Bacterial spores have special significance in foods because they are much more resistant to physical and chemical antimicrobial treatment. Nowadays, there is interest in using natural products such as plant extract for food preservation. In this study, 26 of tropical medicinal plants and spices were screened for their sporicidal activity against the spores of Bacillus cereus. The spores of B. cereus was harvested after incubation at 30°C for 1 week and treated with various plant extracts using the method of Standard Operating Procedure for the AOAC (Association of Official Analytical Chemists) Sporicidal Activity. Glutaraldehyde was used as a positive control. Among them, Indonesian bay leaf (Eugenia polyantha Wight) inactivated more than 3 log of spores/ml of B. cereus (99.99%) at the concentration of 1% and completely killed B. cereus spores at concentration of 2.5%. These results suggest that Indonesian bay leaf extract has strong sporicidal activity against spores of B. cereus.
Piper cubeba L. is traditionally recognised as flavouring ingredient in various types of foods and has been used to marinate meat. Scientifically, it has been reported to possess various valuable nutritional and pharmacological properties including antimicrobial potential. The aim of the present work was to determine the antibacterial activity of ethanolic P. cubeba L. extract against Escherichia coli and its effect on the microbiological quality of raw chicken meat during storage. Disc diffusion assay was done and resulted in 8.40 ± 0.10 mm of inhibition zone. The bacteriostatic and bactericidal effects of the extract were determined at 0.63 ± 0.00 mg/mL and 1.25 ± 0.00 mg/mL of concentration by MIC and MBC methods, respectively. The killing time was recorded at 2 × MIC (1.25 mg/mL) for 4 h. The application of the extract on chicken meat samples showed reduction in TPC and E. coli count with the observed optimum condition at 5.00% concentration stored at -18°C for 14 days based on the consistent reduction. Sensory attributes acceptability evaluation by 9-point hedonic scale showed acceptable score for colour, odour, texture and overall acceptability of the treated raw chicken meat samples. The findings implies that P. cubeba L. can be listed as one of the alternatives to reduce the bacterial load of raw chicken meat prior to cooking which is very important in ensuring food safety as well as reducing the occurrence of foodborne poisoning associated with chicken meat.
Microbial contamination in food system poses risk towards public health. The usage of synthetic
and chemical preservatives to prevent the contamination has become a growing concern due
to the presence of deleterious and harmful substances that can cause environment and health
problems in prolonged exposure. Thus, there are needs to overcome this problem by using
natural products as food preservatives. In this study, the antimicrobial activities of methanolic
Cymbopogon citratus (lemongrass) extracts were tested against five foodborne pathogens,
namely Bacillus cereus, Escherichia coli O157:H7, Klebsiella pneumoniae, Staphylococcus
aureus and Candida albicans. The susceptibility test, minimum inhibitory concentrations
(MIC) and minimum bactericidal concentration (MBC) or minimum fungicidal concentration
(MFC) were conducted using the broth microdilution techniques as described by Clinical and
Laboratory Standard Institute (CLSI). C. citratus extract showed antimicrobial activity against
all tested foodborne pathogens; B. cereus, E. coli O157:H7, K. pneumoniae, S. aureus and C.
albicans with the inhibition zone of 12 mm, 7.5 mm, 11 mm, 10 mm and 9 mm, respectively.
The MIC of C. citratus extract against B. cereus, E. coli O157:H7, K. pneumoniae, S. aureus
and C. albicans was 0.08 mg/ml, 0.63 mg/ml, 0.04 mg/ml, 0.31 mg/ml, and 0.16 mg/ml,
respectively, while the MBC or MFC was 1.25 mg/ml, 2.50 mg/ml, 2.50 mg/ml, 1.25 mg/
ml and 1.25 mg/ml, respectively. Time–kill curves were determined to assess the correlation
between MIC and bactericidal activity of C. citratus extract at concentrations ranging from 0×
MIC to 4× MIC. The bactericidal endpoint for B. cereus, E. coli O157:H7, S. aureus and C.
albicans was at 4× MIC after 2 h, 4× MIC after 2 h, 4× MIC after 30 min and 4× MIC after 4
h, respectively whereas K. pneumoniae was not completely killed after 4 hours of incubation at
4× MIC. The potent antimicrobial activity of C. citratus extract may support its usage as natural
antimicrobial agent
Increase of foodborne diseases has promulgated the development of new natural food additive
with high extraction yield to eliminate food pathogenic organisms. One such possibility is the
use of plant product as antibacterial agents with non-conventional method to enhance the yield.
