Antimicrobial Resistance and Genetic Relationships of Enterococci from Siblings and Non-siblings Caterpillars

Overview

Journal PeerJ

Specialties Biology
Environmental Health
General Medicine

Date 2020 Mar 10

PMID 32149028

Citations 3

Authors

Rosana Huff

Rebeca Inhoque Pereira

Caroline Pissetti

Aldo Mellender de Araujo

Pedro Alves DAzevedo

Jeverson Frazzon

Ana Paula GuedesFrazzon

Affiliations

Soon will be listed here.

Abstract

Background: Studies evaluating bacteria in insects can provide information about host-microorganism-environment interactions. The gut microbial community has a profound effect on different physiological functions of insects. spp. are part of the gut community in humans and other animals, as well as in insects. The presence and antimicrobial resistance profile of enterococci are well studied in different animals; however, data for (Lepidoptera: Nymphalidae) do not yet exist. Therefore, the aims of this study were to evaluate the distribution of enterococcal species, their antimicrobial resistance profile and virulence genes, and the genetic relationships between enterococci isolated from fecal samples from sibling and non-sibling caterpillars collected from different sites in South Brazil.

Methods: Three females were captured (two from a forest fragment and one from an urban area), and kept individually in open-air insectaries. Eggs were collected and caterpillars (siblings and non-siblings) were fed daily with leaves. Fecal samples ( = 12) were collected from fifth-instar caterpillars, inoculated in selective medium, and 15 bacterial colonies were randomly selected from each sample. Enterococci were identified by PCR and MALDI-TOF, analyzed by disk diffusion antimicrobial susceptibility tests, and screened for resistance and virulence genes by PCR. The genetic relationships between the strains were determined using pulsed-field gel electrophoresis (PFGE).

Results: A total of 178 enterococci strains were identified: (74.15%; = 132), (21.34%; = 38), (1.12%; = 2) and sp. (3.37%; = 6). High rates of resistance to rifampicin (56%) and erythromycin (31%) were observed; 120 (67.41%) of the isolates showed resistance to at least one antibiotic and six (3.37%) were multidrug-resistant.None of the erythromycin-resistant strains was positive for the (B) and C genes. The virulence genes , , and were observed in 35%, 7%, and 1% of the strains, respectively. PFGE separated the enterococci into 22 patterns, four being composed of strains from sibling caterpillars.

Conclusion: was the dominant species in fecal samples of fifth-instar caterpillars. Resistant enterococci strains may be related to environmental pollution or the resistome. The PFGE analysis showed genetic relationships between some strains, suggesting that the enterococci isolated from fecal samples of the sibling caterpillars might have come from common sources, e.g., via diet (herbivory) and/or vertical transmission (through the egg surface). Further studies will be conducted to better understand the role of in the microbial community of the gastrointestinal tract of these insects, and the mechanisms involved in acquisition and maintenance of enterococci.

Citing Articles

High nitrogen in maize enriches gut microbiota conferring insecticide tolerance in lepidopteran pest .

Hu L, Sun Z, Xu C, Wang J, Mallik A, Gu C iScience. 2022; 25(1):103726.

PMID: 35072013 PMC: 8762471. DOI: 10.1016/j.isci.2021.103726.

Insects, Rodents, and Pets as Reservoirs, Vectors, and Sentinels of Antimicrobial Resistance.

Gwenzi W, Chaukura N, Muisa-Zikali N, Teta C, Musvuugwa T, Rzymski P Antibiotics (Basel). 2021; 10(1).

PMID: 33445633 PMC: 7826649. DOI: 10.3390/antibiotics10010068.

Multidrug Resistance in Enterococci Isolated From Wild Pampas Foxes () and Geoffroy's Cats () in the Brazilian Pampa Biome.

de Araujo G, Huff R, Favarini M, Mann M, Peters F, Frazzon J Front Vet Sci. 2021; 7:606377.

PMID: 33426025 PMC: 7793794. DOI: 10.3389/fvets.2020.606377.

References

Ferri M, Ranucci E, Romagnoli P, Giaccone V . Antimicrobial resistance: A global emerging threat to public health systems. Crit Rev Food Sci Nutr. 2015; 57(13):2857-2876. DOI: 10.1080/10408398.2015.1077192. View

Teh B, Apel J, Shao Y, Boland W . Colonization of the Intestinal Tract of the Polyphagous Pest Spodoptera littoralis with the GFP-Tagged Indigenous Gut Bacterium Enterococcus mundtii. Front Microbiol. 2016; 7:928. PMC: 4906056. DOI: 10.3389/fmicb.2016.00928. View

Jackson C, Fedorka-Cray P, Barrett J . Use of a genus- and species-specific multiplex PCR for identification of enterococci. J Clin Microbiol. 2004; 42(8):3558-65. PMC: 497640. DOI: 10.1128/JCM.42.8.3558-3565.2004. View

