A specialized DNA extraction method and a SYBR Green quantitative polymerase chain reaction (SyG-qPCR) assay were combined to generate a ready-to-use kit for rapid detection of porcine admixtures in processed meat products. Our qPCR assay utilized repetitive LINE-1 elements specific to the genome of Sus scrofa domesticus (pig) as a target and incorporated internal controls. We improved the genomic DNA extraction method, and reduced extraction times to the minimum. The method was validated for specificity, sensitivity (0.001% w/w) and robustness, and values were compared with those of a commercially available kit. We also tested our method using 121 processed food products and consistently detected amplification only in samples containing pork. Due to its efficiency and cost-effectiveness, our method represents a valuable new method for detecting food adulteration with pork that is superior to existing quality control approaches.
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.
Thiol groups of cysteine (Cys) residues in proteins react with quinones, oxidation products of polyphenols, to form protein-polyphenol adducts. The aim of the present work was to quantify the amount of adduct formed between Cys residues and 4-methylcatechol (4MC) in minced beef. A Cys-4MC adduct standard was electrochemically synthesized and characterized by liquid chromatography-mass spectrometry (LC-MS) as well as NMR spectroscopy. Cys-4MC adducts were quantified after acidic hydrolysis of myofibrillar protein isolates (MPIs) and LC-MS/MS analysis of meat containing either 500 or 1500 ppm 4MC and stored at 4 °C for 7 days under a nitrogen or oxygen atmosphere. The concentrations of Cys-4MC were found to be 2.2 ± 0.3 nmol/mg MPI and 8.1 ± 0.9 nmol/mg MPI in meat containing 500 and 1500 ppm 4MC, respectively, and stored for 7 days under oxygen. The formation of the Cys-4MC adduct resulted in protein thiol loss, and ca. 62% of the thiol loss was estimated to account for the formation of the Cys-4MC adduct for meat containing 1500 ppm 4MC. Furthermore, protein polymerization increased in samples containing 4MC as evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the polymerization was found to originate from protein-polyphenol interactions as evaluated by a blotting assay with staining by nitroblue tetrazolium.
The organic foods’ market is becoming one of the rapidly growing sections in agricultural economies in the world. During the last two decades, food-borne outbreaks associated with fresh produce have rapidly increased. E. coli O57:H7, the caustic agent of acute hemorrhagic diarrhea and abdominal cramps, is mainly associated with meat and poultry product outbreaks but frequent outbreaks linked to the consumption of vegetables have been reported. The aim of this study was to investigate prevalence of E. coli O157:H7 in some organic foods. A total of 230 organic food samples including four-winged bean, tomato, white radish, red cabbage, chinese cabbage, lettuce, cucumber and chicken form retailed groceries and supermarkets in Malaysia were investigated. Low prevalence of E. coli O157:H7 was detected in organic vegetables and chickens. The estimated quantity of E. coli O157:H7 in all samples ranged from 2400 MPN/g. The overall MPN/g estimate of E. coli O157:H7 in the samples from organic groceries was higher than supermarket with the maximum of >2400 MPN/g. Most of the samples from supermarket showed a minimum of
Four formulations were processed into frankfurters with different ratios of mechanically deboned chicken meat (MDCM) and cooked chicken skin (CCS) i.e. 80/0, 70/10, 60/20 and 50/30. The products were evaluated for proximate composition, cholesterol content, colour; 'L' value (lightness) and 'a' value (redness), percentage of cooking loss, physical measurements (shearforce-kgf and folding test), thiobarbituric acid value (TBA) and taste panel evaluation. The increment of CCS in the frankfurters increased the contents of moisture, ash, protein, fat, cholesterol, the lightness ('L' value) and redness ('a' value). After 3 months of frozen storage, the increment continued except for the moisture contents for formulations with 20 and 30% CCS. The lipid oxidation (TBA value) and cooking loss were lowered in formulations with CCS. After 3 months of frozen storage, TBA value decreased, while the cooking loss increased for all the formulations. The addition of CCS increased hardness of the frankfurters but affected folding ability, with formulation with 10% CCS scoring better grade. Sensory evaluation was carried out using 30 untrained panelists to evaluate aroma, colour, appearance, hardness, juiciness, chicken taste, oily taste, rancid taste and overall acceptance of the products. The addition of CCS in the frankfurters at 10 and 20% resulted in products with taste and texture that were acceptable after 3 months of frozen storage.