In this study, cinnamon leaves (Cinnamomum zeylanicum Blume) were subjected to ultrasonic
assisted extraction (UAE) using response surface methodology (RSM) to optimise extraction
yield and total phenolic contents. The effect of two independent factors, extraction temperature
(x1: 25-40°C) and extraction time (x2: 15-45 minutes) were investigated. Optimum extraction
yield and total phenolic contents of cinnamon leaves were 27.49 ± 1.59% and 3987 ± 79.10 mg
GAE/g which were closely as predicted using RSM (28.34%, 4048 mg GAE/g), respectively.
The optimum condition of extraction yield (40°C and 45 minutes) showed the maximum zone
of inhibition against all tested foodborne pathogens (7.33 ± 0.50 to 13.22 ± 0.44 mm), whereas
optimum condition of total phenolic contents (33°C and 31 minutes) showed the lowest zone
inhibition (6.78 ± 0.67 mm to 11.67 ± 1.41 mm). The minimal inhibitory concentration (MIC)
values range from 97.65 to 6250.00 μg/mL and minimal bactericidal concentration (MBC)
values from 6.25 to 50.00 mg/mL. These results indicated that UAE method is excellent in
producing significantly the highest of extraction yield, total phenolic contents and act as a
potential natural antibacterial agent even using low extraction temperature and short time.
The objective of this study was to investigate the effect of nutmeg (Myristica fragrans Houtt.) extract at different concentrations on chemical characteristics of raw beef under frozen storage. Nutmeg extracts at concentrations of 0.25%, 0.65%, 1.25%, 2.50% and 5.00% (g/ml) were used to treat raw beef (2.5 × 2.5 × 1.0 cm; 4 ± 0.5 g) with dilution method. Treated samples were then individually packed in overwrapped trays and stored for 3 weeks at -18 ± 1oC. The effects of the extract on the chemical characteristics such as lipid oxidation, colour, pH, moisture, fat, and protein content of raw beef were evaluated at 0, 4, 7, 10, 14 and 21 days of storage. Lipid oxidation was evaluated based on thiobarbituric acid-reactive substance (TBARS) content. Colour of beef was observed by spectrophotometer in colorimetic parameters CIELabs. Values of pH were measured using pH meter. Moisture, fat and protein content were determined using method by Analysis Association of Official Analytical Chemists (AOAC). The result showed that extract at concentration of 1.25% inhibited TBARS value meaning that extract of 1.25% or more was able to maintain the oxidative stability of beef at -18oC. A 1.25% of extract was also able to maintain the redness (a*) of treated beef compared to untreated during frozen storage. The pH values of all samples beef decreased starting from 10th day of storage. Untreated samples (0.00%) showed the lowest pH values compared to other treated samples at the end day of storage. There was no significant different in term of protein content in all treated or untreated samples. However, fat and moisture content were significantly affected by the concentration of nutmeg extract. Treated beef was able to retain its moisture with only loss of moisture ranging from 0.2% – 2.00% while untreated samples had 5.00% loss of moisture. The fat content of untreated samples (0.00%) showed a reduction of 0.2% of fat content at the end of storage compared to all treated sample with only loss of 0.1% - 0.05%. Overall, nutmeg extract can be used to maintain the chemical characteristics of raw beef during storage for 3 weeks.
