Zoological Research

65-71

Authors: Hao GU, Eben GOODALE, Jin CHEN

The study of mutualistic plant and animal networks is an emerging field of ecological research. We reviewed progress in this field over the past 30 years. While earlier studies mostly focused on network structure, stability, and biodiversity maintenance, recent studies have investigated the conservation implications of mutualistic networks, specifically the influence of invasive species and how networks respond to habitat loss. Current research has also focused on evolutionary questions including phylogenetic signal in networks, impact of networks on the coevolution of interacting partners, and network influences on the evolution of interacting species. We outline some directions for future research, particularly the evolution of specialization in mutualistic networks, and provide concrete recommendations for environmental managers.

79-87

Authors: Qing-Jian LIANG, Lei ZHAO, Jia-Qi WANG, Qian CHEN, Wei-Hong ZHENG, Jin-Song LIU

Food resources play an important role in the regulation of animals' physiology and behavior. We investigated the effect of short-term food restriction on metabolic thermogenesis of Chinese bulbuls (Pycnonotus sinensis) by measuring changes in body mass, body fat, basic metabolic rate (BMR), and organ mass of wild-caught Chinese bulbuls from Wenzhou, China. Short-term food restriction induced a significant decrease in body mass and body fat but body mass returned to normal levels soon after food was no longer restricted. Food restriction caused a significant reduction in BMR after 7 days (P<0.05), which returned to normal levels after food restriction ceased. Log total BMR was positively correlated with log body mass (r²=0.126, P<0.05). The dry masses of livers and the digestive tract were higher in birds that had been subject to temporary food restriction than in control birds and those subject to continual food restriction (P<0.001 and P<0.05, respectively). There was also significant differences in the dry mass of the lungs (P<0.05), heart (P<0.01), and spleen (P<0.05) in birds subject to short-term food restriction compared to control birds and those subject to continual food restriction. BMR was positively correlated with body and organ (heart, kidney and stomach) mass. These results suggest that the Chinese bulbul adjusts to restricted food availability by utilizing its energy reserves, lowering its BMR and changing the weight of various internal organs so as to balance total energy requirements. These may all be survival strategies that allow birds to cope with unpredictable variation in food abundance.

95-102

Authors: Guo-Xiao ZHENG, Jiang-Tao LIN, Wei-Hong ZHENG, Jing CAO, Zhi-Jun ZHAO

Reproduction is the highest energy demand period for small mammals, during which both energy intake and expenditure are increased to cope with elevated energy requirements of offspring growth and somatic protection. Oxidative stress life history theory proposed that reactive oxygen species (ROS) were produced in direct proportion to metabolic rate, resulting in oxidative stress and damage to macromolecules. In the present study, several markers of oxidative stress and antioxidants activities were examined in brain, liver, kidneys, skeletal muscle and small intestine in non-lactating (Non-Lac) and lactating (Lac) KM mice. Uncoupling protein (ucp_s) gene expression was examined in brain, liver and muscle. During peak lactation, gross energy intake was 254% higher in Lac mice than in Non-Lac mice. Levels of H₂O₂ of Lac mice were 17.7% higher in brain (P<0.05), but 21.1% (P<0.01) and 14.5% (P<0.05) lower in liver and small intestine than that of Non-Lac mice. Malonadialdehyde (MDA) levels of Lac mice were significantly higher in brain, but lower in liver, kidneys, muscle and small intestine than that of Non-Lac mice. Activity of glutathione peroxidase (GSH-PX) was significantly decreased in brain and liver in the Lac group compared with that in the Non-Lac group. Total antioxidant capacity (T-AOC) activity of Lac mice was significantly higher in muscle, but lower in kidneys than Non-Lac mice. Ucp₄ and ucp₅ gene expression of brain was 394% and 577% higher in Lac mice than in Non-Lac mice. These findings suggest that KM mice show tissue-dependent changes in both oxidative stress and antioxidants. Activities of antioxidants may be regulated physiologically in response to the elevated ROS production in several tissues during peak lactation. Regulations of brain ucp₄ and ucp₅ gene expression may be involved in the prevention of oxidative damage to the tissue.

