Zoological Research

314-318

Authors: Min WANG, Amy F.T. ARNSTEN

Revealing how molecular mechanisms influence higher brain circuits in primates will be essential for understanding how genetic insults lead to increased risk of cognitive disorders. Traditionally, modulatory influences on higher cortical circuits have been examined using lesion techniques, where a brain region is depleted of a particular transmitter to determine how its loss impacts cognitive function. For example, depletion of catecholamines or acetylcholine from the dorsolateral prefrontal cortex produces striking deficits in working memory abilities. More directed techniques have utilized direct infusions of drug into a specific cortical site to try to circumvent compensatory changes that are common following transmitter depletion. The effects of drug on neuronal firing patterns are often studied using iontophoresis, where a minute amount of drug is moved into the brain using a tiny electrical current, thus minimizing the fluid flow that generally disrupts neuronal recordings. All of these approaches can be compared to systemic drug administration, which remains a key arena for the development of effective therapeutics for human cognitive disorders. Most recently, viral techniques are being developed to be able to manipulate proteins for which there is no developed pharmacology, and to allow optogenetic manipulations in primate cortex. As the association cortices greatly expand in brain evolution, research in nonhuman primates is particularly important for understanding the modulatory regulation of our highest order cognitive operations.

328-336

Authors: Elena V. ANUFRIIEVA, Nickolai V. SHADRIN

Inter-species variability in morphological traits creates a need to know the range of variability of characteristics in the species for taxonomic and ecological tasks. Copepoda Arctodiaptomus salinus, which inhabits water bodies across Eurasia and North Africa, plays a dominant role in plankton of different water bodies-from fresh to hypersaline. This work assesses the intra- and inter-population morphometric variability of A. salinus in the Mediterranean-Black Sea region and discusses some observed regularities. The variability of linear body parameters and proportions was studied. The impacts of salinity, temperature, and population density on morphological characteristics and their variability can manifest themselves in different ways at the intra- and inter-population levels. A significant effect of salinity, pH and temperature on the body proportions was not found. Their intra-population variability is dependent on temperature and salinity. Sexual dimorphism of A. salinus manifests in different linear parameters, proportions, and their variability. There were no effects of temperature, pH and salinity on the female/male parameter ratio. There were significant differences in the body proportions of males and females in different populations. The influence of temperature, salinity, and population density can be attributed to 80%-90% of intra-population variability of A. salinus. However, these factors can explain less than 40% of inter-population differences. Significant differences in the body proportions of males and females from different populations may suggest that some local populations of A. salinus in the Mediterranean-Black Sea region are in the initial stages of differentiation.

342-347

Authors: Patrick Kinyatta MALONZA

Faunal species richness is traditionally assumed to decrease with increasing elevation and decreasing primary productivity. Species richness is reported to peak at mid-elevation. This survey examines the herpetofaunal diversity and distribution in Mt. Kenya (central Kenya) by testing the hypothesis that changes in species richness with elevation relate to elevation-dependent changes in climate. Sampling along transects from an elevation of approximately 1 700 m in Chogoria forest block (wind-ward side) and approximately 2 600 m in Sirimon block (rain shadow zone) upwards in March 2009. This starts from the forest to montane alpine zones. Sampling of reptiles and amphibians uses pitfall traps associated with drift fences, time-limited searches and visual encounter surveys. The results show that herpetofaunal richness differs among three vegetation zones along the elevation gradient. Chogoria has higher biodiversity than Sirimon. More species occur at low and middle elevations and few exist at high elevations. The trends are consistent with expected optimum water and energy variables. The lower alpine montane zone has high species richness but low diversity due to dominance of some high elevations species. Unambiguous data do not support a mid-domain effect (mid-elevation peak) because the observed trend better fits a model in which climatic variables (rainfall and temperature) control species richness, which indirectly measures productivity. It is important to continue protection of all indigenous forests, especially at low to mid elevations. These areas are vulnerable to human destruction yet are home to some endemic species. Firebreaks can limit the spread of the perennial wildfires, especially on the moorlands.

