Cocoa Pulp Research Articles

Isolation and identification of microbial species found in cocoa fermentation as microbial starter culture candidates for cocoa bean fermentation in Colombia.

Microbial activity involved in the cocoa beans fermentation process is essential to maintain and improve the organoleptic and nutritional qualities of chocolate; therefore, the aim of this investigation was to search and select microbial isolates with the potential to improve the quality of cocoa beans. Fermentation experimentswere conducted on farms located in Maceo (Antioquia), San Vicente de Chucurí (Santander), and Rivera and Algeciras (Huila), Colombia. Yeast, lactic acid bacteria (LAB), acetic acid bacteria (AAB), and mesophilic aerobic microorganisms were obtained from different fermentation batches. The growth of these microorganismswas tested in six treatments as follows: 50% cocoa pulp agar (CPA), high concentrations of glucose (10%), ethanol (5%), and acetic acid (7%), an acidic pH of 3.0, and a high temperature of 50oC for 24 h. The isolates with the highest growth were identified by 18S and 16S rRNA gene analysis, revealing a high diversity ofspecies associated with cocoa fermentation, including eight species of yeasts (Debaryomyces hansenii, Meyerozyma guillermondii, Wickerhanomyces anomalus, Pichia guillermondii, Pichia kudriavzevii, Trichosporon asahii, Candida parapsilosis, and Pichia manshurica), six species of LAB (Pediococcus acidilactici, Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus farraginis, Lactobacillus rhamnosus, and Leuconostoc mesenteroides), four species of AAB (Gluconobacter japonicus, Acetobacter tropicalis, Acetobacter pasteurianus, and Acetobacter malorum/tropicalis), and three species of Bacillus spp. (Bacillusaryabhattai /megaterium, Bacillus subtilis, and Bacillus coagulans). In general, microbial populations increased in cocoa batches after 12 h of fermentation and decreased after 84-96 h. All the yeast isolates grew in 10% glucose and CPA, 85.7% in 5% ethanol, and 95% at a pH of 3.0. All the yeast isolates were affectedby 7% acetic acid and incubation at 50oC for 24 h. Eighty-five percent of the LAB grew in 10% glucose, 100% in 5% ethanol, 42.8% in CPA, 64% at a pH of 3.0, and 35.7% grew after being exposed to 50oC for 24 h; all were affected by 7% acetic acid. As for the AAB, 100% grew in 10% glucose, 71% in 7% ethanol, 100% grew in CPA, in 7% acetic acid, and at a pH of 3.0, while 100% were affected by incubation at 50oC. Three yeast isolates, W. anomalus, D. hansenii and M. guillermondii, three LAB isolates, P. acidilactici, L. brevis, and L. plantarum, and three AAB isolates, A. tropicalis, A. pasteurianus and G. japonicus, were selected as promising strains to be used in a microbial starter culture for cocoa bean fermentation to improve the organoleptic quality of cocoa.

Co-culturing fructophilic lactic acid bacteria and yeast enhanced sugar metabolism and aroma formation during cocoa beans fermentation

