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月刊，1984年创刊
主管：内蒙古自治区教育厅
主办：包头医学院
主编：赵云山
特邀主编：高长青
编辑出版：包头医学院学报编辑部
国际刊号：ISSN 1006－740X
国内刊号：CN 15－1182/R
邮发代号：16-292

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2009 Vol. 28, No. 11 Published: 2009-11-25

Article

	613	Isolation and Identification of Bacterial Pathogen Aeromonas hydrophila and Development of the Inactivated Vaccine in Japanese Flounder Paralichthys olivaceus Diseased with Ascides
		SUN Yi-min,LI Nan,SONG Jie,LI Xiao-yue,SHEN Hong-qi,WANG Feng-min,LI Quan-zhen,CAO Jie-ying,ZHAO Bao-hua
		The pathogenic bacteria were isolated from diseased Japanese flounder Paralichthys olivaceus, and identified as Aeromonas hydrophila by biochemical methods and polymerase chain reaction based on the 16s rDNA sequence on GenBank via the homologies of nucleic acids of 99.8% between the isolated strain and the standard strains. The mice and the flounder were immunized with the inactivated vaccine made by this isolated Aeromonas hydrophila via dipping immersion and showed protection rate of 90% and 80% with the incidence of 6%. The antibody level was detected by agglutination reaction and indirect enzyme-linked immunosorbent assay (ELISA) with the antiserum seemed immunogenic crucially. The anatomized results showed that the immunized mice were healthy and the unimmunized mice (as the control) were diseased in liver, indicating that the vaccine was effective.
		2009 Vol. 28 (11): 613-617 [Abstract] ( 69 ) HTML (0 KB) PDF (361 KB) ( 256 )

	618	Effects of Different Diets on Growth, Development and Digestive Enzymes Activity in Tilapia Hybrid Larvae
		QIANG Jun,WANG Hui,LI Rui-wei,DONG Xiao-hui
		The larval hybrid tilapia (Oreochromis niloticus×O. aureus)with initial body weight of (0.006±0.0005) g was fed either artificial diet, rotifers, eggs or rotifers and artificial diet for 28 days with triplication to elucidate the effects of food items on growth, development and activities of the digestive enzymes. The larval tilapia had the best growth (absolute growth rate of 0.150/d and 0.142 g/d and specific growth rate of 0.014%/d and 0.013%/d (P<0.05) when fed rotifers and artificial and artificial diet while the larvae fed the eggs had the worst growth (P<0.05). There were survival of 90.3% and 89.7% in the developed well larvae fed rotifers and artificial or rotifers. The slow developmental larval tilapia fed the eggs had the lowest survival (40.7%). On the activity of the digestive enzymes, different diets had significant effect on activity of the digestive enzymes in the hybrid tilapia larvae (P<0.05). There were the maximal activities of amylase and lipase in the larvae fed the eggs and the maximal activity of pepsin was observed in the larvae fed the rotifers and artificial diets. The minimal activity of pepsin, however, occurred in the larvae fed the eggs.
		2009 Vol. 28 (11): 618-623 [Abstract] ( 56 ) HTML (0 KB) PDF (372 KB) ( 335 )

	624	Effects of Environmental Factors on Growth of Alga Chaetoceros gracilis
		YAN Aiju,WU Hui-xian,XUE Jun-zeng,LIU Yan
		The effects of nitrogen, phosphorus, nitrogen and phosphorus ratio and silicon on growth of alga Chaetoceros gracilis was conducted through the monoculture. The optimal concentration for the alga growth was found to be 5 factor. There was no significant effect of silicon concentration on the growth of the alga(P>0.05), indicating that the silicon is just an influence factor rather than a limiting factor.
		2009 Vol. 28 (11): 624-627 [Abstract] ( 115 ) HTML (0 KB) PDF (286 KB) ( 198 )