109-111

Authors: Yang-Xian ZUO, Zhao-Qing YANG

The validity of Sarcocystis sinensis

115-116

Authors: Chung-Wei YEN

Three Australian leg-flagged Great Knots (Calidris tenuirostris) found on the islet coast of Jinmen (Quimoy) in Fujian, China

1-17

Authors: David M. IRWIN

Ruminant stomach lysozyme is a long established model of adaptive gene evolution. Evolution of stomach lysozyme function required changes in the site of expression of the lysozyme c gene and changes in the enzymatic properties of the enzyme. In ruminant mammals, these changes were associated with a change in the size of the lysozyme c gene family. The recent release of near complete genome sequences from several ruminant species allows a more complete examination of the evolution and diversification of the lysozyme c gene family. Here we characterize the size of the lysozyme c gene family in extant ruminants and demonstrate that their pecoran ruminant ancestor had a family of at least 10 lysozyme c genes, which included at least two pseudogenes. Evolutionary analysis of the ruminant lysozyme c gene sequences demonstrate that each of the four exons of the lysozyme c gene has a unique evolutionary history, indicating that they participated independently in concerted evolution. These analyses also show that episodic changes in the evolutionary constraints on the protein sequences occurred, with lysozyme c genes expressed in the abomasum of the stomach of extant ruminant species showing the greatest levels of selective constraints.

29-33

Authors: Qi-Ping ZHU, Meng-Yao ZHU, Ying-Chao HU, Xue-Ya ZHANG, Guo-Hua DING, Zhi-Hua LIN

In reptiles, habitat selection is the process whereby suitable habitat is selected that optimizes physiological functions and behavioral performance. Here, we used the brown forest skink (Sphenomorphus indicus) as a model animal and examined whether the frequency of active individuals, environmental temperature, illumination of activity area, and habitat type vary with different age classes. We surveyed the number of active individuals and measured environmental variables at Baiyunshan Mountain in Lishui, Zhejiang, China. We found no difference in the activity frequency of adult and juvenile S. indicus; the activity pattern of active individuals was bimodal. The mean environmental temperature selected by adults was higher than that selected by juveniles. The environmental temperature of active areas measured at 0900-1000 h and 1100-1200h was higher than at 1400-1500h; illumination of the active area at 1000-1200h was also higher than at 1400h-1600 h. The number of active individuals, the environmental temperature and illumination of activity areas showed pairwise positive correlation. There was a difference in habitat type between juveniles and adults whereby juveniles prefer rock habitats. We predict that active S. indicus select optimal habitats with different environmental temperatures and types to reach the physiological needs particular to their age classes.

41-47

Authors: Cun-Bao LIU, Bin SHAN, Hong-Mei BAI, Jing TANG, Long-Zong YAN, Yan-Bing MA

Multidrug resistant (MDR) pathogen infections are serious threats to hospitalized patients because of the limited therapeutic options. A novel group of antibiotic candidates, antimicrobial peptides (AMPs), have recently shown powerful activities against both Gram-negative and Gram-positive bacteria. Unfortunately, the viability of using these AMPs in clinical settings remains to be seen, since most still need to be evaluated prior to clinical trials and not all of AMPs are potent against MDR clinical isolates. To find a connection between the characteristics of several of these AMPs and their effects against MDR pathogens, we selected 14 AMPs of animal origin with typical structures and evaluated their in vitro activities against clinical strains of extensive drug-resistant Acinetobacter baumannii, methicillin-resistant Staphylococcus aureus, extended spectrum β-lactamase-producing Pseudomonas aeruginosa and extended spectrum β-lactamase-producing Escherichia coli. Our results showed that these peptides' hydrophilic/hydrophobic characteristics, rather than their secondary structures, may explain their antibacterial effects on these clinical isolates. Peptides that are amphipathic along the longitudinal direction seemed to be effective against Gram-negative pathogens, while peptides with hydrophilic terminals separated by a hydrophobic intermediate section appeared to be effective against both Gram-negative and Gram-positive pathogens. Among these, cathelicidin-BF was found to inhibit all of the Gram-negative pathogens tested at dosages of no more than 16 mg/L, killing a pandrug-resistant A. baumannii strain within 2 h at 4×MICs and 4 h at 2×MICs. Tachyplesin III was also found capable of inhibiting all Gram-negative and Gram-positive pathogens tested at no more than 16 mg/L, and similarly killed the same A. baumannii strain within 4 h at 4×MICs and 2×MICs. These results suggest that both cathelicidin-BF and tachyplesin III are likely viable targets for the development of AMPs for clinical uses.

