2008 Vol. 29, No. 2
Systematics of Perca Species Based on Multivariate Morphometrics and Mitochondrial Cytochrome b Gene Variation Analysis
2008, 29(2): 113-120. doi: 10.3724/SP.J.1141.2008.02113
This paper investigated the classification of the genus Perca based on the 32 multivariate morphometrics and the 1 134 bp sequences of mitochondrial cytochrome b (Cyt b) gene. The result of multivariate morphometric analyses showed that the distance between P. fluviatilis and P. schrenki, between P. schrenki and P. flavescens, between P. fluviatilis and P. flavescens was 0.15, 0.14, 0.09, respectively. Perca fluviatilis and P. flavescens were much more similar in morphology, and there was a remarkable difference between P. schrenki and the two other species. In the scatter-plot figure based on principal components 1 to 2, there was an overlapping section between P. fluviatilis and P. flavescens, but there was no overlapping section between P. schrenki and the two others. In the analysis of mitochondrial cytchrome b gene, the percentage nucleotide sequence divergence between P. fluviatilis and P. schrenki, between P. schrenki and P. flavescens, between P. fluviatilis and P. flavescens was 13.08%, 10.68%, 11.47% respectively. The sequence divergences among the three Perca species were within interspecific divergence. Molecular phylogenetic trees were constructed based on the sequences of 20 samples with maximum parsimony (MP), neighbor-joining (NJ) and Maximum likelihood methods. The topological structures of the three trees were consistent, and they all showed that the relationship between the P. schrenki and P. flavescens was much closer than that between P. fluviatilis and P. flavescens. From the genetic divergence of the Cyt b gene and the isolation in geographic distribution, we further concluded that P. fluviatilis and P. flavescens were different species. The genus Perca therefore, includes three species, P. fluviatilis, P.schrenki and P. flavescens.
Genetic Variation and Diversity of Phrynocephalus vlangalii hongyuanensis in Zoige Wetland Inferred from ND4-tRNAleu Gene
2008, 29(2): 121-126. doi: 10.3724/SP.J.1141.2008.04353
According to the distribution of Phrynocephalus vlangalii hongyuanensis in Zoige Wetland，three geographic units: Zoige Xiaman (XM)，Hongyuan (HY)，both in Sichuan Province and Maqu (MQ) in Gansu Province were defined. We used molecular methods to reveal these unit’s genetic variation and diversity. A 785bp fragment of the mtDNA ND4-tRNAleu was determined from 72 samp1es in seven populations of P. vlangalii hongyuanensis. Seven variable nucleotide sites and nine haplotypes were identified in the 785bp fragments. As a whole，the haplotype diversity was high (0.806±0.024)，but the nucleotide diversity was low (0.00231±0.00016). In a single population，MQa，MQb and XMb had very low genetic diversities，and XMc had a much higher one. The Kimura 2-parameter distances among all the populations were small (0.001-0.005)，and the distance between MQa and XMa was the greatest. Analysis of molecular variance (AMOVA) showed that the three units were distinctly different (P<0.01)，and 62.61% of the total genetic diversity was attributable to variation among units. There were 3 haplotypes shared among XM and HY，and no geographic clustering was observed except MQ from the TCS network. The results from the mismatch distribution analysis and Fu’s Fs test (Fs=－2.21937) implied that there might be a recent population expansion in the XM unit，and this may be the reason why XM had a high haplotype diversity but a low nucleotide diversity. We estimate that the MQ and XMb have lower diversities because of some very recent geographic events，such as the formation of the Yellow river’s upriver and the Zoige Wetland. Although they are distinctly different，not enough time has passed for them to have diverged a great genetic distance.
2008, 29(2): 127-133. doi: 10.3724/SP.J.1141.2008.02127
The sequences of mitochondrial control region (CR) of Macroclemys temminckii, Chelydra serpentina and Platysternon megacephalum were obtained using PCR and sequencing techniques, with gene-specific primers, based on the CR and its flanking sequences from other species. The CR lengths of the three species were 1089 bp, 1124 bp and 1119 bp respectively, and the base composition of A+T were 68.97%, 69.34% and 69.44% respectively. One fragment of (TA)20 microsatellite was found in M. temminckii; one fragment of (TATAT) 13 direct tandem repeats followed by (TA)15 microsatellites were found in C.serpentina; and one fragment of (AGTATGTTAT)4 direct tandem repeats followed by (GTTGTTATATAACATAT) 13 repeats were found in P. megacephalum. The distribution of mtDNA microsatellites in tortoises was discussed based on the CR sequences of the three species and other six tortoises published in GenBank. The result suggested that all the nine tortoises have microsatellites, and there are obvious differences both at the site and the sequences.
