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A new species of Crocodile Newt, genus Tylototriton (Amphibia, Caudata, Salamandridae) from the mountains of Kachin State, northern Myanmar
Than Zaw, Paw Lay, Parinya Pawangkhanant, JVladislav A. Gorin, Nikolay A. Poyarkov, Jr.
Zoological Research    2019, 40 (3): 151-174.   DOI: 10.24272/j.issn.2095-8137.2019.043
Online available: 28 March 2019

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We describe a new species of the genus Tylototriton from Ingyin Taung Mt., Mohnyin Township, Kachin State, Myanmar, based on morphological and molecular evidence. The new species is assigned to the subgenus Tylototriton s. str. and is clearly distinct from all known congeners by the following characters: medium body size; thin, long tail, lacking lateral grooves; rough skin; truncate snout; wide, protruding supratemporal bony ridges on head, beginning at anterior corner of orbit; weak, almost indistinct sagittal ridge; long, thin limbs, broadly overlapping when adpressed along body; distinct, wide, non-segmented vertebral ridge; 13 or 14 rib nodules; brown to dark-brown background coloration with dull orange-brown to yellowish-brown markings on labial regions, parotoids, rib nodules, whole limbs, vent, and ventral tail ridge. We also briefly discuss biogeography and species diversity of the genus Tylototriton in Myanmar.

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Current understanding on the roles of gut microbiota in fish disease and immunity
Jin-Bo Xiong, Li Nie, Jiong Chen
Zoological Research    2019, 40 (2): 70-76.   DOI: 10.24272/j.issn.2095-8137.2018.069
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Intensive aquaculture has increased the severity and frequency of fish diseases. Given the functional importance of gut microbiota in various facets of host physiology, modulation of this microbiota is a feasible strategy to mitigate emerging diseases in aquaculture. To achieve this, a fundamental understanding of the interplay among fish health, microbiota, and invading pathogens is required. This mini-review focuses on current knowledge regarding the associations between fish diseases, dysbiosis of gut microbiota, and immune responses. Furthermore, updated research on fish disease from an ecological perspective is discussed, including colonization resistance imposed by commensals and strategies used by pathogens to overcome resistance. We also propose several directions for future research, such as exploration of the causal links between fish diseases and specific taxa, and identification of universal gut microbial biomarkers for rapid disease diagnosis.
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Involvement of ayu NOD2 in NF-κB and MAPK signaling pathways: Insights into functional conservation of NOD2 in antibacterial innate immunity
Yi Ren, Shui-Fang Liu, Li Nie, Shi-Yu Cai, Jiong Chen
Zoological Research    2019, 40 (2): 77-88.   DOI: 10.24272/j.issn.2095-8137.2018.066
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Nucleotide oligomerization domain 2 (NOD2) is a major cytoplasmic sensor for pathogens and is critical for the clearance of cytosolic bacteria in mammals. However, studies regarding NOD2, especially the initiated signaling pathways, are scarce in teleost species. In this study, we identified a NOD2 molecule (PaNOD2) from ayu (Plecoglossus altivelis). Bioinformatics analysis showed the structure of NOD2 to be highly conserved during vertebrate evolution. Dual-luciferase reporter assays examined the activation of NF-κB signaling and Western blotting analysis detected the phosphorylation of three MAP kinases (p-38, Erk1/2, and JNK1/2). Functional study revealed that, like its mammalian counterparts, PaNOD2 was the receptor of the bacterial cell wall component muramyl dipeptide (MDP), and the leucine-rich repeat motif was responsible for the recognition and binding of PaNOD2 with the ligand. Overexpression of PaNOD2 activated the NF-κB signaling pathway, leading to the upregulation of inflammatory cytokines, including TNF-α and IL-1β in HEK293T cells and ayu head kidney-derived monocytes/macrophages (MO/MΦ). Particularly, we found that PaNOD2 activated the MAPK signaling pathways, as indicated by the increased phosphorylation of p-38, Erk1/2, and JNK1/2, which have not been characterized in any teleost species previously. Our findings proved that the NOD2 molecule and initiated pathways are conserved between mammals and ayu. Therefore, ayu could be used as an animal model to investigate NOD2-based diseases and therapeutic applications.
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Metagenomic comparison of the rectal microbiota between rhesus macaques (Macaca mulatta) and cynomolgus macaques (Macaca fascicularis)
Yan-Fang Cui, Feng-Jie Wang, Lei Yu, Hua-Hu Ye, Gui-Bo Yang
Zoological Research    2019, 40 (2): 89-93.   DOI: 10.24272/j.issn.2095-8137.2018.061
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Rhesus macaques (Macaca mulatta) and cynomolgus macaques (Macaca fascicularis) are frequently used in establishing animal models for human diseases. To determine the differences in gut microbiota between these species, rectal swabs from 20 rhesus macaques and 21 cynomolgus macaques were collected, and the microbial composition was examined by deep sequencing of the 16S rRNA gene. We found that the rectal microbiota of cynomolgus macaques exhibited significantly higher alpha diversity than that of rhesus macaques, although the observed number of operational taxonomic units (OTUs) was almost the same. The dominant taxa at both the phylum and genus levels were similar between the two species, although the relative abundances of these dominant taxa were significantly different between them. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) showed significant differences in the functional components between the microbiota of the two species, in particular the lipopolysaccharide (LPS) synthesis proteins. The above data indicated significant differences in microbial composition and function between these two closely related macaque species, which should be taken into consideration in the future selection of these animals for disease models.
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Taxonomic revision of the genus Mesechinus (Mammalia: Erinaceidae) with description of a new species
Huai-Sen Ai, Kai He, Zhong-Zheng Chen, Jia-Qi Li, Tao Wan, Quan Li, Wen-Hui Nie, Jin-Huan Wang, Wei-Ting Su, Xue-Long Jiang
Zoological Research    2018, 39 (5): 335-347.   DOI: 10.24272/j.issn.2095-8137.2018.034
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Hedgehogs in the genus Mesechinus (Family Erinaceidae), which include two currently recognized species (M. dauuricus and M. hughi), are distributed from northeast Mongolia to the upper Amur Basin in Russia and adjacent areas in northeast and northern China. In recent years, a population of Mesechinus hedgehogs was discovered from Mt. Gaoligong, southwestern Yunnan, China, far from the known distribution range of the genus. Furthermore, these hedgehogs are the only known population to be distributed at elevations higher than 2 100 m and in sympatry with gymnures. To evaluate the taxonomic status of these hedgehogs, we examined specimens representing Mesechinus taxa in China and further conducted morphometric and karyotypic analyses. Our results supported the existence of four species in China. Specifically, we identified the hedgehogs from Mt. Gaoligong as a new species, Mesechinus wangi sp. nov., and recognized M. miodon, previously considered as a synonym of either M. dauuricus or M. hughi, as a distinct species. Interestingly, we observed a supernumerary M4 on all specimens of Mesechinus wangi sp. nov., which is an extremely rare event in the evolution of mammalian dentition.
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Assessment of habitat suitability of the snow leopard (Panthera uncia) in Qomolangma National Nature Reserve based on MaxEnt modeling
De-Feng Bai, Peng-Ju Chen, Luciano Atzeni, Lhaba Cering, Qian Li, Kun Shi
Zoological Research    2018, 39 (6): 373-386.   DOI: 10.24272/j.issn.2095-8137.2018.057
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Habitat evaluation constitutes an important and fundamental step in the management of wildlife populations and conservation policy planning. Geographic information system (GIS) and species presence data provide the means by which such evaluation can be done. Maximum Entropy (MaxEnt) is widely used in habitat suitability modeling due to its power of accuracy and additional descriptive properties. To survey snow leopard populations in Qomolangma (Mt. Everest, QNNR) National Nature Reserve, Tibet, China, we pooled 127 pugmarks, 415 scrape marks, and 127 non-invasive identifications of the animal along line transects and recorded 87 occurrences through camera traps from 2014–2017. We adopted the MaxEnt model to generate a map highlighting the extent of suitable snow leopard habitat in QNNR. Results showed that the accuracy of the MaxEnt model was excellent (mean AUC=0.921). Precipitation in the driest quarter, ruggedness, elevation, maximum temperature of the warmest month, and annual mean temperature were the main environmental factors influencing habitat suitability for snow leopards, with contribution rates of 20.0%, 14.4%, 13.3%, 8.7%, and 8.2% respectively. The suitable habitat area extended for 7001.93 km2, representing 22.72% of the whole reserve. The regions bordering Nepal were the main suitable snow leopard habitats and consisted of three separate habitat patches. Our findings revealed that precipitation, temperature conditions, ruggedness, and elevations of around 4000 m influenced snow leopard preferences at the landscape level in QNNR. We advocate further research and cooperation with Nepal to evaluate habitat connectivity and to explore possible proxies of population isolation among these patches. Furthermore, evaluation of subdivisions within the protection zones of QNNR is necessary to improve conservation strategies and enhance protection. 
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Specific function and modulation of teleost monocytes/macrophages: polarization and phagocytosis
Xin-Jiang Lu, Jiong Chen
Zoological Research    2019, 40 (3): 146-150.   DOI: 10.24272/j.issn.2095-8137.2019.035
Online available: 28 March 2019

