild environments to the greatest extent possible to reduce the impact on their gut microbiota.

Multiple lines of evidence reveal a new species of Krait (Squamata, Elapidae, Bungarus) from Southwestern China and Northern Myanmar.

Kraits of the genus Bungarus Daudin 1803 are widely known venomous snakes distributed from Iran to China and Indonesia. Here, we use a combination of mitochondrial DNA sequence data and morphological data to describe a new species from Yingjiang County, Yunnan Province, China: Bungarus suzhenae sp. nov. Phylogenetically, this species forms a monophyletic lineage sister to the Bungarus candidus/multicinctus/wanghaotingi complex based on cyt b and ND4 genes but forms a sister species pair with the species B. magnimaculatus Wall & Evans, 1901 based on COI gene fragments. Morphologically, B. suzhenae sp. nov. is similar to the B. candidus/multicinctus/wanghaotingi complex but differs from these taxa by a combination of dental morphology, squamation, coloration pattern, as well as hemipenial morphology. A detailed description of the cranial osteology of the new species is given based on micro-CT tomography images. We revised the morphological characters of B. candidus/multicinctus/wanghaotingi complex and verified the validity of three species in this complex. The distribution of these species was revised; the records of B. candidus in China should be attributed to B. wanghaotingi. We also provide an updated key to species of Bungarus.

A new species of the genus Smithophis (Squamata: Serpentes: Natricidae) from southwestern China and northeastern Myanmar.

A new species of natricid snake, Smithophis linearis sp. nov., is described on the basis of a single recently collected specimen from Yingjiang County, Yunnan Province, People's Republic of China, and three historical specimens from Yunnan and from northeastern Myanmar. The new species is assigned to the genus Smithophis on the basis of its single internasal and single prefrontal shields, and on the basis of the results of phylogenetic analysis of mitochondrial cytochrome b DNA sequence data. The new species differs from its congeners in having the following combination of characters: temporal shields present, six or more circumorbital scales, and a distinctive colour pattern comprising regular, narrow, longitudinal dark and pale lines. Morphological and cytochrome b data are consistent with the recognition of Smithophis as distinct from the genus Opisthotropis. A revised key to the identification of the species of Smithophis is provided.

A new pit viper of the genus Trimeresurus (Lacépède, 1804) (Squamata: Viperidae) from Southwest China.

The white-lipped tree viper, Trimeresurus albolabris Gray, is one of the most common species of the genus Trimeresurus with a wide distribution from India eastwards to China and southwards to Indonesia. However, this species was previously split into five geographical clades with significant genetic divergence. Recent surveys in Yingjiang County, Yunnan Province, China resulted in the discovery of one cryptic species of the subgenus Trimeresurus. Combining molecular, morphological and ecological data, we describe it as a new species Trimeresurus caudornatus sp. nov. The new species differs from other Trimeresurus species by a combination of the following characteristics: (1) Head and body generally dark green, postocular stripes absent in both genders, upper labials light green; (2) ventrolateral stripe faint green yellow, present on the first row of DSR in both genders; (3) iris golden yellow in both genders; (4) dorsal tail mostly dark red, lateral and ventral green; an orange red stripe along the ventral part of the tail; (5) DSR 21/22 -21-15, VEN 161-163, SC 52-72; (6) first upper labial partially fused to the nasal. The new species was shown to be a strongly supported monophyletic group (BPP 100%) and sister to T. septentrionalis. The uncorrected pairwise distances of mitochondrial gene Cytb between the new species and other species of the subgenus Trimeresurus, ranging from 0.059 (T. septentrionalis) to 0.134 (T. kanburiensis).

DNA barcoding revises a misidentification on mossy frog: new record and distribution extension of Theloderma corticale Boulenger, 1903 (Amphibia: Anura: Rhacophoridae).

As an endangered animal group, mossy frog (genus Theloderma) has attracted the attention of biologists and wildlife conservationists. Clarifying the taxonomic status and distribution of each species in Theloderma is important to determine the conservation status for each species, establish appropriate conservation strategies and probe the speciation process. Recently, we discovered a medium-sized species of mossy frog of the genus Theloderma in April 2015 during municipal surveys of amphibians in Dayao Mountain of Jinxiu. It was collected from the water-filled tree cavities. However, there remains some uncertainty about the species determination of the mossy frog in the Yinshan station of Dayao Mountain in Guangxi Province, China. Previously, the mossy frog in Guangxi Province was recognized as Th. kwangsiense. In order to clarify the species status of the mossy frog obtained from Guangxi, we sequenced 2414 bp of the 12S and 16S genes in the sample collected from the Dayao Mountain. Combining all the sequence in NCBI, genetic analyses from the data suggest that the sample from the Dayao Mountain is Th. corticale rather than Th. kwangsiense. It is most likely that the most previous studies had wrong species identification. And, it is the first time we use DNA barcoding to prove that the species obtained from Guangxi is a new distribution of Th. corticale.