Livestock Probiotics in China: Quality Analysis and Enterococcus-Associated Antibiotic Resistance Dissemination Risks

Xing Ji , Jiayun Wang , Jun Li , Lili Zhang , Ruicheng Wei , Ran Wang , Tao He

Engineering ››

PDF (5178KB)
Engineering ›› DOI: 10.1016/j.eng.2025.03.032

Livestock Probiotics in China: Quality Analysis and Enterococcus-Associated Antibiotic Resistance Dissemination Risks

Author information +
History +
PDF (5178KB)

Abstract

As a natural alternative to antibiotics, probiotics have considerable potential for use in livestock farming. However, the current use of probiotics in livestock poses potential public health risks due to inadequate regulations, including issues such as the inferior quality and dissemination of antibiotic resistance. In this study, 95 non-duplicate commercial probiotic products for livestock were collected from different regions of China. Our findings revealed that the labeling compliance rate for Lactobacillus was the lowest, at just 11%, and approximately 33.3% of the products were contaminated with opportunistic pathogens containing various virulence and antibiotic-resistance genes (ARGs). Isolates of Bacillus and Enterococcus from the products exhibited diverse clonal types and geographical dispersion, whereas certain Enterococcus exhibited close phylogenetic relationships to clones associated with human infectious diseases. Compared with Bacillus and Lactobacillus, Enterococcus exhibited a higher prevalence of ARGs. Specifically, the oxazolidine-resistance gene optrA, which is located on novel transferable plasmids, was found in one isolate of E. faecium. Using chicken models, we observed that the optrA-positive E. faecium disrupts the normal intestinal microbiota in chickens and alters the abundance of intestinal resistome and mobile genetic elements (MGE). Furthermore, metagenomic analysis revealed that the optrA gene can be transferred via transposon IS1216E to commensal intestinal bacteria, including E. cecorum, E. gallinarum, and L. crispatus species. In summary, our study confirms that the probiotic products used in Chinese livestock production present problems such as non-compliance with good manufacturing practice (GMP) production standards and insufficient elucidation of the molecular genetic background of probiotic strains. The widespread use of low-quality Enterococcus strains containing various ARGs as probiotics could disrupt intestinal homeostasis and serve as a reservoir and source of ARGs. We emphasize the importance of carefully evaluating the use of Enterococcus strains as probiotics to avoid potential negative effects on livestock and human health.

Keywords

Probiotics / Livestock / Quality / Antibiotic-resistance genes / optrA

Cite this article

Download citation ▾
Xing Ji, Jiayun Wang, Jun Li, Lili Zhang, Ruicheng Wei, Ran Wang, Tao He. Livestock Probiotics in China: Quality Analysis and Enterococcus-Associated Antibiotic Resistance Dissemination Risks. Engineering DOI:10.1016/j.eng.2025.03.032

登录浏览全文

4963

注册一个新账户 忘记密码

Acknowledgments

This research was funded by the National Key Research and Development Program 2022YFD1800400 and Jiangsu Provincial Natural Science Foundation Project BK20220746.

Appendix A. Supplementary data

Supplementary data to this article can be found online.

References

[1]

Hill C, Guarner F, Reid G, Gibson GR, Merenstein DJ, Pot B, et al.The international scientific association for probiotics and prebiotics consensus statement on the scope and appropriate use of the term probiotic.Nat Rev Gastroenterol Hepatol 2014; 11(8):506-514.

[2]

Aponte M, Murru N, Shoukat M.Therapeutic, prophylactic, and functional use of probiotics: a current perspective.Front Microbiol 2020; 11:562048.

[3]

Kober AKMH, Riaz MS Rajoka, Mehwish HM, Villena J, Kitazawa H.Immunomodulation potential of probiotics: a novel strategy for improving livestock health, immunity, and productivity.Microorganisms 2022; 10(2):388.

[4]

Britton RA, Hoffmann DE, Khoruts A.Probiotics and the microbiome—how can we help patients make sense of probiotics?.Gastroenterology 2021; 160(2):614-623.

[5]

Ullah M, Raza A, Ye L, Yu Z.Viability and composition validation of commercial probiotic products by selective culturing combined with next-generation sequencing.Microorganisms 2019; 7(7):188.

