ISSN 1820-9955 (Printed Ed.), eISSN 2683-4138 (Online)
STUDY OF BACTERIAL CARRIAGE IN DARKLING BEETLES (ALPHITOBIUS DIAPERINUS) COLLECTED FROM INDUSTRIAL BROILER FLOCKS IN NORTH-EAST OF TUNISIA
PDF

Keywords

Alphitobius diaperinus
broiler chickens
Escherichia coli
resistance to antibiotics
Tunisia

How to Cite

KABOUDI, K., GADRIA, M., CHEBBI, C. and MESSADI, L. (2025) “STUDY OF BACTERIAL CARRIAGE IN DARKLING BEETLES (ALPHITOBIUS DIAPERINUS) COLLECTED FROM INDUSTRIAL BROILER FLOCKS IN NORTH-EAST OF TUNISIA”, Archives of Veterinary Medicine, 18(2). doi: 10.46784/e-avm.v18i2.488.

Abstract

Alphitobius diaperinus is regarded as a highly resilient pest in poultry production systems that use litter-based rearing . It is also known as a potential reservoir of several avian pathogens. The present study aimed to determine darkling beetle carriage of bacterial pathogens in Tunisian poultry houses. A total of 25 industrial broiler flocks naturally infested by darkling beetles and located in the North-East of Tunisia were sampled. A pooled sample of adult beetles (5 g) was collected from five distinct locations within each broiler house. Insects were placed in sterile containers and transported to the laboratory under cooled conditions. All samples were examined for entomological identification. Bacteriological analysis was performed to detect external and internal bacterial carriage, according to the standard protocols. Antimicrobial susceptibility for all Escherichia coli isolates was tested by the disk diffusion method on Mueller–Hinton agar using commercial disks. Antimicrobial susceptibility testing was not performed for the remaining bacterial isolates. Entomological identification showed that all sampled beetles belong to Alphitobius diaperinus species. A total of 108 isolates were detected from the surface (57 isolates) and the interior (51 isolates) of adult insects. Gram-negative (63.89%) (Escherichia coli, Enterobacter spp., Klebsiella pneumoniae, Proteus vulgarus, Citrobacter spp., Pseudomonas spp., Pseudomonas aeruginosa) and coagulase-negative staphylococci (36.11%) (Gram-positive) bacteria were isolated, with the global predominance of coagulase-negative staphylococci, followed by E. coli (23.15%), Pseudomonas spp. (13.89%) and Pseudomonas aeruginosa (10.19%). No Salmonella spp. was detected. Antibiotic resistance profile showed that all Escherichia coli isolates were multi-resistant. High resistance was observed to doxycycline (25/25; 100%), amoxicillin (24/25; 96%), cephalothin (23/25; 92%), streptomycin (22/25; 88%), tetracycline (22/25; 88%), enrofloxacin (22/25; 88%). A high level of resistance was observed to trimethoprim-sulfamethoxazole (13 out of 25 isolates; 52%) and gentamicin  (9/25; 36%). However, a low level of resistance was observed to florfenicol (7/25; 28%) and colistin (3/25; 12%). Our results confirm the potential vector role of beetles in the spread of bacteria pathogens and antibiotic resistance.

https://doi.org/10.46784/e-avm.v18i2.488
PDF

References

Andreasen, C.B. 2020. Staphylococcosis. In: Diseases of poultry, Eds. D.E. Swayne, M. Boulianne, C.M. Logue, L.R. McDougald, V. Nair, D.L. Suarez, S. de Wit, T. Grimes, D. Johnson, M. Kromm, T.Y. Prajitno, I. Rubinoff, G. Zavala, 14th ed. John Wiley & Sons, Ltd, Hoboken, NJ, 995–1003.

Agabou, A., Alloui, N. 2010. Importance of Alphitobius diaperinus (Panzer) as a reservoir for pathogenic bacteria in Algerian broiler houses. Veterinary World, 3, 2, 71-73.