54-58

Authors: Fang DING, Li CHEN, Yong LIU, Feng-Rui WU, Biao DING, Wen-Yong LI, Rong WANG

It is well known that excessive long-term alcohol consumption is harmful, especially in pregnant women. In the present study, the Kunming white mouse was used as an animal model and indirect immunofluorescence was performed to analyze the toxic effects of alcohol on early pre-implantation embryos. H3K9 acetylation immunofluorescence could not be detected in MII oocytes. H3K9 acetylation levels in the treatment group were higher than in the control group during the morula stage, and contrary to results during the blastocyst stage. Other stages showed no obvious differences for in vivo embryos. For in vitro embryos, almost no difference was found between the two experimental groups across all stages, and both groups showed increasing H3K9 acetylation levels (except at the 2-cell stage). This study shows that H3K9 acetylation levels in early pre-implantation embryos are notably impacted by excessive alcohol ingestion by females. These data are the first step in understanding the epigenetic mechanism of alcohol toxicity in early pre-implantation mouse embryos.

465-473

Authors: Hong-Yi ZHENG, Ming-Xu ZHANG, Lin-Tao ZHANG, Xiao-Liang ZHANG, Wei PANG, Long-Bao LYU, Yong-Tang ZHENG

Pig-tailed macaques (Macaca nemistrina group) have been extensively used as non-human primate animal models for various human diseases in recent years, notably for AIDS research due to their sensitivity to HIV-1. Northern pig-tailed macaques (M. leonina) are distributed in China and other surrounding Southeast Asia countries. Although northern pig-tailed macaques have been bred on a large scale as experimental animals since 2012, the reference value of normal levels of leukocytes is not available. To obtain such information, 62 blood samples from male and female healthy northern pig-tailed macaques at different ages were collected. The normal range of major leukocyte subpopulations, such as T lymphocytes, B lymphocytes, natural killer (NK) cells, monocytes, and the expression levels of activation or differentiation related molecules (CD38, HLA-DR, CCR5, CD21, IgD, CD80 and CD86) on lymphocytes were analyzed by flow cytometry. The counts of B cells decreased with age, but those of CD8⁺ T cells and NK cells and the frequency of CD38⁺HLA-DR⁺CD4⁺ T cells were positively correlated with age. The counts of leukocyte subpopulations were higher in males than those in females except for CD4⁺ T cells. Males also showed higher expression levels of IgD and CD21 within B cells. This study provides basic data about the leukocyte subpopulations of northern pig-tailed macaques and compares this species with commonly used Chinese rhesus macaques (M. mulatta), which is meaningful for the biomedical application of northern pig-tailed macaques.

485-491

Authors: Jian-Ping LI, Yun LIAO, Ying ZHANG, Jing-Jing WANG, Li-Chun WANG, Kai FENG, Qi-Han LI, Long-Ding LIU

Coxsackie virus A16 (CA16) is commonly recognized as one of the main human pathogens of hand-foot-mouth disease (HFMD). The clinical manifestations of HFMD include vesicles of hand, foot and mouth in young children and severe inflammatory CNS lesions. In this study, experimentally CA16 infected tree shrews(Tupaia belangeri) were used to investigate CA16 pathogenesis. The results showed that both the body temperature and the percentages of blood neutrophilic granulocytes / monocytes of CA16 infected tree shrews increased at 4-7 days post infection. Dynamic distributions of CA16 in different tissues and stools were found at different infection stages. Moreover, the pathological changes in CNS and other organs were also observed. These findings indicate that tree shrews can be used as a viable animal model to study CA16 infection.

500-509

Authors: Hui-Hua LIU, Cai-Fei HUANG, Xin WANG

Under free field conditions, we used single unit extracellular recording to study the detection of acoustic signals by neurons in the ventral nucleus of the lateral lemniscus (VNLL) in Kunming mouse (Mus musculus). The results indicate two types of firing patterns in VNLL neurons: onset and sustained. The first spike latency (FSL) of onset neurons was shorter than that of sustained neurons. With increasing sound intensity, the FSL of onset neurons remained stable and that of sustained neurons was shortened, indicating that onset neurons are characterized by precise timing. By comparing the values of Q₁₀ and Q₃₀ of the frequency tuning curve, no differences between onset and sustained neurons were found, suggesting that firing pattern and frequency tuning are not correlated. Among the three types of rate-intensity function (RIF) found in VNLL neurons, the proportion of monotonic RIF is the largest, followed by saturated RIF, and non-monotonic RIF. The dynamic range (DR) in onset neurons was shorter than in sustained neurons, indicating different capabilities in intensity tuning of different firing patterns and that these differences are correlated with the type of RIF. Our results also show that the best frequency of VNLL neurons was negatively correlated with depth, supporting the view point that the VNLL has frequency topologic organization.

