Abstract:The “star effect” of flagship species has a good role in promoting biodiversity conservation and national park construction. China attaches great importance to marine conservation, and selecting ideal species as flagship species plays an important role in promoting marine conservation. Sea turtles are marine living fossil species and are listed under level-I protection on the “List of Wildlife under Special State Protection”. They play an important role in the balance and stability of marine biodiversity and ecosystem. The unique appearance features of sea turtles make them highly recognizable, and their lovely image and special life history make them full of charm. The history of sea turtles being used in China is quite long and has very high cultural values. Sea turtles are highly encountered and accessible in coastal areas and have good relationships with local communities. In conclusion, we believe that sea turtles are ideal flagship species for marine conservation.

Abstract:[Objectives] The physiological and ecological characteristics associated with the basal metabolic rate (Rbm) of birds correlate closely with their geographic distribution. In order to understand the ecophysiological characteristics of the Little Curlew Numenius minutus in the Wenzhou area, we measured their body temperature (Tb) and metabolic rate (Rm, metabolic rate was expressed as ml of O2 consumed per h), and calculated their thermal conductance (C) at ambient temperature (Ta) from 5 to 42.5 ℃. [Methods] Metabolic rate (ml/h) was estimated as the rate of oxygen consumption in an open-flow respirometry system (TSE, Germany). Basal metabolic rate was calculated for each individual as the average of 10 lowest consecutive oxygen consumption recordings (about 5 min) (Fig. 1). Thermal conductance [ml/(g.h.℃)] was calculated at each temperature using the formula:C = Rm/(Tb﹣Ta). F value was calculated as F = (Rbm/predicted Rbm)/(C/predicted C). All data were analyzed by SPSS (version 21.0). The effects of ambient temperature on body temperature, metabolic rate, and thermal conductance were analyzed using repeated measures ANOVA. The metabolic rates at different ambient temperatures were directly compared using independent sample t-test. Linear, or exponential, models were fitted where appropriate to describe the relationship between metabolic rate or thermal conductance and ambient temperature. All results were expressed as the mean ± SE, and P< 0.05 was taken to be statistically significant. [Results] There was no significant difference in initial body temperature over an ambient temperature range of 5﹣35 ℃, as Little Curlews could maintain their body temperature at 42.8 ± 0.1 ℃ (Fig. 2). There was, however, a significant variation in final body temperature over the same temperature range. The mean body temperature reached 43.7 ℃ when the ambient temperature was 42.5 ℃ (Fig. 2). The thermal neutral zone was 27.5﹣40.0 ℃, and the meanmetabolic rate within the thermal neutral zone was 221.31 ± 6.01 ml/h, accounting for 141% of the expected value based on the bird’s body mass (Fig. 3). Below the lower critical temperature, metabolic rate increased with declining ambient temperatures, and the relationship between metabolic rate and ambient temperature could be described as Rm (ml/h) = 587.10﹣11.78 Ta (℃). At a 5﹣27.5 ℃ ambient temperature range, the thermal conductance was 0.11 ± 0.00 ml/(g.h.℃), representing 212% of the expected value based on the bird’s body mass (Table 1). However, the thermal conductance increased exponentially from 27.5 ℃ to 42.5 ℃ as described by the equation lg C [ml/(g.h.℃)] = 0.070 + 0.006 Ta (Fig. 4). The F value was 1.21. [Conclusion] The findings suggested that Little Curlews were able to adapt to the environment by implementing a relatively high level of basal metabolic rate, wider thermal neutral zone, high body temperature, and intensity of chemical thermoregulation. These properties would enable the birds to readily acclimate to their environments and survive in relatively cold areas.

