Zoological Research

327-337

Authors: Chatmongkon SUWANNAPOOM, Zhi-Yong YUAN, Nikolay A. POYARKOV Jr., Fang YAN, Somboon KAMTAEJA, Robert W. MURPHY, Jing CHE

We describe a new species of frog in the dicroglossid genus Fejervarya from Ban Monjong, Omkoi District, Chiang Mai Province, northern Thailand. Analysis of DNA sequence data from the mitochondrial gene 16S, advertisement calls, and morphological distinctiveness support recognition of the new species. Matrilineal genealogy suggests that the new population from Chiang Mai is a sister taxon to the South Asian clade that includes F. syhadrensis, F. granosa, and F. pierrei. The new species, Fejervarya chiangmaiensis sp. nov., differs morphologically from its congeners by its relatively small body size and proportions and the presence of dorsal warts and dermal ridges. Discovery of this new species indicates that the biodiversity of amphibians in this region remains underestimated.

347-355

Authors: Jing YANG, Xin-Jiang LU, Fang-Chao CHAI, Jiong CHEN

The piscidin family, which includes potent antimicrobial peptides with broad-spectrum activity, plays an important role in the innate immune system of fish. In this study, we cloned piscidin-5-like type 3 (Lcpis5lt3) in large yellow croaker (Larimichthys crocea). Multiple alignments with other known piscidins revealed amino acid conservation throughout the fish, especially at the signal peptide (22 amino acids). The phylogenetic tree confirmed that Lcpis5lt3 and large yellow croaker piscidin-5-like proteins were grouped together to form a branch. Quantitative real-time PCR revealed that Lcpis5lt3 was expressed in a wide range of tissues, including the brain, muscle, gill, head kidney, intestine, kidney, liver, and spleen. The highest mRNA expression level of Lcpis5lt3 was found in the spleen. After Vibrio alginolyticus infection, mRNA expression was rapidly upregulated in the liver, head kidney, gill, kidney, and intestine at 4, 8, 12, and 24 h post infection (hpi), whereas there were no significant changes in the spleen. The antimicrobial spectrum showed that the synthetic mature peptide of Lcpis5lt3 exhibited different activity in vitro against various bacteria, such as Aeromonas hydrophila, V. anguillarum, V. alginolyticus, V. parahaemolyticus, Staphylococcus aureus, and Listeria monocytogenes. In addition, survival rates from the in vivo assay indicated that the synthetic peptide of Lcpis5lt3 increased the survival rate of large yellow croaker after V. alginolyticus challenge, resulting in a decline in bacterial burden and mRNA expression levels of interleukin-1β, interleukin-10, and tumor necrosis factor-α. These data suggest that Lcpis5lt3 plays an important role in innate immunity in large yellow croaker and might represent a potential therapeutic agent against pathogen invasion.

361-365

Authors: Jia-Li LI, Yong-Tang ZHENG, Xu-Dong ZHAO, Xin-Tian HU

Meeting report: the 4^th symposium on animal models of non-human primates in Kunming, Yunnan, China

263-269

Authors: Yang XU, Si-Wei ZHU, Qing-Wei LI

Lampreys belong to the superclass Cyclostomata and represent the most ancient group of vertebrates. Existing for over 360 million years, they are known as living fossils due to their many evolutionally conserved features. They are not only a keystone species for studying the origin and evolution of vertebrates, but also one of the best models for researching vertebrate embryonic development and organ differentiation. From the perspective of genetic information, the lamprey genome remains primitive compared with that of other higher vertebrates, and possesses abundant functional genes. Through scientific and technological progress, scientists have conducted in-depth studies on the nervous, endocrine, and immune systems of lampreys. Such research has significance for understanding and revealing the origin and evolution of vertebrates, and could contribute to a greater understanding of human diseases and treatments. This review presents the current progress and significance of lamprey research.

