Zoological Research

476-487

Authors: Bao Wang, Lei Chen, Wen Wang

Ruminants (Ruminantia) are among the most successful herbivorous mammals, exhibiting wide-ranging morphological and ecological characteristics (such as headgear and multichambered stomach) and including various key livestock species (e.g., cattle, buffalo, yak, sheep, and goat). Understanding their evolution is of great significance not only in scientific research but also in applications potential for human society. The rapid growth of genomic resources provides unprecedented opportunities to dissect the evolutionary histories and molecular mechanisms underlying the distinct characteristics of ruminants. Here we summarize our current understanding of the genetic, morphological, and ecological diversity of ruminants and provide prospects for future studies.

506-521

Authors: Yu Fan, Mao-Sen Ye, Jin-Yan Zhang, Ling Xu, Dan-Dan Yu, Tian-Le Gu, Yu-Lin Yao, Jia-Qi Chen, Long-Bao Lv, Ping Zheng, Dong-Dong Wu, Guo-Jie Zhang, Yong-Gang Yao

Chinese tree shrews (Tupaia belangeri chinensis) have become an increasingly important experimental animal in biomedical research due to their close relationship to primates. An accurately sequenced and assembled genome is essential for understanding the genetic features and biology of this animal. In this study, we used long-read single-molecule sequencing and high-throughput chromosome conformation capture (Hi-C) technology to obtain a high-qualitychromosome-scale scaffolding of the Chinese tree shrew genome. The new reference genome (KIZ version 2: TS_2.0) resolved problems in presently available tree shrew genomes and enabled accurate identification of large and complex repeat regions, gene structures, and species-specific genomic structural variants. In addition, by sequencing the genomes of six Chinese tree shrew individuals, we produced a comprehensive map of 12.8 M single nucleotide polymorphisms and confirmed that the major histocompatibility complex (MHC) loci and immunoglobulin gene family exhibited high nucleotide diversity in the tree shrew genome. We updated the tree shrew genome database (TreeshrewDB v2.0: http://www.treeshrewdb.org) to include the genome annotation information and genetic variations. The new high-quality reference genome of the Chinese tree shrew and the updated TreeshrewDB will facilitate the use of this animal in many different fields of research.

532-540

Authors: Mei-Ling Zhang, Ming-Li Li, Adeola Oluwakemi Ayoola, Robert W. Murphy, Dong-Dong Wu, Yong Shao

Elucidating the closest living relatives of extant primates is essential for fully understanding important biological processes related to the genomic and phenotypic evolution of primates, especially of humans. However, the phylogenetic placement of these primate relatives remains controversial, with three primary hypotheses currently espoused based on morphological and molecular evidence. In the present study, we used two algorithms to analyze differently partitioned genomic datasets consisting of 45.4 Mb of conserved non-coding elements and 393 kb of concatenated coding sequences to test these hypotheses. We assessed different genomic histories and compared with other molecular studies found solid support for colugos being the closest living relatives of primates. Our phylogeny showed Cercopithecinae to have low levels of nucleotide divergence, especially for Papionini, and gibbons to have a high rate of divergence. The MCMCtree comprehensively updated divergence dates of early evolution of Primatomorpha and Primates.

552-557

Authors: Wan-Sheng Jiang, Jie Li, Xiao-Zhong Lei, Zhou-Rui Wen, Yu-Zhang Han, Jun-Xing Yang, Jian-Bo Chang

A blind fish of Sinocyclocheilus (Cypriniformes: Cyprinidae)was caught in open water in the Three Gorges (Sanxia)reservoir, at a depth of 20 m in the mainstream of YangtzeRiver in Zigui County, Hubei Province, China. This fish can beeasily distinguished from all other congeners by externalmorphological characteristics, and is estimated to havediverged from its sister group about 0.55 million years ago(Ma). The geologically well separated locality of this specieshas expanded the distribution of Sinocyclocheilus cavefishfrom around N25° (latitude) to above N30° . Herein, wedescribe this new species as Sinocyclocheilus sanxiaensis sp.nov., and discuss the possible reasons why the speciesappears, surprisingly, in the Three Gorges reservoir.