Glucose and fructose are the main fermentable sugars in cocoa pulp. During fermentation, glucose is consumed within 48–72 h and fructose only after 120 h, mainly associated with the preferential use of glucose by microorganisms. In the first stage of this study, the complete genome sequence of a lactic acid bacterium with high fructose consumption capacity (Lactobacillus plantarum LPBF35) was reported. The notable genomic features of L. plantarum LPBF35 were the presence of alcohol/acetaldehyde dehydrogenase gene and improved PTS system, confirming its classification as a “facultatively” fructophilic bacterium. Subsequently, this bacterium was introduced into cocoa fermentation process in single and mixed cultures with Pediococcus acidilactici LPBF66 or Pichia fermentans YC5.2. Community composition by Illumina-based amplicon sequencing and viable counts indicated suppression of wild microflora in all treatments. At the beginning of the fermentation processes, cocoa pulp consisted of approximately 73.09 mg/g glucose and 73.64 mg/g fructose. The L. plantarum LPBF35 + P. fermentans YC5.2 process showed the lowest levels of residual sugars after 72 h of fermentation (7.89 and 4.23 mg/g, for fructose and glucose, respectively), followed by L. plantarum LPBF35 + Ped. acidilactici LPBF66 (8.85 and 6.42 mg/g, for fructose and glucose, respectively), single L. plantarum LPBF35 treatment (4.15 and 10.15 mg/g, for fructose and glucose, respectively), and spontaneous process (22.25 and 14.60 mg/g, for fructose and glucose, respectively). The positive interaction between L. plantarum LPBF35 and P. fermentans YC5.2 resulted in an improved formation of primary (ethanol, lactic acid, and acetic acid) and secondary (2-methyl-1-butanol, isoamyl acetate, and ethyl acetate) metabolites during fermentation. The primary metabolites accumulated significantly in cocoa beans fermented by P. fermentans YC5.2 + L. plantarum LPBF35, causing important reactions of color development and key flavor molecules formation. The results of this study suggest that fructophilic lactic acid bacteria and yeast is a microbial consortium that could improve sugar metabolism and aroma formation during cocoa beans fermentation.

Development of a model for the alcoholic fermentation of cocoa beans by a Saccharomyces cerevisiae strain

The aromatic quality of chocolate requires the use of cocoa with high aromatic potential, this being acquired during the fermentation of cocoa beans. Traditional fermentation is still often carried out on a small scale with wild strains of yeasts and acetic bacteria and under poorly controlled conditions leading to cocoa quality ranging from best to worst. This study is the first part of a project aiming to control quality of cocoa to produce high aromatic quality chocolate by using a mixed starter of selected strains of yeast and acetic bacteria and by controlling the conditions of fermentation. To achieve this objective, a mathematical model of the alcoholic fermentation of cocoa beans has been developed. The growth, glucose consumption and ethanol production of Saccharomyces cerevisiae LM strain in synthetic broth were modeled for the most important intrinsic (pH, glucose, ethanol, free nitrogen and oxygen levels) and extrinsic (temperature, oxygen level) fermentation parameters. The model was developed by combining the effects of individual conditions in a multiplicative way using the gamma concept. The model was validated in liquid synthetic medium at two different inoculation levels 104 and 106 CFU/mL with an increase in temperature that recorded during spontaneous fermentations. The model clearly shows that the level of inoculation and the speed of the increase in temperature clearly drive yeast growth, while other factors including pH and ethanol, free nitrogen and oxygen levels have no significant impact on yeast growth.

Potential Applicability of Cocoa Pulp (Theobroma cacao L) as an Adjunct for Beer Production.

The aim of this study was to evaluate the application of cocoa pulp as an adjunct for malt in beer production. The cocoa pulp was analyzed for humidity, proteins, lipids, sugars, total soluble solids, organic acids, and minerals. A study was carried out to reduce the cocoa pulp viscosity by enzymatic depectinization, making its use viable in beer production. The cocoa pulp showed relevant quantities of compounds important in fermentation, such as sugars, acids, and minerals. In fermentation using the adjunct, the proportions of pulp used were 10, 30, and 49%. A significant difference was found between the adjunct and all-malt worts. The 30% cocoa pulp concentration as an adjunct for malt in the fermentation medium contributed the most to the fermentative performance of the yeasts at both 15 and 22°C based on the consumption of apparent extract (°Plato), ethanol production, and cellular growth.

Investigations into the Natural Occurrence of 1-Phenylethyl Acetate (Styrallyl Acetate).

