Abstract
Rhipicephalus microplus tick is the ectoparasite causing the greatest economic losses in the livestock industry. Multi-resistance in ticks is increasing, generating the inefficiency of traditional ixodicides, for which biological control has been proposed as an alternative. In this work, we analyze the histomorphological damage caused by the bacterial strain EC-35 on Rhipicephalus microplus. The ixodicidal effect of EC-35 total protein was evaluated on larval or adult ticks comparing with the commercial ixodicide coumaphos 0.02% as a control. Female ticks were processed using the paraffin-embedding technique and stained with hematoxylin–eosin. Also, the pathogenicity of EC-35 was evaluated by capillary feeding and coelom inoculation tests. The identification of the bacterium was performed using the molecular markers 16S RNA and rpoB, by PCR and sequencing technique, and the evolutionary distance was analyzed by Bayesian phylogenetic inference. No differences were observed in the perimeter and area of larvae treated with EC-35 or Coumaphos. The thickness of the integument decreased a 65% with the EC-35 treatment (6.01 ± 0.6 µm) and of 30% in coumaphos (12.04 ± 1.2 µm) in larvae compared with the control group (18.41 ± 2 µm), while no difference was found in adult ticks. The capillary feeding test and coelom inoculation with EC-35 showed an inhibition of reproductive potential of 99.8 ± 7 and an oviposition Inhibition 97 ± 3.02%. The EC-35 strain was genetically related to Serratia marcescens, concluding that these bacteria caused high mortality, oviposition Inhibition, and integument thinning and drastic loss of histoarchitecture in R. microplus tick larvae.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Availability of data and materials
The authors confirm that the data supporting the findings of this study are available within the article.
References
Aggarwal C, Paul S, Tripathi V, Paul B, Khan MA (2017) Characterization of putative virulence factors of Serratia marcescens strain SEN for pathogenesis in Spodoptera litura. J Invertebr Pathol 143:115–123. https://doi.org/10.1016/j.jip.2016.12.004
Almazán C, Lagunes R, Villar M, Canales M, Rosario-Cruz R, Jongejan F, de la Fuente J (2010) Identification and characterization of Rhipicephalus (Boophilus) microplus candidate protective antigens for the control of cattle tick infestations. Parasitol Res 106(2):471–479. https://doi.org/10.1007/s00436-009-1689-1
Andreotti R, De León AAP, Dowd SE, Guerrero FD, Bendele KG, Scoles GA (2011) Assessment of bacterial diversity in the cattle tick Rhipicephalus (Boophilus) microplus through tag-encoded pyrosequencing. BMC Microbiol 11(1):6. https://doi.org/10.1186/1471-2180-11-6
Bidari F, Shams-Bakhsh M, Mehrabadi M (2017) Isolation and characterization of a Serratia marcescens with insecticidal activity from Polyphylla olivieri (Col: Scarabaeidae). J Appl Entomol. https://doi.org/10.1111/jen.12421
Bradford MM (1976) A rapid and sensitive method for the quantitation microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1006/abio.1976.9999
Bravo A, Sarabia S, Lopez L, Ontiveros H, Abarca C, Ortiz A, Ortiz M, Lina L, Villalobos FJ, Peña G, Nuñez-Valdez ME, Soberón M, Quintero R (1998) Characterization of cry genes in a Mexican Bacillus thuringiensis strain collection. Appl Environ Microbiol 64(12):4965–4972. https://doi.org/10.1128/AEM.64.12.4965-4972.1998
Burritt NL, Foss NJ, Neeno-Eckwall EC, Church JO, Hilger AM, Hildebrand JA, Warshauer DM, Perna NT, Burritt JB (2016) Sepsis and hemocyte loss in honey bees (Apis mellifera) Infected with Serratia marcescens strain sicaria. PLoS ONE 11(12):e0167752. https://doi.org/10.1371/journal.pone.0167752
Castro-Saines E, Hernandez-Ortiz R, Lagunes-Quintanilla R, Peña-Chora G (2021) Characterization of a strain of Serratia sp. with ixodicide activity against the cattle tick Rhipicephalus microplus. Exp Appl Acarol 85(1):101–111. https://doi.org/10.1007/s10493-021-00640-4
Drummond RO, Ernst SE, Trevino JL, Gladney WJ, Graham OH (1973) Boophilus annulatus and B. microplus: Laboratory test of insecticides. J Econ Entomol 66(1):130–133. https://doi.org/10.1093/jee/66.1.130
Dzemo WD, Thekisoe O, Vudriko P (2022) Development of acaricide resistance in tick populations of cattle: A systematic review and meta-analysis. Heliyon 8(1):e08718. https://doi.org/10.1016/j.heliyon.2022.e08718
