1.Studies on macroscopic and microscopic characteristics of Ophiocordyceps xuefengensis.
Hao LIU ; Lan-ping ZHEN ; Ru-cai ZHU ; Shui-han ZHANG ; Hui-yong HUANG
China Journal of Chinese Materia Medica 2015;40(14):2820-2824
The macroscopic characteristics, tissue, caterpillar body wall and powder of Ophiocordyceps xuefengensis in different batch numbers were observed and researched by the macroscopic and microscopic identification methods. The result shows that the morphology, size, abdominal annulations of caterpillar, etc. of 0. xuefengensis are the macroscopic identification characteristics, the caterpillar body surface mycelium, body wall sculpture and crochets on abdominal legs are the microscopic identification characteristics. These characters are stable and regular discriminant features, which are proved to be the identification basis of O. xuefengensis. In addition, The characters such as crochets on abdominal legs arrange in two parallel ellipse rings, the inner crochets are long strip, and the external toes are unciform, are specific.
Animals
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Hypocreales
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cytology
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Moths
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anatomy & histology
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cytology
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microbiology
2.Correlation of extracellular enzymes activity of Candida glabrata clinical isolates with in vivo pathogenicity in Galleria mellonella larvae.
Peng CHENG ; Xiang Ren A ; Xiang Ming MU ; Bo Jie YANG ; Si Si CHAN
Chinese Journal of Preventive Medicine 2023;57(2):229-235
Objective: To explore the relationship between extracellular enzymes activity and virulence of Candida glabrata clinical isolates based on the infection model of Galleria mellonella larvae. Methods: Using experimental research methods, 71 strains of non-repetitive Candida glabrata were collected from Qinghai Provincial People's Hospital from June 2021 to January 2022. Bovine serum protein agar medium, egg yolk agar medium, sheep blood agar medium, Tween-80 agar medium and triglyceride agar medium were used to detect the aspartyl protease activity, phospholipase activity, hemolysis activity, esterase activity and lipase activity of Candida glabrata. Median lethal concentration (LC50) was calculated by using 1.25×108 CFU/ml,2.50×108 CFU/ml,3.75×108 CFU/ml,5.00×108 CFU/ml suspension of Candida glabrata ATCC2001 to infect Galleria mellonella larvae. Histopathological and etiological analysis was performed to determine whether the infection model was successfully established. The clinical isolates of Candida glabrata were configured to infect Galleria mellonella larvae with LC50 concentration to detect the pathogenicity of Galleria mellonella larvae.Spearman test or Pearson test were used to analyze the correlation between the extracellular enzyme activity of Candida glabrata clinical isolates and the pathogenicity of Galleria mellonella larvae. Results: 71 strains of Candida glabrata isolated clinically were detected to have low hemolytic activity after 2 days of culture. Aspartyl protease was detected after 4 days of culture, among which 7 strains (9.86%), 19 strains (26.76%) and 45 strains (63.38%) showed low, medium and high aspartyl protease activity. After 7 days of culture, 71 strains did not detect phospholipase, esterase and lipase activities. Candida glabrata on Galleria mellonella larvae of LC50=2.5×108 CFU/ml Fungal spore were found in the intestinal tissue pathological section of Galleria mellonella larvae in the experimental group, and Candida glabrata was identified by the microbial Mass Spectrometry after culture, while no fungi were found in the pathological section and culture of the control group. Spearman test shows that, there was a linear positive correlation between aspartyl protease activity and the survival rate of Galleria mellonella larvae (r = 0.73, P<0.01), the difference was statistically significant.Pearson test shows that, there was no significant linear relationship between hemolytic activity and survival rate of Galleria mellonella larvae (r = 0.16, P = 0.34), the difference was not statistically significant. Conclusion: The clinical isolates of Candida glabrata in this study had aspartyl protease activity and low hemolytic activity, but no phospholipase, esterase and lipase activity. The activity of aspartyl aspartyl protease of Candida glabrata was positively correlated with the pathogenicity of Galleria mellonella larvae.
