The safety level of microwaved foods remains at vague as this subject was less addressed
scientifically. A study was initiated to address the matter by investigating on the
survivability of Salmonella and Shiga-toxigenic Escherichia coli (STEC) O157 in
microwave heated ready-to-eat (RTE) foods using the Most Probable Number coupled
Polymerase Chain Reaction (MPN-PCR) technique. A total of 329 samples of various
ready-to-eat (RTE) convenience meals were collected around Wilayah Persekutuan Kuala
Lumpur and Selangor regions. Salmonella was positively identified in 66 samples (20.1%,
According to the World Health Organisation (WHO), globally 600 million people suffer
from food-borne diseases (FBD), and 420,000 people die as a result. The European Food
Safety Authority (EFSA) has stated that FBD are linked to the food industry, with the
most common means of transmission being due to poor food handling and hygiene by
food handlers working in the food industry. The aim of this research was to investigate the
effectiveness of mandatory food handler training programmes (FHTP) to prevent FBD in
Malaysia and Ireland. To do this, the FHTP existing in Malaysia and Ireland were
analysed, in addition to the legislation they fall under in each respective country.
Effectiveness was determined by investigating the level of food safety knowledge (FSK)
and food safety practices (FSP) of food handlers in Malaysia and Ireland. A systematic
literature review (SLR) and a narrative literature review (NLR) were conducted for this
research. The SLR was based on the PRISMA diagram, using the Confidence in the
Evidence from Reviews of Qualitative research (CERQual) approach to evaluate the
studies used for this research. A total of 8 Malaysian studies and 1 Irish study were used to
determine the level of FSK and FSP of food handlers in each respective country, to
examine the effectiveness of FHTP. The results of the studies used for this research have
depicted overall good FSP and FSK of food handlers in Malaysia and Ireland; yet trends
continue to show that food handlers are one of the biggest contributors to FBD,
demonstrating that FHTP are not effective in preventing FBD. The findings from this
research highlights that although these trainings can be an effective tool to prevent FBD, if
they are not executed correctly, food handlers will continue to contribute to FBD.
The revolution of agriculture through biotechnology have produced large-scale of genetically
modified crops which brought up a controversy on the safety usage of genetically modified
organisms (GMOs). It has been implemented globally that all GMO products and its derived
ingredients should have regulations on the usage and labelling. Thus, it is necessary to develop
methods that allow rapid screening of GMO products to comply with the regulations. This
study employed a reliable and flexible multiplex polymerase chain reaction (PCR) method for
the rapid detection of transgenic elements in genetically modified soy and maize along with
the soybean LECTIN gene and maize ZEIN gene respectively. The selected four common
transgenic elements were 35S promoter (35S); Agrobacterium tumefaciens nopaline synthase
terminator (NOS); 5-enolypyruvylshikimate-3-phosphate synthase (epsps) gene; and Cry1Ab
delta-endotoxin (cry1Ab) gene. Optimization of the multiplex PCR methods were carried out
by using 1% Roundup ReadyTM Soybean (RRS) as the certified reference material for soybean
that produced fourplex PCR method detecting 35S promoter, NOS terminator, epsps gene and
soybean LECTIN gene and by using 1% MON810 as the certified reference material for maize
that produced triplex PCR method detecting 35S promoter, cry1Ab gene and maize ZEIN gene
prior to screening of the GMO traits in various food products and animal feeds. 1/9 (11.1%) of
the animal feed contained maize and 1/15 (6.7%) of the soybean food products showed positive
results for the detection of GMO transgenic gene. None of the maize food products showed
positive results for GMO transgenic gene. In total, approximately 4% of the food products
and animal feed were positive as GMO. This indicated GMOs have not widely entered the
food chain. However, it is necessary to have an appropriate screening method due to GMOs’
unknown potential risk to humans and to animals. This rapid screening method will provide
leverage in terms of being economically wise, time saving and reliable.