Abstract:[Objectives] The Himantopus himantopus is a widespread species whose breeding sites are mainly distributed in northwest, northeast, north, and east China. Although a lot of researches have observed and recorded the reproductive behavior of H. himantopus, most of them focused on a certain feature of reproductive behavior, and few studies completely observed and recorded the whole breeding cycle and behavior. [Methods] The study was conducted in Qinzhou Bay Balushui Wetland, Fangchenggang City, Guangxi Zhuang Autonomous Region (March to September 2021). The breeding situation in 2021 is shown in Table 1. We observed the breeding behavior of H. himantopus by scanning in 2019 and 2020 to confirm the general breeding sites and seasons. After determining the nest site, the information of state, type, material, and morphological parameters (length, width, and depth) and clutch number of each nest, and size of each egg in every nest were recorded and measured. We selected 6 nests of H. himantopus at random by target animal sampling method to record the behavior of nesting, laying, and incubating the egg. A nest of couple of H. himantopus and the all-event sampling method were used to observe the behavior of mating, brooding, and defense. The time spent at the nest by both sexes was analyzed using t-test, and analyses were completed by SPSS 25.0. During the observation period, male and female adults of both sexes were distinguished by their breeding plumage:males had brightly colored breeding plumage with black eyes, head, nape of neck, back, and wings. Females had no black-colored breeding plumage on the head and neck, but had gray spots behind the eyes (Fig. 1). The observation time was from 7:00 to 19:00, and the reproductive behavior was recorded by infrared cameras at night. We used telescope to monitor each nest from a distance of 10﹣20 m away from the nests. A set of video equipment was also arranged to monitor each nest found at a distance about 1 m away from the nest from 19:00 pm to 7:00 am. The equipment consists of an infrared camera (AM-860 dv-58g., Wuhan, China) and a battery with a capacity of 2 000 mAh. [Results] We observed the reproductive behavior of 39 nests of H. himantopus and found:1) They inhabited salt fields, shrimp ponds, and fishponds while most nests were placed in salt fields. 2) There are four nest types, including nests made of rick, nests made of gravel, nests made of dirt, and nests built on Eleocharis acicularis. These nesting materials consist of rick including Gramineae or Cyperaceae, and some mixture like gravel, shell, and so on. Nest outer diameter was 23.3 ± 10.7 cm, nest inner diameter was 11.2 ± 1.9 cm, nest depth was 1.6 ± 0.5 cm, nest height was 6.5 ± 4.3 cm (n = 39). The nest building period lasted 3 ± 2 d (n = 6) according to the complexity of the nest (Table 1, Fig. 2). 3) The clutch sizes are 2﹣4, and 1 egg was laid with a 1﹣2 d interval and finished within 7 d. 4) The incubation time of females was longer than that of males (female:8 550 ± 245.9 min, male:7 530 ± 263.3 min, P > 0.05), but the difference was not significant, and the incubation period was 25 ± 2 d (n = 6, Fig. 3). 5) In the first and middle stages of brooding (the chicks are 1 to 20 d of age), the female brooding time was nearly twice than that of the male. In the later stages (the chicks over 20 d of age), the time of brooding between the male and the female gradually decreased, and the time of brooding without the parents became longer. The nestling period lasted 26 ± 3 d (n = 6, Fig. 4). 6) For different threat sources, the defense behavior of H. himantopus mainly includes parental defense and group defense (Table 2). [Conclusion] This study is helpful to fully understand the breeding behavior of H. himantopus in the study area and provide a basis for the conservation of H. himantopus.

Abstract:[Objectives] The Great Tit (Parus cinereus) is a passerine bird widely distributed in China, which has strong adaptability to varied environments. In order to understand the breeding ecology of Great Tits in the urban environment, we conducted a systematic survey of Great Tits breeding in the lampposts of Futian Mangrove Ecological Park, Shenzhen, Guangdong. [Methods] From 2020 to 2022, we surveyed Great Tits’ nests in lampposts by using a selfie stick and a mobile phone, and recorded breeding activities at different stages by taking pictures from an appropriate distance. [Results] From 2020 to 2022, the Great Tits began nesting in early March each year, entering and exiting the lampposts from the lowest vent and building their nests between the internal wires and the inner wall of the lampposts (Fig. 1). A total of 43 nests were recorded, and the average clutch size of the Great Tits in the park was 5.0 ± 0.8 (n = 32). Most of their first eggs were laid in late March to early April (n = 20) (Fig. 2). In this study, the average incubation period was 13 ± 1.6 d (n = 18), and the average brooding period was 18.6 ± 1.8 d (n = 19), with a reproductive success rate of 49% (63 fledglings). As Great Tits are territorial, all nests are kept at a certain distance from each other (Fig. 3). The average spacing between the closest nests in 2020 was 94.29 ± 25.68 m (66﹣140 m), while the average spacing in 2021 was 86.69 ± 32.44 m (36﹣146 m), and 85.50 ± 57.71 m (20﹣254 m) in 2022. 12 end-of-breeding nests were collected for nest material analysis. These nests consist of grass leaves, moss, and cotton wool. The 12 nests weigh 70﹣527 g, with an average of 215 ± 171 g (n = 12). [Conclusion] In this study, the breeding parameters of the Great Tits in the lampposts in Shenzhen were compared with the breeding parameters of those in the nest boxes in Hainan, Henan, Hebei, and northeast China (Fig. 4). The average clutch size of the Great Tits in Shenzhen was similar to those in Hainan, and smaller than those in Henan, Hebei, and northeast China. The laying date of the first egg is close to that in Hainan. Higher the latitude, later the first laying date. The hatchability and reproductive success rate were significantly lower in the lamppost than that in the nest boxes. Our study shows that the breeding failures of the Great Tits in the park are mainly due to fallen nests and human disturbance. It is necessary to redesign the lampposts to provide suitable nest sites for the Great Tits in the park. The Great Tits’ breeding behaviors in urban structures reflect the sign of their adaptations to urbanization.