Atere, A., Atere, F.V. 2024. Multidrug resistant Pseudomonas aeruginosa isolated from chicken: A public health concern. Microbiology Research Journal International, 34, 7, 1-7. https://doi.org/10.9734/mrji/2024/v34i71452

Badr, J.M., El Saidy, F.R., Abdelfattah, A.A. 2020. Emergence of multi-drug resistant Pseudomonas aeruginosa in broiler chicks. International Journal of Microbiology and Biotechnology, 5, 2, 41. https://doi.org/10.11648/j.ijmb.20200502.11

Barua, S., Bailey, M., Zhong, K., Iduu, N., Dormitorio, T., Macklin, K., Bourassa, D., Price, S., Hauck, R., Krehling, J., Kitchens, S., Kyriakis, C., Buhr, R.J., Wang, C. 2023. Research Note: Role of darkling beetles (Alphitobius diaperinus) and litter in spreading and maintaining Salmonella Enteritidis and Campylobacter jejuni in chicken flocks. Poultry Science, 102, 11, 103061. https://doi.org/10.1016/j.psj.2023.103061

CLSI (Clinical and Laboratory Standards Institute), 2015. Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Fifth Informational Supplement. CLSI document M100-S25. Wayne, P.A. Clinical and Laboratory Standards Institute.

Crippen, T.L., Sheffield, C.L., Beier, R.C., Nisbet, D.J. 2018. The horizontal transfer of Salmonella between the lesser mealworm (Alphitobius diaperinus) and poultry manure. Zoonoses Public Health, 65, 1 e23–33. https://doi.org/10.1111/zph.12404

Crippen, T.L., Sheffield, C. 2006. External surface disinfection of the lesser mealworm (Coleoptera: Tenebrionidae). Journal of Medical Entomology 43, 5, 916-23. https://doi.org/10.1603/0022-2585(2006)43[916:esdotl]2.0.co;2

Crippen, T.L., Singh, B., Anderson, R.C., Sheffield, C.L., 2022. Management practices affecting lesser mealworm larvae (Alphitobius diaperinus) associated microbial community in a broiler house and after relocating with the litter into pastureland. Frontiers in Microbiology, 13, 875930. https://doi.org/10.3389/fmicb.2022.875930

Cuccini, C., Leo, C., Vitale, M., Frati, F., Carapelli, A., Nardi, F. 2020. Bacterial and fungal diversity in the gut of polystyrene-fed Alphitobius diaperinus (Insecta: Coleoptera). Animal Gene, 17-18, 200109. https://doi.org/10.1016/j.angen.2020.200109

Di Francesco, A., Salvatore, D., Sakhria, S., Bertelloni, F., Catelli, E., Ben Yahia, S., Tlatli, A. 2023. Colistin resistance genes in broiler chickens in Tunisia. Animals, 13, 8, 1409. https://doi.org/10.3390/ani13081409

Donoso, A., Paredes, N., Retamal, P. 2020. Detection of antimicrobial resistant Salmonella enterica strains in larval and adult forms of lesser mealworm (Alphitobius diaperinus) from industrial poultry farms. Frontiers in Veterinary Science, 7, 577848. https://doi.org/10.3389/fvets.2020.577848

EFSA (European Food Safety Authority) NDA panel (EFSA Panel on Nutrition, Novel Foods and Food Allergens), Turck, D., Bohn, T., Castenmiller, J., De Henauw, S., Hirsch-Ernst, K.I., Maciuk, A., Mangelsdorf, I., McArdle, H.J., Naska, A., Pelaez, C., Pentieva, K., Siani, A., Thies, F., Tsabouri, S., Vinceti, M., Cubadda, F., Frenzel, T., Heinonen, M., Marchelli, R., Neuhäuser-Berthold, M., Poulsen, M., Prieto Maradona, M., Schlatter, J.R., van Loveren, H., Ververis, E., Knutsen, H.K. 2022. Scientific Opinion on the safety of frozen and freeze-dried formulations of the lesser mealworm (Alphitobius diaperinus larva) as a Novel food pursuant to Regulation (EU) 2015/2283. EFSA Journal, 20, 7, 7325, 36. https://doi.org/10.2903/j.efsa.2022.7325