257-262

Authors: Xiong GUO, Bo LUO, Ying LIU, Ting-Lei JIANG, Jiang FENG

Identity recognition is one of the most critical social behaviours in a variety of animal species. Microchiropteran bats present a special use case of acoustic communication in the dark. These bats use echolocation pulses for navigating, foraging, and communicating; however, increasing evidence suggests that echolocation pulses also serve as a means of social communication. Compared with echolocation signals, communication calls in bats have rather complex structures and differ substantially by social context. Bat acoustic signals vary broadly in spectrotemporal space among individuals, sexes, colonies and species. This type of information can be gathered from families of vocalizations based on voice characteristics. In this review we summarize the current studies regarding vocal identity recognition in microbats. We also provide recommendations and directions for further work.

285-298

Authors: Lin ZHU, Ai-Hua LEI, Hong-Yi ZHENG, Long-Bao LYU, Zhi-Gang ZHANG, Yong-Tang ZHENG

The complex and dynamic vaginal microbial ecosystem is critical to both health and disease of the host. Studies focusing on how vaginal microbiota influences HIV-1 infection may face limitations in selecting proper animal models. Given that northern pig-tailed macaques (Macaca leonina) are susceptible to HIV-1 infection, they may be an optimal animal model for elucidating the mechanisms by which vaginal microbiota contributes to resistance and susceptibility to HIV-1 infection. However, little is known about the composition and temporal variability of vaginal microbiota of the northern pig-tailed macaque. Here, we present a comprehensive catalog of the composition and temporal dynamics of vaginal microbiota of two healthy northern pig-tailed macaques over 19 weeks using 454-pyrosequencing of 16S rRNA genes. We found remarkably high proportions of a diverse array of anaerobic bacteria associated with bacterial vaginosis. Atopobium and Sneathia were dominant genera, and interestingly, we demonstrated the presence of Lactobacillus-dominated vaginal microbiota. Moreover, longitudinal analysis demonstrated that the temporal dynamics of the vaginal microbiota were considerably individualized. Finally, network analysis revealed that vaginal pH may influence the temporal dynamics of the vaginal microbiota, suggesting that inter-subject variability of vaginal bacterial communities could be mirrored in inter-subject variation in correlation profiles of species with each other and with vaginal pH over time. Our results suggest that the northern pig-tailed macaque could be an ideal animal model for prospective investigation of the mechanisms by which vaginal microbiota influence susceptibility and resistance to HIV-1 infection in the context of highly polymicrobial and Lactobacillus-dominated states.

305-309

Authors: Tian-Zi WANG, Ai SUN, Na WANG, Zhong-Kai CUI, Song-Lin CHEN, Zhen-Xia SHA

An astroglial cell line was established from the brain of half smooth tongue sole (Cynoglossus semilaevis) and was designated as CSAC. CSAC shows the morphological homogeneity of epithelial cells. The cell identity was tested by the presence of glial fibrillary acidic protein (GFAP), which was revealed by RT-PCR and immunofluorescence. The cell line was optimally maintained at 24 °C in minimum essential medium supplemented with HEPES, antibiotics, 20% fetal bovine serum, 2-Mercaptoethanol (2-Me) and basic fibroblast growth factor. Chromosome analysis revealed that the CSAC cells maintained a normal diploid chromosome number (2n=42). The fluorescent signals were observed in CSAC after the cells were transfected with green fluorescent protein (GFP) reporter plasmids. The CSAC cell line may serve as a valuable tool for studies on the potential functions of fish astroglial cells.

183-222

Authors: Yun ZHANG

Venom (toxins) is an important trait evolved along the evolutionary tree of animals. Our knowledges on venoms, such as their origins and loss, the biological relevance and the coevolutionary patterns with other organisms are greatly helpful in understanding many fundamental biological questions, i.e., the environmental adaptation and survival competition, the evolution shaped development and balance of venoms, and the sophisticated correlations among venom, immunity, body power, intelligence, their genetic basis, inherent association, as well as the cost-benefit and trade-offs of biological economy. Lethal animal envenomation can be found worldwide. However, from foe to friend, toxin studies have led lots of important discoveries and exciting avenues in deciphering and fighting human diseases, including the works awarded the Nobel Prize and lots of key clinic therapeutics. According to our survey, so far, only less than 0.1% of the toxins of the venomous animals in China have been explored. We emphasize on the similarities shared by venom and immune systems, as well as the studies of toxin knowledge-based physiological toxin-like proteins/peptides (TLPs). We propose the natural pairing hypothesis. Evolution links toxins with humans. Our mission is to find out the right natural pairings and interactions of our body elements with toxins, and with endogenous toxin-like molecules. Although, in nature, toxins may endanger human lives, but from a philosophical point of view, knowing them well is an effective way to better understand ourselves. So, this is why we study toxins.