Abstract:[Objectives] Electronic fence in preventing and controlling wildlife damages has been widely studied and applied overseas, but it has not been promoted in China. Chunhua Town of Hunchun, Jilin Province was chosen as the study area, and the research on the effect of electronic fences in preventing and controlling Wild Boar (Sus scrofa) damages and the influencing factors has been carried out. [Methods] A total of 40 sample plots were laid out for the study and divided into 12 groups, with a﹣k being the experimental group and l being the control group (Table 2). The number of wires, single pulse intensity, length of control period, crop loss rate, and the number of Wild Boars entering the sample plots were recorded (Table 3). The effectiveness of the control was measured by the length of the control period, the number of Wild Boars entering the sample plot and the crop loss rate. The data were analyzed using the Mann-Whitney U test to determine the significance and variability of the effectiveness of electronic fence. [Results] The length of the control period, the number of Wild Boars entering the sample plots, and the crop loss rate of all experimental groups were significantly different from those of the control group (P < 0.05) (Table 5). There were no significant differences (P > 0.05) in the length of the effective period of control, the number of Wild Boars entering the sample plots, and the rate of crop loss in the sample plots with two fence wires of electronic fence and pulse intensities of 0.3 J, 1.0 J and 2.0 J respectively (Table 4). The effective period of control was 29.2 ± 1.4 d, the number of Wild Boars entering the sample plots was 0.7 ± 1.0 ind and the crop loss rate was 4.28% ± 8.24% for the electronic fence group with three fence wires. The effective period of control was 27.3 ± 3.3 d, the number of Wild Boars entering the sample plots was 1.0 ± 1.3 ind and the crop loss rate was 7.98% ± 14.34% for the electronic fence group with two fence wires. The electronic fence group at the one fence wire had a control period of 23.0 ± 2.3 d, with 2.3 ± 1.5 ind of Wild Boars entering the sample plot and 19.33% ± 8.14% of crop loss (Table 5). [Conclusion] The electronic fence could effectively reduce the damage of Wild Boar to farmland. The pulse intensity of the electronic fence has no significant effect on the prevention and control effect of Wild Boar damage to farmland, and the more the number of coils of the electronic fence, the better the prevention and control effect of Wild Boar damage.

Abstract:[Objectives] Species diversity and geographical distribution are important basis for wildlife research and protection. Therefore, updating the list and fauna of reptiles in Liaoning Province will further lay the foundation for ecological research and protection of the reptiles distributed in the area. [Methods] We updated the list of reptiles in Liaoning Province based on published research literature and recent field investigations. On this basis, according to the composition of reptile species and the distribution range of representative species in various regions, adjustments were made to the reptile geographical flora of Liaoning Province. [Results] Based on the existing research and field investigation of reptiles in Liaoning Province, the checklist and zoogeographic division of the Reptilian Fauna of Liaoning Province were revised. The distribution of Elaphe taeniura in Liaoning Province was further confirmed with the image of E. taeniura from Lingyuan (Fig. 1). Two new snake species are newly recorded in Liaoning Province, which are Ptyas dhumnades (Fig. 2, Fig. 3) and Sibynophis chinensis (Fig. 4). And supplementary description based on an E. carinata specimen from Huludao City (Fig. 5). Up to May 2023, 37 species belonging to 21 genera and 11 families of 2 orders were recorded in Liaoning Province, including 5 species of Testudines in 4 genera, 3 families, 10 species of Lacertilia in 5 genera, 3 families, and 22 species of Serpentes in 12 genera, 5 families (Appendix 1). Reptiles include 3 species listed in category Ⅰ of wild animals under special state protection of China and 6 species listed in category Ⅱ of wild animals under special state protection of China. In addition, 15 species were evaluated as threatened species by China’s Red List of Biodiversity:Vertebrates Volume Ⅲ, Reptiles. According to the latest information on reptile distribution, Liaoning Province was divided into six zoogeographical provinces, which are the eastern hills of Liaoning, the hills of Liaodong Peninsula, Liaohe Plain, the sands and platforms of northern Liaoning, western hills of Liaoning, and the end of the Yanshan Mountains (Fig. 6, Table 1). The end of Yanshan Mountain is the newly added zoogeographical province in this study. And the former hills of the Liaodong Peninsula were expanded. [Conclusion] According to the geographical distribution of reptiles in Liaoning, the end of Yanshan Mountain has the richest species diversity of reptiles, while the sands and platforms of northern Liaoning have the least. In some parts of Liaoning, the habitat of reptiles has been destroyed, leading to a decrease in the number of them. In addition, artificially bred Pelodiscus sinensis entering the wild may damage the population of indigenous P. maackii. Therefore, we suggest establishing more natural reserves and limiting the entry of artificially bred P. sinensis into the wild to protect the diversity of reptiles in Liaoning Province.