Gehring, V.S., Santos, E.D., Mendonça, B.S., Santos, L.R., Rodrigues, L.B., Dickel, E.L., Daroit, L., Pilotto, F. 2020. Alphitobius diaperinus control and physicochemical study of poultry litters treated with quicklime and shallow fermentation. Poultry Science, 99, 4, 2120-2124. https://doi.org/10.1016/j.psj.2019.11.039

Goodwin, M.A., Waltman, W.D. 1996. Transmission of Eimeria, viruses, and bacteria to chicks: darkling beetles (Alphitobius diaperinus) as vectors of pathogens. Journal of Applied Poultry Research, 5, 1, 51-55. https://doi.org/10.1093/japr/5.1.51

Grami, R., Mansour, W., Mehri, W., Bouallègue, O., Boujaâfar, N., Madec, J.Y., Haenni, M. 2016. Impact of food animal trade on the spread of mcr-1-mediated colistin resistance, Tunisia, July 2015. Eurosurveillance, 21, 8, 30144. https://doi.org/10.2807/1560-7917.ES.2016.21.8.30144

Hassen, B., Abbassi, M.S., Ruiz-Ripa, L., Mama, O.M., Hassen, A., Torres, C., Hammami, S. 2020. High prevalence of mcr-1 encoding colistin resistance and first identification of blaCTX-M-55 in ESBL/CMY-2-producing Escherichia coli isolated from chicken faeces and retail meat in Tunisia. International Journal of Food Microbiology, 318, 108478. https://doi.org/10.1016/j.ijfoodmicro.2019.108478

Hazeleger, W.C., Bolder, N.M., Beumer, R.R., Jacobs-Reitsma, W.F. 2008. Darkling beetles (Alphitobius diaperinus) and their larvae as potential vectors for the transfer of Campylobacter jejuni and Salmonella enterica serovar paratyphi B variant Java between successive broiler flocks. Applied and Environmental Microbiology, 74, 22, 6887-6891. https://doi.org/10.1128/AEM.00451-08

Krinsky, W.L. 2002. Beetles (Coleoptera). In Medical and Veterinary Entomology, Eds. G.R. Mullen, L.A. Durden, Academic Press, San Diego, 87-101. https://doi.org/10.1016/B978-0-12-814043-7.00009-1

Maamar, E., Alonso, C.A., Hamzaoui, Z., Dakhli, N., Abbassi, M.S., Ferjani, S., Saidani, M., Boubaker, I.B.B., Torres, C. 2018. Emergence of plasmid-mediated colistin-resistance in CMY-2-producing Escherichia coli of lineage ST2197 in a Tunisian poultry farm. International Journal of Food Microbiology, 23, 269, 60–63. https://doi.org/10.1016/j.ijfoodmicro.2018.01.017

McAllister, J.C., Steelman, C.D., Newberry, L., Skeeles, J.K. 1995. Isolation of infectious bursal disease virus from the lesser mealworm, Alphitobius diaperinus (Panzer). Poultry Science, 74, 1, 45–49. https://doi.org/10.3382/ps.0740045

McAllister, J.C., Steelman, C.D., Skeeles, J.K., Newberry, L.A., Gbur, E.E. 1996. Reservoir competence of the Alphitobius diaperinus (Coleoptera: Tenebrionidae) for Escherichia coli (Eubacteriales: Enterobacteriaceae). Journal of Medical Entomology, 33, 6, 983–9872. https://doi.org/10.1093/jmedent/33.6.983

Nguyen, N., Yang, B.K., Lee, J.S., Yoon, J.U., Hong, K.J. 2019. Infestation status of the darkling beetle (Alphitobius diaperinus) in broiler chicken houses of Korea. Korean Journal of Applied Entomology, 58, 3, 189-196. https://doi.org/10.5656/KSAE.2019.08.0.028

Pfeiffer, D.G., Axtell, R.C. 1980. Coleoptera of poultry manure in caged-layer houses in North Carolina. Environmental Entomology, 9, 1, 21-28.

