The necessity of laboratory analyses to verify the authenticity of halal products

The concern of food safety, authenticity and adulteration has resulted in increased awareness regarding the composition of food products. The identity and source of the ingredients in processed or composite mixtures is not always readily visible. Most frequently, pork meat and its bi-products have been used to substitute other ingredients in food products. Hence, verification that the components are authentic and from sources acceptable to Muslim consumers are indeed essential. Sensitive and reliable methods for detection of halal products adulteration are of paramount important for implementation of halal food labelling, regulations and products quality control. Various techniques have been proposed for the analysis of pork, lard, khamr and gelatine including DNA-based methods, gas chromatography, liquid chromatography, differential scanning calorimetric and fourier transform infrared spectroscopy. Some of the techniques were validated and accredited with MS 17025 to potentially complement the existing verification and monitoring mechanisms for halal products certification

Isolation and Characterisation of Bifidobacterium Spp. from Infant Stools

Infant faecal materials from different feeding regimes (breast fed, breast fed supplemented with mixed diet and bottle fed) were analysed for their distribution of microbes and isolation of bifidobacteria. The predominant bacteria in breast-fed infants after 23 weeks were streptococci and bifidobacteria. The predominant bacteria in breast-fed supplemented with mixed diet after 24 weeks were clostridia, streptococci and bifidobacteria. In the bottle fed infant faeces, bifidobacteria was present in small numbers whereas lactobacilli were one of the most predominant bacteria. 18 isolates of Bifidobacterium in/antis and 10 isolates of Bifidobacterium breve were isolated and tested for their antibacterial activity, survival in low pH conditions, and adhesion to human colon carcinoma cell lines . Three isolates (B. in/antisC040225, B. breve F0526100 and B. breve G012048) exhibited good antibacterial activity. These isolates inhibit the growth of Salmonella enteritidis, Vibrio cholera, Escherichia coli, Bacillus cereus, Pseudomonas aerugmosa and Listeria monocytogenes. The survival rate of bifidobacteria in low pH condition was variable among isolates. All the isolates tested showed good survival in control solution (pH 6.5). None of the isolates tested was able to survive at pH 1.0 after 1 hour incubation. However, at pH 2.0, five isolates (B. infantis D042022, B. infantis F042466, B. infantis F0526100 and B. infantis G001204) survived after 1h incubation but not after 2h incubation at this pH. Four strains (B. infantis D04 2022, B. infantis F0606117, B. infantis F041134 and B. infantis Z040845) possessed good survival at pH 3.0 where viable cells could still be detected after 3 hours incubation. One of the most important criteria of probiotic micro-organism is the ability to adhere and colonise to human intestinal epithelial cells in order to prevent them from being flushed out by the peristaltic movement in the gastrointestinal tract. B. infantis G001204 was found to adhere well to HT29 cell lines in culture. Not all the bifidobacteria isolates tested possessed all the characteristics of probiotic

Species Classification and Molecular Studies of Bile Salt Hydrolase (BSH) Gene in Bifidobacterium Spp

Molecular methods were used to identify and characterise Bifidobacterium isolated from the faeces of breast-fed infants. A Bifidobacterium genus-specific primers based on the V9 variable region of the 168 rDNA was used to identify Bifidobacterium isolates and to distinguish them from other genera. All the Bifidobacterium species tested generated PCR product with the size of approximately 1.35 kb, whereas other genera showed no band with these primers. Furthermore, a sequence analysis of 168 rRNA gene of Bifidobacterium isolates revealed high homology (98% and above) with Bifidobacterium pseudocatenulatum JCM1200. This result indicates that the 16S rRNA gene sequence analysis is a useful and accurate tool for the identification of the genus Bifidobacterium In addition, a set of B. pseudocatenulatum species-specific primer was used as an alternative to identify the species of B. psedocatenulatum using PCR technique. It was found that this set of primer was able to generate PCR product with the size of approximately 278 bp in all the B. pseudocatenulatum isolates as well as the type strain of B. pseudocatenulatum JCM1200. Other species of bifidobacteria gave no band. A protocol for the rapid fingerprinting technique of Bifidobacterium strains and other pro biotic microorganisms based on randomly amplified polymorphic DNA (RAPD) has been developed. Three 10-mer primers (GEN1-60-03, GEN1-60-06 and GEN1-60-07) used generated RAPD patterns with DNA fragments ranging from approximately 0.3 kb to 10.0 kb in size. Examination of the DNA fingerprints using cluster analysis showed a significant genetic diversity among the strains tested. Another fingerprinting technique based on the distribution of dispersed repetitive DNA [enterobacterial repetitive intergenic consensus (ERIC) and repetitive extragenic palindromic (REP)] segments in the genome of Bifidobacterium and other pro biotic bacteria was also examined for the first time using primers derived from the REP and ERIC sequences and PCR. The patterns of the resulting PCR products were analysed on agarose gel and were found to be highly specific for each species. All the B. pseudocatenulatum isolates and B. pseudocatenulatum JCM 1200 type strain presented an approximately 1.5 kb fragment by ERIC and a 800 bp fragment by REP. These two fragments were not detectable in other species of bifidobacteria. This study demonstrates the presence of ERIC and REP-like elements in the genome of bifidobacteria and other probiotic bacteria. PCR technique was also used to detect the presence of bsh gene in Bifidobacterium longum B8536. In this regards, a pair of PCR primers for the rapid detection of bsh gene from B. longum have been synthesised and have revealed the bsh gene of appoximately 970 bp. The bsh gene was cloned and sequenced showing a high homology to bsh gene previously published. The resulting nucleotide sequence encodes a predicted protein of 317 amino acids with a molecular weight of approximately 35 kDa. The bsh gene from B. longum was also cloned and expressed in E. coli BL21-S1 using pRSET -A expression vector. The expressed protein was detected using immunoblot assay