Abstract:[Objectives] Yanbian Keelback SnakeHebius yanbianensis was described based on one specimen, after then, three specimens from Yanbian County in Sichuan Province and three from Yongren County in Yunnan Province were reported. Owing to the limited information of this species, its genetic differentiation, morphological characters, and distribution require to be further studied. [Methods] Based on 20 specimens newly identified from Sichuan and one newly collected from Yunnan, together with seven specimens mentioned above (Table 1 and Fig. 1), we constructed Maximum Likelihood (ML) and Bayesian Inference (BI) phylogenetic trees inferred from Cyt b gene sequence by IQ-TREE 1.6.12 and MrBayes 3.2.1 respectively, calculated genetic distance by uncorrected p-distance in MEGA 11, and conducted morphological measurements and scales counts.[Results] The result of molecular phylogenetic analysis inferred from Cyt b gene sequence (Table 2) showed that H. yanbianensis from Binchuan in Yunnan and Yanbian in Sichuan clustered into one lineage (SHR 99 / UFB 100 / BI 100) (Fig. 2), but the uncorrected p-distance of 1.4% was recognized between them, which is much lower than the interspecific p-distance between 4.5%﹣14.2% of Hebius (Table 3), indicating intraspecific genetic differentiation. The morphological characters of specimens from different locations are generally similar, but there are some variations:(1) total length (TaL) 48.2﹣83.6 (+) cm, tail length (TL) 12.3﹣21.4 (+) cm, tail length/total length (TL/TaL) 25.5%﹣36.2%; (2) supralabials (SL) usually 8 (7 or 8), infralabials (IL) usually 10 (8﹣11), preoculars (PreO) usually 2 (1﹣3),postoculars (PostO) 3, anterior temporals (aTem) usually 2 (1 or 2); (3) dorsal scales (DSR) 19-19-17, ventral scales (VEN) 159﹣175, subcaudal pairs (SC) 67﹣90; (4) anal 1, divided; (5) maxillary teeth (MT) 23﹣25, the last two distinctly enlarging, without diastema; (6) dark brown or tan on the back, dorsal scales inlaid with bright yellow or red-orange spots or patterns, occasionally individual’s coloration composed discontinuity dorsal stripes or faded; (7) milky white or yellow on the venter, occasionally individuals scattering stain spots, on the outer side of the ventral scale each having large black triangle spots composing strong ventrolateral series (Table 4 and Fig. 3 & 4). It can be distinguished from the other 23 Hebius species distributed in China by tail length/total length, dorsal scale rows, ventral scales, subcaudals, dorsal patterns, and ventrolateral stripe (Table 5). The geographical distribution of H. yanbianensis was found not only at Yanbian and Yongren, but also at Miyi, Huili, and Jiulong in Sichuan and Binchuan in Yunnan (Fig. 1). [Conclusion] This study provides external morphological description and conducts molecular systematics of H. yanbianensis from different populations, revealing that the morphological characters of H. yanbianensis are generally stable, and a certain genetic differentiation exists in different geographic populations.

Abstract:[Objectives] Sperm morphology is important in sperm competition and sperm evolution. The complex cloacal glands in both males and females of caudate amphibians with internal fertilization may be responsible for further packaging and modification of spermatogenesis, but little is known about the effects of these processes on sperm morphology and function. [Methods] In this study, the species Liangshantriton taliangensis which practices internal fertilization by transferring a spermatophore was studied. The morphological changes of the features of spermatozoa collected by 3 kinds of sampling methods (testes crashing, spermatophore, and female cloaca flushing) were systematically compared. Using t-test, t’-test, or Mann-Whitney U test, we analyzed the sperm morphological changes, and significant differences were set at P < 0.05. [Results] The sperm from the spermatophore and female cloaca flushing had a distinct neck, while the sperm from the testes had no such structure, but with cytoplasmic masses around the sperm head (Fig. 1). In addition, sperm shortens the acrosome and increases the proportion of sperm tail in the process of being passed out of the body by the testes (Mann-Whitney U test or t-test, P < 0.05) (Table 1), no significant change (Mann-Whitney U test or t-test, P > 0.05) in sperm morphology and measurement during the process from spermatophore to female body except for the further shortening of acrosome (t-test, P < 0.05) (Table 2). [Conclusion] These results suggest that sperm samples from the testis, spermatophore, or female ejaculatory cavity may show morphological and functional differences due to modification processes, which provide great options for the sampling method for the sperm biology or sperm conservation studies.

