Effect of the Reduction of Ochratoxin a and Free Fatty Acids Production in Dry Fermented Cocoa Beans Using Bacillus sp. on the Sensory Quality of Chocolate Produced Thereof | Chapter 1 | Current Perspectives in Agriculture and Food Science Vol. 7

Ochratoxin A (OTA) is a mycotoxin causing serious adverse human and animals health effects while free fatty acids (FFA) are carboxilic acids from hydrolysis of cocoa butter due to the lipolytic filamentous fungi activity. During the primary post-harvest processing notably the storage, the quality of dry fermented cocoa beans is currently altered due to the elevated production of both FFA and OTA ascribed to the growth of both extracellular lipase and mycotoxins producing filamentous fungi species regarding. This study investigated the ability of two Bacillus species to reduce both the OTA and FFA production in dry fermented cocoa beans by inhibiting the growth of producing filamentous fungi strains. Cocoa beans were extracted from cocoa pods of the cultivar ripe pods Amelonado, Ivorian first generation of hybrids (Forastero) and open pollinated progenies harvested at farmer plantations in October 2020. Fourteen Bacillus species isolated from both fermented African locust seeds (9) and cocoa beans (5) were identified using molecular tools. Antifungal activity of these Bacillus species against the growth of OTA producing Aspergillus carbonarius strain was highlighted. Suspensions of each antifungal Bacillus strain cells adjusted to 108 CFU.mL-1 were individually inoculated to 80 kg of fresh cocoa beans extracted from cocoa pods stored during 0, 4, 7 and 10 days. Cocoa beans fermentation was processed with two turnings at 48 and 96 hours of the process for 6 days. Fermented cocoa beans were sun-dried until to 7-8% moisture. FFA and OTA of cocoa beans were quantified by Association of Official Analytical Chemists (AOAC) and high-performance liquid chromatography with fluorescence detection (HPLC-FD) methods respectively. Main results indicated that Bacillus halotolerans and Bacillus thuringiensis strain ATCC 10792 were greater antifungic than Bacillus paramycoides, Bacillus subtilis Subsp subtilis, Bacillus subtilis, Bacillus albus, Bacillus thuringiensis strain IAM 12077. Their inoculation to cocoa beans during spontaneously fermentation totally removed OTA and strongly reduced FFA production. The results of this study indicate that various Bacillus species could be applied as promising biological agents to improve the microbiological quality, to remove OTA from dry fermented cocoa beans and to reduce their FFA content without influencing the organoleptic attributes of the chocolate produced thereof.

Author(s) Details:

Kadjo Adobi Christian,
Laboratoire de Biotechnologie et Microbiologie des aliments, UFR Sciences et Technologies des Aliments, Université Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Houphouet Kouakou Richard,
Laboratoire de Biotechnologie et Microbiologie des aliments, UFR Sciences et Technologies des Aliments, Université Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Kone Koumba Maï,
Département de Formation et de Recherche en Génie Chimique et Agroalimentaire (DFR-GCAA) Institut National Polytechnique Félix HOUPHOUËT-BOIGNY. Bp 1093, Yamoussoukro, Côte d’Ivoire.

Fontana Angélique,
CIRAD, UMR Qualisud, TA B 96/16, 75 Av JF Breton, 34398 Montpellier Cedex 5, France and Qualisud, Univ. Montpellier, Avignon, CIRAD, Institut Agro, La Réunion University, Montpellier, France.

Durand Noël,
CIRAD, UMR Qualisud, TA B 96/16, 75 Av JF Breton, 34398 Montpellier Cedex 5, France and Qualisud, Univ. Montpellier, Avignon, CIRAD, Institut Agro, La Réunion University, Montpellier, France.

Montet Didier,
CIRAD, UMR Qualisud, TA B 96/16, 75 Av JF Breton, 34398 Montpellier Cedex 5, France and Qualisud, Univ. Montpellier, Avignon, CIRAD, Institut Agro, La Réunion University, Montpellier, France.

Guehi Tagro Simplice,
Laboratoire de Biotechnologie et Microbiologie des aliments, UFR Sciences et Technologies des Aliments, Université Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Please see the link here: https://stm.bookpi.org/CPAFS-V7/article/view/13399

Effect of the Improvement of Cocoa Raw Material Quality by Application of Anti-Fungal Bacillus Strains during Fermentation on the Chocolate Sensory Attributes | Chapter 8 | Research Advances and Challenges in Agricultural Sciences Vol. 2

This chapter focuses on evaluating the effect of application of anti-fungal Bacillus strains on cocoa beans fermentation. Contamination with filamentous fungi reduces the quality of cocoa beans and poses a health risk for consumers due to the potential accumulation of mycotoxins. This study involved in developing antifungal Bacillus strains culture to enhance the process of cocoa beans.