Five different brands of Malaysian commercial beef frankfurters were analyzed for quality characteristics. The proximate contents showed significant differences among the samples. The range of moisture content was 63.0-73.9%; the protein content was 10.63-16.43% while the fat content was 1.71-12.22%. The lightness value (L*) of the uncooked frankfurters, which was in the range of 47.02-52.28, was significantly different among the samples. The lightness of the cooked frankfurters, showed a decrease in all the samples compared to the uncooked samples. No significant differences were observed for the folding test; where all samples showed no cracks after they were folded in half. However, significant differences were observed for the texture analysis. The hardness, cohesiveness, chewiness, springiness, gumminess and shear force ranged between 4.59-10.30 kg, 0.26-0.35, 16.15-51.72 kgmm, 12.73-14.79 mm, 1.17-3.49 kg and 1.67-7.08 kg respectively. The results of the study showed that Malaysian commercial beef frankfurters were significantly different in their physicochemical properties.
This study provides a comparative analysis of the effects of pre-slaughter penetrative and non-penetrative stunning and post-slaughter stunning on meat quality attributes in longissimus lumborum (LL) and semitendinosus (ST) muscles in heifers. Ten animals were assigned to each of four treatment groups: i) animals were subjected to conventional Halal slaughter (a clean incision through the structures at the front of the upper neck - the trachea, oesophagus, carotid arteries and jugular veins) and post-cut penetrating mechanical stun within 10 to 20 s of the neck cut (Unstunned; US); ii) high power non-penetrating mechanical stunning followed by the neck cut (HPNP); iii) low power non-penetrating mechanical stunning followed by the neck cut (LPNP); and iv) penetrative stunning using a captive bolt pistol followed by the neck cut (P). For each carcass, muscle samples were removed within 45 min of slaughter, portioned and analysed for pH, cooking loss, water holding capacity (WHC), tenderness (WBS), lipid oxidation (TBARS) and color, over a two week storage period. Stunning did not affect pH and cooking loss. Significant differences in water holding capacity, tenderness, lipid oxidation and color were present at different storage time points. HPNP stunning resulted in lower WHC and color values, particularly lightness (L*), higher TBARS values and peak force values compared with those stunned using LPNP, P and US. These adverse effects on quality were mostly encountered in the ST muscle. In conclusion, the meat quality achieved using P, LPNP and US treatments was comparable, and no treatment stood out as considerably better than another.
The last decade has witnessed dramatic changes in global food consumption patterns mainly because of population growth and economic development. Food substitutions for healthier eating, such as swapping regular servings of meat for protein-rich crops, is an emerging diet trend that may shape the future of food systems and the environment worldwide. To meet the erratic consumer demand in a rapidly changing world where resources become increasingly scarce due largely to anthropogenic activity, the need to develop crops that benefit both human health and the environment has become urgent. Legumes are often considered to be affordable plant-based sources of dietary proteins. Growing legumes provides significant benefits to cropping systems and the environment because of their natural ability to perform symbiotic nitrogen fixation, which enhances both soil fertility and water-use efficiency. In recent years, the focus in legume research has seen a transition from merely improving economically important species such as soybeans to increasingly turning attention to some promising underutilized species whose genetic resources hold the potential to address global challenges such as food security and climate change. Pulse crops have gained in popularity as an affordable source of food or feed; in fact, the United Nations designated 2016 as the International Year of Pulses, proclaiming their critical role in enhancing global food security. Given that many studies have been conducted on numerous underutilized pulse crops across the world, we provide a systematic review of the related literature to identify gaps and opportunities in pulse crop genetics research. We then discuss plausible strategies for developing and using pulse crops to strengthen food and nutrition security in the face of climate and anthropogenic changes.
This work was investigated the protein solubility properties of meat from chicken in different
body part. The effects of fresh and freezing condition were studied on the protein solubility as
a functional property of slaughter and non slaughtering chicken meat. Solubility of proteins
was significantly reduced for slaughtering fresh meat and in contrast, non slaughtering fresh
meat shows the higher protein solubility. On the other hand, frozen storage meat showed the
difference amount of protein solubility between slaughtering and non slaughtering condition
meat. Freezing condition also showed that the different solubility of different body part meat.