Abstract:[Objectives] Capitulum mitella belongs to the Arthropoda, Pollicipedidae, and Capitulum, which is similar in size to the Tortoise’s foot. C. mitella is widely distributed in subtropical and tropical coastal high tide zone, mainly distributed in the sea area south of the Yangtze Estuary. Due to the great environmental differences in the sea areas where they live, different geographical populations of C. mitella will have different environmental adaptability under multiple environmental stresses. This study for the first time applied high-throughput sequencing to C. mitella in the Yangtze River estuary offshore of Zhoushan, Zhejiang Province, the southeast coast of Ningde, Fujian Province, and Zhuhai, Guangdong Province in the South China Sea to compare transcriptomic analysis. Differentially expressed genes (DEGs) among three geographical populations of C. mitella were screened to explore how the different geographic populations adapted to the environment. [Methods] The C. mitella samples were collected from intertidal reefs in Ningde, Fujian Province (120°04′04″ E, 26°52′34″ N), Zhoushan, Zhejiang Province (122°27′04″ E, 30°43′32″ N), and Zhuhai, Guangdong Province (114°02′33″ E, 22°05′55″ N) from September to November 2021. Morphological identification and mitochondrial molecular marker cytochrome c oxidase subunit I (COI) amplification analysis showed that all of them were C. mitella.RNA was extracted for library construction and transcriptome sequencing. Clean reads were obtained after raw data quality control. Trinity (v2.4.0) was used to assemble clean reads to obtain Unigenes, and the functional cluster analysis of Unigenes was conducted. Screening of differentially expressed genes (DEGs) and significant differences in expression of the threshold for P < 0.05 and | log2FoldChange | over 1 using DESeq2 (1.6.3). To analyze the functional characteristics and physiological and biochemical metabolic pathways involved in DEGs, GO and KEGG enrichment analysis of DEGs was conducted using GOseq (1.10.0) and KOBAS (v2.0.12), and the threshold of significant enrichment in DEGs was q < 0.05. Orthologs were identified to analyze the environmental adaptability of different geographical C. mitella populations using OrthoMCL (v2.0.3). Then, the Ka/Ks values of orthologs genes were calculated by the Branch Site model. If Ka/Ks > 1, the genes were evolving rapidly; if Ka/Ks = 1, the genes were under neutral selection; if 0 < Ka/Ks < 1, the gene is selected for purification. GO and KEGG enrichment for genes under positive selection pressure, and gene function and metabolic pathway were annotated, the threshold of significant enrichment was P < 0.05. [Results] In this study, a total of 60.9 Gb of clean reads were obtained from three C. mitella populations, and 61 899 Unigenes were assembled (Table 1). A total of 651 Unigenes were significantly expressed between Zhuhai and Ningde populations, there were 3 738 DEGs between Zhoushan and Ningde populations, and 3 619 DEGs between Zhoushan and Zhuhai populations (Fig. 4). Functional annotation analysis showed that DEGs were significantly enriched in ribosome biogenesis (GO0042254, q = 8.26 × 10^﹣53) and structural constituent of ribosome (GO0003735, q = 9.14 × 10^﹣52) between Zhoushan and Zhuhai populations (Fig. 5). DEGs were further enriched in protein processing in endoplasmic reticulum (ko04141, q = 2.88 × 10^﹣4), antigen processing and presentation (ko04612, q = 1.16 × 10^﹣35) and ECM-receptor interaction (ko04512, q = 4.16 × 10^﹣2) (Fig.6). Among them, genes such as agrin (q = 1.72 × 10^﹣3) and solute carrier family 8 (q = 3.93 × 10^﹣5) may be involved in regulating osmolality balance. Compared with Zhoushan and Zhuhai populations, DEGs function involved in antigen processing and presentation (ko04612) and protein processing in endoplasmic reticulum (ko04141), among which cathepsin B (q = 6.36 × 10^﹣3) and high temperature protein G (q = 9.97 × 10^﹣6) were significantly expressed. Heat shock protein family, including HSP70 (q = 1.45 × 10^﹣6) and HSP40 (q = 4.58 × 10^﹣7) were also significantly expressed in the Zhoushan population (Table 2). Analysis of positive selection pressure among different geographical populations by orthologs screening showed that 7 genes under positive selection pressure were obtained from 254 orthologs including D-aspartate oxidase (Ka/Ks = 2.076), histone deacetylase (Ka/Ks = 1.145), baculoviral IAP resting-containing protein (Ka/Ks = 1.008), GTP-binding nuclear protein Ran (Ka/Ks = 1.001), mediator of RNA polymerase II transcription subunit (Ka/Ks = 2.867), translation initiation factor 2B (Ka/Ks = 1.001), and pyrroline-5-carboxylate reductase (Ka/Ks = 1.131) (Table 3). They are all involved in growth hormone, reproductive, nutritional, nervous, immune system regulation, and neuronal protection. They also participate in physiological processes such as growth and development, tissue differentiation, metabolism, and anti-apoptosis. [Conclusion] This study will help to understand the adaptation mechanism of C. mitella to multiple external environments and provide a basic foundation of genetic diversity, germplasm resources protection, and the development of different geographical populations of C. mitella.