564-574

Authors: Yun-He Wu, Chatmongkon Suwannapoom, Nikolay A. Poyarkov, Jr., Parinya Paawangkhanant, Kai Xu, Jie-Qiong Jin, Robert W. Murphy, Jing Che

Species of Xenophrys are conserved morphologically and live primarily in forests. In Thailand, the genus harbors many cryptic species. Herein we report the collection of specimens from Doi Inthanon, Chiang Mai Province, northern Thailand, which were identified previously as X. minor. Molecular and morphological analyses find that these specimens differ significantly from other known congeners, and therefore we describe a new species. Further, our phylogenetic analyses indicate that X. latidactyla is a junior synonym of X. palpebralespinosa.

580-582

Authors: Fei Wu, De-Jun Kong, Peng-Fei Shan, Jie Wang, Gladys Nyakeru Kungu, Guang-Yi Lu, Xiao-Jun Yang

The green peafowl (Pavo muticus) is one of the most threatened pheasants in the world. In China, it is widely appreciated for its beauty as well as historical and cultural value, but current populations number less than 500 individuals. Recently, Tang and colleagues reported in Science that the green peafowl is likely to become extinct due to the construction of the Jiasajiang Level 1 Hydropower Station within the Red River Upstream District (RRUD) and thus called for a stop to this project (Tang et al., 2019). According to our recent surveys, however, this species is still extant in 22 counties of Yunnan Province, China, among which, only two within the RRUD have been predicted to be affected by floods from the hydropower station. Therefore, the conclusion that the species will likely go extinct in China upon completion of the dam is unwarranted. In fact, construction of the Jiasajiang Level 1 Hydropower Station was stopped in August 2017. The main challenge for green peafowl conservation is that over 65% of the population occurs outside of protected reserves in China. Fortunately, the Chinese government has adopted an Ecological Redline (ERL) strategy to achieve ecological civilization plans, thus bringing new hope to the conservation of green peafowls both inside and outside of protected reserves. As a top conservation priority for China, the government is fully committed to conserving this peafowl.

587-594

Authors: Wendy Li, Zhan-Shan(Sam) Ma

The composition and diversity of the human vaginal microbial community have been investigated intensively due to the diversity-stability relationship (DSR)-based hypothesis for bacterial vaginosis (BV) etiology, which was first proposed in the 1990s and has received renewed interest in recent years. Nevertheless, diversity changes (scaling) across individuals in a cohort or population have not yet been addressed, which is significant both theoretically and practically. Theoretically, biodiversity scaling is the core of biogeography, and practically, inter-subject heterogeneity is critical for understanding the etiology and epidemiology of human microbiome-associated diseases such as BV. Here we applied the diversity-area relationship (DAR), a recent extension to the classic species-area relationship (SAR), to study diversity scaling of the vaginal microbiome by reanalyzing reported data collected from 1 107 postpartum women. The model used here characterized the power-law (or its extension) relationships between accrued diversity and areas (numbers of individuals), upon which four biogeographic profiles were thus defined. Specifically, we established the DAR profile (relationship between diversity scaling parameter and so-termed diversity order (q)), similarly pair-wise diversity overlap (PDO) profile, maximal accrual diversity (MAD) profile, and ratio of individual-level to population-level diversity (RIP) profile. These four profiles offer valuable tools to assess and predict diversity scaling (changes) in the human vaginal microbiome across individuals, as well as to understand the dynamics of vaginal microbiomes in healthy women.

349-357

Authors: Zhen-Zhen Chen, Yu-Yu Niu

Stem cell therapy (SCT) for Parkinson’s disease (PD) has received considerable attention in recent years. Non-human primate (NHP) models of PD have played an instrumental role in the safety and efficacy of emerging PD therapies and facilitated the translation of initiatives for human patients. NHP models of PD include primates with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced parkinsonism, who are responsive to dopamine replacement therapies, similar to human PD patients. Extensive research in SCT has been conducted to better treat the progressive dopaminergic neurodegeneration that underlies PD. For effective application of SCT in PD, however, a number of basic parameters still need to be tested and optimized in NHP models, including preparation and storage of cells for engraftment, methods of transplantation, choice of target sites, and timelines for recovery. In this review, we discuss the current status of NHP models of PD in stem cell research. We also analyze the advances and remaining challenges for successful clinical translation of SCT for this persistent disease.