275-280

Authors: Wan-Sheng JIANG, Tao QIN, Wei-Ying WANG, Ya-Peng ZHAO, Shu-Sen SHU, Wei-Hong SONG, Xiao-Yong CHEN, Jun-Xing YANG

Biological invasion is a pervasive negative force of global change, especially in its effects on sensitive freshwater ecosystems. Even protected areas are usually not immune. Ptychobarbus chungtienensis is a threatened freshwater fish now almost confined to Bita Lake, in the Shangri-La region of Yunnan province, China. Its existence is threatened by the introduction of non-native weatherfishes (Misgurnus anguillicaudatus and Paramisgurnus dabryanus) by an unusual method known as ‘prayer animal release’. Periodic surveys revealed the ratio of invasive weatherfishes to P. chungtienensis has been increasing since the former species was first recorded from the lake in August, 2009. Ptychobarbus chungtienensis shows low genetic diversity in the relict Lake Bita population. Weatherfishes, however, have highly successful survival strategies. The degree of dietary overlap between the species is alarming and perhaps critical if food is found to be a limiting factor.

290-296

Authors: Yu-Yi WU, Zhi-Yu SUN, Yan-Shu GUO

Triplophysa daochengensis sp. nov. is described from the Daocheng River, a northern tributary of the Jinsha River in Sichuan Province, China. The new species can be distinguished from its congeners by the following characters: body smooth and scales absent; lateral line complete; caudal peduncle compressed, depth unchanging; head length equal to caudal-peduncle length; lower jaw shovel-shaped; dorsal-fin origin anterior to pelvic-fin origin and closer to the tip of the snout than to the caudal-fin base, last unbranched ray hard; pelvic-fin tip not reaching anus; posterior chamber of gas bladder absent; intestine of spiral type with three winding coils.

301-306

Authors: Chun-Lei WANG, Zhi-Peng WANG, Jia-Qi WANG, Ming-You Li, Xiao-Wu CHEN

Piwi-interacting RNA (piRNA) plays an important role in the gonadal development and maintenance of Teleostei. In this study, piRNA libraries derived from the adult gonads of Japanese flounder (Paralichthys olivaceus) were generated using next-generation sequencing technology. Using zebrafish piRNAs as a reference, 5 865 unique candidate piRNAs were identified; 289 candidate piRNA clusters (PRCs) were generated from the above piRNAs. Among the isolated candidate PRCs, a total of 38 ovary-specific, 45 ovary-bias, 24 testis-specific, and 131 testis-bias PRCs were found. The relative expression levels of seven PRCs were validated through quantitative reverse transcription-polymerase chain reaction. The results of this study will help facilitate exploration of the development and maintenance of the phenotypic sex mechanism in P. olivaceus.

313-313

Authors: Beuy JOOB, Viroj WIWANITKIT

Authorship misconduct

191-204

Authors: Ming SHAO, Tian-Rui XU, Ce-Shi CHEN

Targeted genome editing technology has been widely used in biomedical studies. The CRISPR-associated RNA-guided endonuclease Cas9 has become a versatile genome editing tool. The CRISPR/Cas9 system is useful for studying gene function through efficient knock-out, knock-in or chromatin modification of the targeted gene loci in various cell types and organisms. It can be applied in a number of fields, such as genetic breeding, disease treatment and gene functional investigation. In this review, we introduce the most recent developments and applications, the challenges, and future directions of Cas9 in generating disease animal model. Derived from the CRISPR adaptive immune system of bacteria, the development trend of Cas9 will inevitably fuel the vital applications from basic research to biotechnology and bio-medicine.

214-219

Authors: Xin LUO, Min LI, Bing SU

In the past three years, RNA-guided Cas9 nuclease from the microbial clustered regularly interspaced short palindromic repeats (CRISPR) adaptive immune system has been used to facilitate efficient genome editing in many model and non-model animals. However, its application in nonhuman primates is still at the early stage, though in view of the similarities in anatomy, physiology, behavior and genetics, closely related nonhuman primates serve as optimal models for human biology and disease studies. In this review, we summarize the current proceedings of gene editing using CRISPR/Cas9 in nonhuman primates.