[6]

Kothari D, Patel S, Kim S.Probiotic supplements might not be universally-effective and safe: a review.Biomed Pharmacother 2019; 111:537-547.

[7]

Vallabhaneni S, Walker TA, Lockhart SR, Ng D, Chiller T, Melchreit R, et al.Notes from the field: fatal gastrointestinal mucormycosis in a premature infant associated with a contaminated dietary supplement—connecticut.MMWR Morb Mortal Wkly Rep 2015; 64(6):155-156.

[8]

Zhu K, Hölzel CS, Cui Y, Mayer R, Wang Y, Dietrich R, et al.Probiotic Bacillus cereus strains, a potential risk for public health in China.Front Microbiol 2016; 7:718.

[9]

Tóth AG, Judge MF, SÁNagy , Papp M, Solymosi N.A survey on antimicrobial resistance genes of frequently used probiotic bacteria, 1901 to 2022.Euro Surveill 2023; 28(14):2200272.

[10]

Radovanovic M, Kekic D, Gajic I, Kabic J, Jovicevic M, Kekic N, et al.Potential influence of antimicrobial resistance gene content in probiotic bacteria on the gut resistome ecosystems.Front Nutr 2023; 10:1054555.

[11]

Gueimonde M, Sánchez B, G. de los Reyes-Gavilán C, Margolles A. Antibiotic resistance in probiotic bacteria. Front Microbiol 2013; 4:202.

[12]

Thumu SCR, Halami PM.Conjugal transfer of erm(B) and multiple tet genes from Lactobacillus spp. to bacterial pathogens in animal gut, in vitro and during food fermentation.Food Res Int 2019; 116:1066-1075.

[13]

Jacobsen L, Wilcks A, Hammer K, Huys G, Gevers D, Andersen SR.Horizontal transfer of tet(M) and erm(B) resistance plasmids from food strains of Lactobacillus plantarum to Enterococcus faecalis JH2-2 in the gastrointestinal tract of gnotobiotic rats.FEMS Microbiol Ecol 2007; 59(1):158-166.

[14]

Fu S, Yang Q, He F, Lan R, Hao J, Ni P, et al.National safety survey of animal-use commercial probiotics and their spillover effects from farm to humans: an emerging threat to public health.Clin Infect Dis 2020; 70(11):2386-2395.

[15]

Sun D, Jiang X, Wu QL, Zhou N.Intragenomic heterogeneity of 16S rRNA genes causes overestimation of prokaryotic diversity.Appl Environ Microbiol 2013; 79(19):5962-5969.

[16]

The European Food Safety Authority (EFS A).Guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance.EFSA J 2012;10(6):2740.

[17]

Inouye M, Dashnow H, Raven L, Schultz MB, Pope BJ, Tomita T, et al.SRST2: rapid genomic surveillance for public health and hospital microbiology labs.Genome Med 2014; 6(11):90.

[18]

Olson RD, Assaf R, Brettin T, Conrad N, Cucinell C, Davis JJ, et al.Introducing the Bacterial and Viral Bioinformatics Resource Center (BV-BRC): a resource combining PATRIC.IRD and ViPR. Nucleic Acids Res 2023; 51(D1):D678-D689.

[19]

Huys G, D K’Haene, Collard J, Swings J.Prevalence and molecular characterization of tetracycline resistance in enterococcus isolates from food.Appl Environ Microbiol 2004; 70(3):1555-1562.

[20]

Bolger AM, Lohse M, Usadel B.Trimmomatic: a flexible trimmer for Illumina sequence data.Bioinformatics 2014; 30(15):2114-2120.

[21]

Lee PH, Shatkay H.BNTagger: improved tagging SNP selection using Bayesian networks.Bioinformatics 2006; 22(14):e211-e219.

[22]

Li D, Liu C, Luo R, Sadakane K, Lam T.MEGAHIT: an ultra-fast single-node solution for large and complex metagenomics assembly via succinct de Bruijn graph.Bioinformatics 2015; 31(10):1674-1676.

