Campylobacter jejuni was found to occur at high prevalence in the raw salad vegetables examined. Previous reports describe cross-contamination involving meat; here we investigated the occurrence of cross-contamination and decontamination events in the domestic kitchen via C. jejuni-contaminated vegetables during salad preparation. This is the first report concerning quantitative cross-contamination and decontamination involving naturally contaminated produce. The study was designed to simulate the real preparation of salad in a household kitchen, starting with washing the vegetables in tap water, then cutting the vegetables on a cutting board, followed by slicing cucumber and blanching (heating in hot water) the vegetables in 85 degrees C water. Vegetables naturally contaminated with C. jejuni were used throughout the simulation to attain realistic quantitative data. The mean of the percent transfer rates for C. jejuni from vegetable to wash water was 30.1 to 38.2%; from wash water to cucumber, it was 26.3 to 47.2%; from vegetables to cutting board, it was 1.6 to 10.3%; and from cutting board to cucumber, it was 22.6 to 73.3%. The data suggest the wash water and plastic cutting board as potential risk factors in C. jejuni transmission to consumers. Washing of the vegetables with tap water caused a 0.4-log reduction of C. jejuni attached to the vegetables (most probable number/gram), while rapid blanching reduced the number of C. jejuni organisms to an undetectable level.
Microbiological quality analysis of freshwater prawns from three sampling sites in Peninsular Malaysia viz: Site 1- Kg. Jumbang, Negri Sembilan; Site 2- Kg. Cangkat Tin, Perak and Site 3- Kg. Cenderiang, Perak for total mesophilic and psychrophilic aerobic counts, proteolytic bacterial counts, histamine producing bacteria, cadaverine producing bacteria and putrescine producing bacteria in the prawns and pond water for the three sites showed that the microbiological quality of freshwater prawns is related to the microflora of pond water in which they were grown. The initial bacterial counts indicated the values were in the range of log 4+ CFU/g for all samples. Total mesophilic and psychrophilic counts of the head regions were higher than that of the body regions for all prawn samples and types of growth media tested. All samples showed an increase in counts with time and temperature of storage up to log 7+ CFU/g for mesophilic counts after 12 hours at ambient, 6 days at 10 ± 2°C and 12 days at iced storage. The samples from Site 2 had relatively higher counts compared to the other two sites which correlated well with the levels determined in the pond water. Similar trends were observed for psychrophilic counts but at lower values for the different types of media studied.
Effects of preservatives on quality changes and shelf life of shrimp during iced storage indicated that boric acid, lactic acid and sodium metabisulphite managed to inhibit psychrophilic bacteria and biogenic amines formation in prawns while maintaining the mesophilic counts at lower levels during iced storage.
Biochemical analysis was carried out for pH profiles, freshness in terms of K-values, amino acids profiles, total volatile bases (TVB), total volatile acids (TVA) and biogenic amines for fresh and preserved Macrobrachium rosenbergii. Results showed that pH profiles of Macrobrachium rosenbergii explain the inability of this parameter to be used to evaluate the quality of Macrobrachium rosenbergii. Thus changes in pH profiles of Macrobrachium rosenbergii should be combined with indicators such as total volatile acids and total volatile bases. Total volatile acids of the shrimps increased steadily in small amounts throughout the storage period. A rapid increase in TVB at 100C was detected due to the increase in total aerobic bacteria at elevated temperatures. The microbial activities caused the decrease in the amino acids arginine, lysine and histidine which correlated well with the increase in the corresponding biogenic amines such as putrescine, cadaverine and histamine respectively. Preservatives used in this study controlled the production of these biogenic amines without significantly altering the pH of preserved shrimp.