So far, the occurrence of the flavor constituent 1-phenylethyl acetate in a natural source has not been unambiguously confirmed. The present work provides the detailed identification of 1-phenylethyl acetate from clove (Syzygium aromaticum (L.) Merr. & L.M. Perry) buds. In addition, headspace solid-phase microextraction-gas chromatography/mass spectrometry (GC/MS) analysis revealed further occurrence of 1-phenylethyl acetate in cocoa pulp and grape hyacinth flowers. A total of 15.2 g of essential oil was recovered from 7.2 kg of clove buds by simultaneous distillation-extraction followed by vacuum distillation. The distillate obtained was fractionated by silica column chromatography, whereby a significant enrichment of 1-phenylethyl acetate was achieved. The fraction containing the target analyte was further purified by preparative high-performance liquid chromatography, resulting in a final purity of ∼93.0%, yielding a total of 1 to 2 mg of 1-phenylethyl acetate. Identification of the isolated compound was achieved by GC/MS, infrared spectroscopy, enantioselective GC, isotope ratio MS, and nuclear magnetic resonance spectroscopy. Enantioselective GC/MS analysis revealed an enantiomeric excess of 60% (1S)-(-)-1-phenylethyl acetate in the isolate. The δ13CV-PDB value of -32.5 ± 0.5‰ was in accordance with that of C3-plants and other constituents found in genuine clove extracts.

Exploring the contribution of fructophilic lactic acid bacteria to cocoa beans fermentation: Isolation, selection and evaluation

Fructophilic lactic acid bacteria (FLAB) are a recently discovered group whose main characteristic is to prefer D-fructose over D-glucose. In this study, laboratory cocoa beans fermentation was analyzed by Illumina-based amplicon sequencing, indicating the presence of potential FLAB of the genera Fructobacillus and Lactobacillus. Eighty efficient fructose-fermenting isolates, obtained from fermenting cocoa pulp beans mass, were identified by 16S rRNA gene sequencing as Pediococcus acidilactici (n = 52), Lactobacillus plantarum (n = 10), Pediococcus pentosaceus (n = 10), Bacillus subtilis (n = 4), and Leuconostoc pseudomesenteroides (n = 4). The growth characteristics of all the 10 L. plantarum strains classified them as “facultatively” fructophilic bacteria, i.e., they grew on glucose without an external electron acceptor but the growth on fructose was faster. Among them, L. plantarum LPBF 35 was characterized by producing a range of aroma-impacting compounds (acetaldehyde, ethyl acetate, nonanal, and octanoic acid), being introduced into a cocoa fermentation process. Although the process started with approximately equal amounts of glucose and fructose, a concomitant, but faster utilization of fructose, was observed in cocoa fermentation conducted with L. plantarum LPBF 35 (with no residual fructose observed) when compared to control fermentation using a glucophilic strain (8.77 mg/g residual fructose) and a spontaneous process (8.38 mg/g residual fructose). L. plantarum LPBF 35 also showed an ideal profile of organic acid metabolism (citric acid consumption and lactic acid production) associated with cocoa fermentation. These results proved new insights on cocoa microbial activity and brings new perspectives on the use of lactic acid bacteria as starter culture.

Functional role of yeasts, lactic acid bacteria and acetic acid bacteria in cocoa fermentation processes.

Cured cocoa beans are obtained through a post-harvest, batchwise process of fermentation and drying carried out on farms in the equatorial zone. Fermentation of cocoa pulp-bean mass is performed mainly in heaps or boxes. It is made possible by a succession of yeast, lactic acid bacteria (LAB) and acetic acid bacteria (AAB) activities. Yeasts ferment the glucose of the cocoa pulp into ethanol, perform pectinolysis and produce flavour compounds, such as (higher) alcohols, aldehydes, organic acids and esters. LAB ferment the glucose, fructose and citric acid of the cocoa pulp into lactic acid, acetic acid, mannitol and pyruvate, generate a microbiologically stable fermentation environment, provide lactate as carbon source for the indispensable growth of AAB, and contribute to the cocoa and chocolate flavours by the production of sugar alcohols, organic acids, (higher) alcohols and aldehydes. AAB oxidize the ethanol into acetic acid, which penetrates into the bean cotyledons to prevent seed germination. Destruction of the subcellular seed structure in turn initiates enzymatic and non-enzymatic conversions inside the cocoa beans, which provides the necessary colour and flavour precursor molecules (hydrophilic peptides, hydrophobic amino acids and reducing sugars) for later roasting of the cured cocoa beans, the first step of the chocolate-making.