Snyman M, Gupta A, Bezuidenhout C, Claassens S, van den Berg J . Gut microbiota of Busseola fusca (Lepidoptera: Noctuidae). World J Microbiol Biotechnol. 2016; 32(7):115. DOI: 10.1007/s11274-016-2066-8. View

Mannu L, Paba A, Daga E, Comunian R, Zanetti S, Dupre I . Comparison of the incidence of virulence determinants and antibiotic resistance between Enterococcus faecium strains of dairy, animal and clinical origin. Int J Food Microbiol. 2003; 88(2-3):291-304. DOI: 10.1016/s0168-1605(03)00191-0. View

Usui M, Shirakawa T, Fukuda A, Tamura Y . The Role of Flies in Disseminating Plasmids with Antimicrobial-Resistance Genes Between Farms. Microb Drug Resist. 2015; 21(5):562-9. DOI: 10.1089/mdr.2015.0033. View

Shao Y, Chen B, Sun C, Ishida K, Hertweck C, Boland W . Symbiont-Derived Antimicrobials Contribute to the Control of the Lepidopteran Gut Microbiota. Cell Chem Biol. 2017; 24(1):66-75. DOI: 10.1016/j.chembiol.2016.11.015. View

Allonsius C, Van Beeck W, De Boeck I, Wittouck S, Lebeer S . The microbiome of the invertebrate model host Galleria mellonella is dominated by Enterococcus. Anim Microbiome. 2021; 1(1):7. PMC: 7807499. DOI: 10.1186/s42523-019-0010-6. View

Schaumburg F, Onwugamba F, Akulenko R, Peters G, Mellmann A, Kock R . A geospatial analysis of flies and the spread of antimicrobial resistant bacteria. Int J Med Microbiol. 2016; 306(7):566-571. DOI: 10.1016/j.ijmm.2016.06.002. View

10.

Murray B, Singh K, Heath J, Sharma B, Weinstock G . Comparison of genomic DNAs of different enterococcal isolates using restriction endonucleases with infrequent recognition sites. J Clin Microbiol. 1990; 28(9):2059-63. PMC: 268103. DOI: 10.1128/jcm.28.9.2059-2063.1990. View

11.

Hammer T, McMillan W, Fierer N . Metamorphosis of a butterfly-associated bacterial community. PLoS One. 2014; 9(1):e86995. PMC: 3900687. DOI: 10.1371/journal.pone.0086995. View

12.

Merrill R, Dasmahapatra K, Davey J, DellAglio D, Hanly J, Huber B . The diversification of Heliconius butterflies: what have we learned in 150 years?. J Evol Biol. 2015; 28(8):1417-38. DOI: 10.1111/jeb.12672. View

13.

Singer A, Shaw H, Rhodes V, Hart A . Review of Antimicrobial Resistance in the Environment and Its Relevance to Environmental Regulators. Front Microbiol. 2016; 7:1728. PMC: 5088501. DOI: 10.3389/fmicb.2016.01728. View

14.

Prichula J, Pereira R, Wachholz G, Cardoso L, Tolfo N, Santestevan N . Resistance to antimicrobial agents among enterococci isolated from fecal samples of wild marine species in the southern coast of Brazil. Mar Pollut Bull. 2016; 105(1):51-7. DOI: 10.1016/j.marpolbul.2016.02.071. View

15.

Sedgley C, Nagel A, Shelburne C, Clewell D, Appelbe O, Molander A . Quantitative real-time PCR detection of oral Enterococcus faecalis in humans. Arch Oral Biol. 2005; 50(6):575-83. DOI: 10.1016/j.archoralbio.2004.10.017. View

16.

Saeedi B, Hallgren A, Jonasson J, Nilsson L, Hanberger H, Isaksson B . Modified pulsed-field gel electrophoresis protocol for typing of enterococci. APMIS. 2003; 110(12):869-74. DOI: 10.1034/j.1600-0463.2002.1101205.x. View

17.

Depardieu F, Perichon B, Courvalin P . Detection of the van alphabet and identification of enterococci and staphylococci at the species level by multiplex PCR. J Clin Microbiol. 2004; 42(12):5857-60. PMC: 535300. DOI: 10.1128/JCM.42.12.5857-5860.2004. View

18.

van Schooten B, Godoy-Vitorino F, McMillan W, Papa R . Conserved microbiota among young butterfly species. PeerJ. 2018; 6:e5502. PMC: 6173163. DOI: 10.7717/peerj.5502. View

19.

Douglas A . Gut microbes alter the walking activity of fruit flies. Nature. 2018; 563(7731):331-332. DOI: 10.1038/d41586-018-07080-y. View

20.

Zhang J, Wang J, Chen L, Yassin A, Kelly P, Butaye P . Housefly (Musca domestica) and Blow Fly (Protophormia terraenovae) as Vectors of Bacteria Carrying Colistin Resistance Genes. Appl Environ Microbiol. 2017; 84(1). PMC: 5734023. DOI: 10.1128/AEM.01736-17. View