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Macrophages exist in most tissues and play a variety of functions in vertebrates. Teleost fish species are found in most aquatic environments throughout the world and are quite diverse for a group of vertebrate animals. Due to whole genome duplication and environmental adaptation, teleost monocytes/macrophages possess a variety of different functions and modulations compared with those of mammals. A deeper understanding of teleost monocytes/macrophages in the immune system will not only help develop teleost-specific methods of disease prevention but will also help improve our understanding of the various immune mechanisms in mammals. In this review, we summarize the differences in polarization and phagocytosis of teleost and mammalian macrophages to improve our understanding of the various immune mechanisms in vertebrates.

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An annotated checklist of mammals of Kenya
Simon Musila, Ara Monadjem, Paul W. Webala, Bruce D. Patterson, Rainer Hutterer, Yvonne A. De Jong, Thomas M. Butynski, Geoffrey Mwangi, Zhong-Zheng Chen, Xue-Long Jiang
Zoological Research    2019, 40 (1): 3-52.   DOI: 10.24272/j.issn.2095-8137.2018.059
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Kenya has a rich mammalian fauna. We reviewed recently published books and papers including the six volumes of Mammals of Africa to develop an up-to-date annotated checklist of all mammals recorded from Kenya. A total of 390 species have been identified in the country, including 106 species of rodents, 104 species of bats, 63 species of even-toed ungulates (including whales and dolphins), 36 species of insectivores and carnivores, 19 species of primates, five species of elephant shrews, four species of hyraxes and odd-toed ungulates, three species of afrosoricids, pangolins, and hares, and one species of aardvark, elephant, sirenian and hedgehog. The number of species in this checklist is expected to increase with additional surveys and as the taxonomic status of small mammals (e.g., bats, shrews and rodents) becomes better understood.
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Effects of age, sex and manual task on hand preference in wild Rhinopithecus roxellana
Wei-Wei Fu, Xiao-Wei Wang, Cheng-Liang Wang, Hai-Tao Zhao, Yi Ren, Bao-Guo Li
Zoological Research    2019, 40 (2): 129-138.   DOI: 10.24272/j.issn.2095-8137.2019.023
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Golden snub-nosed monkeys (Rhinopithecus roxellana), as typical arboreal group-living Old World monkeys, provide an appropriate animal model to research manual laterality and explore the factors affecting hand preference in nonhuman primates. This study investigated hand preference based on 63 subjects and four spontaneous manual tasks (including unimanual and bimanual feeding and grooming), and assessed the effects of age, gender and type of task on handedness in R. roxellana. A population-level left-handedness was found not only in the bimanual coordinated tasks (bimanual feeding and grooming), but also in one unimanual reaching task (unimanual feeding). There were no significant differences between the sexes in either direction or strength of hand preference among any task. However, a significant difference between adults and juveniles was found in the unimanual feeding task. This is the first report on handedness in unimanual and bimanual feeding tasks that require bipedal posture in wild R. roxellana. Furthermore, this study demonstrated spontaneous feeding tasks reported previously only in the quadrupedal posture in this species, supporting the importance of factors such as posture and task complexity in the evolution of primate manual lateralization.
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A transgenic monkey model for the study of human brain evolution
Lei Shi, Bing Su
Zoological Research    2019, 40 (3): 236-238.   DOI: 10.24272/j.issn.2095-8137.2019.031
Online available: 11 April 2019

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Why humans have large brains with higher cognitive abilities is a question long asked by scientists. However, much remains unknown, especially the underlying genetic mechanisms. With the use of a transgenic monkey model, we showed that human-specific sequence changes of a key brain development gene (primary microcephaly1, MCPH1) could result in detectable molecular and cognitive changes resembling human neoteny, a notable characteristic developed during human evolution.

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Marker-assisted selection of YY supermales from a genetically improved farmed tilapia-derived strain
Chao-Hao Chen, Bi-Jun Li, Xiao-Hui Gu, Hao-Ran Lin, Jun-Hong Xia
Zoological Research    2019, 40 (2): 108-112.   DOI: 10.24272/j.issn.2095-8137.2018.071
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Genetically improved farmed tilapia (GIFT) and GIFT-derived strains account for the majority of farmed tilapia worldwide. As male tilapias grow much faster than females, they are often considered more desirable in the aquacultural industry. Sex reversal of females to males using the male sex hormone 17-α-methyltestosterone (MT) is generally used to induce phenotypic males during large-scale production of all male fingerlings. However, the widespread use of large quantities of sex reversal hormone in hatcheries may pose a health risk to workers and ecological threats to surrounding environments. Breeding procedures to produce genetically all-male tilapia with limited or no use of sex hormones are therefore urgently needed. In this study, by applying marker-assisted selection (MAS) for the selection of YY supermales from a GIFT-derived strain, we identified 24 XY pseudofemale and 431 YY supermale tilapias. Further performance evaluation on the progenies of the YY supermales resulted in male rates of 94.1%, 99.5% and 99.6%, respectively, in three populations, and a daily increase in body weight of 1.4 g at 3 months (n=997). Our study established a highly effective MAS procedure in the selection of YY supermales from a GIFT-derived strain. Furthermore, the development of MAS-selected YY supermales will help reduce the utilization of hormones for controlling sex in the tilapia aquaculture.
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Depressed female cynomolgus monkeys (Macaca fascicularis) display a higher second-to-fourth (2D:4D) digit ratio
Wei Li, Ling-Yun Luo, Xun Yang, Yong He, Bin Lian, Chao-Hua Qu, Qing-Yuan Wu, Jian-Guo Zhang, Peng Xie
Zoological Research    2019, 40 (3): 219-225.   DOI: 10.24272/j.issn.2095-8137.2019.022
Online available: 28 March 2019