Cocoa Pulp as Alternative Food Matrix for Probiotic Delivery.

For screening probiotic strains with viability and stability in non-dairy foods for health benefits, we revised all patents relating to probiotics in food. Screening of potential probiotics from Brazilian Minas artisanal cheese and verify their survival in frozen Brazilian cocoa pulp. Isolation and identification of the strains. The potential probiotic characterization involved gastric juice and bile resistance, antibiotic and antimicrobial activity, hydrophobicity, autoaggregation, coaggregation and adhesion assay in HT-29 cells. Organoleptic, viability and stability of probiotic strain in frozen cocoa pulp were evaluated. Fourteen strains of Lactobacillus plantarum (9), Weissella paramesenteroides (3), Lactobacillus fermentum (1), and Leuconostoc mesenteroides (1) were obtained. Most of the strains were resistant to simulated gastric acidity and bile salts. Almost all strains were sensitive to the antibiotics tested, except to ciprofloxacin and vancomycin. About 47% of the strains are potential producers of bacteriocins. High hydrophobicity was observed for four strains. Autoaggregation ranged from 8.3-72.6% and the coaggregation capacity from 5.2-60.2%. All of the assessed strains presented more than 90% of adhesion to HT-29 intestinal cells. The percentage of Salmonella inhibition in HT-29 cells ranged from 4.7-31.1%. No changes in color, aroma, and pH were observed in cocoa pulps after storage at -20 °C for 90 days. Wild strains of acid lactic bacteria from cheese proved to be viable and stable in frozen Brazilian cocoa pulp. This work showed a promising application of L. plantarum isolated strains to be used with frozen cocoa pulp matrix in probiotics food industry.

Flow behavior of cocoa pulp powder containing maltodextrin

Abstract This study aimed to evaluate the influence of maltodextrin addition on the flow properties of cocoa pulp powder obtained by spray and freeze drying. Cocoa pulp samples received 15% and 30% (m m-1) maltodextrin DE20. Two drying methods were used, spray and freeze drying. Powder morphology was evaluated through scanning electron microscopy (SEM). Wall friction angle, bulk density and tapped density were determined. Carr index (CI), Hausner ratio (HR) and flow index (FI) were used to evaluate powder flow. The particles powders obtained by spray drying showed rounded shapes, whereas the freeze dried powder showed irregular shapes. Increased maltodextrin concentration in the samples altered the powder particle size by spray drying and powder particle surface by freeze drying. The powder by spray drying of the sample with 30% of maltodextrin showed smallest wall friction angles, 13.4 to 14.9. The powder by freeze drying of the samples with 15% and 30% of maltodextrin showed wall friction angles between 14.0 and 20.6. Regarding flow, the powders by spray drying containing 30% of maltodextrin showed the best CI, HR and FI, 24.88, 1.33 and 4.88, respectively, being considered an acceptable flow. According to CI, HR and FI values, samples with 15% of maltodextrin produced powder classified as difficult flow in both methods applied. The higher maltodextrin concentration in cocoa pulp, the lower agglomeration in the powder by spray drying and the smoother particles surfaces in the powder by freeze drying. The addition of maltodextrin to the samples, for both drying methods, improve the powder flow and decrease the powder cohesion.