	628	Changes in Main Physicochemical Factors in Water and Assessment of Water Quality during Culture in Higher-level Shrimp Ponds
		ZHANG Yu-bin,ZHANG Jie-xiang,ZHAN Xiao-yan,LI Xue-ying,SUN Xing-li
		The dynamic changes and correlation in main physicochemical factors in water were studied in white leg shrimp Litopenaeus vannamei ponds in Donghaidao of Zhanjiang, and the NQI was adopted to estimate the quality of water in the ponds. The results indicated that DO, pH, temperature and salinity fluctuated gently, but the concentration of COD, DOC, nutrient and chl-a increased relevant to culture. The correlation was complicated among the main physicochemical factors while there was a significantly positive correlation between chll_a and nutrients. The NQI value showed that the eutrophication increased rapidly in the middle and late of culture period, implying serious eutrophication in the shrimp ponds.
		2009 Vol. 28 (11): 628-634 [Abstract] ( 53 ) HTML (0 KB) PDF (433 KB) ( 458 )

	635	Evaluation of Nutritive Quality and Components in Muscle of Brownmarbled Grouper Epinephelus fuscoguttatus
		GUO Yong-jun,XING Ke-zhi,XU Da-wei,CHEN Cheng-xun,WANG Qing-kui,WANG Xiao-xue,ZHANG Shu-sen,YANG Yong-hai
		The contents of crude protein, fat, ash, moisture, and amino acids were determined in the muscle of brownmarbled grouper Epinephelus fuscoguttatus. It was found that the fish had a dressed body weight of 77.3%. There were 18.8% of crude protein, 2.2% of crude fat containing 28.28% of DHA and EPA in total fat, 1.16%of crude ash and 77.1% of moisture in the muscle. The total amount of amino acids (AA) was 18.22% in wet mass, in which the essential amino acids comprised of 6.88%, in the muscle. The total amount of the delicious amino acids was found to be 8.53%. The essential amino acids index (EAAI) was 514 in the muscle, accorded with the FAO/WHO standard.
		2009 Vol. 28 (11): 635-638 [Abstract] ( 56 ) HTML (0 KB) PDF (248 KB) ( 234 )

	639	Sequence Analysis of Mitochondrial Cytochrome b Gene in Four Sardinella Species
		YANG Tian-yan,GAO Tian-xiang,GUO Yan,MENG Wei
		The mitochondrial cytochrome b gene fragment (402 bp) of four Sardinella species were amplified and sequenced with the method of PCR, and the differences in the sequences were analyzed. Nine haplotypes were observed in ten samples. The average contents of T,C,A,and G were 28.9%,28.2%,23.9%,and 19.0%, respectively and the content of A+T was higher than that of G+C which was similar to the sequences of the other fishes. By the Kimura 2-parameter model, the genetic distances were found to change from 0.1616 to 0.2653. Based on sequences of partial cytochrome b gene, a phylogenetic tree by NJ method indicated that the Sardinella zunasi and S. jussieu, S. maderensis and S. aurita had a relatively close relationship, respectively. Molecular evolution rate showed that the divergence of four Sardinella species occurred during the middle and late Miocene about 800 to 1300 million years ago,indicating that the divergence time was earlier than the other fishes.
		2009 Vol. 28 (11): 639-643 [Abstract] ( 51 ) HTML (0 KB) PDF (318 KB) ( 365 )

	644	Changes in Fatty Acid Composition During Early Embryonic Development of Yellow Catfish Pelteobagrus fiulvidraco
		YAO Jun-jie,ZHAO Yun-long,LI Chuan,HE Deng-ju,HU Cong-yun
		The composition of fatty acids in mature eggs and embryos during early development of yellow catfish Pelteobagrus fiulvidraco were determined by biochemical methods. The results showed that the predominant fatty acids, in terms of relative proportion, were C_(16:0), C_(18:0), C_(16:1n-7), C_(18:1n-9),and C_(18:2n-6) both in matureeggs and embryo during early development stage. Besides, large proportion of C_(18:3n-3) was observecl in the. The fatty acid was utilized by various patterns during various embryonic stages. The proportion of the poly-unsaturated fatty acids (HUFA) was high during blastula, gastrula and organogenesis. The monounsaturated fatty acids(MUFA) increased during cleavage and blastula. The saturates fatty acids(SFA) acted as the main energy source during early embryonic development. SFA supplied main energy for metabolism during gastrula, nerve embryo and organogenesis. There were higher proportion of HUFA in fertilized eggs than that in the mature eggs.However, the proportion of SFA increased,indicating that the embryos started the active metabolism after fertilization.
		2009 Vol. 28 (11): 644-647 [Abstract] ( 68 ) HTML (0 KB) PDF (264 KB) ( 309 )