394-403

Authors: Ting Yan, Hui-Hui Zhou

Information flow between the prefrontal and visual cortices is critical for visual behaviors such as visual search. To investigate its mechanisms, we simultaneously recorded spike and local field potential (LFP) signals in the frontal eye field (FEF) and area V4 while monkeys performed a free-gaze visual search task. During free-gaze search, spike-LFP coherence between FEF and V4 was enhanced in the theta rhythm (4–8 Hz) but suppressed in the alpha rhythm (8–13 Hz). Cross-frequency couplings during the Cue period before the search phase were related to monkey performance, with higher FEF theta-V4 gamma coupling and lower FEF alpha-V4 gamma coupling associated with faster search. Finally, feature-based attention during search enhanced spike-LFP coherence between FEF and V4 in the gamma and beta rhythms, whereas overt spatial attention reduced coherence at frequencies up to 30 Hz. These results suggest that oscillatory coupling may play an important role in mediating interactions between the prefrontal and visual cortices during visual search.

416-426

Authors: Lorenzo Alibardi

Ephrin receptors are the most common tyrosine kinase effectors operating during development. Ephrin receptor genes are reported to be up-regulated in the regenerating tail of the Podarcis muralis lizard. Thus, in the current study, we investigated immunolocalization of ephrin receptors in the Podarcis muralis tail during regeneration. Weak immunolabelled bands for ephrin receptors were detected at 15–17 kDa, with a stronger band also detected at 60–65 kDa. Labelled cells and nuclei were seen in the basal layer of the apical wound epidermis and ependyma, two key tissues stimulating tail regeneration. Strong nuclear and cytoplasmic labelling were present in the segmental muscles of the regenerating tail, sparse blood vessels, and perichondrium of regenerating cartilage. The immunolocalization of ephrin receptors in muscle that gives rise to large portions of new tail tissue was correlated with their segmentation. This study suggests that the high localization of ephrin receptors in differentiating epidermis, ependyma, muscle, and cartilaginous cells is connected to the regulation of cell proliferation through the activation of programs for cell differentiation in the proximal regions of the regenerating tail. The lower immunolabelling of ephrin receptors in the apical blastema, where signaling proteins stimulating cell proliferation are instead present, helps maintain the continuous growth of this region.

439-448

Authors: Jing Yang, Guo-Fen Zhu, Jian Jiang, Chang-Lin Xiang, Fu-Li Gao, Wei-Dong Bao

Detailed information on the size and genetic structure of wildlife populations is critical for developing effective conservation strategies, especially for those species that have suffered population decline and fragmentation due to anthropogenic activities. In the present study, we used a non-invasive approach combining fecal pellet sampling with mitochondrial DNA and nuclear DNA microsatellite marker analysis to monitor and compare the population structure of the Chinese goral (Naemorhedus griseus) in Beijing and northeast Inner Mongolia in China. Of the 307 fecal samples confirmed to be from N. griseus, 15 individuals (nine females and six males) were found in the Beijing population and 61 individuals (37 females and 24 males) were found in the Inner Mongolian population. Among these 76 individuals, we identified eight haplotypes and 13 nucleotide polymorphic sites from mtDNA and 45 alleles from 10 microsatellite loci. Spatially structured genetic variation and a significant level of genetic differentiation were observed between the two populations. In both populations, the sex ratios were skewed toward females, indicating high reproductive potential, which is crucial for population recovery and conservation of this patchily distributed vulnerable species. We suggest that managing the two populations as evolutionarily significant units with diverse genetic backgrounds could be an effective solution for present population recovery, with the possible relocation of individuals among different groups to help ensure future goral species prosperity.