Microbial synergy between Pichia kudriazevii YS201 and Bacillus subtilis BS38 improves pulp degradation and aroma production in cocoa pulp simulation medium

Interactions between two major microorganisms from Ivorian cocoa fermentation, namely Bacillus subtilis BS38 and Pichia kudriazevii YS201, were investigated during fermentation in cocoa pulp simulation medium. The strains were mutually inhibitory, with Bacillus being more susceptible to this antagonistic effect than Pichia. However, both strains yielded different pulp-degrading enzymes, namely polygalacturonase (PG) from Pichia and pectate lyase (Pel) from Bacillus, that cooperate to efficiently breakdown pectin and vegetable pulp. The quantification of aromas from microbial cultures using Gas Chromatography-Mass Spectroscopy (GC-MS) coupled with headspace microextration (SPME) method, showed that P. kudriazevii produce mainly alcohols such as ethanol (63.165 g/L), phenylethanol (1.005 g/L), methylbutanol (0.138 g/L) and esters, notably ethyl acetate (0.037 g/L) and isoamyl acetate (0.032 g/L). The volatile fraction produced by Bacillus was dominated by butanediol (5.707 g/L), acetoin (1.933 g/L), phenylethanol (0.035 g/L) and acetic acid (0.034 g/L). In co-culture, Bacillus produced low levels of aroma compounds whereas a moderate decrease in the production of these compounds was observed in the yeasts strain. Thus, the dominant aromas present in the co-culture were mainly those from the yeasts strain; however, a 1.37 fold increase of ethanol production was observed in co-culture indicating a synergy between the strains. This study showed that cooperation between B. subtilis BS38 and P. kudriazevii YS201 leads principally to increasing pulp degradation and ethanol production, known as desirable properties for a well processing of cocoa fermentation.

Physico-chemical Characterization of Blends Based on Milk and Catole Coconut Pulp

Aims: Coconut catole is the fruit of the Syagrus cearenses palm tree, whose pulp is usually eaten in its natural form. The formulation of blends is intended to improve the acceptance of fruit with potential to be exploited. The objective of this work was to evaluate the physico-chemical characteristics of the whole coconut pulp and the blends formulated with the pulp and milk.
 Place and Duration of Study: The coconut catole (Syagrus cearensis) were collected in the surroundings of location 551'30 "S, 35º21'13 "O, in Rio Grande do Norte. The work was carried out from January to February 2019.
 Methodology: The formulations were elaborated by mixing the proportions of 30 and 50% of standardized commercial milk with the whole catole coconut pulp. The products were submitted to the analysis of moisture, water activity (aw), titratable total acidity (TTA), total soluble solids (TSS), TSS/TTA ratio, pH, ascorbic acid, ash, lipids, proteins, color (L*, a*, b*), total sugars, reducers and non-reducers.
 Results: From the results, it was observed that the addition of milk to the pulp resulted in clear samples and that a higher proportion of the milk statistically influenced the results.
 Conclusion: The mixture of couscous coconut pulp and milk gave the product better nutritional characteristics, especially protein content and lipid reduction. The addition of the milk to the cocoa pulp influenced the results statistically, showing that the addition of the milk contributed to enhance the pulp.

Characterization of Cocoa Pulp (Theobroma cacao L) from South Halmahera as an Alternative Feedstock for Bioethanol Production

South Halmahera is one of the cocoa-producing centers in North Maluku, but the pulp of cocoa is less noticed and assumed to be not important in commercial purpose and is largely disposed as agroindustry residue. This study aimed to characterize the cocoa pulp from southern Halmahera for bioethanol production. Survey and observation methods were conducted to obtain bioethanol feedstock from cocoa pulp. Cocoa pulps mucilage is a waste that can be processed into bioethanol products with high economic value. The results of this study indicated that theaverage of 100kg of cocoa beans produced 5-8 litters of mucilage juice residues. The results of this study indicated that cocoa pulp from southern Halmahera had the optimum time for the fermentation process for 12 days. The level of alcohol produced by fermentation was 4.85%, and this level was a low category.

