Molecular detection and characterization of rickettsia species in rodents and shrews from Kilombero district, Tanzania
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Abstract
Background: Rickettsia is a genus of obligatory intracellular bacteria transmitted by arthropods such as lice, ticks, and fleas. Rodents and shrews are well known for harboring a plethora of zoonotic pathogens including rickettsia. The diseases associated with Rickettsia are often misdiagnosed with other febrile infections such as Malaria and typhoid fever. This study aimed to detect and characterize the prevailing Rickettsia species in rodents and shrews from Kilombero district, Tanzania.
Methods: This study was conducted in Kilombero district between June 2022 and March 2023. The rodents and shrews were captured inside the households and outside households or in farms in the selected areas baited with maize bran and peanut butter. Prior to identification, rodents and shrews were anaesthetized by using Halothane was administered at the dose of 2mg/kg. DNA extraction from 204 tissue samples was done using DNeasy Blood and Tissue Kit from Qiagen. Amplification of the Citrate synthase (gltA) gene was done by Nested PCR using two sets of primer sequences in both rounds. PCR products were sequenced twice in the forward and reverse directions, consensus sequences were created and BLAST analysis was done. A phylogenetic tree was constructed using the Neighbor-Joining Method and 1000 bootstrap replications using the same software. The relationship between rodents’ rickettsiosis status to different factors were analyzed using chi-squared test.
Results: The overall molecular prevalence of Rickettsia species in rodents and shrews was: 10.20% (95% CI: 0.0634-0.153; n/N=20/196) and 12.5% (95%: CI: 0.0032-0.5265; n/N=1/8) respectively. Upon sequencing, all sequences were identified as Rickettsia conorii subsp. raoultii with percentage identity ranging from 98% to 100%.
Conclusion: This study confirms that rodents and shrews are infected with Rickettsia conorii in the study area.
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1. Kovalenko N. Morphology of Bacteria, Viruses and Protozoa. Learn Guid 2nd 3rd year English media students Fac Med Fac Dent (Microbiology, Virol Immunol. 2017;1–76.
2. Bermúdez CSE, Troyo A. A review of the genus Rickettsia in Central America. Res Rep Trop Med. 2018 Jun 29; 9:103-112. doi: 10.2147/RRTM.S160951.
3. Mahajan SK. Rickettsial diseases. J Assoc Physicians India. 2012 Jul;60:37-44.
4. Minichová L, Hamšíková Z, Mahríková L, Slovák M, Kocianová E, Kazimírová M, et al. Molecular evidence of Rickettsia spp. in ixodid ticks and rodents in suburban, natural and rural habitats in Slovakia. Parasit Vectors. 2017 Mar 24;10(1):158. doi: 10.1186/s13071-017-2094-8.
5.Valdez KR, Mendell NL, Escárcega-Ávila AM, de la Mora-Covarrubias A, Jiménez-Vega F, Waldrup KA, et al.. Survey of Fleas and Ticks for Rickettsia rickettsii and Rickettsia typhi in the El Paso Community and Other Areas in Texas, New Mexico, and Ciudad Juarez, Mexico. Am J Trop Med Hyg. 2025 Jun 17;113(3):659-665. doi: 10.4269/ajtmh.24-0709.
6. Noden BH, Davidson S, Smith JL, Williams F. First detection of rickettsia typhi and rickettsia felis in fleas collected from client-owned companion animals in the southern great plains. J Med Entomol. 2017 Jul 1;54(4):1093-1097. doi: 10.1093/jme/tjx069.
7. Zahoor A, Abbas RZ, Khurram A, Aslam A, Mateen A, Aslam A, et al. Vector-borne zoonotic diseases. In: Abbas RZ, Hassan MF, Khan A and Mohsin M (eds), Zoonosis, Unique Scientific Publishers, Faisalabad, Pakistan, 2023.Vol. 2: 12-36. https://doi.org/10.47278/book.zoon/2023.49
8. Kumar K, Patnaik R, Kumari H, Sharma N. Acute febrile illness: a systematic review of infectious aetiologies among patients. Journal of Pharmaceutical Negative Results 2022;13 (Special Issue): 5080-5093. DOI:10.47750/pnr.2022.13.S08.667
9. Thomas CA, katakweba AAS, Massawe AW, Makundi RH, Machang RS, Kessy ST. Prevalence of Rickettsia typhi in rodent fleas from areas with and without previous history of plague in Mbulu district, Tanzania. African Journal of Microbiology Research 2020;14(2):65–70. https://doi.org/10.5897/AJMR2019.9198
10. Moorthy GS, Rubach MP, Maze MJ, Refuerzo RP, Shirima GM, Lukambagire AS, et al. Prevalence and risk factors for Q fever, spotted fever group rickettsioses, and typhus group rickettsioses in a pastoralist community of northern Tanzania, 2016-2017. Trop Med Int Health. 2024 May;29(5):365-376. doi: 10.1111/tmi.13980.