233-240

Authors: Huan LI, Ming-Xia ZHANG, Xiao-Jun YANG, Liang-Wei CUI, Rui-Chang QUAN

To fill the gap in breeding biology information about the Red-Whiskered Bulbul (Pycnonotus jocosus), in 2013, we studied the breeding biology of this species in Xishuangbanna, southwest China. The breeding began from February and continued until early August. The breeding strategy of P. jocosus was more flexible and their nests were only built in cultivated landscapes, whereas, no nest building in native tropical rain forest was found. Small open cup nests were built on 50 different plant species, and at heights ranging from 2.1±0.6 m above the ground (n=102). The mean clutch size was 2.53±0.51 eggs (n=40) and the mean egg mass was 2.81±0.25 g (n=60). The average incubation period was 11.1±0.5 days (n=14), and the average nestling period was 11.0±0.8 days (n=31). The overall nest success was 34.22%.The hatching and fledging showed either asynchrony or synchrony. Invertebrate food decreased with nestling age, whereas, plant food increased with nestling age. Moreover, distinct parental roles of the parents in nestling period were found. Compared with other passerine species, P. jocosus spent less time in incubating (58%). The clutch size, incubation and nestling period of the P. jocosus in southwest China were different from those of the P. jocosus in India.

248-254

Authors: Si-Ling FU, Jiang-Lin LI, Jia CHEN, Qiu-Ting WANG, Jian-Jun LI, Xian-Chun WANG

The eggs of oviparous animals are storehouses of maternal proteins required for embryonic development. Identification and molecular characterization of such proteins will provide much insight into the regulation of embryonic development. We previously analyzed soluble proteins in the eggs of the black widow spider (Latrodectus tredecimguttatus), and report here on the extraction and mass spectrometric identification of the egg membrane proteins. Comparison of different lysis solutions indicated that the highest extraction of the membrane proteins was achieved with 3%-4% sodium laurate in 40 mmol/L Tris-HCl buffer containing 4% CHAPS and 2% DTT (pH 7.4). SDS-PAGE combined with nLC-MS/MS identified 39 proteins with membrane-localization annotation, including those with structural, catalytic, and regulatory activities. Nearly half of the identified membrane proteins were metabolic enzymes involved in various cellular processes, particularly energy metabolism and biosynthesis, suggesting that relevant metabolic processes were active during the embryonic development of the eggs. Several identified cell membrane proteins were involved in the special structure formation and function of the egg cell membranes. The present proteomic analysis of the egg membrane proteins provides new insight into the molecular mechanisms of spider embryonic development.

119-132

Authors: Victor Benno MEYER-ROCHOW

Mass aggregations and migrations of millipedes despite numerous attempts to find causes for their occurrences are still an enigma. They have been reported from both southern and northern hemisphere countries, from highlands and lowlands of both tropical and temperate regions and they can involve species belonging to the orders Julida and Spirobolida, Polydesmida and Glomerida. According to the main suggestions put forward in the past, mass occurrences in Diplopoda occur: (1) because of a lack of food and a population increase beyond sustainable levels; (2) for the purpose of reproduction and in order to locate suitable oviposition sites; (3) to find overwintering or aestivation sites; (4) because of habitat disruption and changes in the local environment; (5) as a consequence of weather conditions the year (or winter and spring) before. A recent outbreak (November 2014) of a mass migration of the polydesmid Chamberlinius hualienensis Wang 1956 on the Japanese Izu Island of Hachijojima 300 km to the south of Tokyo gave this author an opportunity to review the existing literature on millipede mass migrations and to carry out additional observations on the phenomenon in the field as well as the laboratory. Hitherto unreported heavy infestations with phoretic deutonymphs of the mite Histiostoma sp. as well as dense populations of internal rhabditid nematodes (Oscheius cf. necromena and an unidentified species of the genus Fictor), suggest that infestations of this kind could be necromenic and either have been a contributing factor for the mass migration or been a consequence of so many individuals occurring together at close proximity. It is concluded that mass migrations and aggregations in millipedes do not have one common cause, but represent phenomena that often are seasonally recurring events and appear identical in their outcome, but which have evolved as responses to different causes in different millipede taxa and therefore need to be examined on a case-to-case basis.

