Objective To isolate culturable intestinal bacteria from Clogmia albipunctata larvae, determine the digestive enzyme-producing activity and drug resistance of the bacteria, and to explore the effects of intestinal bacteria on food digestion and acclimatization of C. albipunctata. Methods The traditional isolation and culture method was used for the isolation and purification of culturable intestinal bacteria from C. albipunctata larvae, and 16S rRNA sequencing was used for identification. The flat transparent circle method was applied to screen for bacteria producing amylase, protease, cellulase, or lipase. The ratio of transparent circle diameter (D) to bacterial colony diameter (d) was calculated to compare the digestive enzyme-producing activities of different bacteria. Statistical analysis was performed using SPSS 20.0 software. The one-way analysis of variance was used for comparison between groups. When the variance is homogeneous, S-N-K multiple comparison was performed; when the variance is not homogeneous, the Dunnett's T3 test was performed. The paper disk diffusion method was used to analyze the drug susceptibility of different culturable intestinal bacteria. Results A total of 12 species of bacteria belonging to 5 genera were isolated from the intestine of C. albipunctata larvae. There were 7 species belonging to the genus Bacillus-B. vietnamensis, B. subtilis subsp., B. thuringiensis, B. aryabhattai, B. cereus, Lysinibacillus fusiformis, and L. pakistanensis. There were 2 species belonging to the genus Enterococcus-E. casseliflavus and E. gallinarum. There was 1 species each belonging to the genus Exiguobacterium, Leucobacter, and Staphylococcus-E. profundum, L. celer subsp., and S. sciuri. Among these bacteria, 8 species showed amylase-producing activity, 6 species showed protease-producing activity, and no bacteria showed cellulase- or lipase-producing activity. There was a significant difference in amylase-producing activity between the 8 species of bacteria (F=8.779, P<0.001). B. vietnamensis, B. subtilis subsp. and E. profundum had relatively high activity of producing amylase. There was a significant difference in protease-producing activity between the 6 species of bacteria (F=23.980, P<0.001). B. vietnamensis and B. subtilis subsp. had relatively high activity of producing protease. In drug susceptibility tests, S. sciuri showed resistance to 6 antibiotics, E. gallinarum were resistant to 5 antibiotics, and B. aryabhattai and E. casseliflavus showed resistance to 4 antibiotics. Conclusion The culturable intestinal bacteria from C. albipunctata larvae have the ability to produce amylase and protease, which may assist in digestion and breakdown of food. The intestinal bacteria generally have certain drug resistance, which can help C. albipunctata larvae survive in a dirty environment.
HE Zhi-wei, DONG Qi-yu, LIU Rui, LI Xin, LIU Xin, XU Cong-wu, ZHAO Xin-qi, WAN Qing, ZHANG Zhong
. Isolation and identification of culturable intestinal bacteria from Clogmia albipunctata larvae and analysis of their digestive enzyme-producing activities[J]. Chinese Journal of Vector Biology and Control, 2021
, 32(2)
: 224
-229
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DOI: 10.11853/j.issn.1003.8280.2021.02.021
[1] 任世恩,南小宁,许淼,等. 野生和人工饲喂条件下甘肃鼢鼠肠道细菌多样性比较[J]. 微生物学报,2020,60(4):826-838. DOI:10.13343/j.cnki.wsxb.20190342.Ren SN,Nan XN,Xu M,et al. Comparison of intestinal bacterial diversity of Gansu zokor under wild and artificial feeding conditions[J]. Acta Microbiol Sin,2020,60(4):826-838. DOI:10.13343/j.cnki.wsxb.20190342.
[2] 张科,苏智鹏,许阳,等. 蟋蟀后肠纤维素降解细菌的分离与鉴定[J]. 生物资源,2020,42(2):228-233. DOI:10.14188/j.ajsh.2020.02.010.Zhang K,Su ZP,Xu Y,et al. Isolation and identification of cellulose-degrading bacteria in the posterior intestine[J]. Biol Resour,2020,42(2):228-233. DOI:10.14188/j.ajsh.2020.02.010.
[3] 高欢欢,覃冬云,代晓彦,等. 肠道细菌弗氏柠檬酸杆菌和产酸克雷伯氏菌对斑翅果蝇生长发育和物质代谢的影响[J]. 昆虫学报,2020,63(4):462-469. DOI:10.16380/j.kcxb.2020. 04.009.Gao HH,Qin DY,Dai XY,et al. Effects of intestinal bacteria Citrobacter freundi and Klebsiella oxytoca on the development and substance metabolism of Drosophila suzukii (Diptera:Drosophilidae)[J]. Acta Entomol Sin,2020,63(4):462-469. DOI:10.16380/j.kcxb.2020.04.009.
