Culture-based screening of selected bacterial indicators in three freeze-dried lots of the polychaete Australonuphis sp. collected on a single date in southern Brazil

Authors

  • Melissa Herrera-Perez Department of Agroindustrial Science and Technology, Federal University of Pelotas, Pelotas, Brazil - Centro de Excelencia en Biotecnología e Investigación Traslacional (CEBIT LATAM), San Vicente de Cañete, Lima, Perú
    • Helenice Gonzalez de Lima Faculty of Veterinary Medicine or College of Veterinary Medicine, Federal University of Pelotas, Pelotas, Brazil https://orcid.org/0000-0003-4567-5119
      • Natacha Deboni Cereser Faculty of Veterinary Medicine or College of Veterinary Medicine, Federal University of Pelotas, Pelotas, Brazil
        • Patricia da Silva Nascente Graduate Program in Microbiology and Parasitology, Federal University of Pelotas, Pelotas, Brazil
          • Camila dos Santos Cardozo Graduate Program in Microbiology and Parasitology, Federal University of Pelotas, Pelotas, Brazil
            • Ivan De Marco Food Technology and Bioprocesses Research Group, Department of Food Science and Technology, Federal University of Santa Catarina (UFSC), Florianópolis, Brazil
              • Cesar Valmor Rombaldi Department of Agroindustrial Science and Technology, Federal University of Pelotas, Pelotas, Brazil

                DOI:

                https://doi.org/10.47193/mafis.4012027010108

                Keywords:

                Marine annelid, feed hygiene, culture-based detection

                Abstract

                Polychaetes have been investigated as aquaculture feed ingredients because of their nutritional characteristics, particularly for broodstock diets of commercially important fish and shrimp species. However, the use of polychaetes from Latin America as feed resources for industrial aquaculture remains poorly documented, and microbiological data are an essential requirement for the development of new aquaculture ingredients. Therefore, this study screened three separately freeze-dried lots of Australonuphis sp., collected on the same date in southern Brazil, for selected bacterial indicators using culture-based ISO methods. From each lot, one 25-g test portion was homogenized in buffered peptone water; aliquots of this initial suspension were used for the enumeration of total coliforms, thermotolerant coliforms, presumptive Escherichia coli and coagulase-positive staphylococci, and the remaining homogenate was pre-enriched for the detection of Salmonella spp. No target microorganism was detected above the relevant method-specific reporting or detection limits in the tested portions. In all three lots, total coliforms were below 5 CFU g-1, thermotolerant coliforms and presumptive E. coli were below 3 MPN g-1, coagulase-positive staphylococci were below 50 CFU g-1, and Salmonella spp. was not detected. These findings provide preliminary microbiological baseline data for the tested lots of freeze-dried Australonuphis sp. only. Results should not be interpreted as evidence of animal-feed regulatory compliance, as the applicable MAPA requirements depend on the regulatory classification and intended use of the material. Further studies including raw-material and multi-season sampling, additional safety parameters, storage stability, digestibility and feeding trials, together with a formal feed-specific regulatory assessment, will be necessary to determine its suitability for future aquaculture applications.

                Downloads

                Download data is not yet available.

                References

                Ahmed N, Thompson S. 2019. The blue dimensions of aquaculture: a global synthesis. Sci Total Environ. 652: 851-861. DOI: https://doi.org/10.1016/j.scitotenv.2018.10.163

                Akbaş Y, Gümüş E, Koçer MAT, Kurtoğlu A, Aydın I, Aktaş Ö, Pak F, Sevgili H. 2026. Impact of partial replacement of fish meal with polychaete meal (Hediste diversicolor) on growth, nutrient digestibility, fatty acid profiles, and fish-in:fish-out ratio in diets of European seabass, Dicentrarchus labrax. Fish Physiol Biochem. 52 (20). DOI: https://doi.org/10.1007/s10695-026-01634-y

                Ang CY, Yong ASK, Azad SA, Lim LS, Zuldin WH, Lal MTM. 2021. Valorization of macroalgae through fermentation for aquafeed production: a review. Fermentation. 7 (4): 304. DOI: https://doi.org/10.3390/fermentation7040304

                Aragão C, Gonçalves AT, Costas B, Azeredo R, Xavier MJ, Engrola S. 2022. Alternative proteins for fish diets: implications beyond growth. Animals. 12 (9): 1211. DOI: https://doi.org/10.3390/ani12091211