528-536

Authors: Ya HE, Zheng-Huan WANG, Xiao-Ming WANG

Cervus sichuanicus is a species of sika deer (Cervus nippon Group). To date, research has mainly focused on quantity surveying and behavior studies, with genetic information on this species currently deficient. To provide scientific evidence to assist in the protection of this species, we collected Sichuan sika deer fecal samples from the Sichuan Tiebu Nature Reserve (TNR) and extracted DNA from those samples. Microsatellite loci of bovine were used for PCR amplification. After GeneScan, the genotype data were used to analyze the genetic diversity and population structure of the Sichuan sika deer in TNR. Results showed that the average expected heterozygosity of the Sichuan sika deer population in TNR was 0.562, equivalent to the average expected heterozygosity of endangered animals, such as Procapra przewalskii. Furthermore, 8 of 9 microsatellite loci significantly deviated from the Hardy-Weinberg equilibrium and two groups existed within the Sichuan sika deer TNR population. This genetic structure may be caused by a group of Manchurian sika deer (Cervus hortulorum) released in TNR.

362-372

Authors: Xiao-Yun SUI, Zhi LU, Yun-Zhi YAN, Yi-Feng CHEN, Yin-Tao JIA

Using seasonally collected data (2009-2010) from 15 sampling sites that represent first- to fifth-order streams within the Qingyi watershed, we examined the spatio-temporal patterns of fish assemblages along two longitudinal gradients to explore the effects of a large dam on fish assemblages at the watershed scale. No significant variation was observed in either species richness or assemblage structure across seasons. Species richness significantly varied according to stream order and gradient. Dam construction appeared to decrease species richness upstream substantially, while a significant decrease between gradients only occurred within fourth-order streams. Along the gradient without the large dam, fish assemblage structures presented distinct separation between two neighboring stream orders, with the exception of fourth-order versus fifth-order streams. However, the gradient disrupted by a large dam displayed the opposite pattern in the spatial variation of fish assemblages related with stream orders. Significant between-gradient differences in fish assemblage structures were only observed within fourth-order streams. Species distributions were determined by local habitat environmental factors, including elevation, substrate, water depth, current discharge, wetted width, and conductivity. Our results suggested that dam construction might alter the longitudinal pattern in fish species richness and assemblage structure in Qingyi Stream, despite the localized nature of the ecological effect of dams.

380-388

Authors: Qiao-Qing XU, Dai-Qin YANG, Rui TUO, Jing WAN, Ming-Xian CHANG, Pin NIE

The Asian swamp eel (Monopterus albus) is one of the most economically important freshwater fish in East Asia, but data on the immune genes of M. albus are scarce compared to other commercially important fish. A better understanding of the eel's immune responses may help in developing strategies for disease management, potentially improving yields and mitigating losses. In mammals, interferon regulatory factors (IRFs) play a vital role in both the innate and adaptive immune system; though among teleosts IRF4 and IRF10 have seldom been studied. In this study, we characterized IRF4 and IRF10 from M. albus (maIRF4 and maIRF10) and found that maIRF4 cDNA consists of 1 716 nucleotides encoding a 451 amino acid (aa) protein, while maIRF10 consists of 1 744 nucleotides including an open reading frame (ORF) of 1 236 nt encoding 411 aa. The maIRF10 gene was constitutively expressed at high levels in a variety of tissues, while maIRF4 showed a very limited expression pattern. Expression of maIRF4 and maIRF10 in head kidney, and spleen tissues was significantly up-regulated from 12 h to 48 h post-stimulation with polyinosinic: polycytidylic acid (poly I:C), lipopolysaccharide (LPS) and a common pathogenic bacteria Aeromonas hydrophila. These results suggest that IRF4 and IRF10 play roles in immune responses to both viral and bacterial infections in M. albus.