The protein solubility of some parts was significantly increased and some were decreased
between the slaughtering and non slaughtering condition.
Protein efficiency ratio (PER) and protein digestibility are important parameters used in protein quality determination. Protein nutritive values of selected protein sources: buffalo meat, casein, soy protein isolate, and tempeh, with sodium caseinate as a reference formulation, were evaluated. Determination of proximate analysis, protein quality and protein digestibility were monitored. Procedures for evaluation of protein quality and digestibility included PER using the rat bioassay and in vivo Apparent Protein Digestibility (APD). The rats fed with buffalo meat had the highest mean increase in body weight (102.73g±8.95) while rats fed with tempeh had the lowest mean for increase in body weight (16.34g±9.11). Although the mean for body weight gained showed significant differences between all treatments (P0.05) found between casein and soy protein isolate for total food intake. For the PER value, buffalo meat had the highest value (2.99), followed by sodium caseinate (2.41), casein (1.93), soy protein isolate (1.52) and tempeh (1.10). The PER value for buffalo meat (2.99) was higher than sodium caseinate (2.41) while the rest of the treatment were comparatively lower than sodium caseinate. For the in vivo apparent protein digestibility, tempeh had the highest value (91.41%±3.76), followed by casein (91.34%±3.15), buffalo meat (90.79%±1.44), soy protein isolate (89.52%±2.96) and sodium caseinate (89.47%±2.31).
The role of selenium (Se) in human health and diseases has been discussed in detail in several recent reviews, with the main conclusion being that selenium deficiency is recognised as a global problem which urgently needs resolution. Since selenium content in plant-based food depends on its availability from soil, the level of this element in food and feeds varies among regions. In general, eggs and meat are considered to be good sources of selenium in human diet. When considering ways to improve human selenium intake, there are several potential options. These include direct supplementation, soil fertilisation and supplementation of food staples such as flour, and production of functional foods. Analysing recent publications related to functional food production, it is evident that selenium-enriched eggs can be used as an important delivery system of this trace mineral for humans. In particular, developments and commercialisation of organic forms of selenium have initiated a new era in the availability of selenium-enriched products. It has been shown that egg selenium content can easily be manipulated to give increased levels, especially when organic selenium is included in hens' diet at levels that provide 0.3-0.5 mg/kg selenium in the feed. As a result, technology for the production of eggs delivering approximately 50% (30-35 microg) of the human selenium RDA have been developed and successfully tested. Currently companies all over the world market selenium-enriched eggs including the UK, Ireland, Mexico, Columbia, Malaysia, Thailand, Australia, Turkey, Russia and the Ukraine. Prices for enriched eggs vary from country to country, typically being similar to free-range eggs. Selenium-enriched chicken, pork and beef can also be produced when using organic selenium in the diet of poultry and farm animals. The scientific, technological and other advantages and limitations of producing designer/modified eggs as functional foods are discussed in this review.
A study was carried out to determine the process parameters and optimization for the hydrolysis of protein precipitate from cockle (Anadara granosa) meat wash water. Precipitation of the protein in the wash water was done using pH manipulation (pH3-8). The precipitate was hydrolyzed using hydrochloric acid (HCl) and optimized for HCl volume, HCl concentration and hydrolysis time using response surface methodology (RSM) based on a central composite rotatable design. Based on the results, hydrolysis of cockle meat wash water precipitate was carried out by precipitation of the wash water at pH4. Optimum condition for the hydrolysis of 2.0 g of cockle meat wash water precipitate was 25 mL of 1 N HCl for 10 h which resulted in nitrogen content (NC) of 0.7% and degree of hydrolysis (DH) of 55%. NC and DH were significantly influenced only by the hydrolysis time.