[4] 丰硕,谢晓晨,黄振东,等. 小菌虫肠道可培养细菌的分离鉴定及产消化酶活性分析[J]. 中国微生态学杂志,2019,31(11):1255-1259. DOI:10.13381/j.cnki.cjm.201911004.Feng S,Xie XC,Huang ZD,et al. The activities of digestive enzymes in culturable bacteria isolated from intestinal tract of Alphitobius laevigatus[J]. Chin J Microecol,2019,31(11):1255-1259. DOI:10.13381/j.cnki.cjm.201911004.
[5] 高绘菊,路国兵,查传勇,等. 家蚕肠道产酶菌的分离与筛选[J]. 蚕业科学,2007,33(2):228-233. DOI:10.3969/j.issn. 0257-4799.2007.02.011.Gao HJ,Lu GB,Cha CY,et al. Isolation and screening of the enzyme-producing bacteria in the intestine of silkworm[J]. Acta Sericol Sin,2007,33(2):228-233. DOI:10.3969/j.issn.0257-4799.2007.02.011.
[6] Bing XL,Gerlach J,Loeb G,et al. Nutrient-dependent impact of microbes on Drosophila suzukii development[J]. mBio,2018,9(2):e02199-17. DOI:10.1128/mBio.02199-17.
[7] 吴晓露,夏晓峰,陈俊晖,等. 取食不同食物对小菜蛾幼虫肠道细菌多样性的影响[J]. 昆虫学报,2019,62(10):1172-1185. DOI:10.16380/j.kcxb.2019.10.006.Wu XL,Xia XF,Chen JH,et al. Effects of different diets on the diversity of larval gut bacteria of the diamondback moth,Plutella xylostella (Lepidoptera:Plutellidae)[J]. Acta Entomol Sin,2019,62(10):1172-1185. DOI:10.16380/j.kcxb.2019.10.006
[8] 朱名超,朱娅,许正敏,等. 泌尿生殖道发现毛蠓幼虫1例[J]. 中国热带医学,2019,19(2):195-196. DOI:10.13604/j.cnki. 46-1064/r.2019.02.23.Zhu MC,Zhu Y,Xu ZM,et al. One case of Psychoda larvae found in the urogenital tract[J]. China Trop Med,2019,19(2):195-196. DOI:10.13604/j.cnki.46-1064/r.2019.02.23.
[9] El-Dib NA,Ali MI,Hamdy DA,et al. Human intestinal myiasis caused by Clogmia albipunctata larvae (Diptera:Psychodidae):first report in Egypt[J]. J Infect Public Health,2020,13(4):661-663. DOI:10.1016/j.jiph.2019.07.023.
[10] Hjaija D,Sawalha SS,Amr ZS,et al. Urinary myiasis caused by Clogmia albipunctata from the Palestinian Territories[J]. Bull Soc Pathol Exot,2018,111(3):148-151. DOI:10.3166/bspe-2018-0037.
[11] El-Dib NA,El Wahab WMA,Hamdy DA,et al. Case report of human urinary myiasis caused by Clogmia albipunctata (Diptera:Psychodidae) with morphological description of larva and pupa[J]. J Arthropod Borne Dis,2017,11(4):533-538.
[12] Mokhtar AS,Braima KAO,Chin HP,et al. Intestinal myiasis in a malaysian patient caused by larvae of Clogmia albipunctatus (Diptera:Psychodidae)[J]. J Med Entomol,2016,53(4):957-960. DOI:10.1093/jme/tjw014.
[13] 黄振东,万晴,薛志静,等. 德国小蠊肠道可培养非厌氧细菌的分离、鉴定与产消化酶活性分析[J]. 中国媒介生物学及控制杂志,2019,30(4):409-413. DOI:10.11853/j.issn.1003.8280. 2019.04.012.Huang ZD,Wan Q,Xue ZJ,et al. Isolation and identification of culturable aerobic bacteria from the intestines of Blattella germanica and the activity of digestive enzymes produced by these bacteria[J]. Chin J Vector Biol Control,2019,30(4):409-413. DOI:10.11853/j.issn.1003.8280.2019.04.012.
[14] Faulde M,Spiesberger M. Role of the moth fly Clogmia albipunctata (Diptera:Psychodinae) as a mechanical vector of bacterial pathogens in German hospitals[J]. J Hosp Infect,2013,83(1):51-60. DOI:10.1016/j.jhin.2012.09.019.
[15] Faulde M,Spiesberger M. Hospital infestations by the moth fly,Clogmia albipunctata (Diptera:Psychodinae),in Germany[J]. J Hosp Infect,2012,81(2):134-136. DOI:10.1016/j.jhin.2012. 04.006.