Fernández-Ruvalcaba M, Peña-Chora G, Romo-Martínez A, Hernández-Velázquez V, de la Parra AB, De La Rosa DP (2010) Evaluation of Bacillus thuringiensis pathogenicity for a strain of the tick, Rhipicephalus microplus, resistant to chemical pesticides. J Insect Sci 10:1–6. https://doi.org/10.1673/031.010.14146
Fukuto TR (1990) Mechanism of action of organophosphorus and carbamate insecticides. Environ Health Perspect 87:245–254. https://doi.org/10.1289/ehp.9087245
Gonsioroski AV, Bezerra IA, Utiumi KU, Driemeier D, Farias SE, da Silva VI, Masuda A (2012) Anti-tick monoclonal antibody applied by artificial capillary feeding in Rhipicephalus (Boophilus) microplus females. Exp Parasitol 130(4):359–363. https://doi.org/10.1016/j.exppara.2012.02.006
Grimont PA, Grimont F (1978) The genus Serratia. Annu Rev Microbiol 32(4):221–248. https://doi.org/10.1146/annurev.mi.32.100178.001253
Hackman RH (1982) Structure and function in tick cuticle. Annu Rev Entomol 27:75–95. https://doi.org/10.1146/annurev.en.27.010182.000451
Hertle R (2005) The family of Serratia type pore forming toxins. Curr Protein Pept Sci 6(4):313–325. https://doi.org/10.2174/1389203054546370
Ishii K, Adachi T, Hamamoto H, Sekimizu K (2014a) Serratia marcescens suppresses host cellular immunity via the production of an adhesion-inhibitory factor against immunosurveillance cells. J Biol Chem 289(9):5876–5888. https://doi.org/10.1074/jbc.M113.544536
Ishii K, Adachi T, Hara T, Hamamoto H, Sekimizu K (2014b) Identification of a Serratia marcescens virulence factor that promotes hemolymph bleeding in the silkworm, Bombyx mori. J Invertebr Pathol 117:61–67. https://doi.org/10.1016/j.jip.2014.02.001
Jeong HU, Mun HY, Oh HK, Kim SB, Yang KY, Kim I, Lee HB (2010) Evaluation of insecticidal activity of a bacterial strain Serratia sp. EML-SE1 against diamondback moth. J Microbiol 48(4):541–545. https://doi.org/10.1007/s12275-010-0221-9
Jongejan F, Uilenberg G (2004) The global importance of ticks. Parasitology 129(Suppl):S3–S14. https://doi.org/10.1017/s0031182004005967
Klafke GM, Sabatini GA, de Albuquerque TA, Martins JR, Kemp DH, Miller RJ, Schumaker TT (2006) Larval immersion tests with ivermectin in populations of the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) from State of Sao Paulo. Brazil Vet Parasitol 142(3–4):386–390. https://doi.org/10.1016/j.vetpar.2006.07.001
Li P, Kwok AH, Jiang J, Ran T, Xu D, Wang W, Leung FC (2015) Comparative genome analyses of Serratia marcescens FS14 reveals its high antagonistic potential. PLoS ONE 10(4):e0123061. https://doi.org/10.1371/journal.pone.0123061
Lormendez CC, Fernandez-Ruvalcaba M, Adames-Mancebo M, Hernandez-Velazquez VM, Zuñiga-Navarrete F, Flores-Ramirez G, Lina-Garcia L, Peña-Chora G (2019) Mass production of a S-layer protein of Bacillus thuringiensis and its toxicity to the cattle tick Rhipicephalus microplus. Sci Rep 9(1):17586. https://doi.org/10.1038/s41598-019-53854-3
Machado-Ferreira E, Vizzoni VF, Piesman J, Gazeta GS, Soares CA (2015) Bacteria associated with Amblyomma cajennense tick eggs. Genet Mol Biol 38(4):477–483. https://doi.org/10.1590/S1415-475738420150040
Mahlen SD (2011) Serratia infections: from military experiments to current practice. Clin Microbiol Rev 24(4):755–791. https://doi.org/10.1128/CMR.00017-11
Miller M et al (2010) Creating the CIPRES science gateway for inference of large phylogenetic trees, vol 1. https://doi.org/10.1109/GCE.2010.5676129, https://ieeexplore.ieee.org/document/5676129?arnumber=5676129
Mollet C, Drancourt M, Raoult D (1997) rpoB sequence analysis as a novel basis for bacterial identification. Mol Microbiol 26(5):1005–1011. https://doi.org/10.1046/j.1365-2958.1997.6382009.x
Nuñez-Valdez ME, Calderón MA, Aranda E, Hernández L, Ramírez-Gama RM, Lina L, Rodríguez-Segura Z, Gutiérrez MDC, Villalobos FJ (2008) Identification of a putative Mexican strain of Serratia entomophila pathogenic against root-damaging larvae of Scarabaeidae (Coleoptera). Appl Environ Microbiol 74(3):802–810. https://doi.org/10.1128/AEM.01074-07
Parte AC, SardàCarbasse J, Meier-Kolthoff JP, Reimer LC, Göker M (2020) List of Prokaryotic names with standing in nomenclature (LPSN) moves to the DSMZ. Int J Syst Evol Microbiol 70(11):5607–5612. https://doi.org/10.1099/ijsem.0.004332