229-236

Authors: Ming LI, Shinn-Yi CHOU

The postpartum period is when a host of changes occur at molecular, cellular, physiological and behavioral levels to prepare female humans for the challenge of maternity. Alteration or prevention of these normal adaptions is thought to contribute to disruptions of emotion regulation, motivation and cognitive abilities that underlie postpartum mental disorders, such as postpartum depression. Despite the high incidence of this disorder, and the detrimental consequences for both mother and child, its etiology and related neurobiological mechanisms remain poorly understood, partially due to the lack of appropriate animal models. In recent decades, there have been a number of attempts to model postpartum depression disorder in rats. In the present review, we first describe clinical symptoms of postpartum depression and discuss known risk factors, including both genetic and environmental factors. Thereafter, we discuss various rat models that have been developed to capture various aspects of this disorder and knowledge gained from such attempts. In doing so, we focus on the theories behind each attempt and the methods used to achieve their goals. Finally, we point out several understudied areas in this field and make suggestions for future directions.

246-251

Authors: Xiao-Liang ZHANG, Jia-Hao SONG, Wei PANG, Yong-Tang ZHENG

Northern pig-tailed macaques (NPMs, Macaca leonina) are susceptible to HIV-1 infection largely due to the loss of HIV-1-restricting factor TRIM5α. However, great impediments still exist in the persistent replication of HIV-1 in vivo, suggesting some viral restriction factors are reserved in this host. The APOBEC3 proteins have demonstrated a capacity to restrict HIV-1 replication, but their inhibitory effects in NPMs remain elusive. In this study, we cloned the NPM A3A-A3H genes, and determined by BLAST searching that their coding sequences (CDSs) showed 99% identity to the corresponding counterparts from rhesus and southern pig-tailed macaques. We further analyzed the anti-HIV-1 activities of the A3A-A3H genes, and found that A3G and A3F had the greatest anti-HIV-1 activity compared with that of other members. The results of this study indicate that A3G and A3F might play critical roles in limiting HIV-1 replication in NPMs in vivo. Furthermore, this research provides valuable information for the optimization of monkey models of HIV-1 infection.

119-125

Authors: Colin GROVES

In this paper, I have introduced the concept of the Evolutionary Species, and shown how it affects the taxonomy of the Artiodactyla of China. The "traditional" taxonomy of the Artiodactyla, which has remained almost unchanged for 100 years, relies on ill-formulated notions of species and subspecies, only slightly modified by the population-thinking of the 1930s. Species are populations (or metapopulations) differentiated by the possession of fixed heritable differences from other such populations (or metapopulations). In the Artiodactyla, there are many more species than "traditionally" recognised; this is by no means a drawback, as it enables the units of biodiversity to be identified in a testable fashion, and brings the taxonomy of large mammals into line with that long practised for small mammals. Species are likely to differentiate where there are natural gaps in the distribution of a genus, such as mountain blocks (for example in the genus Budorcas) or otherwise dissected habitat (for example in the genus Cervus). Natural hybridisation between distinct species is not an uncommon phenomenon, again illustrated well in the genus Cervus, where hybridisation between the elaphus and nippon groups occurs today and evidently occurred in the past, as shown by the distribution of mtDNA.