Response surface methodology (RSM) was used to determine the optimum experimental conditions to extract methylmercury from fish samples for GC analysis. The influence of four variables - acid concentration (3-12 M), cysteine concentration (0.5-2% w/v), solvent volume (3-9 ml) and extraction time (10-30 min) - on recovery of methylmercury was evaluated. The detection limit for methylmercury analysis using a microelectron capture detector was 7 ng g(-1) in fish samples. The mean recovery under optimum conditions was 94%. Experimental data were adequately fitted into a second-order polynomial model with multiple regression coefficients (r(2)) of 0.977. The four variables had a significant effect (p < 0.05) on the recovery of methylmercury from a reference material (BCR-463). Optimum conditions for methylmercury extraction were found using an acid concentration of 12.2 M, cysteine concentration of 2.4%, solvent volume of 1.5 ml and extraction time of 35 min. The validation of the developed method to analyze methylmercury in fish samples exhibited good agreement with mercury content in the samples.
The effect of different drying methods on the degradation of flavonoids in Centella asiatica was evaluated. C. asiatica leaf, root and petiole were dried using air-oven, vacuum oven and freeze drier. Flavonoid was determined utilizing reverse-phase high performance liquid chromatography (RP-HPLC). Results of the study revealed the presence of high concentration of flavonoids in C. asiatica leaf, root and petiole, which include, naringin (4688.8 ± 69 μg/100 g, 3561.3 ± 205 μg/ 100 g, and 978.3 ± 96 μg/ 100 g), rutin (905.6 ± 123 μg/ 100 g, 756.07 ± 95 μg/ 100 g, and 557.25 ± 58 μg/ 100 g), quercetin (3501.1 ± 107 μg/ 100 g, 1086.31 ± 90 μg/ 100 g, and 947.63 ± 83 μg/ 100 g) and catechin (915.87 ± 6.01 μg/ 100 g, 400.6 ± 67 μg/ 100 g, and 250.56 ± 18 μg/ 100g). Luteolin, kaempferol and apigenin on the other hand, were inconsistently present in some parts of C. asiatica. Air-oven treatment resulted in the highest total flavonoids degradation followed by vacuum oven and freeze dried with percent degradation of 97%, 87.6% and 73%, respectively. Catechin and rutin were found to be the most stable flavonoids with percent degradation up to 35%, 66% and 76% for freeze dried, vacuum oven and air oven, respectively.
A new electrochemical biosensor is described for voltammetric detection of gene sequence related to Trichoderma harzianum. The sensor involves immobilization of a 20 base single-stranded probe (ssDNA), which is complementary to a specific gene sequence related to T. harzianum on a gold electrode through specific adsorption. The DNA probe was used to determine the amount of target gene in solution using methylene blue (MB) as the electrochemical indicator. The covalently immobilized probe could selectively hybridize with the target DNA to form a hybrid on the surface despite the bases being attached to the electrode. The changes in the peak currents of methylene blue (MB), an electroactive label, were observed upon hybridization of probe with the target. Peak currents were found to increase in the following order: hybrid-modified AuE and the probe-modified AuE which localized to the affinity of MB. Control experiments with the non-complementary oligonucleotides were performed to assess whether the DNA biosensor responds selectively, via hybridization, to the target. DNA biosensor also able to detect microorganism at the species levels without nucleic acid amplification. The redox current was linearly related to the concentration of target oligonucleotide DNA, ranged from 1-20 ppm. Numerous factors, affecting the probe immobilization, target hybridization and indicator binding reactions are optimized to maximize the sensitivity and reduce the assay time.
PURPOSE: This study was designed to find a reliable Epstein-Barr virus (EBV) immunoglobulin (Ig) G-based diagnostic/screening test for nasopharyngeal carcinoma (NPC) able to demarcate between the NPC-related seropositivity of EBV IgG antibodies and that of other head and neck cancer (HNCA) and control groups. The NPC-associated immunosuppression affects EBV IgA much more than IgG, leading to inconsistent detection of NPC using EBV IgA antibodies.
MATERIALS AND METHODS: One hundred twenty-two HNCA patients, 42 NPC, 66 laryngeal carcinoma, and 14 hypopharyngeal carcinoma and 3 groups of 100 control subjects were enrolled in this study. Enzyme-linked immunosorbent assay (ELISA) was used to find a specific cutoff value for the NPC-related seropositivity of EBV IgG antibodies.