2008, 29(2): 134-138. doi: 10.3724/SP.J.1141.2008.02134
The possibility of ghrelin expressed in sheep oocytes and pre-implantation embryos produced in vivo was investigated in this study. The observed ghrelin immunoreactivity of oocytes and embryos at all stages was predominantly in the cytoplasm. Relative real-time reverse-transciptase (RT)-PCR experiments confirmed that the ghrelin mRNA levels varied depending on the developmental stage, with the highest expression in the blastocyst, metaphase II (MII) oocytes, 2- and 8-cell stages had a significantly higher expression in the germinal vesicle (GV) oocytes, 4-stage and morula. Dynamic changes and the persistent presence of the ghrelin signaling system within oocytes and pre-implantation embryos opens up the possibility of a potential regulatory role of this novel molecule during oocyte maturation and embryonic development.
Expression of Human Semenogelin I-52 and Antibacterial Activity Investigation of Recombinant Peptide
2008, 29(2): 139-144. doi: 10.3724/SP.J.1141.2008.02139
Nucleotide sequence coding for SgI-52 with amino acid residues of 85-136 form mature human semenogelin I was amplified by PCR technique from the cDNA of a human seminal vesicle. The obtained PCR products were inserted into vector pMAL-p2X. The constructed expression vector of pMAL-p2X/SgI-52 was transformed into Escherichia coli DH5α. Fusion protein was expressed in the periplasma of the E. coli DH5αafter IPTG inducement. Recombinant SgI-52 was purified after factor X cleavage and a ultrafiltering process. MALDI-TOF- MS results indicated that the purified recombinant SgI-52 was the target peptide. Recombinant SgI-52 showed antibacterial activities on E. coli ATCC 25922 and E. coli ML-35P with MIC values of 32.45 and 46.30 μg/mL, respectively. Our results and other relevant works suggested that different human semenogelin I degradation fragments during the liquefaction might have various biological functions and deserve to be investigated further.
2008, 29(2): 145-151. doi: 10.3724/SP.J.1141.2008.02145
The relationship between body weight and gonad development of Onchidium struma was studied using a histology method. The relation between the gonadsomatic index (GSI) and hepatopancreas somatic index (HSI) or vitelline somatic index (VSI) were also studied. The composition of germ cells and the percentage difference of phases of germ cells in gonads were analyzed. Spermatogensis and oogenesis of O. struma were observed. The results showed: (1) The development of gonads in O. struma matured as body weight increased. The GSI and HIS or VSI had obvious pertinence respectively（P<0.05）. (2) The oocytes appeared in individuals whose body weight was above 6 g and from this time the oocytes and sperm cells coexisted in the gonads. (3) The spermatozoa were found in all individuals. The main germ cells were the secondary spermatocyte in individuals whose body weight fell under 6 g and those with body weights above 6 g had spermatozoa. The individuals who were above 6 g; the oocytes matured as body weight increased. The main elements of the germ cells were oogoia (57%), exogeous vitellogenic oocyte(69%) in the 6-8g individuals and 10-14 g individuals respectively. The mature oocytes were first found in the 10-14 g individuals. (4) The oogenesis can be divided into 6 stages: oogoia，previtellogeic oocyte，endogenous vitellogeic oocyte，exogeous vitellogenic oocyte，nearly-mature oocyte and mature oocyte (Φ=59.358±3.88 μm). The spermatogensis consisted of five stages: spermatogonia，primary spermatocyte，secondary spermatocyte and spermatozoa (52.44±20.65 μm). We can conclude our research in two points: (1)There is a relationship between body weight and development of gonads. (2) The individual with a body weight above 10 g can be a parent.
2008, 29(2): 152-158.