We report for the first time on the prevalence, antibiotic resistance and RAPD types of Campylobacter species in ducks and duck related environmental samples in Malaysia. Samples were examined by enrichment in Bolton Broth followed by plating onto modified Charcoal Cefoperazone Deoxycholate agar (mCCDA) and/or plating directly onto mCCDA. A total of 643 samples were screened, and the prevalence of Campylobacter spp. in samples from different sources ranged from 0% to 85%. The method of isolation had a significant (P<0.05) effect on the isolation rate. One hundred and sixteen Campylobacter isolates, comprising of 94 Campylobacter jejuni, 19 Campylobacter coli and three Campylobacter lari, were examined for their sensitivity to 13 antibiotics. Majority of the C. jejuni isolates were resistant to cephalothin (99%), tetracycline (96%), suphamethoxazole/trimethoprim (96%), and very few were resistant to gentamicin (5%), chloramphenicol (7%) and erythromycin (1%). All C. coli isolates were resistant to cephalothin, nalidixic acid, norfloxacin and tetracycline but susceptible to chloramphenicol, erythromycin and gentamicin. The three C. lari isolates were resistant to all the antibiotics tested except chloramphenicol and gentamicin (1/3 and 2/3 susceptible, respectively). Genetic diversity of Campylobacter isolates were determined using random amplification of polymorphic DNA (RAPD). C. jejuni and C. coli isolates belong to fifty-eight and twelve RAPD types, respectively.
The aim of the present study was to investigate the prevalence of Salmonella spp., Salmonella Enteritidis and Salmonella Typhimurium in retail beef from different retail markets of Selangor area, as well as, to assess their pathogenic potential and antimicrobial resistance. A total of 240 retail beef meat samples (chuck = 60; rib = 60; round = 60; sirloin = 60) were randomly collected. The multiplex polymerase chain reaction (mPCR) in combination with the most probable number (MPN) method was employed to detect Salmonella spp., S. Enteritidis and S. Typhimurium in the meat samples. The prevalence of Salmonella spp., S. Enteritidis and S. Typhimurium in 240 beef meat samples were 7.50, 1.25, and 0.83%, respectively. The microbial loads of total Salmonella was found in the range of <3 to 15 MPN/g. Eight different serovars of Salmonella were identified among the 23 isolates, and S. Agona was the predominant serovar (26.09%). Interestingly, all the Salmonella isolates were resistant to penicillin, erythromycin and vancomycin, but the sensitivity was observed for tetracycline, gentamicin and amoxicillin/clavulanic acid. All 23 isolates were resistant to at least three antibiotics. Two S. Typhimurium isolates (8.70%) exhibited the highest multiple antibiotic resistance (MAR) index value of 0.56 which shown resistance to nine antibiotics. PCR analysis of virulence genes showed that all Salmonella isolates (100%) were positive for the invA gene. Meanwhile, pefA was only identified in S. Enteritidis and S. Typhimurium. The findings in this study indicate that retail beef products tested were widely contaminated with multi-drug resistant (MDR) Salmonella and various virulence genes are present among the isolated Salmonella serovars.
Listeria spp. are ubiquitous in nature and can be found in various environmental niches such as soil, sewage, river water, plants, and foods, but the most frequently isolated species are Listeria monocytogenes and Listeria innocua. In this study, the presence of Listeria spp. in raw chicken meat and chicken-related products sold in local markets in Klang Valley, Malaysia was investigated. A total of 44 Listeria strains (42 L. innocua and 2 L. welshimeri) were isolated from 106 samples. Antibiotic susceptibility tests of the L. innocua strains revealed a high prevalence of resistance to clindamycin (92.9%), ceftriaxone (76.2%), ampicillin (73.8%), tetracycline (69%), and penicillin G (66.7%). Overall, 31 L. innocua and 1 L. welshimeri strain were multidrug resistant, i.e., nonsusceptible to at least one antimicrobial agent in three or more antibiotic classes. The majority of the L. innocua strains were placed into five AscI pulsogroups, and overall 26 distinct AscI pulsotypes were identified. The detection of multidrug-resistant Listeria strains from different food sources and locations warrants attention because these strains could serve as reservoirs for antimicrobial resistance genes and may facilitate the spread and emergence of other drug-resistant strains.
A survey was carried out to determine the prevalence of Toxoplasma gondii antibodies among cattle farmers and cattle in the Gombak District, Selangor. A total of 79 human and 73 cattle serum samples were tested for Toxoplasma gondii antibodies by the immunofluorescent technique (IFAT). Results of the survey showed that anti-Toxoplasma gondii antibodies were found in 27.8% of the farmers, while in cattle the positive rate was only 3.8%. The prevalence rate obtained in this study did not differ much from the prevalence reported in previous studies. This suggests that the same degree of risk to this infection exists in the community. In view of the relatively low antibody prevalence in cattle, the risk of acquiring this infection from consuming undercooked beef is realtively low. Further survey on larger sample size is needed to validate the observation.