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This research aimed to provide evidence of a relationship between digit ratio and depression status in the cynomolgus monkey (Macaca fascicularis). In stable cynomolgus monkey social groups, we selected 15 depressed monkeys based on depressive-like behavioral criteria and 16 normal control monkeys. All animals were video recorded for two weeks, with the duration and frequency of the core depressive behaviors and 58 other behaviors in 12 behavioral categories then evaluated via behavioral analysis. Finger lengths from the right and left forelimb hands of both groups were measured by X-ray imaging. Finger length and digit ratio comparisons between the two groups were conducted using Student’s t-test. In terms of the duration of each behavior, significant differences emerged in “Huddling” and five other behavioral categories, including Ingestive, Amicable, Parental, Locomotive, and Resting. In addition to the above five behavioral categories, we found that depressed monkeys spent less time in parental and rubbing ‘ and forth behaviors than the control group. Furthermore, the 4th fingers were significantly longer in the left and right hands in the control group relative to the depressed monkeys. The second-to-fourth (2D:4D) digit ratio in the left and right forelimb hands was significantly lower in the control group than that in the depressed group. Our findings revealed significant differences in finger lengths and digit ratios between depressed monkeys and healthy controls, which concords with our view that relatively high fetal testosterone exposure may be a protective factor against developing depressive symptoms (or that low fetal testosterone exposure is a risk factor).

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Developmental expression of three prmt genes in Xenopus
Cheng-Dong Wang, Xiao-Fang Guo, Thomas Chi Bun Wong, Hui Wang, Xu-Feng Qi, Dong-Qing Cai, Yi Deng, Hui Zhao
Zoological Research    2019, 40 (2): 102-107.   DOI: 10.24272/j.issn.2095-8137.2018.064
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Protein arginine methyltransferases (PRMTs) are involved in many cellular processes via the arginine methylation of histone or non-histone proteins. We examined the expression patterns of prmt4, prmt7, and prmt9 during embryogenesis in Xenopus using whole-mount in situ hybridization and quantitative reverse transcription polymerase chain reaction (RT-PCR). Xenopus prmt4 and prmt7 were expressed in the neural crest, brain, and spinal cord, and also detected in the eye, branchial arches, and heart at the tailbud stage. Specific prmt9 signals were not detected in Xenopus embryos until the late tailbud stage when weak expression was observed in the branchial arches. Quantitative RT-PCR indicated that the expressions of prmt4 and prmt7 were up-regulated during the neurula stage, whereas prmt9 maintained its low expression until the late tailbud stage, consistent with the whole-mount in situ hybridization results. Thus, the developmental expression patterns of these three prmt genes in Xenopus embryos provide a basis for further functional study of such genes.
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Identification and characterization of two novel cathelicidins from the frog Odorrana livida
Ruo-Han Qi, Yan Chen, Zhi-Lai Guo, Fen Zhang, Zheng Fang, Kai Huang, Hai-Ning Yu, Yi-Peng Wang
Zoological Research    2019, 40 (2): 94-101.   DOI: 10.24272/j.issn.2095-8137.2018.062
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Antimicrobial peptides (AMPs) are a group of gene-encoded small peptides that play pivotal roles in the host immune system of multicellular organisms. Cathelicidins are an important family of AMPs that exclusively exist in vertebrates. Many cathelicidins have been identified from mammals, birds, reptiles and fish. To date, however, cathelicidins from amphibians are poorly understood. In the present study, two novel cathelicidins (OL-CATH1 and 2) were identified and studied from the odorous frog Odorrana livida. Firstly, the cDNAs encoding the OL-CATHs (780 and 735 bp in length, respectively) were successfully cloned from a lung cDNA library constructed for the frog. Multi-sequence alignment was carried out to analyze differences between the precursors of the OL-CATHs and other representative cathelicidins. Mature peptide sequences of OL-CATH1 and 2 were predicted (33 amino acid residues) and their secondary structures were determined (OL-CATH1 showed a random-coil conformation and OL-CATH2 demonstrated a-helical conformation). Furthermore, OL-CATH1 and 2 were chemically synthesized and their in vitro functions were determined. Antimicrobial and bacterial killing kinetic analyses indicated that OL-CATH2 demonstrated relatively moderate and rapid antimicrobial potency and exhibited strong anti-inflammatory activity. At very low concentrations (10 μg/mL), OL-CATH2 significantly inhibited the lipopolysaccharide (LPS)-induced transcription and production of pro-inflammatory cytokines TNF-a, IL-1b and IL-6 in mouse peritoneal macrophages. In contrast, OL-CATH1 did not exhibit any detectable antimicrobial or anti-inflammatory activities. Overall, identification of these OL-CATHs from O. livida enriches our understanding of the functions of cathelicidins in the amphibian immune system. The potent antimicrobial and anti-inflammatory activities of OL-CATH2 highlight its potential as a novel candidate in anti-infective drug development.
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Diversity and distribution patterns of non-volant small mammals along different elevation gradients on Mt. Kenya, Kenya
Simon Musila, Zhong-Zheng Chen, Quan Li, Richard Yego, Bin Zhang, Kenneth Onditi, Immaculate Muthoni, Shui-Wang He, Samson Omondi, James Mathenge, Esther N. Kioko, Xue-Long Jiang
Zoological Research    2019, 40 (1): 53-60.   DOI: 10.24272/j.issn.2095-8137.2019.004
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The distribution of small mammals in mountainous environments across different elevations can provide important information on the effects of climate change on the dispersal of species. However, few studies conducted on Afromontane ecosystems have compared the altitudinal patterns of small mammal diversity. We investigated the species diversity and abundance of non-volant small mammals (hereafter ‘small mammals’) on Mt. Kenya, the second tallest mountain in Africa, using a standard sampling scheme. Nine sampling transects were established at intervals of 200 m on the eastern (Chogoria) and western (Sirimon) slopes. A total of 1 905 individuals representing 25 species of small mammals were trapped after 12 240 trap-nights. Abundance was highest at mid-elevations on both slopes. However, species richness and distribution patterns differed between the two slopes. More species were recorded on Chogoria (24) than on Sirimon (17). On Chogoria, species richness was higher at mid-high elevations, with a peak at mid-elevation (2 800 m a.s.l.), whereas species richness showed little variation on the Sirimon slope. These results indicate that patterns of species diversity can differ between slopes on the same mountain. In addition, we extensively reviewed literature on Mt. Kenya’s mammals and compiled a comprehensive checklist of 76 mammalian species. However, additional research is required to improve our understanding of small mammal diversity in mountain habitats in Africa.
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Social functions of relaxed open-mouth display in golden snub-nosed monkeys (Rhinopithecus roxellana)
You-Ji Zhang, Yi-Xin Chen, Hao-Chun Chen, Yuan Chen, Hui Yao, Wan-Ji Yang, Xiang-Dong Ruan, Zuo-Fu Xiang
Zoological Research    2019, 40 (2): 113-120.   DOI: 10.24272/j.issn.2095-8137.2018.043
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Relaxed open-mouth display serves important social functions in relation to submission, reconciliation, affiliation and reassurance among non-human primate societies; however, quantitative evidence on this behavior remains insufficient among multi-level social groups. From July to November 2016, we examined four potential functions of the relaxed open-mouth display during pairwise, intra-unit social interactions among 18 free-ranging adult and sub-adult golden snub-nosed monkeys (Rhinopithecus roxellana) who belonged to three one-male, multi-female units (OMU) at Dalongtan, Shennongjia National Park, China. Results showed that: compared with no relaxed open-mouth display, (1) the occurrence of displacement by a dominant individual approaching a subordinate was lower and the distance of the subordinate to the approaching dominant was shorter when the subordinate showed open-mouth display; (2) relaxed open-mouth display reduced the probability of continued attack for victims of aggression and allowed victims to achieve closer proximity to the aggressor during post-conflict periods; (3) relaxed open-mouth display by dominant individuals allowed them to achieve closer proximity to subordinates; and (4) the exchange of relaxed open-mouth display had a greater impact on the outcome of interactions than one individual alone giving this signal. These findings suggest that relaxed open-mouth display serves important functions regarding submission, reconciliation, affiliation and reassurance in coordinating social interactions within OMUs in golden snub-nosed monkeys.
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A new species of the endemic Himalayan genus Liurana (Anura, Ceratobatrachidae) from southeastern Tibet, China, with comments on the distribution, reproductive biology, and conservation of the genus
Ke Jiang, Kai Wang, Yu-Fan Wang, Cheng Li, Jing Che
Zoological Research    2019, 40 (3): 175-184.   DOI: 10.24272/j.issn.2095-8137.2019.025
Online available: 20 March 2019