11. Leulmi H, Socolovschi C, Laudisoit A, Houemenou G, Davoust B, Bitam I, Raoult D, Parola P. Detection of Rickettsia felis, Rickettsia typhi, Bartonella Species and Yersinia pestis in Fleas (Siphonaptera) from Africa. PLoS Negl Trop Dis. 2014 Oct 9;8(10):e3152. doi: 10.1371/journal.pntd.0003152.
12. Issae AR, Katakweba AS, Kicheleri RP, Chengula AA, van Zwetselaar M, Kasanga CJ. Exploring Pathogenic and Zoonotic Bacteria from Wild Rodents, Dogs, and Humans of the Ngorongoro District in Tanzania Using Metagenomics Next-Generation Sequencing. Zoonotic Diseases. 2023; 3(3):226-242. https://doi.org/10.3390/zoonoticdis3030019
13. Moonga LC, Hayashida K, Nakao R, Lisulo M, Kaneko C, Nakamura I, et al. Molecular detection of Rickettsia felis in dogs, rodents and cat fleas in Zambia. Parasit Vectors. 2019 Apr 11;12(1):168. doi: 10.1186/s13071-019-3435-6.
14. Rwela AG. Socio-economic and institutional determinants of membership in Agricultural Marketing Co-operative Societies in Mvomero and Kilombero districts, Tanzania. Development Studies Research, 2023; 10(1): 5095. https://doi.org/10.1080/21665095.2022.2163679
15. Advisory R, Report M. United Republic of Tanzania Ramsar Advisory Mission Report. 2017; 1173.
16. Martin W, Meek AH, Willeberg P, Wayne Martin by S, Meek A. ©1987 S Part 1: Basic Principles Chapter 3: Measurement of Disease Frequency and Production. Vet Epidemiol Princ Methods. 1987.
17. Essbauer S, Hofmann M, Kleinemeier C, Wölfel S, Matthee S. Rickettsia diversity in southern Africa: A small mammal perspective. Ticks Tick Borne Dis. 2018 Feb;9(2):288-301. doi: 10.1016/j.ttbdis.2017.11.002.
18. Mwamengele G, Sabuni C, Mathew C. Seroprevalence and risk factors for brucellosis in cattle and rodents in Kilosa district, Morogoro, Tanzania. J Ideas Heal. 2024;7 (4):1123–30. https://doi.org/10.47108/jidhealth.Vol7.Iss4.363
19. Schlitter DA, Ferguson AW, McDonough MM, Mammals of Africa (Vol. I-VI.), Journal of Mammalogy, Volume 95, Issue 6, December 2014, Pages 1299–1303, https://doi.org/10.1644/14-MAMM-R-122.
20. Kaasalainen U, Kirika PM, Mollel NP, Hemp A, Rikkinen J. The Lichen Genus Sticta (Lobariaceae, Peltigerales) in East African Montane Ecosystems. Journal of Fungi. 2023; 9(2):246. https://doi.org/10.3390/jof9020246.
21. Perles L, Barreto WTG, de Macedo GC, Calchi AC, Bezerra-Santos M, Mendoza-Roldan JA, Otranto D, Herrera HM, Barros-Battesti DM, Machado RZ, André MR. Molecular detection of Babesia spp. and Rickettsia spp. in coatis (Nasua nasua) and associated ticks from midwestern Brazil. Parasitol Res. 2023 May;122(5):1151-1158. doi: 10.1007/s00436-023-07815-5.
22. Palacios-Santana R, Wei L, Fernandez-Santos NA, Rodriguez-Perez MA, Uriegas-Camargo S, Mendell NL, et al. Spotted Fever and Typhus Group Rickettsiae in Dogs and Humans, Mexico, 2022. Emerg Infect Dis. 2023 Jul;29(7):1443-1446. doi: 10.3201/eid2907.230333.