Abstract:[Objectives] This paper reported a new geographical record of the genus Dactylogyrus on the gills of Rutilus rutilus lacustris from Irtysh River (Haba River section) investigation of fish parasites in July 2021. [Methods] Based on careful morphological comparison with those relevant species, significant differences in the copulatory complex and the opisthaptor of the parasite were observed. [Results] In copulatory complex, the parasite has an “8” structure at the base of the tube. Then the diameter of the tube is uniform. The tube extended in a curved shape resembling “6”. About the opisthaptor, the parasite’s inner root of anchor was longer than the length of the outer root, with seven pairs of small marginal hooks. And this Dactylogyrus hasthe “一”-shaped dorsal bar and a butterfly-shaped ventral bar (Fig. 1, 2). The characteristic structure of this parasite differs from other Dactylogyrus that parasitic Rutilus rutiluslacustris in China (Appendix 2). The parasite in this study is small. Compared with D. caballeroi, the morphology of the opisthaptor of the two insects was most similar, but the morphology of the copulatory complex of the two was very different. In comparison with the rest of Dactylogyrus spp, the parasite has a butterfly-shaped ventral bar, D. rarissimus has an inverted “T”-shaped ventral bar, and D. rutili has a “十”-shaped ventral bar. However, D. sphyrna and D. varius have no ventral bar. In the dorsal bar morphology, D. crucifer showed a “一”-shape, while the dorsal bar of D. varius had a “V” shape. After comparison, the morphology of the parasite was highly similar to that of D. crucifer, recorded in Russia. The Bayesian inference and maximum likelihood trees constructed based on the 28S rDNA gene fragments showed that the D. crucifer in this study clustered with D. crucifer in Czech Republic (100/100) (Fig. 3). [Conclusion] The results were combined and the insect was identified as D. crucifer Wagener, 1857, a new record species of Dactylogyrus in China.

Abstract:[Objectives] The research on mammalian sexual reproduction is beneficial to the exploration of human reproduction mechanism, which focuses on reproductive system, germ cells, fertilization cells, external environmental factors, etc. There is little research on mammalian fertility after the elimination of metabolic enzymes from the body. It is known that aldehyde dehydrogenase 2 (ALDH2) is the most important enzyme for aldehyde oxidation, which can metabolize a large number of harmful aldehydes produced in the body. Studies have shown that a large number of reactive oxygen species produced by male sperm during exercise can increase the aldehyde content in sperm. Therefore, we studied the fertility of ALDH2 gene knockout male mice. [Methods] In this study, we analyze the testicular organ coefficient, testicular tissue cell morphology, sperm motility parameters, the number of offspring born after mating, and the ratio of male to female offspring of 52 male C57BL/6 mice of different weeks of age with ALDH2 gene knockout. Mean value comparison, one-way ANOVA, and t-test implemented in SPSS were used to explore the effect of ALDH2 gene knockout on fertility of male mice. [Results] We found that compared with the wild type (WT) mice, the weight and testicular organ coefficient of knockout (KO) mice at 5 and 10 weeks of age and mice at 3 and 10 weeks of age had extremely significant differences (n = 6, P < 0.01) (Table 1), the weight of mice at 3 and 7 weeks of age and testicular organ coefficient of mice at 5 and 7 weeks of age had significant differences (n = 6, P < 0.05) (Table 1). Among them, the cytoplasm of testicular tissue of 10 week old ALDH2 gene knockout male mice became larger (Fig. 2b); the sperm activity rate of knockout mice was significantly reduced (n = 6, P < 0.05) (Table 2); the litter size of ALDH2 transgenic mice was significantly reduced (6.7 ± 0.5) compared with wild type mice (11.0 ± 2.0, n = 3, P < 0.05), and the female to male ratio of ALDH2 gene knockout mice was also significantly reduced (0.8 ± 0.1 vs. 1.8 ± 0.5, n = 3, P < 0.05). [Conclusion] This study provides a basis for revealing the role of ALDH2 gene in male mouse reproduction and provides valuable information for the potential diagnosis and in vitro application of assisted reproductive technology.