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A new species of the genus Liurana Dubois, 1986 is described from Medog County, Tibet, China, based on morphological and molecular data. The new species can be differentiated from all other congeners by the following combination of characters: (1) head wider than long; (2) tympanum distinct and large; (3) hindlimb long, tibiotarsal articulation beyond tip of snout when adpressed; (4) belly with flat tubercles, cloacal region with small tubercles; (5) transverse bands distinctly on dorsal limbs, four bands on thigh and three on tibia; and, (6) dark brown marbled patterns or speckles on white belly. Here, we also discuss the distribution pattern of Liurana in the East Himalaya region, the role of the Yarlung Tsangpo River in the speciation and genetic isolation of congeners, the direct developmental mode of reproduction, and the two different ecotypes of the genus. Lastly, we provide conservation recommendations for the genus in southeastern Tibet.
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Small mammal diversity of Mt. Kenya based on carnivore fecal and surface bone remains
Ogeto Mwebi, Esther Nguta, Veronica Onduso, Ben Nyakundi, Xue-Long Jiang, Esther N. Kioko
Zoological Research    2019, 40 (1): 61-69.   DOI: 10.24272/j.issn.2095-8137.2018.055
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Ecological dynamics and faunal diversity documentation is normally conducted by direct observation and trapping of live animals. However, surveys of carnivore scat prey and surface bone remains, which are relatively inexpensive, can provide complementary data that expand carnivore diet breadth and may improve accuracy regarding inferences of the ecological dynamics of a given ecosystem. We used this inexpensive method to document species diversity variation with elevation on the leeward (Sirimon) and windward (Chogoria) areas of Mt. Kenya. Bone and fecal specimens were opportunistically collected by walking 2 km in opposite directions from transect points selected at 200-m intervals along the elevational gradient of the study areas. We collected a total of 220 carnivore fecal and owl pellet specimens from both study sites, which were mainly deposited by the spotted hyena (Crocuta crocuta), leopard (Panthera pardus), serval (Leptailurus serval), genet (Genetta sp.), and Mackinder’s Cape owl (Bubo capensis mackinderi). Serval scats were the most common, followed by those of the spotted hyena. Scats and bones were found at the lowest density at the lowest elevations, peaked at mid-higher elevations, and then declined at the highest elevations. Based on skeletal analysis only, there were more species in Sirimon (19) than in Chogoria (12). Small fauna (rodents to duiker size bovids) formed the bulk of the identified remains, representing 87.9% of the Sirimon fauna and 90.9% of the Chogoria fauna. The genus Otomys was the dominant prey of the owl and serval in both sites. Three giraffe teeth were found at 3 500 m a.s.l. in Chogoria on the edge of Lake Ellis, suggesting that it is an occasional visitor to such high elevations. This study underscores the value of fecal and bone surveys in understanding the diet and diversity of mammals in ecological ecosystems, but such surveys should be complemented with analysis of hairs found in scats to obtain a more complete list of carnivore prey at Mt. Kenya.
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Purification and characterization of a novel anti-coagulant from the leech Hirudinaria manillensis
Ruo-Mei Cheng, Xiao-Peng Tang, Ai-Lin Long, James Mwangi, Ren Lai, Rui-Pu Sun, Cheng-Bo Long, Zhen-Qing Zhang
Zoological Research    2019, 40 (3): 205-210.   DOI: 10.24272/j.issn.2095-8137.2019.037
Online available: 28 March 2019

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Protease inhibitors have been reported rarely from the leech Hirudinaria manillensis. In this study, we purified a novel protease inhibitor (bdellin-HM-2) with anticoagulant properties from H. manillensis. With a molecular weight of 1.4x104, bdellin-HM-2 was also characterized with three intra-molecular disulfide bridges at the N-terminus and multiple HHXDD and HXDD motifs at the C-terminus. cDNA cloning revealed that the putative nucleotide-encoding protein of bdellin-HM-2 contained 132 amino acids and was encoded by a 399 bp open reading frame (ORF). Sequence alignment showed that bdellin-HM-2 shared similarity with the “non-classical” Kazal-type serine protease inhibitors, but had no inhibitory effect on trypsin, elastase, chymotrypsin, kallikrein, factor XIIa (FXIIa), factor XIa (FXIa), factor Xa (FXa), thrombin, or plasmin. Bdellin-HM-2 showed anticoagulant effects by prolonging the activated partial thromboplastin time (aPTT), indicating a role in enabling H. manillensis to obtain a blood meal from its host. Our results suggest that bdellin-HM-2 may play a crucial role in blood-sucking in this leech species and may be a potential candidate for the development of clinical anti-thrombotic drugs.