Recently, there has been an increasing demand and interest in developing plant extracts as natural food sanitizer, owing to their antimicrobial properties. Hence, the aim of this study was to investigate the effect of salam (Syzygium polyanthum L.) leaves methanolics extract on the number of microflora on chicken meat and shrimp. Salam leaves extract at different concentrations (0.0%, 0.1%, 1.00%) and exposure times (5, and 10 min) used to treat chicken meat and shrimp by using dilution method. Result showed that the total plate count and Staphylococcus aureus had been detected in untreated chicken and shrimp samples with 6.66 ± 0.12, 8.66 ± 0.15 and 7.25 ± 0.21, 6.54 ± 0.21, respectively. However, there was no Escherichia coli, Salmonella spp. and Vibrio cholerae detected in both samples. The number of total plate count (TPC) and S. aureus in chicken meat and shrimp were starting to reduce significantly at 0.01% concentration of salam leaves extract for 5 minutes of exposure time compared to initial count. There was no significantly different between exposure times. The highest reduction in number of microorganism population was at treatment with 1.0% extract for 10 min where TPC was reduced from 6.66 ± 0.12 to 0.00 ± 0.00 log10 CFU/ml, and from 8.66 ± 0.15 to 4.88 ± 0.00 log10 CFU/ml in shrimp while S. aureus reduced from 7.25 ± 0.21 to 3.88 ± 0.01 and from 6.54 ± 0.21 to 4.92 ± 0.04 in chicken and shrimp, respectively. For the sensory acceptability, overall acceptability were accepted by panellists until treatment 0.10% for 5 min and 10 min of soaking time. In conclusion, salam leaves extract might be developed as natural sanitizer for rinsing raw food materials such as chicken meat and shrimp.
The effects of methanolic extract of Javanese turmeric (Curcuma xanthorrhiza Roxb.) at different level of concentrations on the inactivation of Bacillus cereus, Escherichia coli, Pseudomonas spp. and Staphylococcus aureus in oyster mushroom (Pleurotus sajor-caju) were investigated. This study was conducted principally for the achievement on the best combination between the
susceptibility of C. xanthorrhiza extract on natural microflora and foodborne pathogenic bacteria with the sensory acceptability of the soaked oyster mushroom. Three different concentrations (g/ml), 0.05%, 0.50% and 5.00%, of C. xanthorrhiza extract prepared with dilution method were designed as sanitizing agent in treating the oyster mushroom at 5 minutes and 10 minutes.
There was significance reduction in the survival of microbial load between the untreated fresh oyster mushroom and those soaked with 0.05%, 0.50% and 5.00% rhizome extract (P
This study is conducted to investigate the effect of different concentrations of betel leaves extract on color, pH and microbiological in homemade chili bo. The homemade chili bo with different concentrations (0 mg/ml, 0.75 mg/ml, 1.25 mg/ml and 1.75 mg/ml) of betel leaves extract were prepared for analysis. The results showed that the color of chili bo became darker as the concentration of betel leaves extract increased. The extract showed significant in the pH of chili bo after 7 days in which the highest concentration of extract showed the highest value of pH 4.31. The aerobic microbial count was decreased as the concentration of betel leaves extract increased in chili bo. After 7 days of storage, the highest concentration of betel leaves extract showed the highest percentage of reduction (6%), while the control sample showed 2.41% of aerobic reduction. The study also found that the extract contain lesser yeast and mold count (5.22 log CFU/ml) in homemade chili bo compared to the control sample (5.31 log CFU/ml) after 7 days. Betel leaves extract can be considered as natural food preservatives in chili bo to reduce the growth of spoilage microorganism and thus enhance the shelf life of chili bo.