Seven strains of the genus Bacillus, namely Bacillus mycoides, B. protéolyticus, B. albus, B. subtilis, B. paramycoides, B. toyonensis and B. thuringiensis strain ATCC 10792 were used in this study. The antagonistic bacteria of the genus Bacillus were obtained from spontaneous cocoa beans fermentation.

Solid culture media inoculated with bacterial and fungal isolates were cultivated under aerobic conditions at 30ºC for 5 days. All the strain have strong antifungal activity and the cocoa beans in Cote d'Ivoire were inoculated into 180-kg boxes for fermentation in three time-independent replications, each with a spontaneous control fermentation. This chapter revealed that the B. thuringiensis strain ATCC 10792 did not affected the fermentation process and cocoa bean quality. The anti-fungal in vivo assays revealed that the B. thuringiensis strain ATCC 10792 completely inhibited growth of all fungi on the surface of cocoa beans. The result demonstrated that there are no appreciable variations in the sensory assessment of chocolates forall treatments. B. thuringiensis strain ATCC 10792 did not affect the aroma, texture, or appearance of the chocolate, as evidenced by the excellent sensory quality of the control and chocolates made from the inoculated cocoa beans.

B. thuringiensis strain ATCC 10792 isolated from cocoa beans showed the highest inhibitory activity against Aspergillus carbonarius voucher IHEM 661 and contributed in a significant suppress fungal growth when it was cultured in combination with cocoa beans contaminated by molds. About sensory evaluation of chocolates made from the inoculated cocoa beans with B. thuringiensis strain ATCC 10792 and non-inoculated control, consumers did not report a significant preference for either chocolate (p < 0.5). Due to an acceptable sensory property, it can be concluded that B. thuringiensis strain ATCC 10792 could be successfully used in the production of dark chocolate. This observation allows thinking that B. thuringiensis strain ATCC 10792 cultures should be investigated in further studies for an effective biological control of mycotoxins during the traditional processing of cocoa.

Author(s) Details:

Adobi Christian Kadjo,
Laboratoire de Biotechnologie et Microbiologie Alimentaire, Unité de Formation et de Rercherche des Sciences et Technology des Aliments, Université Nangui ABROGOUA, 02 Bp 801 Abidjan 02,Côte d’Ivoire.

Guézéré Corinne Beugre,
Laboratoire de Biotechnologie et Microbiologie Alimentaire, Unité de Formation et de Rercherche des Sciences et Technology des Aliments, Université Nangui ABROGOUA, 02 Bp 801 Abidjan 02,Côte d’Ivoire.

Koumba Maï Kone,
Département de Formation et de Recherche en Génie Chimique et Agro-alimentaire (DFR-GCAA) Institut National Polytechnique Félix HOUPHOUËT-BOIGNY. Bp 1093, Yamoussoukro, Côte d’Ivoire.

Kra Brou Didier Kedjebo,
Laboratoire de Biotechnologie et Microbiologie Alimentaire, Unité de Formation et de Rercherche des Sciences et Technology des Aliments, Université Nangui ABROGOUA, 02 Bp 801 Abidjan 02,Côte d’Ivoire.

Pauline Mounjouenpou,
Laboratoire de Technologie Alimentaire, IRAD, BP 2067 Yaoundé, Cameroon.

Noël Durand,
CIRAD, UMR Qualisud, TA B 96/16, 75 Av JF Breton, 34398 Montpellier Cedex 5, France and Qualisud, Univ. Montpellier, Avignon, CIRAD, Institut Agro, La Réunion University, Montpellier, France.

Angélique Fontana,
CIRAD, UMR Qualisud, TA B 96/16, 75 Av JF Breton, 34398 Montpellier Cedex 5, France and Qualisud, Univ. Montpellier, Avignon, CIRAD, Institut Agro, La Réunion University, Montpellier, France.

Tagro Simplice Guehi,
Laboratoire de Biotechnologie et Microbiologie Alimentaire, Unité de Formation et de Rercherche des Sciences et Technology des Aliments, Université Nangui ABROGOUA, 02 Bp 801 Abidjan 02,Côte d’Ivoire.