Vibrio parahaemolyticus is recognized as a frequent causal agent of human gastroenteritis due to the consumption of raw, undercooked or mishandled seafood in many Asian countries. The number of V. parahaemolyticus cases reported is on the rise, and this becomes a concern to the Asian countries as seafood is favoured by Asians. This study aimed to detect and quantify V. parahaemolyticus in raw oysters and to determine the risk associated with the consumption of raw oysters. A total of 30 oyster samples were collected and analysed in this study. MPN-PCR and MPN-Plating methods were employed and carried out concurrently to determine the prevalence of V. parahaemolyticus in raw oysters. The results showed that the prevalence of total V. parahaemolyticus in oysters was 50.00% (15/30) where the MPN/g range was < 3 – > 11000 MPN/g for MPN-PCR method, and 40.00% (12/30) where the MPN/g range was < 3 – 4300 MPN/g for MPN-Plating method. MPN-PCR method was able to estimate the level of virulence (tdh+ and trh+) V. parahaemolyticus in the raw oysters where 10.00% (3/30) of samples were identified to be in a range of 3 – 30 MPN/g. A microbial risk assessment was conducted based on the enumeration data obtained from MPN-PCR method using @risk. The probability of illness annually was 1.76 X 10-6 with a prediction of 31 cases to occur with respect to the exposed Malaysian population, while the rate per 100,000 people was estimated to be at 0.104. In addition, the antibiogram of V. parahaemolyticus was determined using Kirby Bauer Disk Diffusion Test and the results indicated that the isolates were highly resistant towards Bacitracin (100.00%), Vancomycin (100.00%) and were least resistant to Chloramphenicol (8.70%). The MAR index of the isolates ranged from 0.17 to 0.50. In accordance with the results from this study, the consumption of raw oysters is a risk factor for V. parahaemolyticus infection and proactive actions should be taken to reduce the risk of the pathogen in order to improve public health.
As the society begin to realize the importance of combating antimicrobial resistance, going
back to silver might be the solution. Silver has been known for its potential antimicrobial
activity since ancient times and, the development of nanoparticles has increased its potential
into becoming an antimicrobial agent that can be applied in broad-spectrum. Antimicrobial
resistance has spread into an irrepressible manner which requires drastic action plan as a number
of pathogenic bacteria began to acquire resistance genes. Methicillin Resistant Staphylococcus
aureus (MRSA) is one of the earliest reported resistant clones which is the center of this study.
This study focused on the dissemination and evolution of MRSA on its resistance towards
antibiotics. Disc Diffusion Test was employed to create the antibiograms of MRSA isolates. All
isolates showed resistance towards amoxicillin, ampicillin, cefazolin, oxacillin and penicillin.
In contrast, all isolates were susceptible towards erythromycin. The findings also discovered
isolates that were vancomycin-resistant (66.7%) and vancomycin-intermediate (33.3%). As the
efficacy of antibiotic treatment is at a question, we also investigated on the antimicrobial activity
of colloidal silver in the hope as an alternative treatment. Shiga Toxin producing Escherichia
coli (STEC) and MRSA (ATCC 33591) was tested using modified Quantitative suspension
test for the evaluation of bactericidal activity for chemical disinfectants and antiseptics based
on BS EN 1276:2009. The outcome of this study indicated that the colloidal silver is working
effectively against STEC and MRSA (ATCC 33591), showing killing percentages well above
99.0% at 4 minutes and 8 minutes of contact. Vancomycin-resistant S. aureus (VRSA) and
Vancomycin-intermediate S. aureus (VISA) were also tested and the results indicated that
VISA had higher killing percentages at 4 minutes (99.83%) and 8 minutes (99.85%) compared
to VRSA at 4 minutes (96.72%) and 8 minutes (98.35%). This opens a solution to the rising
problem of antimicrobial resistance.
Vibrio parahaemolyticus is well known to be abundantly distributed in marine, coastal and
estuarine environments. Since 1951, V. parahaemolyticus had been the source of numerous
outbreaks related to contaminated or mishandled seafood. However, V. parahaemolyticus
had been detected on other types of food. This issue has prompted this study to investigate
on the prevalence of V. parahaemolyticus in various food samples and determine the risk
associated with it. The results of the MPN-plating technique of the study indicated that V.