Vibrio parahaemolyticus is a main foodborne disease in seafood and generally seafood is
easily deteriorates in quality of color and flavor. In this study, clove (Syzygium aromaticum)
extract shows potent antibacterial activity against growth of antibiotics resistant Vibrio
parahaemolyticus on seafood samples (cockles and shrimps). Vibrio parahaemolyticus was
artificial contaminates on the samples with 106 CFU/ml. The samples were treated with different
concentration of cloves extract with 10 mg/ml which are 0.5%, 5% and 10% concentration
from methanol food grade extraction in 0 hr, 5 min, 10 min, 15 min, 30 min, 60 min and
120 min. Tab water and deionized water were selected as a negative control. As a result, the
amount of 10 % cloves managed to mitigates the number of V. parahaemolyticus on seafood
samples in 5 minutes and 15 min on both samples. Therefore, our results signify the fact that
cloves can be apply as natural sanitizer which could meet consumer demands for safe and
traditionally consumed either raw without any undesirable effect when applied in the seafood
system industries.
Edible medicinal plants are often used in the treatment of various ailments and spice in traditional food preparation. In this study, 45 of tropical edible medicinal plants extracts from Indonesia, Malaysia, and Thailand were screened for their antimicrobial activity against five standard microorganisms for food preservative namely Aspergillus niger, Candida albicans, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. The methanol extracts of Piper nigrum L. seed, Piper cubeba L. seed, and the root of Ligusticum acutilobum Siebold and Zucc. showed antimicrobial activity against five species of standard microorganisms. Among them, P. cubeba L. extract demonstrated the most susceptible against all tested microorganisms. Minimal inhibitory concentration (MIC) and minimal bactericidal or fungicidal concentration (MBC or MFC) were performed by the broth microdilution techniques as described by the Clinical and Laboratory Standard Institute. MIC values of P. cubeba L. extract to A. niger, C. albicans, E. coli, P. aeruginosa and S. aureus were 12.8, 1.6, 3.2, 6.4, and 1.6 mg/ml, respectively. P. cubeba extract killed A. niger, C. albicans, E. coli, P. aeruginosa and S. aureus with MBC values of 25.6, 3.2, 6.4, 12.8, and 3.2 mg/ml, respectively. The potent antimicrobial activity of P. cubeba L. extract may support its use for natural food preservative.
Klebsiella pneumoniae is a foodborne pathogen associated with pneumoniae. Multiresistance to antibiotics of K. pneumoniae is a significant public health treat. Recently, the use of natural products such as herbs to inhibit the growth of pathogens is increasing. Java turmeric (Curcuma xanthorrhiza Roxb.) has been reported to possess antibacterial activity against foodborne pathogens. Unfortunately, the antibacterial activity of java turmeric extract against the resistance to multiantibiotics of K. pneumoniae has not been investigated. In this study, the antibacterial activity of Java turmeric extract was tested against 24 isolates of resistant K. pneumoniae that was isolated from several vegetables; lettuce, cucumber, tomato and carrot, using the methods recommended by the Clinical and Laboratory Standard Institute (CSLI), including disc diffusion method, minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC) and killing time at concentration 0× MIC, ½× MIC, 1× MIC, 2× MIC and 4× MIC with predetermined time of 0, 0.25 , 0.5, 1, 2 and 4 h. The results showed that Java turmeric extract is susceptible to all resistant K. pneumoniae with inhibition zones ranging from 8.67 ± 0.58 to 10.00 ± 0.00 mm. The MIC and MBC values for the K. pneumoniae isolates against all bacterial isolates was 1.25 and 2.5 mg/ml, respectively. The killing time curve shows the reduction of resistant K. pneumoniae cells is fast acting; > 3 log10 within less than 15 min at 4× MIC (5.0 mg/ml). Finally, the isolates were completely killed at 4× MIC for 15 min. In conclusion, the Java turmeric extracts can be developed as natural antimicrobial agent to inhibit the growth of K. pneumoniae in food system.