Aktivitas Mikrob dalam Pulp Biji Kakao (Theobroma cacao L.) Selama Fermentasi dengan Penambahan Ragi Tape

<em>Microbial activities in cocoa pulp decomposition is key in cocoa beans fermentation. </em>Ragi tape<em> is widely used </em><em>as a source of microbes (starters). The study aimed to investigate the pattern of changes in the number of microbes, substrate degradation activities and primary metabolites production during fermentation with the addition of ragi tape. The study was conducted at the laboratories within the Faculty of Agriculture Technology and Integrated Laboratory of Experiment and Research, UGM from March to October 2018 . Fermentation experiment used 2 kg of fresh cacao beans stored in a plastic box per batch, its temperature was set daily. The amount of microbes was enumerated using total plate count (TPC), whereas substrate degradation and primary metabolites were analyzed using high performance liquid chromatography (HPLC) and gas chromatography (GC). Temperature, pulp and seed pH, and fermentation index were observed. The results showed that adding </em>ragi tape<em> </em><em>for fermentation generated a maximum amount of yeast, lactic acid bacteria (LAB) and acetic acid bacteria (AAB) of 9.03<span style="text-decoration: underline;">+</span>0.85; 9.05<span style="text-decoration: underline;">+</span>0.17; and 9.15<span style="text-decoration: underline;">+</span>0.89 log cfu/g of cocoa pulp respectively, with substrate degradation in the form of sucrose reduced by 97%, glucose 98.6%, fructose 97%, and citric acid 71% on the third day. Maximum production of primary metabolites of ethanol is 27.84<span style="text-decoration: underline;">+</span>21.85 mg/g of cocoa pulp, lactic acid 4.18<span style="text-decoration: underline;">+</span>3.16 mg/g of cocoa pulp and acetic acid 3.38<span style="text-decoration: underline;">+</span>5.43 mg/g of cocoa pulp. Fermentation with the addition of </em>ragi tape<em> accelerates the process to three days with a fermentation index value of 1.05<span style="text-decoration: underline;">+</span>0.06 and seed pH of 5.97<span style="text-decoration: underline;">+</span>0.20.</em>

Nutrient digestibility of complete feed containing cocoa pulp with different fiber sources for local goat

Cocoa pulp can be used as an alternative to molasses in a complete feed formulation with low-quality agricultural byproducts as the main fiber source. The purpose of this study was to determine the nutrient digestibility of complete feed containing cocoa pulp with different fiber sources for local goats. Twelve local male goats, aged 1-1.5-year-old, were placed in individual metabolism cage (1 goat/cage). Then each goat was randomly assigned to get one of three experimental diets according to a completely randomized block design consisting of three treatment diets and four blocks as replications. Initial body weight of the animal was used as the base for grouping. The experimental ration was P1 = complete feed with rice straw as the fiber source, P2 = complete feed with corn cobs as the fiber source, and P3 = complete feed with soybean straw as the fiber source. The three types of complete feed contained 10% of cocoa pulp. The analysis of variance showed that the treatment did not give a significant effect (P>0.05) on the nutrient digestibility of complete feed. In conclusion, the three types of fiber sources, i.e., rice straw, corn cobs, and soybean straw can be used in complete feed formulation containing 10% of cocoa without deteriorating effects on nutrient digestion of complete feed for the local goat.

Performance of local goat fed on complete feed containing cocoa pulp with different fiber sources

Due to the limitation of the forage, especially during the dry season, utilization of complete feed formulated from such agricultural by-product and agro-industry as rice straw, corn cobs, soybean straw, and cocoa pulp is an alternative to solve the problem. The aim of this study was to determine the performance of local goats fed on complete feed containing 10% of cocoa pulp with different fiber sources. Sixteen local male goats, with an average initial body weight of 17.7±4.6 kg, were randomly assigned to one of four treatment diets according to completely randomized block design consisted of four treatments and four blocks as replication. The treatment was T1: Complete feed with rice straw as the fiber source, T2: complete feed with corn cobs as the fiber source, T3: complete feed with soybean straw as the fiber source, and T4: mini elephant grass + concentrate. Analysis of variance indicated that treatment affected DMI (P<0.001) and %DMI/BW (P=0.049) but the treatment did not affect ADG (P = 0.366) and FC (P = 0.468). In conclusion, the use of 10% of pulp cocoa pulp in the formulation of complete feed containing different sources of fiber, i.e., either rice straw corn cobs, or soybean straw is satisfactory in supporting the performance of the local goats.