136-143

Authors: Jing WANG, Guang LI, Guang-Hui QIAN, Jun-Hao HUA, Yi-Quan WANG

The Wnt/β-catenin signaling pathway plays a crucial role in the embryonic development of metazoans. Although the pathway has been studied extensively in many model animals, its function in amphioxus, the most primitive chordate, remains largely uncharacterized. To obtain basic data for functional analysis, we identified and isolated seven genes (Lrp5/6, Dvl, APC, CkIα, CkIδ, Gsk3β, and Gro) of the Wnt/β-catenin signaling pathway from the amphioxus (Branchiostoma floridae) genome. Phylogenetic analysis revealed that amphioxus had fewer members of each gene family than that found in vertebrates. Whole-mount in situ hybridization showed that the genes were maternally expressed and broadly distributed throughout the whole embryo at the cleavage and blastula stages. Among them, Dvl was expressed asymmetrically towards the animal pole, while the others were evenly distributed in all blastomeres. At the mid-gastrula stage, the genes were specifically expressed in the primitive endomesoderm, but displayed different patterns. When the embryo developed into the neurula stage, the gene expressions were mainly detected in either paraxial somites or the tail bud. With the development of the embryo, the expression levels further decreased gradually and remained only in some pharyngeal regions or the tail bud at the larva stage. Our results suggest that the Wnt/β-catenin pathway might be involved in amphioxus somite formation and posterior growth, but not in endomesoderm specification.

151-158

Authors: Salindra K. DAYANANDA, Eben GOODALE, Myung-bok LEE, Jia-Jia LIU, Christos MAMMIDES, Bonifacio O. PASION, Rui-Chang QUAN, J. W. Ferry SLIK, Rachakonda SREEKAR, Kyle W. TOMLINSON, Mika YASUDA

Owls have the potential to be keystone species for conservation in fragmented landscapes, as the absence of these predators could profoundly change community structure. Yet few studies have examined how whole communities of owls respond to fragmentation, especially in the tropics. When evaluating the effect of factors related to fragmentation, such as fragment area and distance to the edge, on these birds, it is also important in heterogeneous landscapes to ask how ‘location factors’ such as the topography, vegetation and soil of the fragment predict their persistence. In Xishuangbanna, southwest China, we established 43 transects (200 m×60 m) within 20 forest fragments to sample nocturnal birds, both visually and aurally. We used a multimodel inference approach to identify the factors that influence owl species richness, and generalized linear mixed models to predict the occurrence probabilities of each species. We found that fragmentation factors dominated location factors, with larger fragments having more species, and four of eight species were significantly more likely to occur in large fragments. Given the potential importance of these birds on regulating small mammal and other animal populations, and thus indirectly affecting seed dispersal, we suggest further protection of large fragments and programs to increase their connectivity to the remaining smaller fragments.

167-175

Authors: Qing-Gang QIAO, Hong-Ji LIANG, Min-Lan BAI, Wei-Hong ZHENG, Jin-Song LIU

Physiological adaptation arises from several fundamental sources of phenotypic variation. Most analyses of metabolic adaptation in birds have focused on the basal metabolic rate (BMR), the lower limit of avian metabolic heat production. In this study, we investigated thermoregulation in three passerine species; the yellow-billed grosbeak Eophona migratoria, white-rumped munia Lonchura striata and black-throated bushtit Aegithalos concinnus, in Wenzhou, China. Metabolic rate was measured using the closed-circuit respirometer containing 3.5 L animal chambers. Body temperature (T_b) was measured during metabolic measurements using a lubricated thermocouple. The minimum thermal conductance of these species was calculated by measuring their T_b and metabolic rates. The yellow-billed grosbeak remained largely normothermic, and the white-rumped munia and black-throated bushtit exhibited variable T_b at ambient temperatures (T_a). Mean metabolic rates within thermal neutral zone were 2.48±0.09 O₂(mL)/g/h for yellow-billed grosbeaks, 3.44±0.16 O₂(mL)/g/h for white-rumped munias, and 3.55±0.20 O₂(mL)/g/h for black-throated bushtits, respectively. Minimum thermal conductance of yellow-billed grosbeak, white-rumped munia and black-throated bushtit were 0.13±0.00, 0.36±0.01, and 0.37±0.01 O₂(mL)/g/h/℃, respectively. The ecophysiological characteristics of these species were:(1) the yellowbilled grosbeak had relatively high T_b and BMR, a low lower critical temperature and thermal conductance, and a metabolic rate that was relatively insensitive to variation in T_a; all of which are typical of cold adapted species and explain its broader geographic distribution; (2) the white-rumped munia and blackthroated bushtit had high thermal conductance, lower critical temperature, and relatively low BMR, all which are adapted to warm environments where there is little selection pressure for metabolic thermogenesis. Taken together, these data illustrate small migratory and resident passerines that exhibit the different characteristics of thermoregulation.