Animals
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Sheep
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Larva/microbiology*
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Virulence
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Candida glabrata
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Agar
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Moths/microbiology*
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Esterases
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Aspartic Acid Proteases
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Lipase
3.Effects of integrated pest control techniques to growth of host larvae Cordyceps sinensis.
Fei LIU ; Xiao-li WU ; Wei ZENG ; De-li ZHANG ; Shi-jiang CHEN ; Ding-hua YIN
China Journal of Chinese Materia Medica 2008;33(23):2741-2744
UNLABELLEDTo study the effects of the integrated pest control techniques on growth of host larvae of Cordyceps sinensis.
METHODThe integrated pest control techniques were compared with conventional techniques to evaluate the effects on growth of host larvae.
RESULT AND CONCLUSIONThe results showed that the techniques had broken the balance of the microbial living in the material, produced effective inhibition on the pests, raised the survival rate and promoted the growth of the host larvae at the same time.
Animals ; Cordyceps ; physiology ; Larva ; growth & development ; microbiology ; Moths ; growth & development ; microbiology ; Pest Control, Biological ; methods
4.Study on the biology of adults parasite of Cordyceps sinensis, Hepialus biruensis.
Shi-jiang CHEN ; Din-hua YIN ; Guo-yue ZHONG ; Tian-fu HUANG
China Journal of Chinese Materia Medica 2002;27(12):893-895
OBJECTIVETo supply the basis of the breeding the host of parasite of Cordyceps sinensis for it's natural culturing, by studying the biology of the host of parasite of C. sinensis, Hepialus biruensis.
METHODThe host of parasite was collected in field, eclosion, mating and spawning in the different conditions were made, and it's biology and the regularity of the growth and development were observed.
RESULT AND CONCLUSIONIn the natural condition, H. biruensis has the rate of eclosion, 42.3%, and the sex ratio, 1:0.86, and it's lifespan is 2-5 days. It has the rate of eclosion, 67.4%, in the constant temperature. At the 18 degrees C, H. biruensis has the highest rate of mating and spawning, 92% and 98%, and its rate of mating and spawning varies in the different temperature conditions. The male moth cherishes average 814.4 eggs and lays 607.2 eggs. In the open country, the optimum air relative humidity for mating and laying eggs is 75%-90%, while indoors, the optimum air relative humidity is 78%.
Animals ; Cordyceps ; Ecosystem ; Female ; Humidity ; Male ; Materia Medica ; Moths ; anatomy & histology ; microbiology ; physiology ; Oviposition ; Temperature
5.Effect of wound to growth of larva of host to Ophiocordyceps sinensis during artificial breeding.
Fei LIU ; De-li ZHANG ; Wei ZENG ; Li LI ; Qing-ming LUO ; Yong-qin TU ; Shi-jiang CHEN ; Ding-hua YIN
China Journal of Chinese Materia Medica 2015;40(2):210-212
To clear the effect of the wound to the growth of the larva of the host to the Ophiocordyceps sinensis, the wounds of same severity at the same position were made artificially to the larva and which were artificial fed at the same environment and condition. The results indicated that, over the winter, the survival rate of the wounded of the infection larva was lower than that of the healthy larva, but the weight had no significant difference between the wounded and the healthy larva. The survival rate of the wounded of the no infection larva was lower than that of the healthy larva, but except with black skin, the wounded larva with offwhite and dusty red had no influence on the variety of the weight. In summery, wound had no advantage to the survival rate, but had no influence to the weight. The result had provided theoretical basis to the reforming of the system of the artificial culture O. sinensis.
Animals
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Body Weight
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Breeding
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methods
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Hypocreales
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growth & development
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Larva
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Moths
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growth & development
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microbiology
6.Relevance analysis of amino acid contents in cultivated Cordyceps and cultivation materials.
Xiaoli WU ; Fei LIU ; Wei ZENG ; Guoyue ZHONG ; Yongqin TU ; Shijiang CHEN
China Journal of Chinese Materia Medica 2010;35(2):142-144
OBJECTIVETo clarify the relevance of amino acid content in cultivated Cordyceps and cultivation materials.
