Isolation and Characterization of Bacteria Present in the Gut of Dead Females Glossina morsitan Submorsitans Using Culture Dependent and Independent Methods
Journal of Advances in Biology & Biotechnology, Volume 25, Issue 8,
Page 1-11
DOI:
10.9734/jabb/2022/v25i8589
Abstract
This study aimed to isolate and characterize bacteria present in the gut of death female Glossina morsitan submorsitans using culture dependent and independent methods. Thirty-four fresh dead female flies were obtained from the insectary of Nigerian Institute for Trypanosomiasis Research (NITR), Kaduna. The flies were surface sterilized and the mid-guts were dissected under aseptic conditions. The gut bacterial cultures were done for pregnant and non-pregnant dead female flies (6weeks- 15weeks old) a total of 20 isolates. The culture dependent methods identified six bacterial genera namely, Serratia species, Staphylococcus species, Escherichia species, Myroides species, Providencia species and Alkaligenes species. Molecular analysis only identified four (4) out of the six (6) isolates; as 4(50%) Serratia mercesens, 1(12.5%) Alcaligenes faecalis, 2(25%) Myroides odoratimus and 1(12.5%) Providencia. This is not surprising because the microorganisms were not included in the targeted spectrum of the Polymerase Chain Reaction. Our study recorded for the first time the presence of Myroides odoratimus in the gut of G. M. submorsitans in Nigeria. Some of these bacteria are pathogenic to the flies and may be the cause of mortality of laboratory reared flies. We recommend intensive research on tse tse gut bacterial flora to give insight into their use as potential vector bio-control tool.
- Bacteria
- glossina
- isolation
- characterization
How to Cite
References
Lindh j, Jenny M, Michael J. The tsetse fly Glossina fuscipes harbours a surprising diversity of bacteria other than symbiont. Antonie Leeuwenhoek. 2011;7720:1.
Mbewe NJ, Sole CL, Masiga DK, Yusuf AA. Efficiencies of stationary sampling tools for tsetse fly Glossina fuscipes in western Kenya. Act Trop. 2021;223-235.
Lindh JM, Lehane MJ. The tsetse fly Glossina fuscipes fuscipes (Diptera:Glossina) harbours a surprising diversity of bacteria other than symbionts. Antonie Leeuwenhoek. 2011;99:711-20.
Aksoy E, Telleria EL, Echodu R, Wu Y, Okedi LM, Weiss BL et al. Analysis of multiple tsetse fly populations in Uganda reveals limited diversity and species-specific gut microbiota. Appl Environ Microbiol. 2014;80(14):4301-12.
DOI: 10.1128/AEM.00079-14, PMID 24814785.
Colman DR, Toolson EC, Takacs-Vesbach CD. Do diet and taxonomy influence insect gut bacterial communities? Mol Ecol. 2012;21(20):5124-37.
DOI: 10.1111/j.1365-294X.2012.05752.x, PMID 22978555.
Geiger A, Fardeau ML, Njiokou F, Ollivier B. Glossina spp. Gut bacterial flora and their putative role fly-hosted trypanosome development. Cell Infect Microbiol. 2013;3:1-8.
Poinar GO, Wassink HJ, Leegwater-van der Linden ME, van der Geest LP. Serratia marcescens as a pathogen of tsetse flies. Acta Trop. 1979;36(3):223-7. PMID 43085.
Simo G, Njiokou F, Mbida Mbida JA, Njitchouang GR, Herder S, Asonganyi T, et al. Tsetse fly host preference from sleeping sickness foci in Cameroon: epidemiological implications. Infect Genet Evol. 2008; 8(1):34-9.
DOI: 10.1016/j.meegid.2007.09.005, PMID 17977803.
Farikou O, Njiokou F, Simo G, Asonganyi T, Cuny G, Geiger A. Tsetse flies blood meal modification and trypanosome identification in two sleeping sickness foci in the forest of southern Cameroon. Acta Trop. 2010;116(1):81-8.
DOI: 10.1016/j.actatropica.2010.06.002, PMID 20541513.
Bukachi SA, Wandibba S, Nyamongo IK. The socio-economic burden of human African trypanosomiasis and the coping strategies of households in the South Western Kenya foci. PLOS Negl Trop Dis. 2017;11(10):e0006002.
DOI: 10.1371/journal.pntd.0006002, PMID 29073144.
Ahmed AB. Laboratory rearing of tsetse flies. NITR annual scientific report. 1990;73.
Abubakar A, Abdullahi RA, Jibril HZ, Samdi SM, Mohammed H, Sabo E, et al. Patterns of wild pupae emergence from 2000-2009 at the insectary of the Department of Vector and Parasitology Studies, Nigerian Institute for Parasitology Research (NITR), Kaduna, Nigeria. Curr Resour J Biol Sci. 2010;2(6):407-9.
FAO. La situation mondiale de l’alimentation et de l’agriculture. Edition collection FAO. Agriculture; 2002.
Bouyer J, Yoni W, Bila C, Desquesnes M, Guerrini L [technical guide]. The Dissection of Tsetse flies. 2000;1:1-16.
Hedges AJ. Estimating the precision of serial dilutions and viable bacterial counts.642. Int J Food Microbiol. 2002;76(3):207-14.