The comparison of goat rumen fermentation given the cocoa pulp-based complete feed and corn cob as fiber source

Cocoa pulp is one of the agricultural waste which its use is still less, cocoa pulp which owns a high glucose content can be utilized by cattle into energy, while corn cob is rich of rough fiber. This research aims to evaluate the condition of rumen fermentationin goats which has been given complete cocoa pulp-based feed with corn cob fiber sources. This research was conducted with completely randomized model programs, using goats to the age of 1 - 1.5 years as many as eight tails in place of the individual metabolic cages for 2 months. Treatment of complete feed is feed kompit P1 cocoa pulp with corn cob fiber resources while P2 is added to the concentrate on elephant grass, though the data using Test T. rumen fluid sampling is performed twice which are before cattle get fed 0 hours and 4 hours after get fed. From the results of research, found that the overall of the value of 6.97 pH 0 hours P1 and P2 6.92 hiccup on for 4 hours after the news feed P1 and P2 7.12 7.40. The overall value of NH3 0 hours P1 P2 30.20 mg to 24.55 mg and 65.77 mg 4 hours P1, P2 54.72 and for the average value of VFA 0 hours P1 P2 13.2 mm and 16.4 mm for 4 hours 17.3 mm P1, P2 19.8 mm. Conclusions Of analysis results in use of variance pulp-based complete feed use cocoa in comparison with the concentrates are added to the elephant grass showed the effect was not significant (P> 0.05) pH values, NH3 and VFA close relation to a source of protein feed used and easy or not The protein in degradation in the rumen.

Estimation of microbial protein supply on goat fed on complete feed containing cocoa pulp with different fiber sources

Supplementation both energy and protein of goats can support optimal microbial growth so that utilization of fibrous feed can be optimized. The aim of this research is to observe the effect of goats which fed complete feed containing 10% cocoa pulp and various fiber sources toward estimation of microbial protein supply (MPS) and efficiency of microbial protein supply (EMPS). Twelve local male goats, aged 1-1.5 years old, were randomly assigned to one of four treatments according to completely randomized block design consisted of 3 treatments and 4 blocks as replications. The treatments were P1: Complete feed containing 50% rice straw, P2: Complete feed containing 50% corn cobs and P3: Complete feed containing 50% soybean straw. The results showed the estimated of MPS was P1 = 16.25 g/d P2 = 14.05 g/d, and P3 = 24.57 g/d, respectively. The EMPS per kg organic matter fermented in rumen was P1 = 86.5 g/kg BOFR, P2 = 73.0 g/kg BOFR, and P3 = 150.25 g/kg BOFR. Analysis of variances indicated that treatment did not affect (P>0.05) MPS and EMPS. In conclusion, based on estimated MPS and EMPS, the three types of fiber could be used as the main fiber sources in formulating complete feed containing cocoa pulp for local goats.

Screening of lactic acid bacteria and yeast strains to select adapted anti-fungal co-cultures for cocoa bean fermentation