METHODThe content of amino acid was determined with L-8800 amino acid analyzer, and the relevance of amino acid content was analyzed with SPSS.
RESULT AND CONCLUSIONExcept mycelium of the C. sinensis or the blood-lymph of the larva, the cultivated Cordyceps and the main relevant cultivation materials had detected to contain all kinds of amino acids. Except among the mycelium, the blood-lymph of the larva, the part of the larva or of the stroma of cultivated Cordyceps, there was distinct relevance of amino acid contents in cultivated Cordyceps and the cultivation materials (P<0.01).
Amino Acids ; analysis ; metabolism ; Animals ; Cordyceps ; chemistry ; metabolism ; Larva ; chemistry ; microbiology ; Moths ; chemistry ; microbiology ; Mycelium ; chemistry ; metabolism
7.Pathogenicity of bacterium, Xenorhabdus nematophila isolated from entomopathogenic nematode (Steinernema carpocapsae) and its secretion against Galleria mellonella larvae.
Ali Nawaz MAHAR ; Muhammad MUNIR ; Sami ELAWAD ; Simon Richard GOWEN ; Nigel Graham Meckenzi HAGUE
Journal of Zhejiang University. Science. B 2005;6(6):457-463
The entomopathogenic bacterium, Xenorhabdus nematophila was isolated from the hemolymph of Galleria mellonella infected with Steinernema carpocapsae. The bacterial cells and its metabolic secretions have been found lethal to the Galleria larvae. Toxic secretion in broth caused 95% mortality within 4 d of application whereas the bacterial cells caused 93% mortality after 6 d. When filter and sand substrates were compared, the later one was observed as appropriate. Similarly, bacterial cells and secretion in broth were more effective at 14% moisture and 25 degrees C temperature treatments. Maximum insect mortality (100%) was observed when bacterial concentration of 4x10(6) cells/ml was used. Similarly, maximum bacterial cells in broth (95%) were penetrated into the insect body within 2 h of their application. However, when stored bacterial toxic secretion was applied to the insects its efficacy declined. On the other hand, when the same toxic secretion was dried and then dissolved either in broth or water was proved to be effective. The present study showed that the bacterium, X. nematophila or its toxic secretion can be used as an important component of integrated pest management against Galleria.
Animals
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Bacterial Proteins
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pharmacology
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Bacterial Toxins
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pharmacology
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Larva
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drug effects
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microbiology
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Moths
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drug effects
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microbiology
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Nematoda
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microbiology
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Pest Control, Biological
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methods
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Survival Analysis
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Survival Rate
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Xenorhabdus
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metabolism
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pathogenicity
8.Study on effect of artificial CsB and its components on bile acid metabolism in rats with liver fibrosis and its mechanism.
Xing ZHANG ; Feng-Hua LI ; Ping LIU ; Jia LIU ; Shuang WANG
China Journal of Chinese Materia Medica 2013;38(22):3943-3948
Bile acid is a type of metabolite degraded from cholesterol in liver. Its accumulation in liver could cause liver diseases, liver damage and liver fibrosis. In this experiment, dimethyl nitrosamine (DMN) liver fibrosis was established in rats. The rats were delivered into the normal group, the model group and four treated groups. After the four-week modeling, the treated groups were orally administered with drugs for 2 weeks, whereas the model and normal groups were given equal amount of sterile water at the same time. In the experiment, serum bile acid was taken the as marker, and liver function indexes and changes in bile acid metabolism were detected and observed to identify liver damage-related bile acid targets. It was the first time to evaluate the reverse effect of artificial CsB and its components on liver fibrosis in rats with bile acid metabolic level, and discuss its potential mechanism. The main study contents and results are as follows: a quantitative analysis was made on totally 17 endogenous bile acids, including taurocholic acid conjugated bile acid, glycine conjugated bile acid and free bile acid, and a liver damage evaluation was made for the model according to the detection of serum biochemical indexes and the pathological biopsy. After modeling, ALT, AST activity and TBil content significantly increased, whereas Alb significantly decreased. According to the pathological biopsy HE staining, the model group showed damage in normal hepatic lobule structure, liver cell edema and connective tissue proliferation in portal area; The treated groups showed mitigation in pathological changes to varying degrees. Cordyceps sinensis and its components may impact the bile acid metabolism in rats by activating HDCA, TCA, TCDCA, TLCA, TUDCA, UDCA, THDCA metabolim-related receptors or blocking relevant signaling pathway.