142-151

Authors: Feng-Chao LANG, Xin LI, Olga VLADMIROVA, Zhuo-Ran LI, Gui-Jun CHEN, Yu XIAO, Li-Hong LI, Dan-Feng LU, Hong-Bo HAN, Ju-Min ZHOU

Herpes simplex virus type 1 (HSV-1) enters productive infection after infecting epithelial cells, where it controls the host nucleus to make viral proteins, starts viral DNA synthesis and assembles infectious virions. In this process, replicating viral genomes are organized into replication centers to facilitate viral growth. HSV-1 is known to use host factors, including host chromatin and host transcription regulators, to transcribe its genes; however, the invading virus also encounters host defense and stress responses to inhibit viral growth. Recently, we found that HSV-1 replication centers recruit host factor CTCF but exclude γH2A.X. Thus, HSV-1 replication centers may selectively recruit cellular factors needed for viral growth, while excluding host factors that are deleterious for viral transcription or replication. Here we report that the viral replication centers selectively excluded modified histone H3, including heterochromatin mark H3K9me3, H3S10P and active chromatin mark H3K4me3, but not unmodified H3. We found a dynamic association between the viral replication centers and host RNA polymerase II. The centers also recruited components of the DNA damage response pathway, including 53BP1, BRCA1 and host antiviral protein SP100. Importantly, we found that ATM kinase was needed for the recruitment of CTCF to the viral centers. These results suggest that the HSV-1 replication centers took advantage of host signaling pathways to actively recruit or exclude host factors to benefit viral growth.

161-166

Authors: Lin ZHANG, Xu-He HUANG, Ping-Ping ZHOU, Guo-Long YU, Jin YAN, Bing QIN, Xin-Ge YAN, Li-Mei DIAO, Peng LIN, Yi-Qun KUANG

Chemokine receptors CXCR4 and CCR5 are indispensable co-receptors for HIV-1 entry into host cells. In our previous study, we identified that dopamine receptor-interacting protein 78 (DRiP78) and Na⁺-H⁺ exchanger regulatory factor 1 (NHERF1) are the CXCR4 and CCR5 homo- or hetero-dimer-interacting proteins. DRiP78 and NHERF1 are able to influence the co-receptor internalization and intracellular trafficking. Over-expression of NHERF1 affects the ligands or HIV-1 gp120-induced CCR5 internalization and HIV-1 production. It is reasonable to speculate that DRiP78 and NHERF1, as well as the signaling pathways involved in viral replication, would probably affect HIV-1 replication through regulating the co-receptors. In this present study, we designed two short hairpin RNAs (shRNAs) targeting the DRiP78 and NHERF1, respectively, and constructed the pLenti6/BLOCK-iT-DEST lentiviral plasmids expressing DRiP78 or NHERF1 shRNA. The packaged lentiviruses were used to transduce the widely-applied HIV-1 model cell line GHOST(3). Then, cells with stable knockdown were established through selecting transduced cells with Blasticidin. This study, for the first time, reported the establishment of the GHOST(3) with DRiP78 and NHERF1 knockdown, which is the first stable cell line with HIV-1 co-receptor-interacting molecular defects.

174-177

Authors: Wen-Bin JIANG, Ma HAKIM, Lei LUO, Bo-Wen LI, Shi-Long YANG, Yu-Zhu SONG, Ren LAI, Qiu-Min LU

As a group of intestinal hormones and neurotransmitters, cholecystokinins (CCKs) regulate and affect pancreatic enzyme secretion, gastrointestinal motility, pain hypersensitivity, digestion and satiety, and generally contain a DYMGWMDFG sequence at the C-terminus. Many CCKs have been reported in mammals. However, only a few have been reported in amphibians, such as Hyla nigrovittata, Xenopus laevis, and Rana catesbeiana, with none reported in urodele amphibians like newts and salamanders. Here, a CCK called CCK-TV was identified and characterized from the skin of the salamander Tylototriton verrucosus. This CCK contained an amino acid sequence of DYMGWMDF-NH₂ as seen in other CCKs. A cDNA encoding the CCK precursor containing 129 amino acid residues was cloned from the cDNA library of T. verrucosus skin. The CCK-TV had the potential to induce the contraction of smooth muscle strips isolated from porcine gallbladder, eliciting contraction at a concentration of 5.0x10^-11 mol/L and inducing maximal contraction at a concentration of 2.0x10^-6 mol/L. The EC₅₀ was 13.6 nmol/L. To the best of our knowledge, this is the first report to identify the presence of a CCK in an urodele amphibian.

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