DOI: 10.1016/s0168-1605(02)00022-3, PMID 12051477.
Kadhem HS, Abed Ali AA, Hassan OM. Isolation and identification of bacteria isolated from different parts of cell phones. World J Exp Biosci. 2016;4:29-31.
Van Den Abbeele J, Claes Y, van Bockstaele D, Le Ray D, Coosemans M. Trypanosoma brucei spp. development in the tsetse fly: characterization of the post-mesocyclic stages in the foregut and proboscis development in the steste fly. Parasitology. 1999;118(5):469-78.
DOI: 10.1017/s0031182099004217, PMID 10363280.
Kayser FH. Haemophilus and pasteurella. medical microbiology. 10th ed. Stuttgart: Georg Thieme Verlag. 2005;300-1.
WHO. World Health Organization: manual of basic techniques for health laboratory. 2nd ed. Geneva: World Health Organization; 2003.
Yumoto I, Ichihashi D, Iwata H, Istokovics A, Ichise N, Matsuyama H, et al. Purification and characterization of a catalase from the facultatively psychrophilic bacterium Vibrio rumoiensis S-1(T) exhibiting high catalase activity. J Bacteriol. 2000;182(7):1903-9.
DOI: 10.1128/JB.182.7.1903-1909.2000, PMID 10714995.
Vashist H, Sharma D, Gupta A. A review of commonly used biochemical tests for bacteria. Innovare J Life Sci. 2013;1(1):1-7.
Lehman D. Triple sugar iron agar protocols. Vol. 3. American Society for Microbiology. 2005;1-7.
Dahlén G, Hassan H, Blomqvist S, Carlén A. Rapid urease test (RUT) for evaluation of urease activity in oral bacteria in vitro and in supragingival dental plaque ex vivo. BMC Oral Health. 2018;18(1):89.
DOI: 10.1186/s12903-018-0541-3, PMID 29776416.
Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual. 2nd ed. Vol. 3. Cold Spring Harbor: Cold Spring Harbor Laboratory Press. 1989;1625.
Courtois S, Frostegård A, Göransson P, Depret G, Jeannin P, Simonet P. Quantification of bacterial subgroups in soil: comparison of DNA extracted directly from the soil or cells previously released by density gradient centrifugation. Environ Microbiol. 2001;3(7):431-9.
DOI: 10.1046/j.1462-2920.2001.00208.x, PMID 11553233.
Shi W, Syrenne R, Sun JZ, Yuan JS. Molecular approaches to study the insect gut symbiotic microbiota at the ’omics’ age. Insect Sci. 2010;17(3):199-219.
DOI: 10.1111/j.1744-7917.2010.01340.x
Nam YD, Jung MJ, Roh SW, Kim MS, Bae JW. Comparative Analysis of Korean Human Gut microbiota by barcoded Pyrosequencing. PLOS ONE. 2011;6(7):e22109.
DOI: 10.1371/journal.pone.0022109, PMID 21829445.
Akman L, Yamashita A, Watanabe H, Oshima K, Shiba T, Hattori M et al. Genome sequence of the endocellular obligate symbiont of tsets flies, Wigglesworthia glossinidia. Nat Genet. 2002;32(3):402-7.
DOI: 10.1038/ng986, PMID 12219091.
Griffith BC, Weiss BL, Aksoy E, Mireji PO, Auma JE, Wamwiri FN, et al. Analysis of the gut-specific microbiome from field-captured tsetse flies, and its potential relevance to host trypanosome vector competence. BMC Microbiol. 2018; 18(Suppl 1):146.
DOI: 10.1186/s12866-018-1284-7, PMID 30470178.
Grif K, Heller I, Prodinger WM, Lechleitner K, Lass-Flörl C, Orth D. Improvement of detection of bacterial pathogens in normally sterile body sites with a focus on orthopedic samples by use of a commercial 16S rRNA broad-range PCR and sequence analysis. J Clin Microbiol. 2012;50(7):2250-4.
DOI: 10.1128/JCM.00362-12, PMID 22553237.
Pumpuni CB, Demaio J, Kent M, Davis JR, J, Beier C. Bacterial population dynamics in three Anopheline species: an impact on Plasmodium sporogonic development. Am J Trop Med Hyg. 1996; 54(2):214-8.
Straif SC, Mbogo CNM, Toure AM, Walker ED, Kaufman M, Toure YT, et al. Midgut bacteria in Anopheles gambiae and A. funestus (Diptera: Culicidae) from Kenya and Mali. J Med Entomol. 1998;35(3): 222-6.
DOI: 10.1093/jmedent/35.3.222, PMID 9615538.
Geiger A, Fardeau ML, Grebaut P, Vatunga G, Josénando T, Herder S, et al. First isolation of Enterobacter, Enterococcus, and Acinetobacter species as inhabitants of the tsetse fly (Glossina palpalis palpalis) midgut. Infect Genet Evol. 2009;9(6):1364-70.
DOI: 10.1016/j.meegid.2009.09.013.
Geiger A, Fardeau ML, Falsen E, Ollivier B, Cuny G. Serratia glossinae species.not, isolated from the midgut of the tsetse fly Glossina palpalis gambiensis. Int J Syst Evol Microbiol. 2010;60(6):1261-5.
DOI: 10.1099/ijs.0.013441-0, PMID 19667382.
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