Animals
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Bile Acids and Salts
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metabolism
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Biological Factors
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administration & dosage
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Cordyceps
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chemistry
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physiology
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Humans
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Liver Cirrhosis
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drug therapy
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metabolism
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Male
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Moths
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chemistry
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microbiology
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Rats
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Rats, Wistar
9.Study on biological character of Hepialus introduced from Yunnan province.
Fei LIU ; Xiaoli WU ; Deli ZHANG ; Qingming LUO ; Shijiang CHEN ; Dinghua YIN ; Wei ZENG
China Journal of Chinese Materia Medica 2009;34(4):379-381
OBJECTIVETo study the biological character of Hepialus introduced from Yunnan province.
METHODH. jianchuanensis introduced from Yunnan was bred artificially to observe its biological character.
RESULTThe pupa duration was about 40 d, pupa emergence lasted for one day, female to male ratio was about 1.2:1, the mating peak was 19:30-20:30, the adults duration was about 25 d, the average life time of female was 5-6 d and that of male 6-7 d, the average number of the producing egg was about 300, the average weight of the 100 eggs was 10.33 mg. Lasting period of the larva is 350 d and the infancy larva showed the character of the polyphagy and cold endurance.
CONCLUSIONResults indicate that there are some differences in the whole duration, weight of the 100 eggs and the number of the eggs left in the abdomen between H. jianchuanensis and others bred at the Kangding base.
Animals ; Arachnid Vectors ; microbiology ; physiology ; China ; Cold Temperature ; Feeding Behavior ; physiology ; Female ; Larva ; anatomy & histology ; physiology ; Male ; Moths ; anatomy & histology ; physiology ; Oviposition ; physiology ; Ovum ; physiology ; Pupa ; physiology
10.Relationship between structure and function of loops from Bacillus thuringiensis insecticidal crystal protein Cry1Ba.
Guangjun WANG ; Jie ZHANG ; Donghui SUN ; Fuping SONG ; Dafang HUANG
Chinese Journal of Biotechnology 2008;24(9):1631-1636
To indicate the relationship between structure and function of loops from Bacillus thuringiensis insecticidal crystal protein Cry1Ba, and the influence of amino acids mutation on toxicity against diamond back moth Plutella xylostella, five mutations at the loops of Cry1Ba were constructed by overlapping primer PCR, and expressed in E. coli BL21 (DE3). Bioassay results showed that the toxicity of mutation M1 (loop1: 340WSNTR344-deletion), compared with that of Cry1Ba (LC50 0.96 microg/mL), decreased significantly with LC50 35.51 microg/mL. And the toxicity of mutation M2 (402Y-G), M3 (400GIYLEP405-PSAV), M4 (400GIYLEPIH407-ILGS) was also reduced to some extent respectively. Only M5 (mutation at loop3: 472LQSRV476 - AGAVYTL) showed slightly increased activity against P. xylostella, but not significantly (LC50 0.81 microg/mL). Referring to the structures of Cry1Ba which was predicted using Swiss-Model software, and bioassay data, we can conclude that loop1 and loop2 play a important role on determining the activity of Cry1Ba against P. xylostella.
Animals
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Bacillus thuringiensis
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genetics
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metabolism
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Bacterial Proteins
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chemistry
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genetics
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Endotoxins
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chemistry
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genetics
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Escherichia coli
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genetics
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metabolism
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Hemolysin Proteins
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chemistry
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genetics
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Models, Molecular
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Moths
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microbiology
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Mutation
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Protein Structure, Secondary
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Structure-Activity Relationship