RESULTS: NPC group showed higher serum level of EBV IgG antibodies than control and other HNCA groups (P < .05). However, the traditional cutoff value, mean + 2 SDs of control subjects, failed to demarcate the seropositives of NPC patients from those of healthy population (P > .05). The new cutoff value, mean + 2 SDs of the seropositives group of control subjects who had already been grouped by the traditional cutoff value, proved successful. It succeeded to demarcate between the NPC-related EBV IgG seropositivity and that issued from the persistent, latent, or reactivated EBV infection in the population (P < .05). The sensitivity/specificity of NPC detection by the new cutoff-based ELISA kit, 76.19% and 86%, was close or higher than that of EBV IgA antibodies.
CONCLUSION: EBV IgG-based ELISA could be used for the diagnosis of NPC using a new cutoff threshold that excludes the population baseline of EBV IgG seropositivity.
The activities of lipase produced by five lipases-producing thermophilic bacteria strains (SY1, SY5, SY6, SY7 and SY9) isolated from Selayang Hot Spring in the western part of Peninsular Malaysia were analyzed and compared. SY7 and SY9 had considerably higher lipolytic activities than those of SY1, SY5 and SY6. Thermostabilities of lipase produced by all strains were determined after heating at 80°C for 30 minutes. Strain SY7 retained the highest lipolytic activity of 77%, while others had infinitesimally low thermostability (retaining less than 34% of their original activity) at the same temperature and time. SY7 was chosen for further characterization because it showed exceptionally high lipase activity and thermostability. It was identified as belonging to Bacillus species by the conventional Gram-staining technique, Biochemical tests and Biolog Microstation system. By using 16S rRNA gene sequencing, strain SY7 generated the same expected PCR product with molecular weight of 1500 base pair. It displayed 98% sequence similarity to Bacillus cereus strain J-1 16S rRNA gene partial sequence with accession number: AY305275 and has been deposited in the database of Genbank.
Bacteria with amine oxidase activity have become a particular interest to reduce biogenic amines concentration in food products such as meat and fish sausages. However, little information is available regarding the application of these bacteria in fish sauce. Hence, our study was aimed to investigate the effect of such starter cultures in reducing biogenic amines accumulation during fish sauce fermentation. Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05 isolated from fish sauce which possess amine oxidase activity were used as starter cultures in this study. Fermentation was held for 120 days at 35 °C. The pH value increased in all samples, while salt concentration remained constant throughout fermentation. Aerobic bacteria count was significantly lower (p < 0.05) in the control than in inoculated samples as a result of starter cultures addition. However, it decreased during fermentation due to the growth inhibition by high salt concentration. Proteolytic bacterial count decreased during fermentation with no significant difference (p > 0.05) among samples. These bacteria hydrolyzed protein in anchovy to produce free amino acid precursors for amines formation by decarboxylase bacteria. The presence of biogenic amines producing bacteria in this study was considered to be indigenous from raw material or contamination during fermentation, since our cultures were negative histamine producers. Amino acid histidine, arginine, lysine and tyrosine concentration decreased at different rates during fermentation as they were converted into their respective amines. In general, biogenic amines concentration namely histamine, putrescine, cadaverine and tyramine increased throughout fermentation. However, their concentrations were markedly higher (p < 0.05) in the control (without starter cultures) as compared to the samples treated with starter cultures. Histamine concentration was reduced by 27.7% and 15.4% by Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively. Both cultures could also reduce other amines during fermentation. After 120 days of fermentation, the overall biogenic amines concentration was 15.9% and 12.5% less in samples inoculated with Staphylococcus carnosus FS19 and Bacillus amyloliquefaciens FS05, respectively, as compared to control samples. These findings emphasized that application of starter cultures with amines oxidase activity in fish sauce fermentation was found to be effective in reducing biogenic amines accumulation.