[23]

Kang DD, Li F, Kirton E, Thomas A, Egan R, An H, et al.MetaBAT 2: an adaptive binning algorithm for robust and efficient genome reconstruction from metagenome assemblies.PeerJ 2019; 7:e7359.

[24]

Parks DH, Chuvochina M, Rinke C, Mussig AJ, Chaumeil P, Hugenholtz P.GTDB: an ongoing census of bacterial and archaeal diversity through a phylogenetically consistent, rank normalized and complete genome-based taxonomy.Nucleic Acids Res 2022; 50(D1):D785-D794.

[25]

Hyatt D, Chen G, Locascio PF, Land ML, Larimer FW, Hauser LJ.Prodigal: prokaryotic gene recognition and translation initiation site identification.BMC Bioinformatics 2010; 11(1):119.

[26]

Lu J, Rincon N, Wood DE, Breitwieser FP, Pockrandt C, Langmead B, et al.Metagenome analysis using the Kraken software suite.Nat Protoc 2022; 17(12):2815-2839.

[27]

Maia MS, Domingos MM, de JFB São José.Viability of probiotic microorganisms and the effect of their addition to fruit and vegetable juices.Microorganisms 2023; 11(5):1335.

[28]

Hu YJ, Cowling BJ.Reducing antibiotic use in livestock.China. Bull World Health Organ 2020; 98(5):360-361.

[29]

Zamojska D, Nowak A, Nowak I, Macierzy Eńska-Piotrowska.Probiotics and postbiotics as substitutes of antibiotics in farm animals: a review.Animals (Basel) 2021; 11(12):3431.

[30]

Liang D, Wu F, Zhou D, Tan B, Chen T.Commercial probiotic products in public health: current status and potential limitations.Crit Rev Food Sci Nutr 2024; 64(19):6455-6476.

[31]

Cui Y, Märtlbauer E, Dietrich R, Luo H, Ding S, Zhu K.Multifaceted toxin profile, an approach toward a better understanding of probiotic Bacillus cereus.Crit Rev Toxicol 2019; 49(4):342-356.

[32]

Sheppard SK.Strain wars and the evolution of opportunistic pathogens.Curr Opin Microbiol 2022; 67:102138.

[33]

Merenstein D, Pot B, Leyer G, Ouwehand AC, Preidis GA, Elkins CA, et al.Emerging issues in probiotic safety: 2023 perspectives.Gut Microbes 2023; 15(1):2185034.

[34]

Wang X, Yang Y, Huycke MM.Risks associated with enterococci as probiotics.Food Res Int 2020; 129:108788.

[35]

Groussin M, Poyet M, Sistiaga A, Kearney SM, Moniz K, Noel M, et al.Elevated rates of horizontal gene transfer in the industrialized human microbiome. Cell 2021;184(8):2053–67.e18.

[36]

Keogh D, Tay WH, Ho YY, Dale JL, Chen S, Umashankar S, et al.Enterococcal metabolite cues facilitate interspecies niche modulation and polymicrobial infection.Cell Host Microbe 2016; 20(4):493-503.

[37]

Xu W, Yuan G, Fang Y, Liu X, Ma X, Zhu K.Coumarin glycosides reverse Enterococci-facilitated enteric infections.Research (Was DC) 2024; 7:374.

[38]

Ding D, Wang B, Zhang X, Zhang J, Zhang H, Liu X, et al.The spread of antibiotic resistance to humans and potential protection strategies.Ecotoxicol Environ Saf 2023; 254:114734.

[39]

Montassier E, Vald Rés-Mas, Batard E, Zmora N, Dori-Bachash M, Suez J, et al.Probiotics impact the antibiotic resistance gene reservoir along the human GI tract in a person-specific and antibiotic-dependent manner.Nat Microbiol 2021; 6(8):1043-1054.

[40]

Shan X, Li X, Wang N, Schwarz S, Zhang S, Li D, et al.Studies on the role of IS1216E in the formation and dissemination of poxtA-carrying plasmids in an Enterococcus faecium clade A1 isolate.J Antimicrob Chemother 2020; 75(11):3126-3130.

AI Summary AI Mindmap
PDF (5178KB)

547

Accesses

0

Citation

Detail

Sections
Recommended

AI思维导图

/