                Argudín MÁ, Mendoza MC, Rodicio MR. 2010. Food poisoning and Staphylococcus aureus enterotoxins. Toxins. 2 (7): 1751-1773. DOI: https://doi.org/10.3390/toxins2071751

                Bagordo F, Brigida S, Grassi T, Caputo MC, Apollonio F, De Carlo L, Savino AF, Triggiano F, Turturro AC, De Donno A, et al. 2024. Factors influencing microbial contamination of groundwater: a systematic review of field-scale studies. Microorganisms. 12 (5): 913. DOI: https://doi.org/10.3390/microorganisms12050913

                Balqadi AA, Al-Gahwari YA, Ali HP, Awdha AF, Bakr HS, Bin Frijan SA. 2025. Effects of the partial replacing of fish meal with marine polychaetes (Nereis sp.) on the nutritional value of Macrobrachium rosenbergii shrimp. Asian J Biol Sci. 18 (2): 418-422. DOI: https://doi.org/10.3923/ajbs.2025.418.422

                Carrera-Parra LF. 2021. Onuphidae Kinberg, 1865. In: De León-González JÁ, Bastida-Zavala JR, Carrera-Parra LF, García-Garza ME, Salazar-Vallejo SI, Solís-Weiss V, Tovar-Hernández MA, editors. Anélidos marinos de México y América tropical. Monterrey: Universidad Autónoma de Nuevo León. p. 513-531.

                Cao W, Passot S, Irlinger F, Fonseca F. 2024. Investigation of freezing and freeze-drying for preserving and re-using a whole microbial cheese community. Foods. 13 (12): 1809. DOI: https://doi.org/10.3390/foods13121809

                Chen F, Leng Y, Lu Q, Zhou W. 2021. The application of microalgae biomass and bio-products as aquafeed for aquaculture. Algal Res. 60: 102541. DOI: https://doi.org/10.1016/j.algal.2021.102541

                Foster-Nyarko E, Pallen MJ. 2022. The microbial ecology of Escherichia coli in the vertebrate gut. FEMS Microbiol Rev. 46 (2): fuac008. DOI: https://doi.org/10.1093/femsre/fuac008

                Glencross BD, Huyben D, Schrama JW. 2020. The application of single-cell ingredients in aquaculture feeds-a review. Fishes. 5 (3): 22. DOI: https://doi.org/10.3390/fishes5030022

                González-Machado C, Alonso-Calleja C, Capita R. 2024. Prevalence and types of methicillin-resistant Staphylococcus aureus (MRSA) in meat and meat products from retail outlets and in samples of animal origin collected in farms, slaughterhouses and meat processing facilities: a review. Food Microbiology. 123: 104580. DOI: https://doi.org/10.1016/j.fm.2024.104580

                Hammad AM, Eltahan A, Hassan HA, Abbas NH, Hussien H, Shimamoto T. 2022. Loads of coliforms and fecal coliforms and characterization of thermotolerant Escherichia coli in fresh raw milk cheese. Foods. 11 (3): 332. DOI: https://doi.org/10.3390/foods11030332

                Herrera-Perez M. 2026. Carnadas empleadas en la pesca deportiva en el Perú. Biotempo. 23 (1): 55-62. DOI: https://doi.org/10.31381/biotempo.v1i23.8466

                Herrera-Perez M, Ferrando A. 2026. Polychaetes aquaculture: global successes and opportunities for development in Latin America. Aquacult Fish Fish. 6 (2): e70208. DOI: https://doi.org/10.1002/aff2.70208

                Hussain SM, Bano AA, Ali S, Rizwan M, Adrees M, Zahoor AF, Sarker PK, Hussain M, Arsalan MZU, Yong JWH, Naeem A. 2024. Substitution of fishmeal: Highlights of potential plant protein sources for aquaculture sustainability. Heliyon. 10 (4): e26573. DOI: https://doi.org/10.1016/j.heliyon.2024.e26573

                [ICMSF] International Commission on Microbiological Specifications for Foods. 2018. Microorganisms in foods 7: microbiological testing in food safety management. 2nd ed. Cham: Springer. 479 p. DOI: https://doi.org/10.1007/978-3-319-68460-4

                [ISO] International Organization for Standardization. 2005. ISO 7251:2005. Microbiology of food and animal feeding stuffs - horizontal method for the detection and enumeration of presumptive Escherichia coli - most probable number technique. Geneva: ISO. Amendment 1:2023 (ISO 7251:2005/Amd 1:2023). Inclusion of performance testing of culture media and reagents.