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Patterns of human-wildlife conflict and compensation practices around Daxueshan Nature Reserve, China
Cheng Huang, Xue-You Li, Liu-Jun Shi, Xue-Long Jiang
Zoological Research    2018, 39 (6): 406-412.   DOI: 10.24272/j.issn.2095-8137.2018.056
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Understanding the spatial patterns of human-wildlife conflict is essential to inform management decisions to encourage coexistence, but it is constrained by the lack of spatially-explicit data. We collected spatially-implicit data of human-wildlife conflicts from 2009–2015 around Daxueshan Nature Reserve, Yunnan, China, and investigated the patterns and drivers of these conflicts. A questionnaire was also designed to capture local resident attitudes toward insurance-based compensation for the losses caused by targeted wildlife. We found that the Asiatic black bear (Ursus thibetanus) was the most conflict-prone animal around the reserve, followed by the rhesus macaque (Macaca mulatta) and Southeast Asian sambar (Cervus equinus). Conflicts were unevenly distributed among seasons, villages, and communities, with several grids identified as conflict hotspots. Poisson models revealed that human-bear conflicts were negatively related to distance to the reserve and proportion of forest, but positively correlated to the proportion of cropland. Binomial models showed that communities affected by crop depredation were positively correlated with the proportion of cropland and negatively correlated with distance to the reserve, whereas communities affected by livestock depredation were negatively correlated with the proportion of cropland. The insurance-based scheme has compensated over 90% of losses, to the satisfaction of 90.6% of respondents. Our results suggest that human-bear conflict could be potentially reduced by eliminating food crops near the reserve boundary and livestock grazing at conflict hotspots. In addition, the insurance-based scheme could be replicated at a broader scale with improvement in loss assessment.
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Effects of C-terminal amidation and heptapeptide ring on the biological activities and advanced structure of amurin-9KY, a novel antimicrobial peptide identified from the brown frog, Rana kunyuensis
Fen Zhang, Zhi-Lai Guo, Yan Chen, Li Li, Hai-Ning Yu, Yi-Peng Wang
Zoological Research    2019, 40 (3): 198-204.   DOI: 10.24272/j.issn.2095-8137.2018.070
Online available: 15 August 2018

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Rana kunyuensis is a species of brown frog that lives exclusively on Kunyu Mountain, Yantai, China. In the current study, a 279-bp cDNA sequence encoding a novel antimicrobial peptide (AMP), designated as amurin-9KY, was cloned from synthesized double-strand skin cDNA of R. kunyuensis. The amurin-9KY precursor was composed of 62 amino acid (aa) residues, whereas the mature peptide was composed of 14 aa and contained two cysteines forming a C-terminal heptapeptide ring (Rana box domain) and an amidated C-terminus. These structural characters represent a novel amphibian AMP family. Although amurin-9KY exhibited high similarity to the already identified amurin-9AM from R. amurensis, little is known about the structures and activities of amurin-9 family AMPs so far. Therefore, amurin-9KY and its three derivatives (amurin-9KY1–3) were designed and synthesized. The structures and activities were examined to evaluate the influence of C-terminal amidation and the heptapeptide ring on the activities and structure of amurin-9KY. Results indicated that C-terminal amidation was essential for antimicrobial  activity, whereas both C-terminal amidation and the heptapeptide ring played roles in the low hemolytic activity. Circular dichroism (CD) spectra showed that the four peptides adopted an a-helical conformation in THF/H2O (v/v 1:1) solution, but a random coil in aqueous solution. Elimination of the C-terminal heptapeptide ring generated two free cysteine residues with unpaired thiol groups, which greatly increased the concentration-dependent anti-oxidant activity. Scanning electron microscopy (SEM) was also performed to determine the possible bactericidal mechanisms.
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Rhinogobius immaculatus, a new species of freshwater goby (Teleostei: Gobiidae) from the Qiantang River, China
Fan Li, Shan Li, Jia-Kuan Chen
Zoological Research    2018, 39 (6): 396-405.   DOI: 10.24272/j.issn.2095-8137.2018.065
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A new freshwater goby, Rhinogobius immaculatus sp. nov., is described here from the Qiantang River in China. It is distinguished from all congeners by the following combination of characters: second dorsal-fin rays I, 7–9; anal-fin rays I, 6–8; pectoral-fin rays 14–15; longitudinal scales 29–31; transverse scales 7–9; predorsal scales 2–5; vertebrae 27 (rarely 28); preopercular canal absent or with two pores; a red oblique stripe below eye in males; branchiostegal membrane mostly reddish-orange, with 3–6 irregular discrete or connected red blotches on posterior branchiostegal membrane and lower operculum in males; caudal-fin base with a median black spot; and no black blotch on anterior part of first dorsal fin in males.
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Passive eye movements induced by electromagnetic force (EMF) in rats
Yue Yu, Jun Huang, Chun-Ming Zhang, Tian-Wen Chen, David S. Sandlin, Shao-Xun Wang, Alberto A. Arteaga, Jerome Allison, Yang Ou, Susan Warren, Paul May, Hong Zhu, Wu Zhou
Zoological Research    2019, 40 (3): 211-218.   DOI: 10.24272/j.issn.2095-8137.2019.024
Online available: 30 January 2019

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Accurate information on eye position in the orbit is available from visual feedback, efference copy of the oculomotor commands and proprioceptive signals from the extraocular muscles (EOM). Whereas visual feedback and oculomotor commands have been extensively studied, central processing of EOM proprioceptive signals remains to be elucidated. A challenge to the field is to develop an approach to induce passive eye movements without physically contacting the eyes. A novel method was developed to generate passive eye movements in rats. A small rare-earth magnet disk (0.7 mm diameter, 0.5 mm thickness) was attached to the surface of a rat’s eyeball. A metal rod (5 mm diameter) wrapped with an electromagnetic (EM) coil was placed near the magnet (8–15 mm). By passing currents to the EM coil, electromagnetic force (EMF) was generated and acted upon the magnet and induced passive eye movements. The EMF induced well-defined passive eye movements, whose directions were dependent on current polarity and amplitudes and peak velocities were dependent on current intensity and duration. Peak velocities of the EMF-induced eye movements were linearly related to amplitudes, exhibiting main sequence relationships similar to that of saccades in awake rats and eye movements induced by electrical microstimulation of the abducens nucleus in anesthetized rats. Histological examination showed that repetitive EMF stimulations did not appear to result in damages in the EOM fibers. These results validated the EMF approach as a novel tool to investigate EOM proprioceptive signals and their roles in visual localization and gaze control.
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High egg rejection rate in a Chinese population of grey-backed thrush (Turdus hortulorum)
Can-Chao Yang, Long-Wu Wang, Wei Liang, Anders Møller
Zoological Research    2019, 40 (3): 226-230.   DOI: 10.24272/j.issn.2095-8137.2019.039
Online available: 28 March 2019

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Several previous studies have indicated that nest sanitation behavior is a general adaptation in altricial birds, with egg recognition capacity evolving as a specific response to interspecific brood parasitism (IBP). However, a recent study suggested an alternative hypothesis, concluding that conspecific brood parasitism (CBP) selects for egg rejection in thrushes, with IBP as a by-product. In the present study, we used a spectrophotometer to quantify egg coloration and egg mimicry and performed artificial parasitism experiments in the grey-backed thrush (Turdus hortulorum). We showed that individuals of this species rejected 100% of 12 foreign eggs, without IBP or CBP detected. In a review of previous studies, we also discuss possible explanations for the high egg rejection rate in the grey-backed thrush and suggest areas for future study.