398-403

Authors: Yu-Cheng JI, Hui ZHUGE, Shan-Shan ZHANG, Shu-Fang ZHANG, Xian-Yu Yang

In this study, to clarify the bioactive polypeptides included in the skins and secretions of Bufo, we screened the Japanese toad (Bufo japonicus formosus) skin cDNA library by colony polymerase chain reaction (PCR), and obtained a transcript of 1 075 bp consisting of 1 37 bp 5' untranslated region (UTR), 515 bp 3' UTR and a 423 bp open reading frame (ORF) encoding a polypeptide of 140 amino acid residues (GenBank accession number: KF359945). Homolog analysis showed a 70%-96% homology with sterol carrier protein-2 (SCP-2) present in other animals, which is implicated in lipid metabolism of other organisms. The gene SCP-2 of Chinese toad (B. gargarizans) was cloned from a first strand cDNA of Bufo skin (GenBank accession number: KF381341) via PCR, whose encoding polypeptide has only one amino acid difference from that of Japanese toad. Tissue distribution analysis showed that SCP-2 expressed in all organs tested, though in the liver and spleen it manifested lower expression than in other organs. These findings might indicate SCP-2 being one of the active ingredients in toad skin. These findings may in turn have implications for further drug development from traditional Chinese medicine sources.

411-419

Authors: Peng LI, Cai-Hong JIN, San JIANG, Miao-Miao LI, Zi-Lu WANG, Hui ZHU, Cui-Yun CHEN, Tian-Miao HUA

The influence of intracortical inhibition on the response adaptation of visual cortical neurons remains in debate. To clarify this issue, in the present study the influence of surround suppression evoked through the local inhibitory interneurons on the adaptation effects of neurons in the primary visual cortex (V1) were observed. Moreover, the adaptations of V1 neurons to both the high-contrast visual stimuli presented in the classical receptive field (CRF) and to the costimulation presented in the CRF and the surrounding nonclassical receptive field (nCRF) were compared. The intensities of surround suppression were modulated with different sized grating stimuli. The results showed that the response adaptation of V1 neurons decreased significantly with the increase of surround suppression and this adaptation decrease was due to the reduction of the initial response of V1 neurons to visual stimuli. However, the plateau response during adaptation showed no significant changes. These findings indicate that the adaptation effects of V1 neurons may not be directly affected by surround suppression, but may be dynamically regulated by a negative feedback network and be finely adjusted by its initial spiking response to stimulus. This adaptive regulation is not only energy efficient for the central nervous system, but also beneficially acts to maintain the homeostasis of neuronal response to long-presenting visual signals.

426-435

Authors: Jian-Li WANG, Bei WANG, Wen CHEN

C57BL/6J and BALB/cJ mice display significant differences in sociability and response to drugs, but the phenotypic variability of their susceptibility to cocaine is still not well known. In this study, the differences between these two mice strains in the persistence of cocaine-induced conditioned place preference (CPP), as well as the locomotion and social behaviors after the 24-hour withdrawal from a four-day cocaine (20 mg/kg/day) administration were investigated. The results showed that the cocaine-induced CPP persisted over two weeks in C57BL/6J mice, while it diminished within one week among BALB/cJ mice. After 24-hours of cocaine withdrawal, high levels of locomotion as well as low levels of social interaction and aggressive behavior were found in C57BL/6J mice, but no significant changes were found in BALB/cJ mice, indicating that cocaine-induced CPP persistence, locomotion and social behavior are not consistent between these two strains, and that overall C57BL/6J mice are more susceptible to cocaine than BALB/cJ mice at the tested doses.

256-261

Authors: Liang JING, Qi-Xin ZHOU, Lin XU

The social transmission of food preferences (STFP) is a behavioural task of olfactory memory, in which an observer rat learns safe food odours from a demonstrator rat, and shows preference for this odour in a subsequent choice test. However, previous studies have failed to detect the transmission of information about food of potential danger and food aversion using STFP test. In this study, we tested how demonstrators' health affects the exchange of odour information and whether observers can learn danger information from an unhealthy demonstrator. As expected, the observer rat formed an odour preference after interacting with a demonstrator rat that had just eaten food containing a new odour, however, odour preference rather than aversion was also formed after interacting with a demonstrator rat injected with LiCl (used to induce gastric malaise). Furthermore, anaesthetized demonstrator rats and half-anaesthetized demonstrator rats, which showed obvious motor deficits suggesting an unhealthy state, also socially transmitted food preferences to observers. These results suggest that the social transmission of food preferences task is independent of a demonstrators' health, and that information about dangerous foods cannot be transmitted using this behavioural task.