	648	Effects of Dietary Bacterium Bacillus thuringiensis on Immune Response of Common Carp
		YIN Hai-cheng,ZHAO Hong-yue
		The common carp Cyprinus carpio immunized with Aacillus hydrophila (groups A_0～A_3) and un-immunized (groups B_0～B_3) were fed the diets containing bacterium Bacillus thuringiensis at a rate of 1.0×10~(11), 3.0×10~(11), and 5.0×10~(11)cfu/kg to investigate the effect of the bacterium on the immune response of the common carp in Yellow River. The phagocytic activities of leucocytes, lysozyme activities, and agglutinating antibody titer were determined 0, 15, 30, and 40 days after the feeding trial. The results showed that the fish fed the diets containing different levels of Bacillus thuringiensis had significantly enhanced phagocytic activities of leukocytes, and lysozyme activities(P < 0.05), increasing with the increment in the levels of the bacterium. There were no significant differences in phagocytic activities of leukocytes for the immunized and un-immunized fish (P>0.05), but significant differences in the lysozyme activities(P<0.05). The significant differences in the phagocytic activities of leucocytes and lysozyme activities were not observed 10 days after the fish stopped feeding the diets(P>0.05). The different concentrations of Bacillus thuringiensis improved the agglutinating antibody titer and the survival of the fish after the fish challenged with live Aeromonas hydrophila. The immunized fish had the maximal relative percent survival (RPS) when fed the diets containing the Bacillus thuringiensis at a rate of 5.0×10~(11)cfu/kg feed.
		2009 Vol. 28 (11): 648-652 [Abstract] ( 55 ) HTML (0 KB) PDF (327 KB) ( 224 )

	653	The Electrophoresis Patterns of Mantle Protein in Clam Meretrix meretrix with Different Stripes
		LI Chun-yan,LI Tai-wu,SU Xiu-rong
		The electrophoresis patterns of mantle protein were compared by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) in clam Meretrix meretrix with different stripes collected in Guangxi coast to optimize the clam seed. The results showed that most of the protein electrophoresis of the mantle had the same bands, while a few clams with different stripes had different contents and types of protein, and these differences were not found to be related to the stripes.
		2009 Vol. 28 (11): 653-655 [Abstract] ( 58 ) HTML (0 KB) PDF (309 KB) ( 275 )

	656	The Cause of Massive Mortality of Clam Meretrix meretrix Linnaeus in Qidong ity,Jiangsu Province
		GUO Chuang,CHEN Jing,ZOU Yong,HUANG Chun-gui,CHEN Hui
		Seawater samples were collected from the area where the high mortality of clam Meretrix meretrix occurred in Qidong city, Jiangsu province, and the tissues and organs in the infected clam such as mantle, branchia, foot and digestive deverticula were analyzed by chemical method, bacteria isolation and ultrathin sections. The results indicated that the chemical pollution and bacteria were not the cause of the high mortality;the ultrathin sections observed with transmission electron microscope showed that there were some inclusion bodies with viruses and other dispersed viruses in the mantle. These spherical viruses with the sizes of 60~80 nm in diameter and lack of capsula. Also, severe cytopathological changes including condensing of nuclei, dissolving of mitochondria,desquamation of microvilli and vesicling of other organells were observed obviously in other infected tissues.
		2009 Vol. 28 (11): 656-658 [Abstract] ( 64 ) HTML (0 KB) PDF (308 KB) ( 245 )

	659	Anti-bacterial Activities of Extracts from Kelp Laminaria japonica
		MU Kai-feng,WU Yong-pei,YANG Fang,YANG Qiu-ming
		The anti-bacterial activities of extracts from kelp Laminaria japonica against four microorganism species Bacillus subtilis, Escherichia coli, Aspergillus niger, and Cerevisiae fermented were studied, and the extraction conditions were optimized. The extract from the kelp showed anti-bacterial activity only against Bacillus subtilis. Good extraction and anti-bacterial activities were observed in 95% methanol under the optimal conditions of pH 7 at 30 ℃ for 8 h based on the inhibition circles.
		2009 Vol. 28 (11): 659-662 [Abstract] ( 61 ) HTML (0 KB) PDF (244 KB) ( 335 )