Nowadays consumer is more demand on natural foodstuff instead of synthetic product due to their concern on health. The objective of this study is to investigate the effect of C. caudatus extract on the number of microflora in oyster mushroom at different concentration of C. caudatus extract and exposure time using dilution method. The results showed that the number of microorganisms (Log10 CFU/g) in oyster mushroom in term of Total Plate Count (TPC), Bacillus cereus, Escherichia coli and Staphylococcus aureus were 6.13 ± 0.04, 6.15 ± 0.09, 5.97 ± 0.04, and 6.46 ± 0.00, respectively. The effect of C. caudatus extract on microflora in oyster mushroom at concentrations of 0.00%, 0.05%, 0.5%, and 5.0% with exposure time of 0, 5, 10, and 15 min demonstrated that the reduction number of microflora in oyster mushroom was dependent on the concentration of C. caudatus extract and exposure times. The number of TPC (Log10 CFU/g) in oyster mushroom was significantly reduced after treated with C. caudatus extract at concentration of 0.05% for 15 min; 6.13 ± 0.04 reduced to 2.62 ± 0.07. Moreover, B. cereus (Log10 CFU/g) in oyster mushroom was significantly reduced by treatment of C. caudatus extract at concentration of 0.05% for 5 min; 6.15 ± 0.09 reduced to 3.77 ± 0.15. Meanwhile, the number of E. coli (Log10 CFU/g) in oyster mushroom was significantly reduced at concentration of 0.05% for 10 min; 5.97 ± 0.04 reduced to 3.21 ± 0.13. Lastly, the survival number of S. aureus in oyster mushroom was significantly reduced after treated with C. caudatus extract at concentration of 0.05% for 15 min; 6.46 ± 0.00 reduced to 4.83 ± 0.07. In conclusion, C. caudatus extract has potentiality to be developed as natural sanitizer for rinsing raw food materials such as oyster mushroom.
The filamentous spoilage fungi in vegetables can lead to significant impact in food and economic loss. In order to overcome this problem, chemical fungicide has been implemented in vegetable farming and processing but it causes problems towards environment and food safety. Thus, the utilization of natural products such as plants extracts, which exhibit antimicrobial and antifungal activity, is more acceptable to solve this problem. The aim of this study is to investigate the antifungal activity of Boesenbergia rotunda extract against ten filamentous spoilage fungi isolated from five vegetables. The extract was used to treat fungal isolates from vegetables; CRb 002 (Penicillium sp.), CHa 009 (Aspergillus sp.), TMa 001 (Geotrichum sp.), TMa 002 (Aspergillus sp), ONb 001 (Aspergillus sp.), WBb 003 and WBb 004 (Fusarium sp.) WBb 007 (unidentified), WBb 008 (Aureobasidium sp.) and WBb 010 (Penicillium sp.). The results showed that the yield of the extract of B. rotunda using ethanol (95%) was 11.42% (w/v). The 10% of B. rotunda extract exhibited antifungal activities against ten filamentous fungi after 5 days treatment with growth reduction of 41.56%, 30.68%, 86.20%, 50.62%, 26.67%, 47.44%, 50.74%, 36.39%, 42.86%, and 39.39% for WBb 008, WBb 004, WBb 007, WBb 003, CRb 002, WBb 010, CHa 009, TMa 001, ONb 001, and TMa 002, respectively. B. rotunda extract showed highest antifungal activity against fungi isolated from winged bean (WBb 007) with percentage reduction in growth was 86.20%, while the lowest activity was against fungi isolated from the carrot (CRb 002) with 26.67% reduction in growth. Generally, the TPC of fungi in the vegetable samples were reduced after treatment with 5% of B. rotunda extract at 5 min and 10 min of exposure time. The results suggested that B. rotunda extract has high potential to become natural food preservative which can reduce the fungi spoilage of vegetables.
Plants have been used recently to eliminate bacterial growth in food products. This study was undertaken to test the in vitro sanitizing effect of crude extract from bitter gourd (BG) fruit on the growth of native microorganisms in raw chicken leg meat. Hot air dried BG and extrudate extracts at 1% concentration and exposure times of (5, 10 and 15 min) were used to treat the samples using dilution method. Results showed that BG extrudate had a slightly stronger bactericidal activity against the microflora than the B.G. hot air drying treatment, especially, on E. coli at all exposure time. Overall, there is no significant difference between the treatments; Total Plate Count (TPC), Escherichia coli, Bacillus cereus, Staphylococcus aureus. The best reduction time of microflora by hot air dried extract was at (15 min) except for B. cereus was at (5 min) and for extrudate extract was at (5 min) except for E. coli was at (10 min). In conclusion, bitter gourd extract could be used as an important natural sanitizer for rinsing raw food matrials such chicken meat.