Streptococcus bovis (S. bovis) bacteria are associated with colorectal cancer and adenoma. S. bovis is currently named S. gallolyticus. 25 to 80% of patients with S. bovis/gallolyticus bacteremia have concomitant colorectal tumors. Colonic neoplasia may arise years after the presentation of bacteremia or infectious endocarditis of S. bovis/gallolyticus. The presence of S. bovis/gallolyticus bacteremia and/or endocarditis is also related to the presence of villous or tubular-villous adenomas in the large intestine. In addition, serological relationship of S. gallolyticus with colorectal tumors and direct colonization of S. gallolyticus in tissues of colorectal tumors were found. However, this association is still under controversy and has long been underestimated. Moreover, the etiological versus non-etiological nature of this associationis not settled yet. Therefore, by covering the most of up to date studies, this review attempts to clarify the nature and the core of S. bovis/gallolyicus association with colorectal tumors and analyze the possible underlying mechanisms.
Biological amines are nitrogenous compounds that occur naturally in wide variety of food. Histamine, putrescine, cadavarine, tyramine, spermine, spermidine, tryptamine and β-phenylethylamine are the biogenic amines that are normally present in foods. Although the biogenic amines play some important physiological functions but high level of amines can cause toxicological effects. High amount of amines can be produced by bacteria during amino acids decarboxylation and have been identified as one of the important agent causing seafood intoxication. Temperature is the major factor for controlling the biogenic amines formation in food. The effects of other alternatives are also discussed including salting, packaging, irradiation, high pressure processing and the use of starter culture. A variety of techniques can be combined together to control the microbial growth and enzyme activity during processing and storage for better shelf life extension and food safety.
To explore new approaches of phage-based bio-process of specifically pathogenic Escherichia coli bacteria in food products within a short period. One hundred and forty highly lytic designed coliphages were used. Escherichia coli naturally contaminated and Enterohemorrhagic Escherichia coli experimentally inoculated samples of lettuce, cabbage, meat, and egg were used. In addition, experimentally produced biofilms of E. coli were tested. A phage concentration of 10(3) PFU/ml was used for food products immersion, and for spraying of food products, 10(5) PFU/ml of a phage cocktail was used by applying a 20-s optimal dipping time in a phage cocktail. Food samples were cut into pieces and were either sprayed with or held in a bag immersed in lambda buffer containing a cocktail of 140 phages. Phage bio-processing was successful in eliminating completely E. coli in all processed samples after 48 h storage at 4°C. Partial elimination of E. coli was observed in earlier storage periods (7 and 18 h) at 24° and 37°C. Moreover, E. coli biofilms were reduced >3 log cycles upon using the current phage bio-processing. The use of a phage cocktail of 140 highly lytic designed phages proved highly effective in suppressing E. coli contaminating food products. Proper decontamination/prevention methods of pathogenic E. coli achieved in this study can replace the current chemically less effective decontamination methods.
By-products from different animal sources are currently being utilised for beneficial purposes. Chicken processing plants all over the world generate large amount of solid by-products in form of heads, legs, bones, viscera and feather. These wastes are often processed into livestock feed, fertilizers and pet foods or totally discarded. Inappropriate disposal of these wastes causes environmental pollution, diseases and loss of useful biological resources like protein, enzymes and lipids. Utilisation methods that make use of these biological components for producing value added products rather than the direct use of the actual waste material might be another viable option for dealing with these wastes. This line of thought has consequently led to researches on these wastes as sources of protein hydrolysates, enzymes and polyunsaturated fatty acids. Due to the multi-applications of protein hydrolysates in various branches of science and industry, and the large body of literature reporting the conversion of animal wastes to hydrolysates, a large section of this review was devoted to this subject. Thus, this review reports the known functional and bioactive properties of hydrolysates derived from chicken by-products as well their utilisation as source of peptone in microbiological media. Methods of producing these hydrolysates including their microbiological safety are discussed. Based on the few references available in the literature, the potential of some chicken by-product as sources of proteases and polyunsaturated fatty acids are pointed out along with some other future applications.
The transformation of an animal into pieces fit for human consumption is a very important operation. Rather than argue about halal slaughter without stunning being inhumane or stunning being controversial from the Islamic point of view, we discuss slaughter, stunning and animal welfare considering both Islamic and animal welfare legislation requirements. With the world Muslim population close to two billion, the provision of halal meat for the Muslim community is important both ethically and economically. However, from the animal welfare standard point of view, a number of issues have been raised about halal slaughter without stunning, particularly, about stressful methods of restraint and the latency of the onset of unconsciousness. This paper sets out to, discuss the methods of stunning that are acceptable by Islamic authorities, highlight the requirements for stunning to be acceptable in Islam and suggest practical ways to improve the humanness of slaughter.