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Species delimitation based on diagnosis and monophyly, and its importance for advancing mammalian taxonomy
Eliécer E. Gutiérrez, Guilherme S. T. Garbino
Zoological Research    2018, 39 (5): 301-308.   DOI: 10.24272/j.issn.2095-8137.2018.037
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A recently proposed taxonomic classification of extant ungulates sparked a series of publications that criticize the Phylogenetic Species Concept (PSC) claiming it to be a particularly poor species concept. These opinions reiteratively stated that (1) the two fundamental elements of the "PSC", i.e., monophyly and diagnosability, do not offer objective criteria as to where the line between species should be drawn; and (2) that extirpation of populations can lead to artificial diagnosability and spurious recognitions of species. This sudden eruption of criticism against the PSC is misleading. Problems attributed to the PSC are common to most approaches and concepts that modern systematists employ to establish species boundaries. The controversial taxonomic propositions that sparked criticism against the PSC are indeed highly problematic, not because of the species concept upon which they are based, but because no evidence (whatsoever) has become public to support a substantial portion of the proposed classification. We herein discuss these topics using examples from mammals. Numerous areas of biological research rest upon taxonomic accuracy (including conservation biology and biomedical research); hence, it is necessary to clarify what are (and what are not) the real sources of taxonomic inaccuracy.

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A new genus of Asiatic short-tailed shrew (Soricidae, Eulipotyphla) based on molecular and morphological comparisons
Kai He, Xing Chen, Peng Chen, Shui-Wang He, Feng Cheng, Xue-Long Jiang, Kevin Campbell
Zoological Research    2018, 39 (5): 321-334.   DOI: 10.24272/j.issn.2095-8137.2018.058
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Blarinellini is a tribe of soricine shrews comprised of nine fossil genera and one extant genus. Blarinelline shrews were once widely distributed throughout Eurasia and North America, though only members of the Asiatic short-tailed shrew genus Blarinella currently persist (mostly in southwestern China and adjacent areas). Only three forms of Blarinella have been recognized as either species or subspecies. However, recent molecular studies indicated a strikingly deep divergence within the genus, implying the existence of a distinct genus-level lineage. We sequenced the complete mitochondrial genomes and one nuclear gene of three Asiatic short-tailed and two North American shrews and analyzed them morphometrically and morphologically. Our molecular analyses revealed that specimens ascribed to B. griselda formed two deeply diverged lineages, one a close relative to B. quadraticauda, whereas the other — comprised of topotype specimens from southern Gansu — diverged from other Blarinella in the middle Miocene (ca. 18.2 million years ago (Ma), 95% confidence interval=13.4–23.6 Ma). Although the skulls were similarly shaped in both lineages, we observed several diagnostic characteristics, including the shape of the upper P4. In consideration of the molecular and morphological evidence, we recognize B. griselda as the sole species of a new genus, namely, Pantherina gen. nov. Interestingly, some characteristics of Pantherina griselda are more similar to fossil genera, suggesting it represents an evolutionarily more primitive form than Blarinella. Recognition of this new genus sheds light on the systematics and evolutionary history of the tribe Blarinellini throughout Eurasia and North America. 
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Interchange between grooming and infant handling in female Tibetan macaques (Macaca thibetana)
Qi Jiang, Dong-Po Xia, Xi Wang, Dao Zhang, Bing-Hua Sun, Jin-Hua Li
Zoological Research    2019, 40 (2): 139-145.   DOI: 10.24272/j.issn.2095-8137.2018.049
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In some nonhuman primates, infants function as a social tool that can bridge relationships among group members. Infants are a desired commodity for group members, and mothers control access to them. The biological market theory suggests that grooming is widespread and represents a commodity that can be exchanged for infant handling. As a limited resource, however, the extent to which infants are interchanged between mothers (females with an infant) and non-mothers (potential handlers, females without an infant) remains unclear. In this study, we collected behavioral data to investigate the relationship between grooming and infant handling in free-ranging Tibetan macaques (Macaca thibetana) at Mt. Huangshan, China. Our results showed that females with infants received more grooming than females without infants. After her infant was handled, mother females received more grooming than they did during daily grooming interactions. However, with the increasing number of infants within the social group, both the grooming that mothers received and the grooming that non-mothers invested for handling infants decreased. We also found that non-mothers invested more time in grooming to gain access to younger infants than older infants. Our results provide evidence that infants are social commodities for both mother and non-mother females. Mothers use infants for obtain grooming and non-mothers use grooming to gain access to infants. The current study implies a bidirectional and complex interchange pattern between grooming and infant handling to compensate for the dyadic grooming disparity in non-human primates.
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Home range variation of two different-sized groups of golden snub-nosed monkeys (Rhinopithecus roxellana) in Shennongjia, China: implications for feeding competition
Peng-Lai Fan, Yi-Ming Li, Craig B. Stanford, Fang Li, Ze-Tian Liu, Kai-Hua Yang, Xue-Cong Liu
Zoological Research    2019, 40 (2): 121-128.   DOI: 10.24272/j.issn.2095-8137.2018.044
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Knowledge on the home range size of a species or population is important for understanding its behavioral and social ecology and improving the effectiveness of conservation strategies. We studied the home range size of two different-sized groups of golden snub-nosed monkeys (Rhinopithecus roxellana) in Shennongjia, China. The larger group (236 individuals) had a home range of 22.5 km2 from September 2007 to July 2008, whereas the smaller group (62 individuals) occupied a home range of 12.4 km2 from November 2008 to July 2009. Both groups exhibited considerable seasonal variation in their home range size, which was likely due to seasonal changes in food availability and distribution. The home range in any given season (winter, spring, summer, or winter+spring+summer) of the larger group was larger than that of the smaller group. As the two groups were studied in the same area, with the confounding effects of food availability thus minimized, the positive relationship between home range size and group size suggested that scramble feeding competition increased within the larger group.
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Maternal gene Ooep may participate in homologous recombination-mediated DNA double-strand break repair in mouse oocytes
Da-Jian He, Lin Wang, Zhi-Bi Zhang, Kun Guo, Jing-Zheng Li, Xie-Chao He, Qing-Hua Cui, Ping Zheng
Zoological Research    2018, 39 (6): 387-395.   DOI: 10.24272/j.issn.2095-8137.2018.067
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DNA damage in oocytes can cause infertility and birth defects. DNA double-strand breaks (DSBs) are highly deleterious and can substantially impair genome integrity. Homologous recombination (HR)-mediated DNA DSB repair plays dominant roles in safeguarding oocyte quantity and quality. However, little is known regarding the key players of the HR repair pathway in oocytes. Here, we identified oocyte-specific gene Ooep as a novel key component of the HR repair pathway in mouse oocytes. OOEP was required for efficient ataxia telangiectasia mutated (ATM) kinase activation and Rad51 recombinase(RAD51)focal accumulation at DNA DSBs. Ooep null oocytes were defective in DNA DSB repair and prone to apoptosis upon exogenous DNA damage insults. Moreover, Ooep null oocytes exhibited delayed meiotic maturation. Therefore, OOEP played roles in preserving oocyte quantity and quality by maintaining genome stability. Ooep expression decreased with the advance of maternal age, suggesting its involvement in maternal aging. 
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How many species of Apodemus and Rattus occur in China? A survey based on mitochondrial cyt b and morphological analyses
Shao-Ying Liu, Kai He, Shun-De Chen, Wei Jin, Robert W. Murphy, Ming-Kun Tang, Rui Liao, Feng-Jun Li
Zoological Research    2018, 39 (5): 309-320.   DOI: 10.24272/j.issn.2095-8137.2018.053
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Apodemus (mice) and Rattus (rats) are the top rodent reservoirs for zoonoses in China, yet little is known about their diversity. We reexamined the alpha diversity of these two genera based on a new collection of specimens from China and their cyt b sequences in GenBank. We also tested whether species could be identified using external and craniodental measurements exclusively. Measurements from 147 specimens of Apodemus and 236 specimens of Rattus were used for morphological comparisons. We analysed 74 cyt b sequences of Apodemus and 100 cyt b sequences of Rattus to facilitate phylogenetic estimations. Results demonstrated that nine species of Apodemus and seven species of Rattus, plus a new subspecies of Rattus nitidus, are distributed in China. Principal component analysis using external and craniodental measurements revealed that measurements alone could not separate the recognized species. The occurrence of Rattus pyctoris in China remains uncertain.
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Duplication and diversification of insulin genes in ray-finned fish
David M. Irwin
Zoological Research    2019, 40 (3): 185-197.   DOI: 10.24272/j.issn.2095-8137.2018.052
Online available: 20 August 2018