The purpose of this study was to investigate the activity of xanthorrhizol isolated from Curcuma xanthorrhiza Roxb. on Candida albicans biofilms at adherent, intermediate, and mature phase of growth. C. albicans biofilms were formed in flat-bottom 96-well microtiter plates. The biofilms of C. albicans at different phases of development were exposed to xanthorrhizol at different concentrations (0.5 µg/mL-256 µg/mL) for 24 h. The metabolic activity of cells within the biofilms was quantified using the XTT reduction assay. Sessile minimum inhibitory concentrations (SMICs) were determined at 50% and 80% reduction in the biofilm OD₄₉₀ compared to the control wells. The SMIC₅₀ and SMIC₈₀ of xanthorrhizol against 18 C. albicans biofilms were 4--16 µg/mL and 8--32 µg/mL, respectively. The results demonstrated that the activity of xanthorrhizol in reducing C. albicans biofilms OD₄₉₀ was dependent on the concentration and the phase of growth of biofilm. Xanthorrhizol at concentration of 8 µg/mL completely reduced in biofilm referring to XTT-colorimetric readings at adherent phase, whereas 32 µg/mL of xanthorrhizol reduced 87.95% and 67.48 % of biofilm referring to XTT-colorimetric readings at intermediate and mature phases, respectively. Xanthorrhizol displayed potent activity against C. albicans biofilms in vitro and therefore might have potential therapeutic implication for biofilm-associated candidal infections.
A novel way to reduce rambutan wastage is to ferment the fruit and valorise the seed post-fer- mentation into other food products and ingredients. Hence, the objective of this study was to investigate the physicochemical properties of rambutan seed during solid-state fermentation of the fruit. Peeled rambutan fruits were subjected to natural fermentation for ten days at 30°C. The environmental temperature, relative humidity, internal and external temperatures of the fermentation mass were measured daily. After ten days of fermentation, the seeds had higher cut test score (867.5), fermentation index (1.527), and a* value (8.20 for non-dried seeds and
9.93 for dried seeds), and lower L* (51.90 for non-dried seeds and 49.22 for dried seeds) and b* (30.52 for non-dried seeds and 30.12 for dried seeds) values; as compared to the non-fer- mented seeds (cut test score, 0.0; fermentation index, 0.856; L*, a*, and b* values, 64.52, 2.25, and 42.07 for non-dried seeds, respectively, and 61.03, 3.23 and 36.70 for dried seeds, respectively). During this time, pH, total soluble solids, fructose, glucose, sucrose, citric acid, and tartaric acid contents of the seeds decreased by 46, 44, 59, 61, 100, 85, and 100%, respec- tively, while the titratable acidity, lactic acid, acetic acid, and ascorbic acid contents of the seeds increased by 5.5, 7.8, 6.0, and 2.2-fold, respectively. Results showed that eight days of fermentation are adequate to produce well-fermented rambutan seeds that could be further processed into a cocoa powder-like product by roasting the fermented fruits in a manner similar to that of cocoa bean roasting.
An awareness of Escherichia coli as a foodborne pathogen and illness causing bacterium has been increased among consumers. Moreover, there is demand for natural product in order to reduce synthetic product that can cause toxic to the human. In this study, antibacterial activity, in term of MIC, MBC and killing-time curve of methanolic extract of Boesenbergia rotunda have been tested against a standard E. coli ATCC 25922 and two E. coli isolated from milk products using Clinical and Laboratory Standard Institute (CLSI) methods. The results show that B. rotunda extract was susceptible to all E. coli strains. The MIC and MBC values of B. rotunda extract against E. coli ranged 0.019 mg/mL 2.5 mg/mL and 0.039 mg/mL – 5.0 µg/mL, respectively. Killing-time curves were constructed at concentrations of 0x MIC, 1/2x MIC, 1x MIC, and 2x MIC. All E. coli strains can be killed with concentration of 2x MIC after 2 hours. The results show that B. rotunda extract has potential antibacterial activity against E. coli.