272-279

Authors: Li WEI, Wei-Wei SHAO, Guo-Hua DING, Xiao-Li FAN, Miao-Ling YU, Zhi-Hua LIN

We studied acute and joint toxicity of three different agrochemicals (chlorantraniliprole, flubendiamide-abamectin and penoxsulam) to Chinese tiger frog (Hoplobatrachus chinensis) tadpoles with the method of stability water tests. Results showed that the three agrochemicals increased tadpole mortality. For acute toxicity, the LC₅₀ values after 24, 48 and 72 h of chlorantraniliprole, flubendiamide-abamectin and penoxsulam exposure were 5.37, 4.90 and 4.68 mg/L; 0.035, 0.025 and 0.021 mg/L; 1.74, 1.45 and 1.29 mg/L, respectively. The safety concentrations (SC) of chlorantraniliprole, flubendiamide-abamectin and penoxsulam to the tadpoles were 1.23, 0.30 and 0.003 mg/L, respectively. Based on these findings, chlorantraniliprole and penoxsulam were moderately toxic, while flubendiamide-abamectin was highly toxic. All pairwise joint toxicity tests showed moderate toxicity. The LC₅₀ values after 24, 48 and 72 h of exposure were 7.08, 6.61 and 6.03 mg/L for chlorantraniliprole+penoxsulam, with corresponding values of 2.455, 2.328 and 2.183 mg/L for chlorantraniliprole+flubendiamide-abamectin, and 1.132, 1.084 and 1.050 mg/L for penoxsulam+flubendiamide-abamectin, with safe concentrations of 1.73, 0.63 and 0.30 mg/L, respectively. For toxic evaluations of pairwise combinations of the three agrochemicals, only the joint toxicity of chlorantraniliprole and flubendiamide-abamectin after 24 h was found to be synergistic, whereas all other tests were antagonistic. Our findings provide valuable information on the toxic effects of agrochemicals on amphibians and how various types of agrochemicals can be reasonably used in agricultural areas.

287-293

Authors: Zhi-Gui ZHANG, De-Yan GE

In the present study, postnatal ontogenetic size and shape changes in the cranium of two lagomorph species, the plateau pika (Ochotona curzoniae) and woolly hare (Lepus oiostolus), were investigated by geometric morphometrics. The ontogenetic size and shape changes of their cranium exhibited different growth patterns in response to similar environmental pressures on the Qinghai-Tibetan Plateau. The overall size change in the cranium of the plateau pika was slower than that of the woolly hare. The percentage of ontogenetic shape variance explained by size in the woolly hare was greater than that in the plateau pika. The overall shape of the cranium was narrowed in both species, and morphological components in relation to neural maturity showed negative allometry, while those responsible for muscular development showed isometric or positive allometry. The most remarkable shape variations in the plateau pika were associated with food acquisition (temporalis development), though other remarkable shape variations in the incisive and palatal foramen in the ventral view were also observed. The most important shape change in the woolly hare was demonstrated by the elongation of the nasal bones, expansion of the supra-orbital process and shape variation of the neurocranium.

300-306

Authors: Yu-Mei TIAN, Jie CHEN, Yang TAO, Xia-Yun JIANG, Shu-Ming ZOU

Insulin-like growth factor-binding protein 1 (IGFBP-1), a hypoxia-induced protein, is a member of the IGFBP family that regulates vertebrate growth and development. In this study, full-length IGFBP-1a cDNA was cloned from a hypoxia-sensitive Cyprinidae fish species, the blunt snout bream (Megalobrama amblycephala). IGFBP-1a was expressed in various organs of adult blunt snout bream, including strongly in the liver and weakly in the gonads. Under hypoxia, IGFBP-1a mRNA levels increased sharply in the skin, liver, kidney, spleen, intestine and heart tissues of juvenile blunt snout bream, but recovered to normal levels after 24-hour exposure to normal dissolved oxygen. In blunt snout bream embryos, IGFBP-1a mRNA was expressed at very low levels at both four and eight hours post-fertilization, and strongly at later stages. Embryonic growth and development rates decreased significantly in embryos injected with IGFBP-1a mRNA. The average body length of IGFBP-1a-overexpressed embryos was 82.4% of that of the control group, and somite numbers decreased to 85.2%. These findings suggest that hypoxia-induced IGFBP-1a may inhibit growth in this species under hypoxic conditions.

