Md. Abdul Kader

Gelatin Controversies in Food, Pharmaceuticals and Personal Care Products: Authentication Methods, Current Status and Future Challenges.

ABSTRACT Gelatin is a highly purified animal protein of pig, cow, and fish origins and is extensively used in food, pharmaceuticals, and personal care products. However, the acceptability of gelatin products greatly depends on the animal sources of the gelatin. Porcine and bovine gelatins have attractive features but limited acceptance because of religious prohibitions and potential zoonotic threats, whereas fish gelatin is welcomed in all religions and cultures. Thus, source authentication is a must for gelatin products but it is greatly challenging due to the breakdown of both protein and DNA biomarkers in processed gelatins. Therefore, several methods have been proposed for gelatin identification, but a comprehensive and systematic document that includes all of the techniques does not exist. This up-to-date review addresses this research gap and presents, in an accessible format, the major gelatin source authentication techniques, which are primarily nucleic acid and protein based. Instead of presenting these methods in paragraph form which needs much attention in reading, the major methods are schematically depicted, and their comparative features are tabulated. Future technologies are forecasted, and challenges are outlined. Overall, this review paper has the merit to serve as a reference guide for the production and application of gelatin in academia and industry and will act as a platform for the development of improved methods for gelatin authentication.

Lab-on-a-Chip-Based PCR-RFLP Assay for the Detection of Malayan Box Turtle (Cuora amboinensis) in the Food Chain and Traditional Chinese Medicines

The Malayan box turtle (Cuora amboinensis) (MBT) is a vulnerable and protected turtle species, but it is a lucrative item in the illegal wildlife trade because of its great appeal as an exotic food item and in traditional medicine. Although several polymerase chain reaction (PCR) assays to identify MBT by various routes have been documented, their applicability for forensic authentication remains inconclusive due to the long length of the amplicon targets, which are easily broken down by natural decomposition, environmental stresses or physiochemical treatments during food processing. To address this research gap, we developed, for the first time, a species-specific PCR-restriction fragment length polymorphism (RFLP) assay with a very short target length (120 bp) to detect MBT in the food chain; this authentication ensured better security and reliability through molecular fingerprints. The PCR-amplified product was digested with Bfa1 endonuclease, and distinctive restriction fingerprints (72, 43 and 5 bp) for MBT were found upon separation in a microfluidic chip-based automated electrophoresis system, which enhances the resolution of short oligos. The chances of any false negative identifications were eliminated through the use of a universal endogenous control for eukaryotes, and the limit of detection was 0.0001 ng DNA or 0.01% of the meat under admixed states. Finally, the optimized PCR-RFLP assay was validated for the screening of raw and processed commercial meatballs, burgers and frankfurters, which are very popular in most countries. The optimized PCR-RFLP assay was further used to screen MBT materials in 153 traditional Chinese medicines of 17 different brands and 62 of them were found MBT positive; wherein the ingredients were not declared in product labels. Overall, the novel assay demonstrated sufficient merit for use in any forensic and/or archaeological authentication of MBT, even under a state of decomposition.

Quantitative Tetraplex Real-Time Polymerase Chain Reaction Assay with TaqMan Probes Discriminates Cattle, Buffalo, and Porcine Materials in Food Chain

Cattle, buffalo, and porcine materials are widely adulterated, and their quantification might safeguard health, religious, economic, and social sanctity. Recently, conventional polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) assays have been documented but they are just suitable for identification, cannot quantify adulterations. We described here a quantitative tetraplex real-time PCR assay with TaqMan Probes to quantify contributions from cattle, buffalo, and porcine materials simultaneously. Amplicon-sizes were very short (106-, 90-, and 146-bp for cattle, buffalo, and porcine) because longer targets could be broken down, bringing serious ambiguity in molecular diagnostics. False negative detection was eliminated through an endogenous control (141-bp site of eukaryotic 18S rRNA). Analysis of 27 frankfurters and 27 meatballs reflected 84-115% target recovery at 0.1-10% adulterations. Finally, a test of 36 commercial products revealed 71% beef frankfurters, 100% meatballs, and 85% burgers contained buffalo adulteration, but no porcine was found in beef products.

Effects of Animal Proteins in Practical Diets on Growth and Economic Performance of Climbing Perch, Anabas testudineus (Bloch)

A ten-week feeding trial was carried out to evaluate animal proteins, either alone or in combination (1:1:1), on the growth performances and economic benefit of climbing perch, Anabas testudineus. Four iso-proteinous (40%) diets were prepared, using fishmeal, protein concentrate, and meat and bone meal as major protein source in diets 1, 2 and 3, respectively, while diet 4 consisted of an equal proportion (1:1:1) of the above protein sources. Triplicate groups of fish (0.53 ± 0.02 g) were randomly stocked in 12 earthen ponds at a stocking density of 40,000 ha−1. Fish fed diet 4 had significantly (P < 0.05) higher percent weight gain and SGR % per day−1 compared to the fish fed diets 1, 2, or 3. Significantly higher apparent protein efficiency ratio (1.25) and lower apparent feed conversion ratio (2.00) were also observed in fish fed diet 4. Total yield (1,123 kg ha−1) and net profit (Tk. 116,569 ha−1) were also higher in fish fed diet 4. It can be concluded that the combination of fishmeal, protein concentrate, and meat and bone meal is more effective for climbing perch than any of the three protein sources alone.

The use of fermented soybean meal and squid by-product blend as a substitute for fish meal in practical diets for climbing perch, Anabas testudienus (Bloch, 1792), pond culture: Growth performance and economics analysis

ABSTRACT A feeding trial was conducted to evaluate the effects of different ratios of fish meal (FM) to a fermented blend (FB: fermented soybean meal and squid by-product blend) on growth and economic performance of climbing perch, Anabas testudineus (Bloch, 1792), in earthen ponds. Five diets were prepared where FB was substituted for 0 (D1), 25 (D2), 50 (D3), 75 (D4), or 100% FM protein (D5), while a commercial diet (D6) was used for comparison. Triplicate groups of fish (1.03 ± 0.07 g) were fed twice daily up to satiation for 70 days in earthen ponds. No significant (P > 0.05) differences were found in growth performance of fish fed D1, D2, D3, and D6 diets, while fish growth decreased significantly (P < 0.05) in D4 and D5 groups. Feed intake decreased (P < 0.05) with the increasing levels of FB with no significant difference in the feed conversion ratio, which ranged from 1.98 to 2.08. The fish survival decreased significantly in D5 alone. While considering the overall production, the total yield and economic return were found to be highest in D2 (64176.7 TK ha−1) and lowest in D5 (14418.3 Tk. ha−1). Based on the present experimental condition, it is concluded that FB can economically be included in aquafeed as a substitute for up to 50% of FM protein from the diets of climbing perch.