Patil CD, Patil SV, Salunke BK, Salunkhe RB (2012) Insecticidal potency of bacterial species Bacillus thuringiensis SV2 and Serratia nematodiphila SV6 against larvae of mosquito species Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus. Parasitol Res 110(5):1841–1847. https://doi.org/10.1007/s00436-011-2708-6
Petersen LM, Tisa LS (2013) Friend or foe? A review of the mechanisms that drive Serratia towards diverse lifestyles. Can J Microbiol 59(9):627–640. https://doi.org/10.1139/cjm-2013-0343
Pineda-Castellanos ML, Rodríguez-Segura Z, Villalobos FJ, Hernández L, Lina L, Nuñez-Valdez ME (2015) Pathogenicity of Isolates of Serratia marcescens towards larvae of the Scarab Phyllophaga Blanchardi (Coleoptera). Pathogens 4(2):210–228. https://doi.org/10.3390/pathogens4020210
Raymann K, Coon KL, Shaffer Z, Salisbury S, Moran NA (2019) Correction for Raymann et al., “Pathogenicity of Serratia marcescens Strains in honey bees.” Mbio 10(1):e02855-e2918. https://doi.org/10.1128/mBio.02855-18
Rodriguez-Vivas RI, Jonsson NN, Bhushan C (2018) Strategies for the control of Rhipicephalus microplus ticks in a world of conventional acaricide and macrocyclic lactone resistance. Parasitol Res 117(1):3–29. https://doi.org/10.1007/s00436-017-5677-6
Romo-Martínez A, Fernández-Ruvalcaba M, Hernández-Velázquez VM, Peña-Chora G, Lina-García LP, Osorio-Miranda J (2013) Evaluation of natural origin products for the control of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) on cattle artificially infested. Basic Res J Agri Sci Rev ISSN 2(3):2315–6880
Segura JA, Isaza JP, Botero LE, Alzate JF, Gutiérrez LA (2020) Assessment of bacterial diversity of Rhipicephalus microplus ticks from two livestock agroecosystems in Antioquia. Colombia Plos One 15(7):e0234005. https://doi.org/10.1371/journal.pone.0234005
Soenens A, Imperial J (2020) Biocontrol capabilities of the genus Serratia. Phytochem Rev 19:577–587. https://doi.org/10.1007/s11101-019-09657-5
Soufiane B, Côté JC (2009) Discrimination among Bacillus thuringiensis H serotypes, serovars and strains based on 16S rRNA, gyrB and aroE gene sequence analyses. Antonie Van Leeuwenhoek 95(1):33–45. https://doi.org/10.1007/s10482-008-9285-4
Tu S, Qiu X, Cao L, Han R, Zhang Y, Liu X (2010) Expression and characterization of the chitinases from Serratia marcescens GEI strain for the control of Varroa destructor, a honey bee parasite. J Invertebr Pathol 104(2):75–82. https://doi.org/10.1016/j.jip.2010.02.002
Veliz EA, Martínez-Hidalgo P, Hirsch AM (2017) Chitinase-producing bacteria and their role in biocontrol. AIMS Microbiol 3(3):689–705. https://doi.org/10.3934/microbiol.2017.3.689
Acknowledgements
ECS acknowledges the PhD fellowship awarded by the National Council for Science and Technology (CONACyT 811487). Authors acknowledge the technical support from M.C. Erika Segura Salinas.
Funding
The authors did not receive support from any organization for the submitted work.
Author information
Authors and Affiliations
Contributions
Conceptualization: GPC, IFP; Methodology: ECS; Formal analysis and investigation: ECS, GPC, RHO, RLQ, IFP; Writing—original draft preparation: ECS, CHC; Writing—review and editing: ECS, CHC; Resources: All authors; Supervision: GPC, IFP, RHO. All authors read and approved the final manuscript.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Conflict of interest
The authors have no relevant financial or non-financial interests to disclose.
Ethical approval
No approval of research ethics committees was required to accomplish the goals of this study because experimental work was conducted with an unregulated invertebrate species (ticks). Regarding the production of ticks on cattle, it was approved by the Animal Experimentation and Ethics Committee of the Nacional Center of Disciplinary Research in Animal Health and Safety of the National Institute for Forestry, Agricultural and Livestock Research (Castro-Saines et al 2021).
Consent to participate
This research does not involve humans.
Consent for publication
All the authors give consent for the publication of the content in this manuscript.
Additional information
Communicated by Yusuf Akhter.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Castro-Saines, E., Peña-Chora, G., Hallal-Calleros, C. et al. Histometric and morphological damage caused by Serratia marcescens to the tick Rhipicephalus microplus (Acari: Ixodidae). Arch Microbiol 204, 677 (2022). https://doi.org/10.1007/s00203-022-03275-0
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00203-022-03275-0