Please see the link here: https://stm.bookpi.org/RACAS-V2/article/view/13055

Link between the Lipolytic Fungi Contamination and the Production of Free Fatty Acids in Dry Fermented Cocoa Beans | Chapter 5 | Research Advances in Microbiology and Biotechnology Vol. 9

Background: Cocoa butter is manufactured as the main by-product from dry fermented cocoa beans. The biochemical composition of this cocoa by-product schows triacylglycerols, di- and mono-glycerides, phospholipids and glycolipids and unsaponifiable matter. During primary post-harvest processing, cocoa beans harbor lipase producing mold species contamination could trigger the accumulation of FFA. High free fatty acids (FFA) content of dry fermented cocoa beans currentely alter physical properties of both fat fraction and chocolate.

Aims: This study investigated the improvement of the quality of raw cocoa material sourced from Côte d’Ivoire regarding its FFA concentration.

Methods: Fifty dry fermented cocoa beans samples were randomly collected during October 2018 from Aniassué, Akoupé and Yakassé-Attobrou, 3 big cocoa producing regions located in Southeast of Côte d'Ivoire. Dry fermented cocoa beans were sampled on-farm level in the same cocoa producing regions. Contaminating filamentous fungi strains were isolated using Sabouraud chloramphenicol agar incubated at 25°C for 7 days. Morphological identification of fungi isolates was preformed from younger culture. Biochemical study for the determination of lipolytic properties of fungi was carried out using Cazpeck-Dox Agar medium enriched with olive oil as carbon source and rhodamine B as indicator of extracellular lipase production. Extraction, PCR amplification using a specific transcribed spacer primer (ITS4/ITS5) and sequencing of DNA from fungal species were performed for molecular identification.

Results: Approximately 154 fungal isolates belonging to 8 genera were found in tested cocoa beans sourced from Côte d’Ivoire. Only Rhizopus oryzae, Absidia corymbifera, Mucor heimalis, Aspergillus niger, Aspergillus tubingensis, Aspergillus aculeatus, Aspergillus flavus, Aspergillus tamari, Aspergillus fumigatus, Nigrospora sphaerica, Curvularia geniculate and Penicillium chrysogenum species exhibited greater extracellular lipase producing ability. Absidia corymbifera, Aspergillus niger, Aspergillus fumigatus and Rhizopus orysae were mostly frequent and predominant species contaminated Ivorian cocoa beans. Dry fermented cocoa beans inoculated with Rhizopus oryzae, Absidia corymbifera or Aspergillus niger recorded higher FFA content above 1.75%.

Conclusion: Reducing of the growth of the fungi species growth during cocoa primary post-harvest processing could sustainably produce raw cocoa material with less FFA content. This study clearly indicated that the control of lipolytic fungi contamination in dry fermented cocoa beans could reduce FFA content and sustainably improve the quality of cocoa beans sourced from Côte d’Ivoire, cocoa butter and chocolate produced thereof.

Author(s) Details:

Tape Joëlle Stéphanie,
Laboratoire de Biotechnologie et Microbiologie des Aliments, UFR Sciences et Technologies des Aliments, Université Nangui ABROGOUA, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Houphouët Kouakou Richard,
Laboratoire de Biotechnologie et Microbiologie des Aliments, UFR Sciences et Technologies des Aliments, Université Nangui ABROGOUA, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Kone Koumba Maï,
Institut National Polytechnique Félix Houphouët-Boigny, Bp 1093, Yamoussoukro, Côte d’Ivoire.

Mounjouenpou Pauline,
Laboratoire de Technologie Alimentaire, IRAD. BP 2067 Yaoundé, Cameron.

Guibert Benjamin,
CIRAD, UMR Qualisud, TA B 96/16, 75 Av JF Breton, 34398 Montpellier Cedex 5, France and Qualisud, Univ. Montpellier, Avignon, CIRAD, Institut Agro, Université de La Réunion, Montpellier, France.

Metayer-Piro Isabelle,
CIRAD, UMR Qualisud, TA B 96/16, 75 Av JF Breton, 34398 Montpellier Cedex 5, France and Qualisud, Univ. Montpellier, Avignon, CIRAD, Institut Agro, Université de La Réunion, Montpellier, France.