Fermented fish and meat samples were purchased from supermarket and wet market for microbiological analysis of Listeria species and Listeria monocytogenes contamination. Listeria species were isolated from 17 (73.9%) of 23 samples of imported frozen beef, 10 (43.5%) of the 23 samples of local beef and 14 (56%) of the 25 samples of fermented fish from wet market. Listeria monocytogenes occurred in 15 (75%) of the frozen beef samples, 6 (30.4%) of the 23 samples of local meat and 3 (12%) of the 25 samples from fermented fish. Listeria species was not isolated from any of the 23 samples of imported frozen beef from supermarket and from the 5 samples of buffalo meat examined. This highlights the possibility of Listeria spp or L. monocytogenes to persist in meat and fermented fish in wet market and raises the problem of illness due to the handling and consumption of Listeria-contaminated meat or fermented fish are likely as evidence by the high contamination rates of samples sold at the wet market.
A total of 112 burger patties (35 beef burger patties, 39 chicken burger patties and 38 fish burger patties) which are commercially available at retail level were investigated for the presence and number of Listeria monocytogenes. These samples were analyzed using MPN-PCR method and conventional culturing methods. L. monocytogenes was detected in 33.3% of chicken burger patties, 22.9% of beef patties, and 10.5% of fish patty samples. From all contaminated raw burger patties, the estimated count of L. monocytogenes was ranged from 3 to 75 MPN/g. The results suggest that burger act as a potential source of listeriosis if the contaminated burger patty is consumed without adequate cooking. The risk associated with consumption of these samples was found to be high particularly for processed food at retail level in Malaysia. Therefore, food manufacturers play an important role in monitoring the manufacturing process and conduct a periodical surveillance on microbiological quality assessment on the processing plants. Besides, there is a need to increase awareness of consumers and food handlers to practice proper cooking of the burger patties before the point of consumption, to reduce the risk of listeria infection.
Kelantan is a chiefly agrarian state with abundant small-holder ruminant farms in the East Coast economic Region of Malaysia. Ectoparasitism affects small ruminant production in Malaysia. It often causes reduction in meat quality and milk production which affect the farmers' income. To date, no report for the prevalence of ectoparasitism on small ruminant in Kelantan compared to other state in Malaysia. This study aims to determine the prevalence and associated risk factor of ruminant ectoparasitism in Kelantan. Ectoparasites were collected by manual picking and skin scrapping from 462 sheep and goats in Kelantan between April and September 2017 (during dry season). 60% of the sampled animals were infested with at least one species of the ectoparasites. In this study, lice and ticks were the most prevalent ectoparasites on small ruminant, which were 43.64% and 22.98%, respectively. The high biotic potential of lice population on host might be one of the factors they become the most prevalent species found on the animals. There was no significant relationship between ectoparasitism prevalence and species of small ruminants (χ2 = 1.12, p = 0.293). However, there was significant variations in prevalence between the regions where the animals were sampled from (χ2 = 30.25, p = 0.002) and farm management system for both species. This present study provides baseline epidemiological data on the prevalence of ectoparasitism in small ruminant. This information is useful for the formulation of prevention and control measures in order to enhance ruminant productivity in Kelantan.
A total of 216 chicken offal samples (chicken liver = 72; chicken heart = 72; chicken gizzard = 72) from wet markets and hypermarkets in Selangor, Malaysia, were examined for the presence and density of Listeria monocytogenes by using a combination of the most probable number and PCR method. The prevalence of L. monocytogenes in 216 chicken offal samples examined was 26.39%, and among the positive samples, the chicken gizzard showed the highest percentage at 33.33% compared with chicken liver (25.00%) and chicken heart (20.83%). The microbial load of L. monocytogenes in chicken offal samples ranged from <3 to 93.0 most probable number per gram. The presence of L. monocytogenes in chicken offal samples may indicate that chicken offal can act as a possible vehicle for the occurrence of foodborne listeriosis. Hence, there is a need to investigate the biosafety level of chicken offal in Malaysia.