313-318

Authors: Fang ZHAO, Jun-Ying MA, Hui-Xia CAI, Jian-Ping SU, Zhi-Bin HOU, Tong-Zuo ZHANG, Gong-Hua LIN

Cestode larvae spend one phase of their two-phase life cycle in the viscera of rodents, but cases of cestodes infecting subterranean rodents have only been rarely observed. To experimentally gain some insight into this phenomenon, we captured approximately 300 plateau zokors (Eospalax baileyi), a typical subterranean rodent inhabiting the Qinghai-Tibet Plateau, and examined their livers for the presence of cysts. Totally, we collected five cysts, and using a mitochondrial gene (cox1) and two nuclear genes (pepck and pold) as genetic markers, we were able to analyze the taxonomy of the cysts. Both the maximum likelihood and Bayesian methods showed that the cysts share a monophyly with Taenia mustelae, while Kimura 2-parameter distances and number of different sites between our sequences and T. mustelae were far less than those found between the examined sequences and other Taeniidae species. These results, alongside supporting paraffin section histology, imply that the cysts found in plateau zokors can be regarded as larvae of T. mustelae, illustrating that zokors are a newly discovered intermediate host record of this parasite.

326-331

Authors: Xin-Yi LI, Qian LI, Yu-Hui ZHANG

In the current study, 5-nydroxytryptamine (5-HT) and gastrin (GAS) cells in the digestive canals of Rana chensinensis tadpoles at different developmental stages were investigated by immunohistochemistry. Results showed that the 5-HT cells were only detected in the duodenum before metamorphosis began, and were extensively distributed in the stomach, duodenum, small intestine, and rectum thereafter, with the highest counts found in the duodenum and rectum when metamorphosis was completed. The GAS cells were only distributed in the stomach and duodenum, and only rarely detected in the duodenum before metamorphosis began, but increased in the stomach during metamorphosis and showed zonal distribution in the gastric mucosa when metamorphosis was completed. Metamorphosis is a critical period for amphibians, during which structural and functional physiological adaptations are required to transition from aquatic to terrestrial environments. During metamorphosis, the differentiations of 5-HT cells in the gastrointestinal canals of tadpoles could facilitate mucus secretion regulation, improve digestive canal lubrication, and help water-shortage food digestion in terrestrial environments. Conversely, GAS cell differentiations during metamorphosis might contribute to the digestive and absorptive function transition from herbivore to omnivore.

338-345

Authors: Muhammad Naeem AWAN, Hassan ALI, David Charles LEE

The cheer pheasant Catreus wallichi is a globally threatened species that inhabits the western Himalayas. Though it is well established that the species is threatened and its numbers declining, updated definitive estimates are lacking, so in 2011, we conducted a survey to assess the density, population size, and threats to the species in Jhelum valley, Azad Kashmir, which holds the largest known population of cheer pheasants in Pakistan. We conducted dawn call count surveys at 17 points clustered in three survey zones of the valley, 11 of which had earlier been used for a 2002-2003 survey of the birds. Over the course of our survey, 113 birds were recorded. Mean density of cheer pheasant in the valley was estimated at 11.8±6.47 pairs per km², with significant differences in terms of both counts and estimated density of cheer were significantly different across the three survey zones, with the highest in the Chinari region and the lowest, that is the area with no recorded sightings of the pheasants, in Gari Doppata. The total breeding population of cheer pheasants is estimated to be some 2 490 pairs, though this does not consider the actual area of occupancy in the study area. On the whole, more cheer pheasants were recorded in this survey than from the same points in 2002-2003, indicating some success in population growth. Unfortunately, increasing human settlement, fires, livestock grazing, hunting, and the collection of non-timber forest products continue to threaten the population of cheer in the Jhelum valley. To mitigate these potential impacts, some degree of site protection should be required for the conservation of cheer pheasants in Pakistan, and more effective monitoring of the species is clearly needed.

350-352

Authors: Qin LIU, Guang-Hui ZHONG, Shi-Ze LI, Jing-Cai L&220;, Peng GUO

Three Asian green pit vipers were collected in August 2013 during a field trip in Fanjin Mt. National Conservation Area, Guizhou. These specimens were identified as Sinovipera sichuanensis, based on subsequent examination and comparison. This is a new record of the genus Sinovipera and S. sichuanensis in Guizhou, and the first time that male specimens have been collected in the field.