Halal food authenticity: does it matter to you?

To the Muslims, the claim for Halal is not debatable. The current Halal food industry as required by 1.8 billion Muslims is estimated to be worth USD 1.6 trillion by 2018. In this respect, the Halal food industry is projected to be the fastest growing and most lucrative business segment in the very near future in view of the rapid increase in the Muslim population in the world. This enormous business potential will be further augmented by the preference, on the part of non-Muslims for the “safe, wholesome and clean” food concept referred to as the Halalan toyyiban. The Manufacture of Halal products, especially food requires an uncompromised understanding and knowledge of the Islamic laws and regulations along with the advancement in food processing and complex ingredients in the entire supply chain. However, unethical practices and fraud of the Halal logo is becoming a regular occurrence and poses a threat to the authenticity of halal products. Thus, authentication using specific, sensitive, easy-to-use and reliable scientific techniques based on DNA, proteins, lipids and metabolomics should be performed to complement the conventional audit-based certification process

Effect of lactic fermentation on the antioxidant capacity of Malaysian herbal teas

This study evaluated and compared the antioxidant capacity between freshly prepared and lactic fermented Malaysian herbal teas. Herbal teas are rich in antioxidants. Fermentation has been known to be the oldest and cost effective method with the ability to preserve or improve food nutritional qualities. Information on the antioxidant capacity of lactic fermented food or beverage is still lacking. Hence, the objective of this study is to determine the changes in the antioxidant properties of Malaysian herbal teas after being subjected to lactic fermentation. Commercially available local herbal teas were used for this study. Herbal teas such as “Allspice”, “Scaphium”, “Gora” and “Cinnamon” were purchased from the local store in Malaysia and were subjected to 24-hour lactic fermentation. Lactic fermented herbal teas were analyzed for their total phenolic, total flavonoid and antioxidant properties via DPPH, FRAP, and β-carotene linoleate bleaching assay. All lactic fermented herbal teas exhibited higher phenolic contents, flavonoid contents and antioxidant properties compared to the freshly-prepared herbal teas with majority showing significant changes (p < 0.05) in FRAP and β-carotene bleaching assay. Lactic fermented herbal teas also showed an increase in antioxidant capacity in DPPH assay, however non-significant changes were observed

Evaluation of electric and magnetic fields distribution and SAR induced in 3D models of water containers by radiofrequency radiation using FDTD and FEM simulation techniques

In this study, two software packages using different numerical techniques FEKO 6.3 with Finite-Element Method (FEM) and XFDTD 7 with Finite Difference Time Domain Method (FDTD) were used to assess exposure of 3D models of square, rectangular, and pyramidal shaped water containers to electromagnetic waves at 300, 900, and 2400 MHz frequencies. Using the FEM simulation technique, the peak electric field of 25, 4.5, and 2 V/m at 300 MHz and 15.75, 1.5, and 1.75 V/m at 900 MHz were observed in pyramidal, rectangular, and square shaped 3D container models, respectively. The FDTD simulation method confirmed a peak electric field of 12.782, 10.907, and 10.625 V/m at 2400 MHz in the pyramidal, square, and rectangular shaped 3D models, respectively. The study demonstrated an exceptionally high level of electric field in the water in the two identical pyramid shaped 3D models analyzed using the two different simulation techniques. Both FEM and FDTD simulation techniques indicated variations in the distribution of electric, magnetic fields, and specific absorption rate of water stored inside the 3D container models. The study successfully demonstrated that shape and dimensions of 3D models significantly influence the electric and magnetic fields inside packaged materials; thus, specific absorption rates in the stored water vary according to the shape and dimensions of the packaging materials.Comment: 22 pages, 30 figures and 2 table

Purification of pectinase from mango (Mangifera indica L. cv. Chokanan) waste using an aqueous organic phase system: a potential low cost source of the enzyme