23. Tsui PY, Tsai KH, Weng MH, Hung YW, Liu YT, Hu KY, Lien JC, Lin PR, Shaio MF, Wang HC, Ji DD. Molecular detection and characterization of spotted fever group rickettsiae in Taiwan. Am J Trop Med Hyg. 2007 Nov;77(5):883-90. Erratum in: Am J Trop Med Hyg. 2008 Jul;79(1):140. Wang, Hsis-Chieh [corrected to Wang, Hsi-Chieh].
24. Tamura K, Stecher G, Kumar S. MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Mol Biol Evol. 2021 Jun 25;38(7):3022-3027. doi: 10.1093/molbev/msab120.
25. Laudisoit A, Falay D, Amundala N, Akaibe D, de Bellocq JG, Van Houtte N, et al. High prevalence of Rickettsia typhi and Bartonella species in rats and fleas, Kisangani, Democratic Republic of the Congo. Am J Trop Med Hyg. 2014 Mar;90(3):463-8. doi: 10.4269/ajtmh.13-0216.
26. Chitanga S, Chibesa K, Sichibalo K, Mubemba B, Nalubamba KS, Muleya W, et al. Molecular Detection and Characterization of Rickettsia Species in Ixodid Ticks Collected From Cattle in Southern Zambia. Front Vet Sci. 2021 Jun 7;8:684487. doi: 10.3389/fvets.2021.684487.
27. Rio P, Caldarelli M, Miccoli E, Guazzarotti G, Gasbarrini A, Gambassi G, Cianci R. Sex Differences in Immune Responses to Infectious Diseases: The Role of Genetics, Hormones, and Aging. Diseases. 2025 Jun 7;13(6):179. doi: 10.3390/diseases13060179.
28. Rakotonanahary RJ, Harrison A, Maina AN, Jiang J, Richards AL, Rajerison M, Telfer S. Molecular and serological evidence of flea-associated typhus group and spotted fever group rickettsial infections in Madagascar. Parasit Vectors. 2017 Mar 4;10(1):125. doi: 10.1186/s13071-017-2061-4.
29. Nallan K, Ayyavu V, Ayyanar E, Thirupathi B, Gupta B, Devaraju P, et al. Molecular Evidence of Rickettsia conorii subsp. raoultii and Rickettsia felis in Haemaphysalis intermedia Ticks in Sirumalai, Eastern Ghats, Tamil Nadu, South India. Microorganisms. 2023 Jun 30;11(7):1713. doi: 10.3390/microorganisms11071713.
30. Cossu CA, Cassini R, Bhoora RV, Menandro ML, Oosthuizen MC, Collins NE, Wentzel J, Quan M, Fagir DM, van Heerden H. Occurrence and molecular prevalence of Anaplasmataceae, Rickettsiaceae and Coxiellaceae in African wildlife: A systematic review and meta-analysis. Prev Vet Med. 2024 Sep;230:106257. doi: 10.1016/j.prevetmed.2024.106257.
31. Maina AN. Sero-epidemiology and molecular characterization of Rickettsiae infecting humans, selected animals and arthropod vectors in Asembo, western Kenya,. 2012;2007–10.
32. Heglasová I, Víchová B, Kraljik J, Mošanský L, Miklisová D, Stanko M. Molecular evidence and diversity of the spotted-fever group Rickettsia spp. in small mammals from natural, suburban and urban areas of Eastern Slovakia. Ticks Tick Borne Dis. 2018 Sep;9(6):1400-1406. doi: 10.1016/j.ttbdis.2018.06.011.
33. Chen L, Cai Y, Zhou G, Shi X, Su J, Chen G, Lin K. Rapid Sanger sequencing of the 16S rRNA gene for identification of some common pathogens. PLoS One. 2014 Feb 14;9(2):e88886. doi: 10.1371/journal.pone.0088886.
34. Zhu WJ, Ye RZ, Tian D, Wang N, Gao WY, Wang BH, et al. The first direct detection of spotted fever group Rickettsia spp. diversity in ticks from Ningxia, northwestern China. PLoS Negl Trop Dis. 2025 Jan 2;19(1):e0012729. doi: 10.1371/journal.pntd.0012729.
35. Rungrojn A, Chaisiri K, Paladsing Y, Morand S, Junjhon J, Blacksell SD, Ekchariyawat P. Prevalence and Molecular Characterization of Rickettsia spp. from Wild Small Mammals in Public Parks and Urban Areas of Bangkok Metropolitan, Thailand. Trop Med Infect Dis. 2021 Nov 11;6(4):199. doi: 10.3390/tropicalmed6040199.