Modified atmosphere packaging (MAP) has become a popular method for packaging foods as it can extend the shelf life of food with minimal quality defect. Oxygen, nitrogen and carbon dioxide are the common gases used in MAP, Oxygen and carbon dioxide inclusive as only these two gaseous have the preservative effects on the packed food product. Their effect on microbial changes of any food product throughout storage period is highly depend on type of the product and packaging materials, appropriate gas composition, storage temperature, the ratio between gas and product volume, and hygienic manner during processing and packaging. MAP with highest percentage of carbon dioxide is proven to be more effective than vacuum packaging in inhibiting the growth of spoilage and pathogenic bacteria in many fishery products. This article reviews the consequences of MAP towards microbial changes in fishery products.
Nipa sap or air nira is a sweet natural beverage obtained from a type of palm tree, Nypa fruticans.
It is readily and spontaneously fermented resulting in the development of alcoholic fermentation products. Objective of this study is to determine the volatile compounds (VOCs) responsible for the aroma in fresh and fermented nipa sap. The sap was left for natural fermentation at 30ºC for 63 days. VOCs of the sap were analysed using static headspace gas chromatography-mass spectrometry (GC-MS). Fresh nipa sap contained ethanol (83.43%), diacetyl (0.59%), and esters
(15.97%). Fermented nipa sap contained alcohols (91.16 – 98.29%), esters (1.18 – 8.14%), acetoin (0.02 – 0.7%), diacetyl (0.04 – 0.06%), and acetic acid (0.13 – 0.68%). Concentration of ethanol in fresh nipa sap increased from 0.11% (v/v) to 6.63% (v/v) during the fermentation, and slightly decreased to 5.73% (v/v) at day 63. No higher alcohols were detected in the fresh nipa sap. Concentration of 1-propanol and 2-methylpropanol were constant throughout the fermentation with average of 0.004 to 0.006% (v/v) and 0.0001 to 0.0009% (v/v), respectively. 3-methylbutanol increased during the fermentation process. The highest concentration (0.001% v/v) was recorded at day 35. This study has shown differences in VOCs types between fresh and fermented nipa sap.
This article summarises the current methods for total malachite green (MG) detection which is known as a sum of MG and leuco-malachite green (LMG) that has been used extensively in aquaculture as fungicide, dye color in textile and other purposes in food industries. LMG is a reducing form of MG, where the MG is easily reduced due to the photo-oxidative de-methylation process. Nevertheless, the use of MG had become an issue due to its toxicity effects. Many analytical instruments such as HPLC, LC—MS/MS, GC—MS, and spectrometry have been widely used for detection of MG. However, these methods require long time sample preparation and analysis, expensive, use hazardous reagents and indirect measurements. Hence, other analytical methods which are more sensitive, safe, rapid, inexpensive and portable are required. Alternatively, biosensors promise a more sensitive and rapid detection method for MG and LMG.
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.
Biogenic amines formation in Indian mackerel of tropical region was investigated during storage at ambient (25-29 °C) and ice temperature (0 °C) in relation with changes of amino acids content and amines forming bacteria. All amines increased significantly during storage at two temperatures except for spermidine and spermine. Histamine concentration of 363.5 ppm was detected after 16 h stored at ambient temperature. Aerobic plate count of fish stored at ambient temperature reached 6.98 log CFU g(-1) after 16 h, close to the upper limit (7 log CFU g(-1)) suggested by International Commission on the Microbiological Specifications for Foods (ICMSF). However, proper icing procedure retarded the formation of histamine effectively, resulting only 8.31 ppm after 16 days of ice storage. Aerobic plate count of 5.99 and 7.72 log CFU g(-1) were recorded for fish stored in ice after 16 days and ambient temperature after 20 h, respectively. Histamine exhibited high correlation with histidine (r(2) = -0.963, P 0.05). As storage time progressed, the amines forming bacteria grew significantly except for that stored in ice.