The physiological function of cerebellum declines during ageing, but its neural mechanism is not still clear. Therefore, we applied an immunohistochemical method to exhibit the Glu and GABA immunoreactive (Glu-IR and GABA-IR) cells, to investigate aged changes of Glu/GABA expression in the cerebellar cortex of young adult and old cats, and to discuss the possible impact of these changes. Under the Olympus BX-51 light microscope, the density of the immunoreactive cells and its gray value in the molecular layer, Purkinje layer and granular layer were counted respectively with Image-Pro Express Software 5.1. Compared with young cats, the density and average grey value of Glu immunoreactive cells increased significantly in the Purkinje and granular layer of old cats (P﹤0.01). The density and intensity of GABA immunoreactive neurons declined significantly in the molecular and Purkinje layers of old cats (P﹤0.01). The density of GABA immunoreactive neurons showed no evident differences in the granular layer between young and old cats (P﹥0.05), but the intensity of GABA immunoreactive neurons was visibly weaker in the granular layer of old cats (P﹤0.01). The results indicated that changes of Glu/GABA expression in the cerebellar cortex during ageing may be one of the important factors mediating the functional degeneration of accurate timing and neurons lost in old individuals.
Residues of Organochlorine Pesticides in the Feathers of Oriental White Stork and Siberian White Crane
2008, 29(2): 159-164.
We collected 51 feather samples from the breasts, tails and wings of Oriental White Storks（Ciconia boyciana）and Siberian White Cranes（Grus leucogeranus）at Hefei Wild Animal Park in May, 2007. Environment-Determination of methylmercury-Gas chromatography was used to determine the residues of op’-DDT，pp’-DDD，pp’-DDE，pp’-DDT，α-BHC，β- BHC，γ- BHC，δ- BHC and Hexachlorobenzene. The results showed that pp’-DDD，pp’-DDE，pp’-DDT，β-BHC， δ-BHC were detected in feathers of both Oriental White Storks and Siberian White Cranes，in which pp’-DDD had the greatest proportion，and reached 0.8936，0.8353 and 0.7516 μg/g (dry weight) respectively in the breast contour feathers，pluma and tail feathers of Oriental White Storks. The residues were 0.5685，0.5077 and 0.4657 μg/g (dry weight) in the feathers of Siberian White Cranes. There was not a significant difference in the residues of pp’-DDD and pp’-DDT in the breast contour feathers，pluma and tail feathers between both birds，but residues of pp’-DDD in the breast contour feathers and tail feathers of the Oriental White Stork was significantly higher than that of the Siberian White Crane.
Fish Production, Water Quality and Bacteriological Parameters of Koi Carp Ponds Under Live-food and Manure Based Management Regimes
2008, 29(2): 165-173. doi: 10.3724/SP.J.1141.2008.02165
To test the effectiveness of introducing live zooplankton against direct manuring in ornamental fish ponds upon their survival and production, larvae of koi carp, Cyprinus carpio L., were cultured for 11 weeks in earthen ponds maintained according to four management regimes: (1) live zooplankton fed to carp larvae (LF); (2) direct fertilization with poultry manure (PM); (3) direct fertilization with cowdung (CD); and (4) a control treatment (C). There were three replicates for each treatment. The growth of heterotrophic bacteria and pathogenic microorganisms like Aeromonas sp. and Pseudomonas sp. were also examined in response to pond management. Values of dissolved oxygen were significantly higher (P<0.05) in the water of LF ponds, compared to other treatments, while the PM and CD treatments recorded were significantly higher (P<0.05) values of PO4 – P, NH4 – N, NO3 – N, NO2 – N, specific conductivity, alkalinity, and BOD, compared to the LF and C treatments. The percentages of organic carbon and total nitrogen in the bottom sediments were higher in the PM and CD treatments compared to LF (P<0.05). Average counts of heterotrophic bacteria in the water of PM and CD ponds were significantly higher than other treatments (P<0.05). The development of Aeromonas sp. and Pseudomonas sp. were significantly higher (P<0.05) in the PM and CD treatments. Weight gain of koi carp stocked in LF was significantly higher (P<0.05) than that of fish in the other treatments. There was a significant difference in the survival rate of koi carp among the treatments ranging from 67.21% in C to 90.11% in LF. The results suggest that raising koi carp larvae in ponds and feeding them exogenously with zooplankton would support high rates of survival and production through maintenance of better water quality and greater abundance of zooplankton in the system. Significantly lower abundance of Aeromonas sp. and Pseudomonas sp. in the LF treatment considerably lowered any possibility of occurrence of bacterial disease.
