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Identification of the Bacteria and Their Metabolic Activities Associated with the Microbial Spoilage of Custard Cream Desserts

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Identification of the Bacteria and Their Metabolic Activities Associated with the Microbial Spoilage of Custard Cream Desserts Identification of the bacteria and their metabolic activities associated with the microbial spoilage of custard cream desserts Clarisse Techer, Sophie Jan, Anne Thierry, Marie-Bernadette Maillard, Noel Grosset, Olivier Galet, Valérie Breton, Michel Gautier, Florence Baron To cite this version: Clarisse Techer, Sophie Jan, Anne Thierry, Marie-Bernadette Maillard, Noel Grosset, et al.. Identifi- cation of the bacteria and their metabolic activities associated with the microbial spoilage of custard cream desserts. Food Microbiology, Elsevier, 2020, 86, pp.103317. 10.1016/j.fm.2019.103317. hal- 02283796 HAL Id: hal-02283796 https://hal-agrocampus-ouest.archives-ouvertes.fr/hal-02283796 Submitted on 17 Sep 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Food Microbiology 86 (2020) 103317 Contents lists available at ScienceDirect Food Microbiology journal homepage: www.elsevier.com/locate/fm Identification of the bacteria and their metabolic activities associated with T the microbial spoilage of custard cream desserts ∗ Clarisse Techera, , Sophie Janb, Anne Thierryb, Marie-Bernadette Maillardb, Noël Grossetb, Olivier Galetc, Valérie Bretonc, Michel Gautierb, Florence Baronb a Mixscience, 2 Avenue de Ker Lann, 35712, Bruz, France b UMR1253 Science et Technologie du Lait et de l’Œuf, Agrocampus Ouest, INRA, F-35042, Rennes, France c Ovoteam, 13 Rue Porte de Chammay, 53300, Ambrières-les-Vallées, France ARTICLE INFO ABSTRACT Keywords: The famous French dessert “ile flottante” consists of a sweet egg white foam floating on a vanilla custard cream, B. cereus group which contains highly nutritive raw materials, including milk, sugar and egg. Spoilage issues are therefore a key Egg product concern for the manufacturers. This study explored the bacterial diversity of 64 spoiled custard cream desserts Spoilage marker manufactured by 2 French companies. B. cereus group bacteria, coagulase negative Staphylococcus, Enterococcus Volatilome and Leuconostoc spp. were isolated from spoiled products. Thirty-one bacterial isolates representative of the main spoilage species were tested for their spoilage abilities. Significant growth and pH decrease were observed regardless of species. While off-odours were detected with B. cereus group and staphylococci, yoghurt odours were detected with Enterococcus spp. and Leuconostoc spp. B. cereus group bacteria produced various esters and several compounds derived from amino acid and sugar metabolism. Most Staphylococci produced phenolic compounds. Enterococcus spp. and Leuconostoc spp. isolates produced high levels of compounds derived from sugar metabolism. Each type of spoilage bacteria was associated with a specific volatile profile and lactic acid was identified as a potential marker of spoilage of custard cream-based desserts. These findings provide valuable information for manufacturers to improve food spoilage detection and prevention of chilled desserts made with milk and egg. 1. Introduction et al., 2001). Some acids, such as lactic and succinic acids, have been associated with food spoilage and can serve as spoilage indicators for Spoilage is defined as any change in a food product that renders it foods such as milk or egg products (Alamprese et al., 2004; Cousin, unacceptable to the consumer from a sensory point of view (Sperber, 1982; Dainty, 1996; Nassos et al., 1988; Stijve and Diserens, 1987). 2009). Microbial growth and metabolic activities are the most im- The spoilage of custard cream has been poorly documented despite portant causes of food spoilage (Sperber, 2009). Microbial con- its extensive use in the formulation of various kinds of desserts parti- taminants can use available substrates for growth and produce a range cularly susceptible to spoilage. French custard cream, composed of of metabolic by-products, such as organic acids and volatile com- milk, sucrose, egg products and minor components such as vanilla or pounds. These activities may result in changes in food appearance and starch is widely used in cream puffs, pastries, cakes or chilled dairy texture, in pH modifications, and/or in the formation of undesirable desserts. Custard cream is potentially a highly nutritive growth medium odours or flavours. Spoilage issues are highly variable depending onthe that can support the growth of bacteria and the production of meta- type of food, extent of processing and storage conditions (Nassos et al., bolites that compromise shelf-life, even at chilled temperatures. 