parahaemolyticus was detected in seafood (33.3%, 95% Confidence Interval [CI] 31.9 – 34.8 ,
94 – 290 MPN/g) and vegetables (10.0%, 95% CI 9.7 – 10.3 , 9.2 – 23 MPN/g) while negative
V. parahaemolyticus was detected in fruits (0.0%, 95% CI 0 – 1,
Escherichia coli O157:H7 is a major food-borne pathogen that has resulted in numerous
outbreaks around the world. Widespread distribution of the organism in various ecological
niches impedes the control measures. This study aimed to detect and quantify E. coli O157:H7
in beef sold in wet markets and hypermarkets in Malaysia and to determine the risk of E. coli
O157:H7 infection linked to consumption of beef. The rfbO157 and flicH7 primers targeted on
somatic antigen (O157) and flagellar antigen (H7) respectively of E. coli O157:H7 was used for
the MPN-PCR method. A total of 99 beef samples were collected from local wet markets and
hypermarkets. The highest E. coli O157:H7 contamination rate was observed in beef samples
collected from wet markets (89.50%), whereas the contamination rate in hyper market A and B
were compratively low (35.35 and 20% respectively). However, the microbial load was highest
in the beef samples from hypermarket A (1100 MPN/g) while E. coli O157:H7 bacterial load
in beef samples from hypermarket B and wet market ranged from 3 to 93 MPN/g and 3 to 240
MPN/g, respectively. Using the Quantitative Microbial Risk Assessment (QMRA) approach
the risk was estimated incorporating the findings of the prevalence study and predictions
based on home storage, cooking and consumption patterns. Three different exposure pathways
were investigated to estimate the risk associated with contaminated beef and Monte Carlo
simulation was used to determine the level of uncertainty. The developed model predicated that
consumption of contaminated beef can be accountable for 1.83E+06 E. coli O157:H7 cases per
year in Malaysia. The reliability of the model, data gaps and further research needs, is discussed.
Through continuous improvement Quantitative Microbial Risk Assessment provides valuable
insight into controlling and prevention strategies.
Cross contamination is one of the most important contributing factors in foodborne illness
originating in household environments. The objective of this research was to determine the
transfer between naturally contaminated chicken liver and leg to cutting board, hand glove,
knife and cucumber, during slicing. The microorganism tested was Campylobacter jejuni and
the results showed that the pathogen transferred to all utensils, at different transfer rate, despite
the low level of the naturally contaminating pathogen. With unknown concentration bacteria in
the naturally contaminated samples, a proportion of the utensils were still contaminated with C.
jejuni and not surprisingly, when the sample were contaminated with higher concentrations of
the pathogen, a higher proportion of the utensils had detectable C. jejuni cells present, though
in many cases cross contamination seems to be a random event. Transfer of the naturally
contaminating C. jejuni from the chicken liver and leg to the utensils were
The continued and increasing development of antimicrobial resistant bacteria among the
foodborne pathogens had caused worldwide to be alarmed. Being the earliest to develop
antimicrobial resistance, Staphylococcus aureus is constantly monitored for any new
resistance development. The resistance development is often linked to wastewater and the
treatment plants where the pressure of antibiotic is the highest. Hence, this study
investigated on the prevalence of high antimicrobial resistant S. aureus in the wastewater
eluted from a poultry slaughterhouse. A total of thirty wastewater samples were collected
from a poultry slaughterhouse in Semenyih, Selangor. Most probable number (MPN)-
plating method was employed to enumerate the S. aureus count in the wastewater. The
results indicated that S. aureus was highly present whereby all samples (100%) were
positive and the concentration ranged between 11 – 2.1 x 104 MPN/ml. Isolated S. aureus
strains were screened for their antimicrobial susceptibility using the Kirby-Bauer Disk
Diffusion Test method to classify their antimicrobial resistance eleven antibiotics. The
MAR index measured was between 0.18 and 0.91, inferring that the strains are highly
antimicrobial resistance. All S. aureus strains were 100% resistant to ampicillin (25 µg)
and cefazolin (30 µg). 94.1% of the strains were resistant to penicillin (10 µg) which
phenotypically indicated these strains are Methicillin-resistant S. aureus (MRSA).
Notably, 17.6% of the strains developed resistance to vancomycin and was categorized as
Vancomycin-resistant S. aureus (VRSA). There is a need to take drastic preventive
measures to control the resistance development in S. aureus to conserve public health.