456-465

Authors: Kai Wang, Jia-Wei Wu, Ke Jiang, Jin-Min Chen, Ben-Fu Miao, Cameron D. Siler, Jing Che

Despite continuous studies on the cryptic diversity of the Diploderma flaviceps complex in Southwest China for the pastdecade, little attention has been given to other widespreadcongeners in China. Combining both morphological and phylogenetic data, we describe a new species of Diplodermafrom populations identified previously as D. dymondi in thelower Yalong River Basin in southern Sichuan Province. Thenew species is morphologically most similar to D. dymondiand D. varcoae, but it can be differentiated by a considerablegenetic divergence and a suite of morphological characters,including having taller nuchal crest scales, smaller tympana,and a distinct oral coloration. Additionally, we discuss otherputative species complexes within the genus Diploderma inChina.

241-243

Authors: Bing Dong, Rui-Chang Quan, Zhan-Qi Chen

Prolonged milk provisioning and extended parental care for nutritionally independent offspring, previously considered to only co-occur in long-lived mammals (Clutton-Brock, 1991; Royle et al., 2012), were recently reported in the reproduction of the milking spider, Toxeus magnus (Chen et al. 2018). Newly hatched T. magnus spiderlings require 53 days to develop to maturity, with an average adult body length of 6.6 mm. The mother provides milk droplets to her newly hatched spiderlings until they develop into subadults (~38 days old), during which their body lengths increase from 0.9 mm at birth to 5.3 mm at weaning. Although spiderlings can forage for themselves at around 20 days old, they remain in the breeding nest for weeks after maturity.

277-292

Authors: Eduard Galoyan, Alisa Bolshakova, Manush Abrahamyan, Ruzanna Petrosyan, Valeria Komarova

Valentin’s rock lizard (Darevskia valentini) is suggested to be the parent for several parthenogenetic species (e.g., D. armeniaca, D. bendimahiensis, D. sapphirina, and D. unisexualis) that evolved through hybridization. Complex evolutionary processes (including reticulate evolution) are occurring within the areas where Valentin’s rock lizard coexists with these and other rock lizards. Hence, a detailed biological specification of this species is important for understanding how vertebrates evolve. Valentin’s rock lizard is a long-lived (up to 9 years), small diurnal lizard with larger females than males, which is unlike other species of the genus. Their relatively large eggs and early reproduction period, which occurs just after emergence from winter shelters, are adaptations for living in a high elevation climate (higher than 2 000 m a.s.l.). Their body temperatures (31–32 °С) are comparable to body temperatures of rock lizards living in milder climates, though female body temperature is more dependent on substrate temperature and basking due to their lower activity than that found in males. Population density fluctuates from several individuals to several hundred per hectare and is not affected by parthenogen coexistence, although hybrids do occur in sexually biased populations where males are more common than females. The male home range is larger than that of females, though these home ranges broadly overlap. Prey is not limited in the mountain meadows and Valentin’s rock lizards feed on a great variety of arthropods. Infanticide occurs in high-density populations.

305-316

Authors: De-Li Xu,Meng-Meng Xu,De-Hua Wang

Ambient temperature is an important factor influencing many physiological processes, including antioxidant defense and immunity. In the present study, we tested the hypothesis that antioxidant defense and immunity are suppressed by high and low temperature treatment in Brandt’s voles (Lasiopodomys brandtii). Thirty male voles were randomly assigned into different temperature groups (4, 23, and 32 °C, n=10 for each group), with the treatment course lasting for 27 d. Results showed that low temperature increased gross energy intake (GEI) and liver, heart, and kidney mass, but decreased body fat mass and dry carcass mass. With the decline in temperature, hydrogen peroxide (H2O2) concentration, which is indicative of reactive oxygen species (ROS) levels, increased in the liver, decreased in the heart, and was unchanged in the kidney, testis, and small intestine. Lipid peroxidation indicated by malonaldehyde (MDA) content in the liver, heart, kidney, testis, and small intestine did not differ among groups, implying that high and low temperature did not cause oxidative damage. Similarly, superoxide dismutase (SOD) and catalase (CAT) activities and total antioxidant capacity (T-AOC) in the five tissues did not respond to low or high temperature, except for elevation of CAT activity in the testis upon cold exposure. Bacteria killing capacity, which is indicative of innate immunity, was nearly suppressed in the 4 °C group in contrast to the 23 °C group, whereas spleen mass and white blood cells were unaffected by temperature treatment. The levels of testosterone, but not corticosterone, were influenced by temperature treatment, though neither were correlated with innate immunity, H2O2 and MDA levels, or SOD, CAT, and T-AOC activity in any detected tissues. Overall, these results showed that temperature had different influences on oxidative stress, antioxidant enzymes, and immunity, which depended on the tissues and parameters tested. Up-regulation or maintenance of antioxidant defense might be an important mechanism for voles to survive highly variable environmental temperatures.