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Insulin is a key hormone for the regulation of metabolism in vertebrates. Insulin is produced by pancreatic islet cells in response to elevated glucose levels and leads to the uptake of glucose by tissues such as liver and adipose tissue to store energy. Insulin also has additional functions in regulating development. Previous work has shown that the proglucagon gene, which encodes hormones counter regulating insulin, is duplicated in teleost fish, and that the peptide hormones encoded by these genes have diversified in function. I sought to determine whether similar processes have occurred to insulin genes in these species. Searches of fish genomes revealed an unexpected diversity of insulin genes. A triplication of the insulin gene occurred at the origin of teleost fish, however one of these three genes, insc, has been lost in most teleost fish lineages. The two other insulin genes, insa and insb, have been retained but show differing levels of selective constraint suggesting that they might have diversified in function. Intriguingly, a duplicate copy of the insa gene, which I named insab, is found in many fish. The coding sequence encoded by insab genes is under weak selective constraint, with its predicted protein sequences losing their potential to be processed into a two-peptide hormone. However, these sequences have retained perfectly conserved cystine residues, suggesting that they maintain insulin’s three-dimensional structure and therefore might modulate the processing and secretion of insulin produced by the other genes.
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Tail regeneration reduction in lizards after repetitive amputation or cauterization reflects an increase of immune cells in blastemas
Lorenzo Alibardi
Zoological Research    2018, 39 (6): 413-423.   DOI: 10.24272/j.issn.2095-8137.2018.050
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Lizards are key amniote models for studying organ regeneration. During tail regeneration in lizards, blastemas contain sparse granulocytes, macrophages, and lymphocytes among the prevalent mesenchymal cells. Using transmission electron microscopy to examine scarring blastemas after third and fourth sequential tail amputations, the number of granulocytes, macrophages, and lymphocytes increased at 3–4 weeks in comparison to the first regeneration. An increase in granulocytes and agranulocytes also occurred within a week after blastema cauterization during the process of scarring. Blood at the third and fourth regeneration also showed a significant increase in white blood cells compared with that under normal conditions and at the first regeneration. The extracellular matrix of the scarring blastema, especially after cauterization, was denser than that in the normal blastema and numerous white blood cells and fibroblasts were surrounded by electron-pale, fine fibrinoid material mixed with variable collagen fibrils. In addition to previous studies, the present observations support the hypothesis that an increase in inflammation and immune reactions determine scarring rather than regeneration. These new findings verify that an immune reaction against mesenchymal and epidermal cells of the regenerative blastema is one of the main causes for the failure of organ regeneration in amniotes.
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Receptor variability-driven evolution of snake toxins
Xian-Hong Ji, Shang-Fei Zhang, Bin Gao, Shun-Yi Zhu
Zoological Research    2018, 39 (6): 431-436.   DOI: 10.24272/j.issn.2095-8137.2018.063
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Three-finger toxins (TFTs) are well-recognized non-enzymatic venom proteins found in snakes. However, although TFTs exhibit accelerated evolution, the drivers of this evolution remain poorly understood. The structural complexes between long-chain α-neurotoxins, a subfamily of TFTs, and their nicotinic acetylcholine receptor targets have been determined in previous research, providing an opportunity to address such questions. In the current study, we observed several previously identified positively selected sites (PSSs) and the highly variable C-terminal loop of these toxins at the toxin/receptor interface. Of interest, analysis of the molecular adaptation of the toxin-recognition regions in the corresponding receptors provided no statistical evidence for positive selection. However, these regions accumulated abundant amino acid variations in the receptors from the prey of snakes, suggesting that accelerated substitution of TFTs could be a consequence of adaptation to these variations. To the best of our knowledge, this atypical evolution, initially discovered in scorpions, is reported in snake toxins for the first time and may be applicable for the evolution of toxins from other venomous animals.
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Genomic and transcriptomic analyses reveal selection of genes for puberty in Bama Xiang pigs
Yang Yang, Adeniyi C. Adeola, Hai-Bing Xie, Ya-Ping Zhang
Zoological Research    2018, 39 (6): 424-430.   DOI: 10.24272/j.issn.2095-8137.2018.068
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The Bama Xiang pig (BMX) is a famous early-maturing Chinese indigenous breed with a two-end black coat. To uncover the genetic basis of the BMX phenotype, we conducted comparative genomic analyses between BMX and East Asian wild boars and Laiwu pigs, respectively. Genes under positive selection were enriched in pathways associated with gonadal hormone and melanin synthesis, consistent with the phenotypic changes observed during development in BMX pigs. We also performed differentially expressed gene analysis based on RNA-seq data from pituitary tissues of BMX and Large White pigs. The CTTNBP2NL, FRS2, KANK4, and KATNAL1 genes were under selection and exhibited expressional changes in the pituitary tissue, which may affect BMX pig puberty. Our study demonstrated the positive selection of early maturity in the development of BMX pigs and advances our knowledge on the role of regulatory elements in puberty evolution in pigs. 
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Threshold for maximal electroshock seizures (MEST) at three developmental stages in young mice
Cheng Xiang, Zhi-Na Li, Tian-Zhuang Huang, Jing-Hui Li, Lei Yang, Jing-Kuan Wei
Zoological Research    2019, 40 (3): 231-235.   DOI: 10.24272/j.issn.2095-8137.2019.038
Online available: 28 March 2019

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Early brain development after birth is extremely dynamic, suggesting that potential functional changes occur during this period. In this study, the maximal electroshock seizure threshold (MEST) was used to explore the electrophysiological variation among three developmental stages in young mice (no more than 5 weeks old). The induced electroshock seizure (ES) behavior of early postnatal mice (1–2-weeks old) differed from that during weaning (3 weeks old) and early puberty (4–5-weeks old). Thus, we further explored their respective characteristic responses to the ES parameters. When the stimulation current (SC) was limited to 4.0 mA, only the 1–2-week-old mice were induced to exhibit ES behavior at voltages of 30 V and 40 V, indicating they were more sensitive to maximal electroshock seizure (MES) (response to lower voltage). Surprisingly, however, they showed substantially lower mortality than the older groups under higher voltage conditions (60, 100, 160, and 200 V), suggesting better tolerance to the SC. We also found that when the current limit decreased to 3.5 mA, the 4–5-week-olds mice exhibited stable ES behavior with low mortality, while for 3-week-olds mice, the SC limit required to be reduced to 1.5 mA. In conclusion, our findings showed that neural sensitivity to MES was significantly different in young mice before puberty. Thus, greater attention should be given to distinguishing the developmental period of mice, especially in electrophysiological examination.