	663	Tetragenus Active Bacterial Agent for Degradation of Ammonia Nitrogen and Nitrite
		SHI Da-lin,HE Yi-jin,SUN Mei,PAN Liang-kun,LIU Huai,HU Ling,ZHOU Qun-lan,CHEN Qiu-hong,ZHANG Wei-na,XU Pao,KUANG Qun
		The active bacterial tetragenus agent with four available microbes was added to the water in aquaculture ponds to decrease the contentss of ammonia nitrogen and nitrite in the ponds. The results showed that the contents of ammonium nitrogen and nitrite were decreased by the microbes. The more microbes was added, the faster ammonium nitrogen and nitrite decreased. The rank of elimination of the ammonium nitrogen by the microbes was ranged in a descendant order as the following:Lactobacillus ＞ photosynthetic bacteria ＞nitrobacteria＞ Bacillus natto. For nitrite, the rank of elimination of the ammonium nitrogen by the microbes was ranged in a descendant order as the following:nitrobacteria ＞ bacillus natto ＞photosynthetic bacteria ＞ Lactobacillus. 52%, 80% and 74%of the ammonium nitrogen were eliminated 5 days after the active bacteria tetragenus agent was added at a rate of 1.5×10~(-7), 3.0×10~(-7), and 4.5×10~(-7) g/ml, respectively, significantly higher than that by the single microbes at the same dose.
		2009 Vol. 28 (11): 663-666 [Abstract] ( 75 ) HTML (0 KB) PDF (242 KB) ( 273 )

	667	Optimization of Alginate-degrading Bacterial Fermentation Process for Alginate Lyase Production
		HOU Bao-bing,LIU Shu-lai,ZHANG Jian-you,BAO Hua-fang,DING Yu-ting
		The enzyme production of a bacterium strain with relatively high alginate lyase activity known as Acinetobacter X8 isolated from decaying part of seeweed Sargassum fusiform was investigated under liquid fermentation. The optimized liquid fermentation medium(m/V) was comprised of 0.6 % alginate, 0.7 % peptone, 0.2 % K_2HPO_4, and 2 % NaCl at initial pH of 7.0. After fermentation for 16 h at 150 r/min and 25 ℃, the yield of alginate lyase was found to be up to 157.32 U/ml, 159.01% higher than that under the ordinary conditions. However, the production process was inhibited by Mg~2, Ca~(2+) and Fe~(2+).
		2009 Vol. 28 (11): 667-670 [Abstract] ( 73 ) HTML (0 KB) PDF (292 KB) ( 514 )

	671	Prokaryotic Expression,Purification and Polyclonal Antibody Preparation of Antifreeze Protein AFPⅢ
		HAO Feng-xia,HU Wen-ge,FU Wei-chao,KANG Zhuang-li,ZHANG Gui-ling
		Gene recombination technology was used to link AFPⅢ gene and prokaryotic expression vector pET32a(+) and then the recombinant plasmid was transformed into Escherichia coli(BL21)and the positive clones were identified by restriction enzyme digestion, PCR and sequencing. Expression AFPⅢ was induced with IPTG and the expression product was purified, which the purified AFPⅢ was used to immunize mouse to obtain the antiserum.The specificity of the antibodies was examined by Western blotting.The purified antigen was found to have antigenicity by ELISA and the prokaryotic expression vector pET32a-AFPⅢ to be successfully constructed. A fusion protein with a molecular weight of about 26 kD was obtained after induction with IPTG and affinity chromatography.The anti-AFPⅢ antibody was obtained from the immunized mouse.The results of Western blotting indicated that the polyclonal antibody had high specificity to AFPⅢ, providing a foundation for further research on the biological function of AFPⅢ and expression of AFPⅢ in transgenic fish tissues.
		2009 Vol. 28 (11): 671-674 [Abstract] ( 62 ) HTML (0 KB) PDF (354 KB) ( 355 )