Abstract:[Objectives] In order to study the effects of different doses of pregnant mare serum gonadotropin (PMSG) on ovarian morphology and early embryo development in vitro after superovulation in rabbits (Oryctolagus cuniculus f. domesticus). [Methods] Twenty-four rabbits were randomly divided into four groups, with 6 rabbits in each group. The control group was not treated. The latter three groups were injected with 50 IU, 70 IU and 90 IU PMSG , and 100 IU human chorionic gonadotropin (HCG) for superovulation. After dissection, the morphology and tissue parameters of ovaries were measured. Embryos were flushed from the left and right Fallopian tubes into Petri dishes by using 5 ml washing solution and then transferred to the cell culture room for in vitro culture and observation. The results were expressed as “Mean ± standard deviation, and the data were analyzed by one-way ANOVA with SPSS 25.0. P < 0.01 means the difference is extremely significant, P < 0.05 means the difference is significant, and P ≥ 0.05 means the difference is not significant. [Results] The results showed that compared with the control group, the weight, width, and thickness of ovaries in the PMSG-treated group increased extremely significantly (P < 0.01), the length of ovaries increased significantly (P < 0.05), the surface of ovaries became congested, and the number of follicles increased with the increase of PMSG injection dose (Fig. 1, Table 1). In addition, with the increase of PMSG injection dose, the diameters of secondary follicles and their enclosed oocytes, and the diameter of tertiary follicles decreased (P < 0.01), while the diameter of oocytes in tertiary follicles increased, with a significant difference compared with the control group (P < 0.05, Fig. 3, Table 3). The diameter of tertiary follicles and the dominance rate of follicles in the 70 IU group were significantly higher than those in the control group, 50 IU group, and 90 IU group (P < 0.05, Table 4). When the embryos were cultured in vitro, compared with the control group, PMSG treated group had a significantly higher embryo deformity rate (P < 0.01), significantly lower 2-cell rate (P < 0.01), and significantly lower 4-cell rate (P < 0.05, Fig. 2, Table 2) due to the increase of PMSG injection dose. [Conclusion] 90 IU PMSG superovulation can cause adverse effects, while 70 IU PMSG can increase the numbers of follicles at all levels and the dominance rate, and promote the development of oocytes in tertiary follicles, but reduce the diameters of follicles at all levels.

Abstract:During the investigation on amphibians in October 2022, 5 female specimens of Rana (Anura:Ranidae) were collected from Xianliang Town, Qingyuan County, Zhejiang Province, China. We compared their features by morphology and phylogeny and identified them as R. longicrus. This represents the first record of this species in Zhejiang Province. 5 female samples collected from Qingyuan County were measured with electronic vernier calipers. The measurements were:snout-vent length, head length, head width, snout length, internasal space, interorbital space, diameter of eye, length of lower arm and hand, tibia length, foot length, and hindlimb length. We compared these morphological characteristics with the specimens from Fujian, Jiangxi, Guangdong, and Taiwan, China. In this study, phenol/chloroform extraction was used to extract genomic DNA from the hindlimb muscle tissue of Brown Frogs collected in Qingyuan County, Zhejiang Province. 16S rRNA gene was amplified, and the primers were 5′-CGC CTG TTT ACC AAA AAC AT-3′ and 5′-CCG GTC TGA ACT CAG ATC ACG T-3′. Based on 16S rRNA gene fragments, we used Mrbayes-3.1.2 to construct Bayesian tree for phylogenetic analysis and used p-distance in MEGA 7 to calculate the genetic distance between species of Rana. All the new sequences in this research have been uploaded to the GenBank database. The 16S rRNA sequences of 16 species of Rana distributed in East China and Glandirana tientaiensis were obtained from GenBank (Table 2).The morphological characteristics are consistent between the specimens collected in this study and the topotype of R. longicrus. Our specimens have a medium snout-vent length (40.4 to 55.2 mm); head length is larger than the head width; some sparse granules can form a V-shaped figure on the shoulder; dorsolateral fold obvious and connect with the crotch; arms and legs with dark brown bands; tibiotarsal articulation beyond snout. In addition, the Bayesian analyses based on 16S rRNA strongly support the specimens we collected form a monophyletic clade with the topotype specimens (Fig 2), and the genetic distance is less than 0.005% between our specimens and topotype (Table 3).The specimens collected in Qingyuan County are determined to be R. longicrus, which is the first record of this species in Zhejiang Province. In this study, we further updated the distribution range of R. longicrus.