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Identification and characterization of short tandem repeats in the Tibetan macaque genome based on resequencing data
San-Xu Liu, Wei Hou, Xue-Yan Zhang, Chang-Jun Peng, Bi-Song Yue, Zhen-Xin Fan, Jing Li
Zoological Research    2018, 39 (4): 291-300.   DOI: 10.24272/j.issn.2095-8137.2018.047
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The Tibetan macaque, which is endemic to China, is currently listed as a Near Endangered primate species by the International Union for Conservation of Nature (IUCN). Short tandem repeats (STRs) refer to repetitive elements of genome sequence that range in length from 1–6 bp. They are found in many organisms and are widely applied in population genetic studies. To clarify the distribution characteristics of genome-wide STRs and understand their variation among Tibetan macaques, we conducted a genome-wide survey of STRs with next-generation sequencing of five macaque samples. A total of 1 077 790 perfect STRs were mined from our assembly, with an N50 of 4 966 bp. Mono-nucleotide repeats were the most abundant, followed by tetra- and di-nucleotide repeats. Analysis of GC content and repeats showed consistent results with other macaques. Furthermore, using STR analysis software (lobSTR), we found that the proportion of base pair deletions in the STRs was greater than that of insertions in the five Tibetan macaque individuals (P<0.05, t-test). We also found a greater number of homozygous STRs than heterozygous STRs (P<0.05, t-test), with the Emei and Jianyang Tibetan macaques showing more heterozygous loci than Huangshan Tibetan macaques. The proportion of insertions and mean variation of alleles in the Emei and Jianyang individuals were slightly higher than those in the Huangshan individuals, thus revealing differences in STR allele size between the two populations. The polymorphic STR loci identified based on the reference genome showed good amplification efficiency and could be used to study population genetics in Tibetan macaques. The neighbor-joining tree classified the five macaques into two different branches according to their geographical origin, indicating high genetic differentiation between the Huangshan and Sichuan populations. We elucidated the distribution characteristics of STRs in the Tibetan macaque genome and provided an effective method for screening polymorphic STRs. Our results also lay a foundation for future genetic variation studies of macaques.
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Why China is important in advancing the field of primatology
Paul A. Garber
Zoological Research    2018, 39 (4): 241-243.   DOI: 10.24272/j.issn.2095-8137.2018.012
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Over the past few decades, field studies conducted by Chinese primatologists have contributed significant new theoretical and empirical insights into the behavior, ecology, biology, genetics, and conservation of lorises, macaques, langurs, snub-nosed monkeys, and gibbons. With the recent establishment and inaugural meeting of the China Primatological Society in 2017, China has emerged as a leading nation in primate research. Several research teams have conducted long-term studies despite the difficult challenges of habituating and observing wild primates inhabiting mountainous temperate forests, and the fact that some 80% of China’s 25–27 primate species are considered vulnerable, endangered, or critically endangered and are distributed in small isolated subpopulations. In going forward, it is recommended that primatologists in China increase their focus on seasonal differences in the social, ecological, physiological, and nutritional challenges primates face in exploiting high altitude and cold temperate forests. In addition, provisioning as a habitation tool should be minimized or eliminated, as it is difficult to control for its effects on group dynamics, patterns of habitat utilization, and feeding ecology. Finally in the next decade, Chinese primatologists should consider expanding the taxonomic diversity of species studied by conducting research in other parts of Asia, Africa, and the Neotropics.
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 Impacts of late Quaternary environmental change on the long-tailed ground squirrel (Urocitellus undulatus) in Mongolia
Bryan S. McLean, Batsaikhan Nyamsuren, Andrey Tchabovsky, Joseph A. Cook
Zoological Research    2018, 39 (5): 364-372.   DOI: 10.24272/j.issn.2095-8137.2018.042
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Impacts of Quaternary environmental changes on mammal faunas of central Asia remain poorly understood due to a lack of geographically comprehensive phylogeographic sampling for most species. To help address this knowledge gap, we conducted the most extensive molecular analysis to date of the long-tailed ground squirrel (Urocitellus undulatus Pallas 1778) in Mongolia, a country that comprises the southern core of this species’ range. Drawing on material from recent collaborative field expeditions, we genotyped 128 individuals at 2 mitochondrial genes (cytochrome b and cytochrome oxidase I; 1 797 bp total). Phylogenetic inference supports the existence of two deeply divergent infraspecific lineages (corresponding to subspecies U. u. undulatus and U. u. eversmanni), a result in agreement with previous molecular investigations but discordant with patterns of range-wide craniometric and external phenotypic variation. In the widespread western eversmanni lineage, we recovered geographically-associated clades from the: (a) Khangai, (b) Mongolian Altai, and (c) Govi Altai mountain ranges. Phylogeographic structure in U. u. eversmanni is consistent with an isolation-by-distance model; however, genetic distances are significantly lower than among subspecies, and intra-clade relationships are largely unresolved. The latter patterns, as well as the relatively higher nucleotide polymorphism of populations from the Great Lakes Depression of northwestern Mongolia, suggest a history of range shifts into these lowland areas in response to Pleistocene glaciation and environmental change, followed by upslope movements and mitochondrial lineage sorting with Holocene aridification. Our study illuminates possible historical mechanisms responsible for U. undulatus genetic structure and contributes to a framework for ongoing exploration of mammalian response to past and present climate change in central Asia.

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Zoological Research    2019, 40 (3): 0-0.  
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Female choice impacts residential male takeover in golden snub-nosed monkeys (Rhinopithecus roxellana)
Gu Fang, Jing Chen, Ru-Liang Pan, Xiao-Guang Qi, Bao-Guo Li
Zoological Research    2018, 39 (4): 266-271.   DOI: 10.24272/j.issn.2095-8137.2018.035
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In primate species with social systems consisting of one-male breeding units (OMUs), resident male takeover represents a major challenge to individual reproductive success and mating strategies. The golden snub-nosed monkey (Rhinopithecus roxellana) is characterized by large multilevel societies (MLS) comprised of several OMUs and all-male units (AMUs); however, the factors and mechanisms associated with resident male takeover, which offer important insight into primate reproduction and social strategies, are still poorly understood. Based on 5-year monitoring data from a free-ranging herd of golden monkeys from the Qinling Mountains in China, we categorized three phases of an OMU, that is, a rising phase, developing phase, and declining phase. The rising and declining phases were unstable periods in which male takeover in an OMU might occur. Factors causing takeover, such as leader male rank, fighting ability, reproduction rate, and affiliation (proximity, allogrooming), were analyzed for males and females and for different OMUs. Results indicated that the new resident male’s fighting ability was lower than that of the former resident male in 23 cases. After replacement, the rank order of the new resident male significantly declined. Females involved in a takeover increased their distance from the resident male and decreased mating frequency during the three months prior to takeover. Females with infants under one-year-old had a marked effect on the specific time of takeover occurrence. These results suggested that female choice was the main factor deciding whether a takeover attempt was successful. Furthermore, rather than male conflict, females more often initiated and affected takeover and outcome, implying that the social status and competitive ability of the males played lesser roles during takeover.
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