	675	Construction and Analysis of Suppression Subtractive Hybridization Library of Dinoflagellate Alexandrium catenella
		SHI Xi-zhi,LIU Bing,HE Jing-jing,WANG Meng-qian,GUO Hao,SU Xiu-rong,LI Tai-wu
		To isolate the bioluminescence specific expressed genes of dinoflagellate Alexandrium catenella(A. catenella), suppression subtractive hybridization was performed to construct the differential expressing cDNA libraries of Alexandrium catenella. A total of 500 cDNA clones were picked. PCR amplification revealed that the length of inserted fragments was mainly from 250 to 1000 base pairs. Ten randomly selected clones after PCR screening were sequenced and analysed in GenBank with Blast search. Two ESTs shared significant identity with the luciferin binding protein and Ribulose-1,5-bisphosphate carboxylase/oxygenase and the other ESTs represent novel genes of Alexandrium catenella with no significant homology in GenBank. These results had provided the foundation for cloning new specific genes of Alexandrium catenella and further studying and establishing the novel detection method for specific dinoflagellata.
		2009 Vol. 28 (11): 675-677 [Abstract] ( 60 ) HTML (0 KB) PDF (243 KB) ( 205 )

	678	The Effects of Feed Formulations on Growth and Benefit for Chinese Mitten Carb Eriocheir sinensis Culture in Paddy Fields
		LI Jing-wei,LI Wen-kuan,XIAO Zu-guo,YU Yong-qing,LI Chang-jun
		Four feed formulations containing 38.06%, 33.43%, 28.48% and 25.32% of protein were fed to Chinese mitten crab Eriocheir sinensis cultured in paddy fields for 90 days. The results showed that when the diet contains protein 38.06 %, fat 6.01 % and calcium- phosphorus ratio 1.35, the growth was the best with the highest profit, and that higher nutritional level promoted, improved the quality of the crab and increased profit the sharply.
		2009 Vol. 28 (11): 678-682 [Abstract] ( 66 ) HTML (0 KB) PDF (272 KB) ( 263 )

	683	Effect of Different Diets on Growth Performance and Survival of Northern Snakehead (Channa argus) and its Hybrid in a Pond
		WANG Yu-xi,FENG Xiao-yu,LI Xing-xian,GUO Shui-rong,XIE Nan
		Northern snakehead (Channa argus) and its hybrid were stocked into a pond and fed formulated feed or frozen fish separately to evaluate the effect of different diets on growth performance and survival. During the experiment survival of 80.9%, yield of 5.2 kg/m~2 and feed conversion ratio of 1.367 were found in the hybrid culture pond in which the water was exchanged twice during the experiment while survival of 53.9%, yield of 1.8 kg/m~2 and feed conversion ratio of 4.29 were observed in the northern snakehead pond in which the water was exchanged 6 times.
		2009 Vol. 28 (11): 683-686 [Abstract] ( 71 ) HTML (0 KB) PDF (236 KB) ( 331 )

	687	Determination of Endosulfan and Deltamethrin Residues in Fishery Foods by Gas Chromatography
		ZHANG Jin-hu
		The determination method of endosulfan and deltamethrin pesticide residues in fishery products was established by gas chromatography. The sample was extracted by acetonitrile plus water, degreased by n-hexane, and purified by adsorption. The detection limit was 0.004 mg/kg for endosulfan, and 0.01mg/kg for deltamethrin under the optimal conditions of pretreatment and determination. There were recoveries from 64.6% to 88.9%, and the relative standard deviations from 4.1% to 9.5% for the two pesticides at the level of 1, 2, and 5 times of determination limit, indicating that the method was characterized by accurate, rapid and high efficiency, and low cost within the limit for import and export.
		2009 Vol. 28 (11): 687-690 [Abstract] ( 69 ) HTML (0 KB) PDF (245 KB) ( 464 )