Abstract:An unidentified snake specimen was collected from a heap of sticks and dry leaves in Xiangyindu Town, Yongxing County, Hunan Province, China (113°01′18″ E，26°11′87″ N, 174 m) during a biodiversity survey in early August 2022. Morphological characteristics comparison revealed that the collected specimen was generally consistent with the morphological characteristics of the Calamariaarcana from Guangdong Province, China. Bayesian and maximum likelihood phylogenetic analyses based on the mitochondrial gene fragment Cyt b showed a high support monophyletic group between the collected specimen with the C. arcana (PP 1.00, BS 100), and the uncorrected pairwise sequence divergence was 1%. Thus, based on morphological analysis and phylogenetic comparison, this specimen was finally identified as C. arcana, which is a new snake record in Hunan Province.

Abstract:During a field investigation in Xintian Town, Pengshui Miao, and Tujia Autonomous County, Chongqing in July 2022, two Odorrana specimens were found (Fig. 1). The purpose of this study is to identify the species using morphological and phylogenetic methods, as well as to ascertain its distribution in Chongqing.A comparative analysis of the morphological characteristics of the species discovered from Chongqing and O. wuchuanensis from Guizhou and Hubei was conducted. For phylogenetic analyses, mitochondrial 16S rRNA fragments were amplified, and the 16S rRNA gene information of 11 Odorrana species from China and abroad was obtained from the GenBank database (Table 1). Corresponding sequences of one Pseudorana weiningensis and one Nidirana daunchina were also downloaded and used as outgroups. The maximum likelihood phylogenetic tree was constructed in IQ-TREE, the Bayesian tree construction was carried out in MrBayes 3.2.1, and the best-fit model was selected according to the Bayesian inference criteria (BIC) computed with JModelTest 0.1.1. Morphological characteristics appear to differ little between those specimens of Chongqing, Guizhou, and Hubei (Fig. 2). The snout vent length of the male species discovered in Pengshui was 81.3 mm, which was longer than the maximum snout vent length of the type specimen and O. wuchuanensis from Jianshi County, Hubei Province. The female of the species had a snout vent length of 87.1 mm, which was within the range of the type specimen’s body snout vent length but smaller than the female O. wuchuanensis from the Jianshi County (Table 2). Genetic analysis based on the mitochondrial 16S rRNA gene revealed that the specimens found in Pengshui County of Chongqing clustered with O. wuchuanensis from Wuchuan County of Guizhou Province with a high support (Fig. 3). The genetic distance between O. wuchuanensis in Wuchuan County, Guizhou Province, and the specimens found in Pengshui County is 0 (Table 3). According to a thorough comparison with morphological analysis and phylogenetic analysis, the specimens discovered in Pengshui were identified as O. wuchuanensis, which is a new record of amphibian distribution in Chongqing.

Abstract:[Objectives] Plateau Pika (Ochotona curzoniae) is an endemic and key species in the alpine meadow ecosystem, and its behaviors of digging, foraging, and moving directly affect the health and stability of the alpine meadow. However, it is difficult to achieve continuous behavioral observation for several days to obtain all the data required for the biological rhythm observation of Plateau Pika. Due to the shortcoming of the traditional method,a Plateau Pika monitoring system is constructed by integrating image automatic acquisition, optimizing digital image and processing algorithm, and intelligent recognition technology, which realize non-contact, continuous, and digital monitoring of Plateau Pika. [Methods] According to the need for Plateau Pika activity rhythm monitoring, the experimental layout was designed with an area of 25 m × 30 m in Henan Mongolian Autonomous County, Qinghai Province. The Plateau Pika monitoring system is designed, and the optimized and improved frame difference method is proposed. [Results] Through the monitoring of the experimental area, the video image data of the recorded Plateau Pika is obtained. The Plateau Pika monitoring system meets the basic application requirements of observing Plateau Pika activities in the natural environment. The improved frame difference method is used to realize the detection of moving targets based on the complex image. The average accuracy rate, recall rate, and weighted average were 87.20%, 83.37%, and 0.85 (Table 2). The peak of ground activity of Plateau Pika appeared at 10:00 and 16:00, and it was inactive at night (Fig. 6), so it belonged to diurnal animals. [Conclusion] These results can meet the target detection requirements of Plateau Pika’s monitoring system, recording the activity rhythm and frequency, which provides new research ideas and effective technical means for Plateau Pika behavior research.

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