Listeria monocytogenes (L. monocytogenes) is a gram positive food-borne pathogen that is able to form biofilm on food factory surfaces. Formation of biofilm makes the bacteria much more resistance to environmental stresses such as disinfectant. The extracellular polymeric matrix (biofilm structure) which is mostly comprised of sticky extracellular polysaccharides (EPS) and proteins can protect bacteria in a harsh condition. The efficiency of four disinfectants on removing L. monocytogenes biofilm was investigated. Five concentration levels (100, 50, 25, 12.5, and 6.25%) of disinfectants were tested. In the microtitre assay, the optical density at 595 nm CV-OD595 value, was used to measure the amount of remained biofilm after 24 h. Results showed that disinfectants did not have significant effect on removing L. monocytogenes biofilm. Formation of L. monocytogenes biofilm significantly decreased the efficiency of disinfectants. Biofilm produced by strain number 9 showed higher resistance to disinfectant. Low concentrations (
A total of 85 food handlers participated in this study to determine the hygienic status of their hands in primary schools located in the state of Selangor (Malaysia). Overall findings revealed that the fecal contamination and personal hygiene of the food handlers were well maintained with the range of mean bacterial counts from 0.18 to 0.47 log10 Colony Forming Units/cm2 during the three intervals of hand swabbing (before, during and after) preparation of ready-to-eat foods. However, the general indication of the microbiological quality (Aerobic Plate Count) was out of the standard (range of mean bacterial counts from 1.39 to 1.56 log10 Colony Forming Units/cm2) based on previous literature. This study highlighted that the food handler’s adherence to Good Manufacturing Practice and Sanitation Standard Operating Procedures was insufficient and suggested that attention should be emphasized on their practices at the intervals of school recess: before, during and after the preparation of ready-to-eat foods. In addition, there is also a need in the implementation of an effective HACCP program in Malaysia school foodservice operations.
Two cocoa bean fermentation methods (spontaneous fermentation and the use of starter culture) for 7 days fermentation were compared in terms of safety and quality fermented beans. Candida sp. was used as a starter culture in this study. The safety of the fermented cocoa beans were measured by the growth colonies of pathogenic microorganisms namely Bacillus cereus, Escherichia coli, Salmonella sp., Staphylococcus aureus, and Pseudomonas sp., on Bacillus cereus agar, eosin-methylene blue (EMB) agar, xylose lysine deoxycholate (XLD) agar, Baird-Parker agar (BPA), and Pseudomonas agar, respectively. B. cereus, E. coli and Salmonella sp. were early present in both fermentations. Candida sp.-fermentation showed detection of B. cereus at 5.34 log10 CFU/g and absence after 24 hours of fermentation while in spontaneous-fermentation B. cereus was too few to count. Moreover, the log10 E. coli number in Candida sp.-fermentation and spontaneous-fermentation were reduced from 5.72 to 3.66 and from 7.15 to 4.46 on day 1 to day 3, respectively. There were no presences of pathogenic microorganisms on day 5 and day 7 for both fermentations. In term of quality, proximate analysis of spontaneous-fermentation resulted that the content of moisture, ash, fat, crude protein, crude fibre and carbohydrate was 56.47%, 2.32%, 3.17%, 7.02%, 28.14% and 2.88%, meanwhile for the Candida sp.-fermentation was 53.96%, 2.19%, 3.44%, 8.25%, 25.46% and 6.70%, respectively. This study showed that both fermentations are considered to be safe and there is no significant difference in proximate value in fermented cocoa beans from spontaneous-fermentation and Candida sp.-fermentation.