186-188

Authors: Marco ENDRUWEIT

The natural distributional range of the cobitid loach Lepidocephalichthys alkaia is extended into Yunnan Province, China. The modified sexually dimorphic pectoral fin in males of L. alkaia is described.

65-65

Authors: Ying-Xiang WANG

In Memory of Professor Ying-Xiang WANG

75-83

Authors: Jia-Qi WANG, Jia-Jia WANG, Xu-Jian WU, Wei-Hong ZHENG, Jin-Song LIU

Environmental cues play important roles in the regulation of an animal's physiology and behavior. One such cue, photoperiod, plays an important role in the seasonal acclimatization of birds. It has been demonstrated that an animal's body mass, basal metabolic rate (BMR), and energy intake, are all affected by photoperiod. The present study was designed to examine photoperiod induced changes in the body mass, metabolism and metabolic organs of the silky starling, Sturnus sericeus. Captive silky starlings increased their body mass and BMR during four weeks of acclimation to a short photoperiod. Birds acclimated to a short photoperiod also increased the mass of certain organs (liver, gizzard and small intestine), and both gross energy intake (GEI) and digestible energy intake (DEI), relative to those acclimated to a long photoperiod. Furthermore, BMR was positively correlated with body mass, liver mass, GEI and DEI. These results suggest that silky starlings increase metabolic thermogenesis when exposed to a short photoperiod by increasing their body and metabolic organ mass, and their GEI and DEI. These findings support the hypothesis that bird species from temperate climates typically display high phenotypic flexibility in thermogenic capacity.

90-95

Authors: Yuan-Wei ZHANG, Xiao-Fu PAN, Xiao-Ai WANG, Wan-Sheng JIANG, Qian LIU, Jun-Xing YANG

The effects of osmotic pressure, temperature and stocking density on medusae survival of Craspedacusta sowerbii were examined. The medusae were shown to be sensitive to the variations of osmotic pressure. And the survival time was <90 h at 34 mOsm/L and it declined rapidly with rising osmotic pressure. The peak survival time of >200 h was recorded at 0.2 mOsm/L. Comparing with 27℃ and 32℃ treatments, 23℃ treatment yielded lower activities at a range of 8-13/min. However, there was a longer survival time. A non-linear relationship existed between survival time and stocking density. Lower density resulted in larger body size. And sexual reproduction resumed after breeding for >22 days. Newly-formed polyps and medusae appeared subsequently but only in the higher-density groups of 10, 14 and 18 ind./L. It suggested that the number of newly-formed polyps and medusae was highly dependent on stocking density. That is, a higher stocking density produced more organisms. However, newly-formed medusae died within one month and none grew a diameter of >5 mm.

103-109

Authors: Xin ZHENG, Tian-Qi ZHOU, Tao WAN, Anabel PERDICES, Jin-Quan YANG, Xin-Sheng TANG, Zheng-Ping WANG, Li-Qun HUANG, Song HUANG, Shun-Ping HE

Six main mitochondrial DNA (mtDNA) lineages have been described in minnow (Zacco platypus) samples obtained from northern, western and southern China. Perdices et al. (2004) predicted that further sampling of other tributaries might discover more lineages of this species. In this study, we collected 26 Zacco platypus individuals in the Huangshan area of eastern China and determined the cytochrome b (cytb) sequence variations. Combined with reported data in GenBank, we identified ten matrilines (Zacco A-J) in a total of 169 samples, with relatively high molecular divergence found among them. The Huangshan population had the greatest genetic variation among all sampled regions and hosted six of the ten matrilines. Our results highlight the significance of the Huangshan area for the conservation of Zacco platypus.