1988). Food spoilage mechanisms and chemical indicators are well- Spoilage of commercial custard creams was reported to be associated described for several foodstuffs such as fish and meat or derived pro- with the presence of bacilli mainly belonging to the B. cereus group, ducts (Dainty, 1996; Nychas et al., 2008). Various volatile compounds staphylococci, lactic acid bacteria, and psychrotrophic Gram-negative have been proposed for early detection of the spoilage risk in various rods (Arakawa et al., 2008). food products (Derevel et al., 1994) and for the characterization of Food quality control is usually achieved using traditional micro- bacteria involved in food spoilage issues (Ercolini et al., 2009; Joffraud biological analyses that target specific bacterial groups that are ∗ Corresponding author. Mixscience, Agrocampus Ouest, INRA, F-35042, Rennes, France. E-mail address: [email protected] (C. Techer). https://doi.org/10.1016/j.fm.2019.103317 Received 21 February 2019; Received in revised form 26 July 2019; Accepted 1 September 2019 Available online 05 September 2019 0740-0020/ © 2019 Elsevier Ltd. All rights reserved. C. Techer, et al. Food Microbiology 86 (2020) 103317 detected using selective agars. However, this approach does not always Frozen isolates were thawed and twice propagated in BHI-YE at provide sufficiently informative data due to ambiguity associated with 30 °C for 24 h under aerobic conditions, without agitation. Each in- specific microbiological methods and a lack of correlation with spoilage oculum was diluted in TS in order to reach an initial population of 3 log potential. More robust, sensitive approaches are thus required for a CFU/mL in 5 mL sterile commercial custard cream in 15 mL Falcon better control of spoilage phenomena. Moreover, it may be preferable tubes (Sarstedt, Nümbreccht, Germany). Initial concentrations were to target metabolites associated with spoilage events rather than the measured using a plate-counting micro-method (Baron et al., 2006) on microorganisms themselves. BHI-YE agar, which was incubated for 24 h at 30 °C. The inoculated The purpose of the present study was to fully characterize the custard cream samples were placed in an incubator at 10 °C and held for bacterial spoilage association and relevant metabolites in spoiling 21 days under aerobic conditions, without agitation. The experiments custard cream to identify relevant spoilage markers. This work re- were carried out at chilled temperature in order to get closer re- presents a first step in the development of efficient detection andpre- frigeration conditions as specified in the French standard for shelf-life vention methods to support industry efforts to reduce spoilage events in validation of perishable and refrigerated food (Afnor NF V 01–003, chilled dessert manufacturing and distribution. 2010) and to mimic the conditions of the ageing tests usually carried out in the chilled industry (Techer et al., 2015). 2. Material and methods Microbiological, sensory, and biochemical analyses were performed at the end of the storage period. Total aerobic population was measured 2.1. Origin of spoiled custard cream based desserts using the plate-counting micro-method described above. A total of 64 spoiled custard cream-based desserts from two man- 2.5. Sensory analyses ufacturers referred to as A (n = 36) and B (n = 28) were analysed. The desserts were produced from May 2012 to August 2013, and were Sensory analyses were performed by an untrained panel of 4 in- considered spoiled after “accelerated shelf-life tests” performed by in- dividuals who were tasked with the detection of changes in odour or cubation of samples at 30 °C for 24 h and 10 °C for 21 d. The criteria appearance of custard cream using a free vocabulary. Main defects used by the manufacturers to qualify desserts as spoiled were based on evaluated by judges were odours, which were classified into three pH modification and/or sensorial changes (mainly odours). Whena groups (acceptable odours, off-odours and yogurt-type odours), and the spoilage event was detected, the dessert was kept refrigerated (< 4 °C) presence (e.g. thickening, phase shift) or absence of visual defects ex- and was transported to our laboratory at refrigeration temperature pressed as (+) or (−). (< 4 °C) for analysis within two days of reception. 2.6. pH measurements 2.2. Measurement of total aerobic microbial populations pH was measured before and after custard cream inoculation and Total mesophilic aerobic populations were measured by pour plate incubation (pH meter
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