Contamination with filamentous fungi during cocoa bean fermentation and drying reduces the quality of cocoa beans and poses a health risk for consumers due to the potential accumulation of mycotoxins. The aim of this study was to develop anti-fungal lactic acid bacteria (LAB)-yeast co-cultures by selecting anti-fungal strains best adapted to the cocoa bean fermentation process from 362 LAB and 384 yeast strains isolated from cocoa bean post-harvest processes. The applied multiphasic screening approach included anti-fungal activity tests in vitro and in vivo and assessment of the carbon metabolism and stress tolerance of the anti-fungal strains in a cocoa pulp simulation medium. The anti-fungal strains, Lactobacillus fermentum M017, Lb. fermentum 223, Hanseniaspora opuntiae H17, and Saccharomyces cerevisiae H290, were selected based on their high fungal growth inhibition capacity and their well-adapted metabolism. Up to seven filamentous fungal strains of the genera Aspergillus, Penicillium, and Gibberella were inhibited on average by 63 and 75% of the maximal inhibition zone by M017 and 223, respectively, and by 25 and 31% by the strains H17 and H290, respectively. Both Lb. fermentum strains converted the medium's glucose, fructose, and citric acid into 20.4–23.0 g/l of mannitol, 3.9–6.2 g/l acetic acid, and 8.6–10.3 g/l lactic acid, whereas the two yeast strains metabolized glucose and fructose to produce 7.4–18.4 g/l of ethanol. The Lb. fermentum strains were further characterized as particularly tolerant towards ethanol, acetic acid, and heat stress and both yeast strains tolerated high amounts of ethanol and lactic acid in the medium. Finally, the anti-fungal in vivo assays revealed that the two Lb. fermentum strains completely inhibited growth of the citrinin-producing strain, P. citrinum S005, and the potentially fumonisin-producing strain, G. moniliformis S003, on the surface of cocoa beans. Furthermore, growth of the aflatoxin-producer A. flavus S075 was inhibited after 10-14 days by all four selected anti-fungal strains, i.e. Lb. fermentum M017, Lb. fermentum 223, H. opuntiae H17, and Sacc. cerevisiae H290, at 51–95% when applied as single cultures and at 100% when the strains were combined into four co-cultures, each composed of a Lb. fermentum and one of the two yeast strains. As a conclusion, these four LAB-yeast co-cultures are recommended for future applications to limit the growth of filamentous fungi and the concomitant mycotoxin production during the fermentation of cocoa beans.

PEMANFAATAN LIMBAH PULP KAKAO MENJADI NATA DE CACAO

This community service aims to empower the unproductive community in Wonosari Village through the utilization of cocoa pulp waste into Nata De Cacao. This method of community service is divided into three stages: preparation stage, implementation stage, and final stage. The result of community service is the community has handled the utilization of cocoa bean pulp waste into Nata De Cacao. The society's understanding of the making of Nata De Cacao is quite high. But after we do the evaluation, people still need guidance in making Nata De Cacao. People love the Nata De Cacao that we make when conducting training on the utilization of cacao bean pulp waste. After we deliver the training, the community is well received and their thinking starts to open with the utilization of cacao pulp waste into Nata De Cacao can reduce environmental pollution and can be used as an opportunity for entrepreneurship. Therefore, with the utilization of this pulp waste can change the behavior of unproductive communities to be productive.

Initial assessment on the use of cocoa pulp in complete feed formulation: in vitro dry matter and organic matter digestibility

Cocoa pulp is a by-product from cocoa industry which is produced in large quantity, but very limited study has been carried out in utilizing it as energy source in animal feed. The purpose of this study was to assess the in vitro dry matter (IVDMD) and in vitro organic matter digestibility (IVOMD) of complete feed containing different levels of cocoa pulp. The experiment was carried out according to completely randomised design consisting of four treatments and three replications. The treatments were P0 = Complete feed containing 0% cocoa pulp, P1 = Complete feed containing 5% cocoa pulp, P2 = Complete feed containing 10% cocoa pulp, and P3 = Complete feed containing 15% cocoa pulp on dry matter basis. The results of the study indicated that the average IVDMD was 567, 538, 566, and 526 g kg-1 DM, while the average IVOMD was 522, 491, 502, and 461 g/kg DM, respectively for treatment P0, P1, P2, and P3. Statistical analysis indicated that increasing levels of coca pulp in the feed significantly affected (P<0.05) the IVDMD and IVOMD of the feed. In conclusion, cocoa pulp is potential to be used up to 10% in complete feed with corn cobs as the fibre source.