Montet Didier,
CIRAD, UMR Qualisud, TA B 96/16, 75 Av JF Breton, 34398 Montpellier Cedex 5, France and Qualisud, Univ. Montpellier, Avignon, CIRAD, Institut Agro, Université de La Réunion, Montpellier, France.

Guehi Tagro Simplice,
Laboratoire de Biotechnologie et Microbiologie des Aliments, UFR Sciences et Technologies des Aliments, Université Nangui ABROGOUA, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Please see the link here: https://stm.bookpi.org/RAMB/article/view/12956

High Free Fatty Acids (FFA) Content in Fermented and dry Cocoa Beans with Various Cocoa Producing Geographical Location of Côte d’Ivoire in the Climate Change Context | Chapter 4 | Emerging Issues in Agricultural Sciences Vol. 7

 Aims: Cocoa beans garnered from Côte d’Ivoire faces more and more to the high FFA chapters. Yet, generation of FFA levels degrades advantageous rheological properties of cocoa fat and chocolate. This study examined the factors that potentially advance the increase of high FFA elements of larger object of Ivorian cocoa beans.Methodology: Samples: 240 cocoa beans collections were sampled from cocoa growers of some big dark producing domains for their FFA contents analysis. Also, influence of basic post-harvest processing in the way that pod’s sanitary status, pod hole delay, fermentation event and dry cocoa beans storage under miscellaneous relative humidity (RH) categorized 80-90% on FFA’s formation of Ivorian cocoa beans was examined. Data interpretations were acted according to the Fisher’s test.Results: Main results showed that dry dark beans recorded FFA content variable from 0.99 to 14% in function of the geographical origin. Beside, 53% of proven cocoa grain samples exhibited FFA content up to 1.75%. Poor clean status of pods and 7-days simmered cocoa beans recorded FFA content above 1.75%. Dry burnished color beans stored at RH above 85% advanced more the FFA’s formation (14%) than those stored under lower RH principles. 53% of tested burnished color beans samples presented FFA contents until 1.75%. Controlled primary post-harvest deal with highlighted that poor clean status of burnished color pods promotes the formation of FFA. Interacting weak sanitary rank of pods with long fermentation exasperated occurrence of FFA aggregation above 1.75%. However, the fermentation duration of dark beans from healthy pods had no effect on the changes of FFA content. The FFA content of beans increases better with both the event of cocoa effervescence and storage of dry cocoa beans at RH above 85% until FFA content above 1.75%.Conclusion: So, in order to produce ripened and dry cocoa beans containing depressed and suitable FFA divisions in the context of climate change, alleviation strategies need to mix good crop conditions regarding postharvest handle and to improve advantageous ones.

Author(s) Details:

Kouakou Richard Houphouet,
Laboratoire de Biotechnologie et Microbiologie des Aliments, UFR des Sciences et Technologies des Aliments, Université Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Joëlle Stéphanie Tape,
Laboratoire de Biotechnologie et Microbiologie des Aliments, UFR des Sciences et Technologies des Aliments, Université Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Kouadio Ignace Kouassi,
Laboratoire de Biotechnologie et Valorisation des Agro-Ressources, UFR des Sciences Biologiques, Université Péléforo Gon Coulibaly, BP 1328 Korhogo, Côte d’Ivoire.

Konan Mathurin Yao,
UFR des Sciences de la Nature, Université Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Kra Brou Dider Kedjebo,
Laboratoire de Biotechnologie et Microbiologie des Aliments, UFR des Sciences et Technologies des Aliments, Université Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Pauline Mounjouenpou,
Laboratoire de Technologie Alimentaire, IRAD, BP 2067 Yaoundé, Cameroon.

Noêl Durand,
CIRAD, UMR Qualisud, TA B 96/16, 75 Av JF Breton, 34398 Montpellier Cedex 5, France and Qualisud, Univ Montpellier, CIRAD, Université d’Avignon, Université de la Réunion, Montpellier SupAgro, Montpellier, France.

Didier Montet,
CIRAD, UMR Qualisud, TA B 96/16, 75 Av JF Breton, 34398 Montpellier Cedex 5, France and Qualisud, Univ Montpellier, CIRAD, Université d’Avignon, Université de la Réunion, Montpellier SupAgro, Montpellier, France.

Tagro Simplice Guehi,
Laboratoire de Biotechnologie et Microbiologie des Aliments, UFR des Sciences et Technologies des Aliments, Université Nangui Abrogoua, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Please see the link here: https://stm.bookpi.org/EIAS-V7/article/view/11849