324-330

Authors: Ping Wang, Lei Shi, Peng Guo

In this study, a total of 106 individuals of Oreocryptophis porphyraceus from mainland China were morphologically examined and recorded. Differences between populations were compared by combining data from this study and other published research. The skulls of three specimens representing three proposed subspecies (i.e., O. p. pulchra, O. p. vaillanti, and O. p. hainana) were examined by computed tomography (CT) scanning. Both external morphological characters and skull comparisons consistently showed significant differences between the studied populations. Based on these data, we suggest that at least four subspecies of O. porphyraceus should be recognized in mainland China: i.e., O. p. porphyraceus, O. p. pulchra, O. p. vaillanti, and O. p. hainana. However, the taxonomical arrangement of the central Chinese populations with intermediate morphology remain unresolved.

337-342

Authors: Jian-Peng Chen, Wei Pang, Zi-Wei Zhao, Yan-Hui Bi, Xiao-Wu Chen

Hemorrhagic septicemia is an acute, highly fatal disease that affects goldfish (Carassius auratus). To gain a better understanding of related immune genes, the transcriptomes of the skin and head kidney of goldfish suffering hemorrhagic septicemia were sequenced, assembled, and characterized. Based on functional annotation, an extensive and diverse catalog of expressed genes were identified in both the skin and head kidney. As two different organs, pair-wise comparison identified 122/77 unigenes up/down-regulated (two-fold change with P<0.05) in the skin and head kidney. Most genes of the immune pathways were expressed and isolated in both skin and head kidney, including interferon (IFN) transcription factors 1–10 and Toll-like receptors (TLRs). Interferon regulatory factor 3 (IRF3), a key IFN transcription factor, was up-regulated at the transcriptional level by polyriboinosinic: polyribocytidylic acid (poly I:C) challenge and regulated the IFN response by increasing the activity of IFN-β and IFN-stimulated response element (ISRE)-containing promoter. This study will benefit the identification and understanding of novel genes that play important roles in the immunological reactions of fish suffering from hemorrhagic septicemia.

151-174

Authors: Than Zaw, Paw Lay, Parinya Pawangkhanant, JVladislav A. Gorin, Nikolay A. Poyarkov, Jr.

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.

185 - 197

Authors: David M. Irwin

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.

205-210

Authors: Ruo-Mei Cheng, Xiao-Peng Tang, Ai-Lin Long, James Mwangi, Ren Lai, Rui-Pu Sun, Cheng-Bo Long, Zhen-Qing Zhang

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.

219-225

Authors: Wei Li, Ling-Yun Luo, Xun Yang, Yong He, Bin Lian, Chao-Hua Qu, Qing-Yuan Wu, Jian-Guo Zhang, Peng Xie

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).

231-235

Authors: Cheng Xiang,Zhi-Na Li,Tian-Zhuang Huang,Jing-Hui Li,Lei Yang,Jing-Kuan Wei

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.

77-88

Authors: Yi Ren, Shui-Fang Liu, Li Nie, Shi-Yu Cai, Jiong Chen

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.

94-101

Authors: Ruo-Han Qi, Yan Chen, Zhi-Lai Guo, Fen Zhang, Zheng Fang, Kai Huang, Hai-Ning Yu, Yi-Peng Wang

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.

108-112

Authors: Chao-Hao Chen, Bi-Jun Li, Xiao-Hui Gu, Hao-Ran Lin, Jun-Hong Xia

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.

121-128

Authors: Peng-Lai Fan, Yi-Ming Li, Craig B. Stanford, Fang Li, Ze-Tian Liu, Kai-Hua Yang, Xue-Cong Liu

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.

139-145

Authors: Qi Jiang, Dong-Po Xia, Xi Wang, Dao Zhang, Bing-Hua Sun, Jin-Hua Li

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.