As a novel method of purification, an aqueous organic phase system (AOPS) was employed to purify pectinase from mango waste. The effect of different parameters, such as the alcohol concentration (ethanol, 1-propanol, and 2-propanol), the salt type and concentration (ammonium sulfate, potassium phosphate and sodium citrate), the feed stock crude load, the aqueous phase pH and NaCl concentration, were investigated in the recovery of pectinase from mango peel. The partition coefficient (K), selectivity (S), purification factor (PF) and yield (Y, %) were investigated in this study as important parameters for the evaluation of enzyme recovery. The desirable partition efficiency for pectinase purification was achieved in an AOPS of 19% (w/w) ethanol and 22% (w/w) potassium phosphate in the presence of 5% (w/w) NaCl at pH 7.0. Based on the system, the purification factor of pectinase was enhanced 11.7, with a high yield of 97.1%

Characterization of pectinase from mango (Mangifera indica Cv. chokanan) peel

Today, pectinase has emerged as an integral part of the food and feed industries. Plant peel could be a potential source of pectinase, which has been extracted and purified from mango (Mangifera indica cv. Chokanan) peel using the aqueous two-phase system (ATPS). In the present study, the effects of temperature, pH and metal ions on the stability and activity of pectinase were investigated. In addition, the molecular weight of this enzyme was determined as 31 kDa with SDS-PAGE. Pectinase showed the highest enzyme activity at 60ºC for 30 min after incubation at different temperatures (20 to 80ºC). Also, this enzyme has been shown to be thermostable because more than 90% of residual enzyme activity was retained at temperatures of 20 to 60ºC for 30 min. Pectinase was incubated in different pH from 3 to 9 and the highest enzyme activity was achieved at pH 8. Furthermore, the enzyme was stable at pH 5 to 9 after enzyme incubation at different pH for 24 h at 4ºC. Activity of the enzyme was significantly decreased at pH 3 and 9 due to the protein denaturation. Pectinase activated by Ca2+ showed that this cation has an important effect on activity and stability of the enzyme; but Li+, Na+ and K+ had no effect on its activity. Also, the reduction in the activity of pectinase was observed in the presence of Fe2+, Cu2+, Mn2+, Zn2+ and Al3+. Therefore, pectinase extracted from mango peel has potential applications in various industries like food and feed because it is thermostable under high temperatures in either alkaline medium or when there is the presence of metal ions

Purification of a novel protease enzyme from kesinai plant (Streblus asper) leaves using a surfactant–salt aqueous micellar two-phase system: a potential low cost source of enzyme and purification method

Serine protease from kesinai leaves was purified for the first time by a surfactant–polymer aqueous micellar two-phase system. The effectiveness of different types and concentrations of non-ionic surfactants (Pluronic series and X-114) on the partitioning behaviour of the protease was evaluated. The results showed that the enzyme preferentially partitioned into the bottom surfactant-rich phase, while the hydrophilic amino acid preferred the top aqueous phase. This distribution of the enzyme is due to the hydrophobic interaction of the serine protease with the hydrophobic lid of the micelle core in the bottom phase. The influence of different types of salts (K2SO4, KH2PO4, KCl and KNO3) on the purification and selectivity of the enzyme was determined. The protease partitioning in the bottom phase increased in the presence of KNO3, which confirmed that the salt was able to improve the protein solubility in bottom phase and increase the hydrophobic interaction between the two phases. In addition, the protease from the bottom phase was re-extracted to a new aqueous phase solution to remove and recycle the surfactant. Addition of potassium thiocyanate led to the partitioning of the enzyme in top aqueous phase due to high ionic strength of SCN−, which forced the lighter micellar phase toward the upper position of the system. A high purification factor (10.3) and yield of 92 % of the enzyme were achieved in a solution of 31 % of Pluronic L61 using 0.3 % KNO3 and 50 % crude feedstock at pH 7.0

Characterization of polyphenol oxidase from mango (Mangifera indica L. cv. Chokanan) peel

Plant polyphenol oxidase showed positive effect in the production of coca, black tea and flavonoid-derived colorants and antioxidants. High activity and stability in a wide range of pH and temperature of plant enzyme make it suitable and also inexpensive for use in industry. For these reasons, there is growing interest in seeking more plant sources of polyphenol oxidase. Mango (Mangifera indica L. cv. Chokanan) peel can be a potential source of polyphenol oxidase, which has been extracted and purified from peel of mango using the aqueous two-phase system (ATPS). In the present study, the effects of different temperatures, pH, inhibitors and metal ions on the stability and activity of polyphenol oxidase from mango peel were investigated. In addition, the molecular weight of this enzyme was estimated at 133 kDa by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The highest enzyme activity of polyphenol oxidase to catalyze catechol in sodium phosphate buffer was achieved at 55°C at pH 5.5. Furthermore, the enzyme was stable at temperatures of 10 to 60°C and pH 3 to 6. Beta-mercaptoethanol, ascorbic acid, l-cysteine and pyrogallol were effective inhibitors of the enzyme. Also, activity of polyphenol oxidase was increased in the presence of some metal ions such as Ca2+, Mg2+ and Cu2+ which implies that the enzyme involved metal ions. Therefore, polyphenol oxidase extracted from mango (Mangifera indica L. cv. Chokanan) peel has potential applications in various industries because it is thermostable under high temperatures in either acidic medium, or when there is the presence of metal ions