2008, 29(2): 174-180. doi: 10.3724/SP.J.1141.2008.02174
The spring diet of Hoolock Gibbons (Hoolock hoolock) was observed by tracking them at Nankang of Mt. Gaoligong from March 3rd to April 14th, 2007. Their feeding behavior could be divided into eating fruit, drinking juice and eating leaves. Feeding behavior, food species, part of food to be eaten and feeding sites of the gibbons were recorded by scanning samples. In their active regions, nineteen used plots (20 m×20 m) around the feeding tree as well as the same number of available plots by random sample were set. Data of six factors involving feeding tree and food resource were collected from the plots. Food species and their quantity, selective index and selectivity of feeding spots were calculated. The results showed that in spring at Nankang the gibbons ate 10 botanic species, three of them for fruits, one for juice and six for young leaves. Among 10 food species, only fruits of Schima khasiana and young leaves of Maglietia haokeri were under selected and the other eight species were over selected. The result of a Mann-Whitney U test showed that the food resource and food species were significantly different between the used and available plots. There were more food resources and food species in the used plots. Fruits were the main food resource of the gibbons in spring at Nankang and young leaves were the supplement. The over selected food was not always at a high feeding frequency; in contrast, the food with a high feeding frequency was not always the over selected food resource. The gibbons often selected fruits that could be eaten easily and young leaves containing less ash. The fruit was also the main water resource. Hoolock Gibbons usually chose regions full in food resources and food species, so they could spend less energy looking for food, and at the same time choose from several species which satisfied their nutritional needs.
Altitudinal Range of Black-and-white Snub-nosed Monkeys (Rhinopithecus bieti ) at Baima Snow Mountain, China
2008, 29(2): 181-188. doi: 10.3724/SP.J.1141.2008.02181
Faeces were counted along horizontal, 5.0 m wide strips at altitude intervals of 100 m to examine the altitudinal ranging pattern of a band of black-and-white snub-nosed monkeys (Rhinopithecus bieti) at Nanren (99o04’E, 28o34’N, Baima Snow Mountain Nature Reverse, Yunnan, China) at four areas in each season between 2000-2001. Faeces were also counted along vertical, 2.5 m wide strips in one subvalley and on one subridge in each area. Monkeys used an altitudinal range of 3500-4300 m, but preferred occupying the upper forest belt between 3900 and 4200 m asl year-round, being at the highest altitude in summer, the lowest altitude in spring, and middle altitudes between autumn and winter (lower in winter than autumn). Moreover, there were secondary peaks of lower altitude use (3700 m) in spring and winter. In addition, more faeces were distributed in subvalleys than on subridges in winter, this suggests that monkeys tend to spend more time in subvalleys where there is less wind and fluctuating temperatures. The significant correlation between faecal density and lichen load might indicate that altitudinal distribution of food resources may act as an important factor affecting the monkeys’ range. The monkeys might migrate to lower altitudes to eat young sprouts and leaves in spring, and to shelter from snowstorms in winter, possibly resulting in the secondary peaks at lower altitudes.
2008, 29(2): 189-194. doi: 10.3724/SP.J.1141.2008.02189
Using morphometry of ungulate pellets to determine sex and age is the preferred method in field studies on ecological differences between two sexes, especially sexual segregation of ungulates. This study examined the morphometry of 4,006 pellets from ≤145 Eld’s Deer (Cervus eldi hainanus) collected in Hainan Datian National Nature Reserve in March 2006. The deer were divided into five sex-age categories as adult male, adult female, sub-adult male, sub-adult female, and calf. They were analyzed using four direct measurement indices (dry weight, length, width, and volume) and two indirect indices (length-to-width ratio, and ellipsoid-shape index) by use of step-wise discriminant and fuzzy cluster analyses. Discriminant analysis could correctly distinguish 76.17% of adult males, 42.22% of adult females, 34.94% of sub-adult males, 40.46% of sub-adult females, and 79.34% of calves. Fuzzy clustering analysis enabled to correctly distinguish 19.48% of adult males, 20.02% of adult females, 37.37% of sub-adult males, 42.82% of sub-adult females, and 91.50% of calves. Therefore, it is more credible to determine calves and secondarily adult males by use of pellet morphometry. We advise that pellet morphometry should be used for determining sex and age categories in surveys at the population level rather than at the individual level.