                [ISO] International Organization for Standardization. 2006a. ISO 4832:2006. Microbiology of food and animal feeding stuffs - horizontal method for the enumeration of coliforms - colony-count technique. Geneva: ISO.

                [ISO] International Organization for Standardization. 2006b. ISO 4831:2006. Microbiology of food and animal feeding stuffs - horizontal method for the detection and enumeration of coliforms - most probable number technique. Geneva: ISO.

                [ISO] International Organization for Standardization. 2017. ISO 6579-1:2017. Microbiology of the food chain - horizontal method for the detection, enumeration and serotyping of Salmonella. Part 1: detection of Salmonella spp. Geneva: ISO. Amendment 1:2020 (ISO 6579-1:2017/Amd 1:2020). Broader range of incubation temperatures, amendment to the status of Annex D, and correction of the composition of MSRV and SC.

                [ISO] International Organization for Standardization. 2021. ISO 6888-1:2021. Microbiology of the food chain - horizontal method for the enumeration of coagulase-positive staphylococci (Staphylococcus aureus and other species). Part 1: method using Baird-Parker agar medium. Geneva: ISO.

                Lana PC. 1991. Onuphidae (Annelida: Polychaeta) from southeastern Brazil. Bull Mar Sci. 48 (2): 280-295.

                Lee K, Koyama K, Kawai K, Koseki S. 2021. Why does Cronobacter sakazakii survive for a long time in dry environments? Contribution of the glass transition of dried bacterial cells. Microbiol Spectrum. 9 (3). DOI: https://doi.org/10.1128/spectrum.01384-21

                Léguillier V, Pinamonti D, Chang CM, Mukherjee G, Himanshu, Cossetini A, Manzano M, Anba-Mondoloni J, Malet-Villemagne J, Vidic J. 2024. A review and meta-analysis of Staphylococcus aureus prevalence in foods. Microbe. 4: 100131. DOI: https://doi.org/10.1016/j.microb.2024.100131

                Li Q, Dou L, Zhang Y, Luo L, Yang H, Wen K, Yu X, Shen J, Wang Z. 2024. A comprehensive review on the detection of Staphylococcus aureus enterotoxins in food samples. Compr Rev Food Sci Food Saf. 23: e13264. DOI: https://doi.org/10.1111/1541-4337.13264

                Liu S, Roopesh MS, Tang J, Wu Q, Qin W. 2022. Recent development in low-moisture foods: Microbial safety and thermal process. Food Res Int. 155: 111072. DOI: https://doi.org/10.1016/j.foodres.2022.111072

                Lupatsch I. 2014. Assessing the potential of polychaete meal in shrimp feeds. International Aquafeed. May-June. p. 10-12.

                Ma J, Dai J, Cao C, Su L, Cao M, He Y, Li M, Zhang Z, Chen J, Cui S, Yang B. 2024. Prevalence, serotype, antimicrobial susceptibility, contamination factors, and control methods of Salmonella spp. in retail fresh fruits and vegetables: a systematic review and meta-analysis. Compr Rev Food Sci Food Saf. 23 (4): e13407. DOI: https://doi.org/10.1111/1541-4337.13407

                Mahulette F. 2020. Traditionally catching and processing of Laor in Moluccas Islands. Local Wisdom. 12 (2): 99-110. DOI: https://doi.org/10.26905/lw.v12i2.4078

                Majowicz SE, Musto J, Scallan E, Angulo FJ, Kirk M, O’Brien SJ, Jones TF, Fazil A, Hoekstra RM. 2010. The global burden of nontyphoidal Salmonella gastroenteritis. Clin Infect Dis. 50 (6): 882-889. DOI: https://doi.org/10.1086/650733

                Martin NH, Evanowski RL, Wiedmann M. 2023. Invited review: Redefining raw milk quality-evaluation of raw milk microbiological parameters to ensure high-quality processed dairy products. J Dairy Sci. 106 (3): 1502-1517. DOI: https://doi.org/10.3168/jds.2022-22416

                Martins C, Rodrigo AP, Cabrita L, Leal MC, Pinteus S, Alves C, Gaspar H, Pedrosa R. 2019. The complexity of porphyrin-like pigments in a marine annelid sheds new light on haem metabolism in aquatic invertebrates. Sci Rep. 9: 12930. DOI: https://doi.org/10.1038/s41598-019-49433-1

                Milana M, van Asselt ED, van der Fels-Klerx HJ. 2024. The chemical and microbiological safety of emerging alternative protein sources and derived analogues: a review. Compr Rev Food Sci Food Saf. 23: e13377. DOI: https://doi.org/10.1111/1541-4337.13377