	691	Effects of Snails Feeding Rates on Size, Yield and Survival of Chinese Mitten Crab Eriocheir sinensis
		DONG Jiang-shui,CHEN Hong-jun,WANG Xin-hua
		Juvenile (140 individuals/kg) Chinese mitten crab Eriocheir sinensis were randomly stocked into seven ponds of each 0.67 hm~2 with depth of 1.5 m at Cangxi 2814 fish farm in Gaochun county at a rate of 6000 ind/hm~2, and fed snails with feeding rate of 750, 1070, 2250, 4500, 5440, 5630 and 6750 kg/hm~2 to evaluate the effects of feeding rates on size, yield and survival of the crab. The test ponds were also polycultured with freshwater shrimp (Macrobrachium japonicum), mandarinfish (Siniperca chuatsi) and a few silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) juveniles. The results showed that the relationship between the crab yield (Z) and snail amounts (x) was expressed as Z=528.138+0.228x-0.000029x~2 (R~2=0.959) (P<0.05);the relationship between the crab size (Y) and snail amount (x) was expressed as Y=155.368+0.014x-0.0000017x~2(R~2=0.934)(P<0.05);the relationship between the crab survival (L) and snail feeding rate (x) was equated as L=52.208+0.018x-0.0000023x~2(R~2=0.957)(P<0.05). The optimal snail feeding rate was suggested to be 4500 kg/hm~2.
		2009 Vol. 28 (11): 691-694 [Abstract] ( 62 ) HTML (0 KB) PDF (246 KB) ( 432 )

	695	Rapid Diagnoses and Treatment of Diseases Found in Leech Whitmania Pigra
		CAI Li-juan,ZHOU Li-gen,LIN Qi-cun
		One pathogenic bacterium was isolated from diseased leech Whitmania pigra using aseptic technique, and identified by VITEK-32 automatic microorganism identification and susceptibility system based on the physiological and biochemical characteristics.Drug sensitive tests to Enterobacter cloacae were investigated for seven types of antibiotics (Compound sulfamethoxazole,Gentamicin, Norfloxacin, Amikacin Sulfate, Tetracycline, Enrofloxacin,and Doxycycline).The results showed that the bacterium E. cloacae was found to be the pathogenic microorganism and to be sensitive to Norfloxacin. The disease was controlled by feeding the diet containing Norfloxacin at a dose of 50 mg/kg for 5 successive days,together with chlorine dioxide disinfectant,and the symptoms disappeared three days after administration.
		2009 Vol. 28 (11): 695-697 [Abstract] ( 79 ) HTML (0 KB) PDF (171 KB) ( 275 )

	698	The Effects of Salinity on Oxygen Consumption and Ammonia Excretion in Juvenile Crayfish Procambarus clarkii
		LI Ting-gu
		The effects of salinity on oxygen consumption and ammonia excretion were studied in juvenile crayfish Procambarus clarkii with initial body weight of (1.82±0.21) g and body length of (3.13±0.30) cm at the salinity of 0, 2, 4, 6, 8, 10, and 12 at 25 ℃. The minimal oxygen consumption and ammonia excretion were found at a salinity of 2, indicating that the iso-osmotic point of the juveniles was around 2. The rates of oxygen consumption and ammonia excretion increased with increase in salinity. Turkey HSD test revealed that there were no significant differences of oxygen consumption and ammonia excretion in the juveniles exposed to salinity of 0, 2, 4, and 6.
		2009 Vol. 28 (11): 698-700 [Abstract] ( 71 ) HTML (0 KB) PDF (188 KB) ( 255 )

	701	Recent Advances on Immunoglobulin in Fish
		ZHANG Li-ying,ZHAO Meng

		2009 Vol. 28 (11): 701-705 [Abstract] ( 57 ) HTML (0 KB) PDF (359 KB) ( 776 )

	706	Research Advances of Largemouth Bronze Gudgeon,Coreius guichenoti
		CHEN Chun-na,HUANG Ying-ying

		2009 Vol. 28 (11): 706-709 [Abstract] ( 53 ) HTML (0 KB) PDF (271 KB) ( 295 )

	710	Effects of the Environmental Factors on Breeding of Procambarus clarkii
		REN Xin-lin,LING Wu-hai,JI Cui-ping

		2009 Vol. 28 (11): 710-712 [Abstract] ( 58 ) HTML (0 KB) PDF (235 KB) ( 395 )