Rawat, N., Sabu, B., Jamwal, R., Devi, P.P., Yadav, K., Raina, H.S., Rajagopal, R. 2023. Understanding the role of insects in the acquisition and transmission of antibiotic resistance. Science of the Total Environment, 858, 159805. https://doi.org/10.1016/j.scitotenv.2022.159805

Roche, A.J., Cox, N.A., Richardson, L.J., Buhr, R.J., Cason, J.A., Fairchild, B.D., Hinkle, N.C., 2009. Transmission of Salmonella to broilers by contaminated larval and adult lesser mealworms, Alphitobius diaperinus (Coleoptera: Tenebrionidae). Poultry Science, 88, 1, 44-48. https://doi.org/10.3382/ps.2008-00235

Rueda, L.M., Axtell, R.C. 1997. Arthropods in litter of poultry (broiler chicken and turkey) houses. Journal of Agriculture Entomology, 14, 1, 81–91.

Schawaller, W., Grimm, R. 2014. The genus Alphitobius Stephens (Coleoptera, Tenebrionidae, Alphitobiini) in Africa and adjacent islands. ZooKeys, 415, 169–90. https://doi.org/10.3897/zookeys.415.6676

Segabinazi, S.D., Flôres, M.L., Barcelos, A.S., Jacobsen, G., Eltz, R.D. 2005. Bactérias da família Enterobacteriaceae em Alphitobius diaperinus oriundos de granjas avícolas dos Estados do Rio Grande do Sul e Santa Catarina, Brasil. Acta Scientiae Veterinariae, 33, 1, 51-55. https://doi.org/10.22456/1679-9216.14449

Skewes, P.A., Monroe, J.L. 1991. Research note: the effects of darkling beetles on broiler performance. Poultry Science, 70, 4, 1034-1036. https://doi.org/10.3382/ps.0701034

Skov, M.N., Spencer, A.G., Hald, B., Petersen, L., Nauery, B., Carstensen, B., Madsen, M. 2004. The role of litter beetles as potential reservoir for Salmonella enterica and thermophilic Campylobacter spp. between broiler flocks. Avian Diseases, 48, 1, 9-18. https://doi.org/10.1637/5698

Smith, R., Hauck, R., Macklin, K., Price, S., Dormitorio, T., Wang, C. 2021. A review of the lesser mealworm beetle (Alphitobius diaperinus) as a reservoir for poultry bacterial pathogens and antimicrobial resistance. World’s Poultry Science Journal, 78, 1, 197–214. https://doi.org/10.1080/00439339.2022.2003172

Tamburro, M., Sammarco, M., Trematerra, P., Colacci, M. 2022. Alphitobius diaperinus panzer (insecta, coleoptera) in a single house of a broiler production facility as a potential source of pathogenic bacteria for broilers and humans. Letters in Applied Microbiology, 74, 6, 883–892. https://doi.org/10.1111/lam.13679

Tayh, G., Nsibi, F., Chemli, K., Daâloul-Jedidi, M., Abbes, O., Messadi, L. 2024. Occurrence, antibiotic resistance and molecular characterisation of Shiga toxin-producing Escherichia coli isolated from broiler chickens in Tunisia. British Poultry Science, 65, 6, 751-761. https://doi.org/10.1080/00071668.2024.2368906

Terra, M.T., Macklin, K.S., Burleson, M., Jeon, A., Beckmann, J.F., Hauck, R. 2023. Mapping the poultry insectome in and around broiler breeder pullet farms identifies new potential Dipteran vectors of Histomonas meleagridis. Parasite Vectors, 16, 1, 244. https://doi.org/10.1186/s13071-023-05833-x

Watson, D.W., Guy, J.S., Stringham, S.M. 2000. Limited transmission of turkey coronavirus in young turkeys by adult Alphitobius diaperinus (Coleoptera: Tenebrionidae). Journal of Medical Entomology, 37, 3, 480–483. https://doi.org/10.1093/jmedent/37.3.480

Wynants, E., Crauwels, S., Verreth, C., Gianotten, N., Lievens, B., Claes, J., Van Campenhout, L. 2018. Microbial dynamics during production of lesser mealworms (Alphitobius diaperinus) for human consumption at industrial scale. Food Microbiology, 70, 181-191. https://doi.org/10.1016/j.fm.2017.09.012

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Copyright (c) 2025 Archives of Veterinary Medicine