                Monteiro M, Rimoldi S, Costa RS, Kousoulaki K, Hasan I, Valente LMP, Terova G. 2023. Polychaete (Alitta virens) meal inclusion as a dietary strategy for modulating gut microbiota of European seabass (Dicentrarchus labrax). Front Immunol. 14: 1266947. DOI: https://doi.org/10.3389/fimmu.2023.1266947

                Morasi RM, Rall VLM, Dantas STA, Alonso VPPP, Silva NCCC. 2022. Salmonella spp. in low water activity food: occurrence, survival mechanisms, and thermoresistance. J Food Scie. 87 (6): 2310-2323. DOI: https://doi.org/10.1111/1750-3841.16152

                Muli BM, Kihia C, Fulanda BM, Munguti J, Muthumbi A. 2016. Effects of polychaete-based diets on growth and survival of tiger shrimp juveniles. Fifth Reforum Working Document Series. 14 (2): 907-911.

                Ndraha N, Lin HY, Hsiao HI, Lin HJ. 2024. Managing the microbiological safety of tilapia from farm to consumer. Compr Rev Food Sci Food Saf. 23 (5): e70023. DOI: https://doi.org/10.1111/1541-4337.70023

                Rana YS, Chen L, Jiao Y, Johnson LM, Snyder AB. 2024. A meta-analysis of microbial thermal inactivation in low moisture foods. Food Microbiol. 121: 104515. DOI: https://doi.org/10.1016/j.fm.2024.104515

                Rosario AILSS, Castro VS, Santos LF, Lisboa RC, Vallim DC, Silva MCA, Figueiredo EES, Conte-Junior CA, Costa MP. 2021. Shiga toxin-producing Escherichia coli isolated from pasteurized dairy products from Bahia, Brazil. J Dairy Sci. 104 (6): 6535-6547. DOI: https://doi.org/10.3168/jds.2020-19511

                Serra V, Pastorelli G, Tedesco DEA, Turin L, Guerrini A. 2024. Alternative protein sources in aquafeed: current scenario and future perspectives. Vet Anim Sci. 25: 100381. DOI: https://doi.org/10.1016/j.vas.2024.100381

                Silva JN, Baliza MD, Freitas F, Cruz ES, Camilo VMA, Silva IMM. 2021. Genotypical characterization of thermotolerant coliforms isolated from food produced by a solidarity economic venture of Bahia (Brazil). Braz J Bioly. 81 (1): 189-194. DOI: https://doi.org/10.1590/1519-6984.226833

                Snyder AB, Martin N, Wiedmann M. 2024. Microbial food spoilage: impact, causative agents and control strategies. Nat Rev Microbiol. 22: 528-542. DOI: https://doi.org/10.1038/s41579-024-01037-x

                Uwineza A, Zhang X. 2026. Application of freeze-drying technology in the food industry: a review. Foods. 15 (4): 790. DOI: https://doi.org/10.3390/foods15040790

                [WHO] World Health Organization. 2015. WHO estimates of the global burden of foodborne diseases: foodborne disease burden epidemiology reference group 2007-2015. Geneva: WHO. https://apps.who.int/iris/handle/10665/199350.

                [WOAH] World Organisation for Animal Health. 2024. Terrestrial animal health code. The control of hazards of animal health and public health importance in animal feed. Paris: WOAH.

                Xing L, Cheng M, Wang S, Jiang J, Li T, Zhang X, Yang J, Tian Y, Liu W. 2025. Methicillin-resistant Staphylococcus aureus contamination in meat and meat products: a systematic review and meta-analysis. Front Microbiol. 16: 1636622. DOI: https://doi.org/10.3389/fmicb.2025.1636622

                Zaffalon GB, Decol CM, Santos EP, Almeida Filho ES. 2025. Avaliação da qualidade microbiológica e físico-química do leite cru comercializado informalmente por rede social. Cienc Anim Bras. 26: e82514P. DOI: https://doi.org/10.1590/1809-6891v26e-82514P

                Downloads

                Published

                2026-10-29

                How to Cite

                Herrera-Perez, M., Gonzalez de Lima, H., Deboni Cereser, N., da Silva Nascente, P., dos Santos Cardozo, C., De Marco, I., & Valmor Rombaldi, C. (2026). Culture-based screening of selected bacterial indicators in three freeze-dried lots of the polychaete Australonuphis sp. collected on a single date in southern Brazil. Marine and Fishery Sciences (MAFIS), 40(1). https://doi.org/10.47193/mafis.4012027010108