Diurnal Time Budget and Behavior Rhythm of Wintering Black-necked Crane (Grus nigricollis) at Dashanbao in Yunnan
2008, 29(2): 195-202. doi: 10.3724/SP.J.1141.2008.02195
From October 2006 to April 2007, wintering behaviors and the daily cycle of Black-necked Cranes (Grus nigricollis) were observed with instantaneous and scan sampling methods at Dashanbao National Nature Reserve, Zhaotong, Yunnan. During the wintering period, to meet wintering energy needs Black-necked Cranes spent most of their time feeding, which was about 53.05±4.93%. Other behaviors such as vigilance, searching, maintenance, walking, flight, and fight were 18.75±2.65%, 10.38±1.34%, 10.32±4.93%, 4.90±1.59%, 1.70±0.38%, 0.55±0.41%, 0.36±0.25%, respectively. Our results indicated that roost departure time had positive correlation with sunrise time, while roost arrival time had negative correlation with sunset time. One-Way ANOVA analysis showed that the eight wintering behaviors of Black-necked Cranes had significant differences between every one-hour period (09:00－18:00) and over the five month period (2006-11－2007-03). There were two feeding peaks in the morning and afternoon, with one maintenance peak at noon. Partial analyses showed that environmental temperature affected wintering behaviors independently, while environmental moisture had the opposite effect on behaviors dependent on environmental moisture. When the temperature was high, feeding was low and maintenance was high. When the temperature was low, maintenance was high and feeding was low.
2008, 29(2): 203-211. doi: 10.3724/SP.J.1141.2008.02203
Bird diversity was surveyed in five urban parks of Guangzhou from January 1999 to April 2000, and from July 2006 to June 2007, using a transect line method. A total of sixty-four bird species were recorded. Japanese White-eye (Zosterops japonica), Chinese Bulbul (Pycnonotus sinensis), and Black-crowned Night Heron (Nycticorax nyctinorax) were dominant species. Residents, winter visitors and summer visitors accounted for 64.1%, 26.6% and 7.8% of recorded bird species, respectively. The greatest number of species was recorded in September (31), the least was recorded in July (20) and November (20). The mean encounter rates of birds was 65±;5 ind./h (±SE), with the highest in March (98±29 ind./h) and lowest in January (35±11 ind./h). There was fluctuation, but it did not differ significantly between months (F 11,48=1.35, P= 0.226). There were a greater number of migratory species in April, September and December. Encounter rates of migratory birds significantly differed between months (F 11，48=3.098，P=0.003). Bird richness differed among the five parks and significantly and positively correlated with the park area (R=0.905, P=0.035), with S=11.02 A0.28 (S：bird species richness, A：park area). This meant that a greater number of bird species occurred in larger parks. Parks with an area of about 65 hm2 were better for avian diversity conservation and land use in Guangzhou.
Spatial Patterns for the Distribution of Winter Waterbirds in the Aquaculture Ponds of Chongming Dongtan，Shanghai
2008, 29(2): 212-218. doi: 10.3724/SP.J.1141.2008.02212
The spatial utility pattern of birds reflects their selection of habitats. Based on field surveys, spatial statistics and landscape pattern analyses，we investigated the distribution pattern of winter waterbirds in the aquaculture ponds of Chongming Dongtan，Shanghai. The results showed that ducks had a different pattern from that of waders. The distribution pattern of ducks revealed a significantly positive autocorrelation and clustered distribution，while that of waders showed a non-autocorrelation and random distribution. As a supplementary habitat，the aquaculture ponds play an important role for winter waterbirds. There are，however，differences in selecting habitats among groups of winter waterbirds. Ducks prefer ponds with well-developed reed vegetation，a relatively smaller water area and deeper water，while waders favor larger water areas and shallower water. The requirement of habitats for different groups of winter waterbirds should be taken into account when planning land use and managing a natural reserve.
2008, 29(2): 219-222. doi: 10.3724/SP.J.1141.2008.02219
A new horn toad Megophrys sangzhiensis sp. nov. (Holotype: CIB 200078, adult male, SVL 54.7 mm) has been recognized based on collections from Sangzhi County, Hunan Province in China. It was previously classified as M. caudoprocta Shen, in 1994 as both had a tail projecting above the vent. This new species can be distinguished from the latter by the following characteristics: Body is smaller in size, SVL 54.7 mm; a smaller tubercle is present on the outer edge of the upper eyelids; chest and belly is yellowish with dark brown or orange red spots; vomerine ridge is slender and not dilated at its posterior region, without vomerine teeth; larger and sparse black nuptial spinules are found on the first two fingers of males with vocal sac. The new species differs from M. spinata Liu Hu, in 1973 by having a tail projecting above the vent at the ventral view, smaller and denser black nuptial spinules are found on the first two fingers in males. The size and density of the nuptial spinules on the first two fingers of the new species presents at a middle degree between those of M. caudoprocta and M. spinata.
2008, 29(2): 223-224. doi: 10.3724/SP.J.1141.2008.02223
Hubei; Xiantao; Black Stork(Ciconia nigra); New wintering area