To date, cholera has cycle the world seven times through the seven pandemic cycles that has
affected tens of millions of people. The objective of this study was to determine the presence
and density as well as the antibiotic resistance profile of Vibrio cholerae isolated from catfish
(Pangasius hypohthalamus). From the combination of the Most Probable Number-Polymerase
Chain Reaction-plating on TCBS agar methods, V. cholerae was detected in 32 samples and
V. cholerae O139 was detected in 7 samples, with a density ranging between
This goal of this study was to investigate the presence of Vibrio cholerae in street food,
namely satar and otak-otak, using Loop-Mediated Isothermal Amplification (LAMP),
multiplex Polymerase Chain Reaction (mPCR) and conventional plating on Thiosulphate
Citrate Bile-Salt Sucrose (TCBS) agar methods. A total of 78 satar and 35 otak-otak were
purchased from different districts of Terengganu (Besut, Setiu, Kuala Terengganu and
Kemaman). V. cholerae was found in satar with LAMP (10.3%), mPCR (10.3%) and
plating (0%). No V. cholerae was found in otak-otak using the three methods. This might
be due to V. cholerae able to survive in satar after grilling due to its thickness which may
contribute to undercooking. This study concluded that low presence of V. cholerae in satar
and otak-otak can be detected by molecular methods but not the conventional plating
method. LAMP assay is a useful tool for rapid detection of pathogens in food due to its
simplicity, highly sensitive and visual interpretation capability. Though the prevalence of
V. cholerae was low in the samples, proper handling of this food will help in reducing the
risk of acquiring infection from V. cholerae in contaminated samples.
Foodborne illness is a global burden that impacts a country politically, economically and
socio-economically. The severity of the burden can be unmeasurable as foodborne illness
is often an underestimated problem. In order to enlighten the burden, appropriate food
safety control measures should be taken. This study aimed to optimize a multiplex
Polymerase Chain Reaction (mPCR) detection method to identify foodborne pathogens
simultaneously. Six foodborne pathogens namely, Salmonella spp., Escherichia coli O157,
Vibrio parahaemolyticus, Vibrio cholerae, Listeria monocytogenes and Campylobacter
spp., were targeted in the mPCR detection method. Each mPCR parameter was tested and
the outcome was analysed to obtain a successful mPCR protocol to detect the targeted
foodborne pathogens. The amplified PCR products showed that the optimized mPCR
protocol will be a potential rapid diagnostic tool in foodborne pathogen detection.
Foodborne pathogens have become a constant threat to the consumer and food industry.
Reduce efficacy of antibiotics with emergence of resistant bacteria has limited the opportunities
for controlling pathogenic bacteria in food commodities and treating foodborne infections.
Bacteriophages can be a promising alternative for alleviate the risk of transmitting pathogenic
bacteria via food commodities. Therefore, this research was conducted to find distribution of
bacteriophages in diverse niches in order to identify suitable sources for isolating bacteriophages
to use controlling foodborne pathogens. Firstly bacterial strains were screened for lysogenic and
selected suitable host bacterial strains were used for isolating and determining bacteriophage titer
in fresh raw food and environmental samples. Eighteen different lytic bacteriophages effective
against Campylobacter, S. aureus, L. monocytogenes and E. coli were isolated from this study.
Bacteriophages titer was determined within range of 102
to 1010 PFU/mL and bacteriophages
were most frequently isolated from chicken (60%) samples. The isolated bacteriophages could
be potential candidates for controlling foodborne diseases.
Food safety in Malaysia is not considered an issue yet. From the previous year (2005-
2015) records, the incidence rate of food poisoning had been fluctuating and despite that,
cases continue to occur especially among school students. As a developing nation, it is
high-time that Malaysia begins to emphasize on food safety to reduce the burden of
foodborne illness in the socio-economic development of the country, and at the same time,
gain benefits in terms of economic returns and trade through food safety enforcement.
Most importantly, public health is achieved through food safety implementation and
accentuation. The current standing point of the Malaysia’s food safety is discussed in this
review. In addition, the review will also discuss the role of academicians as intervention
contributions in tackling food safety issues. The review is hoped to provide valuable and
concentrated information and knowledge to readers in the light to drive Malaysia into
ensuring safer food for the public.
Irrespective of its health effects, street foods are very popular with the consumers. The main
purpose of this research was to study the biosafety of Escherichia coli in popiah, a Malaysian
street food sold at a roadside food stall and a restaurant in Sri Serdang, Selangor, Malaysia,
using the combination of the most probable number (MPN)-Polymerase Chain Reaction
(PCR) assay-plating on Eosin Methylene Blue (EMB) agar methods. Using these biomolecular
methods, E. coli was detected in 12/15 (80%) and 11/15 (73%) of the collected samples from
the roadside food stall and the restaurant respectively. The incidence of stx virulence-associated
genes was detected in 1/15 (7%) among the E. coli isolated from samples taken from the
roadside food stall while the E. coli isolated from the restaurant was 3/15 (20%). The density
of E. coli ranged from 1100 MPN/g and the density of E. coli positive with stx genes
was
Bacteriophages are ubiquitous in our world, mainly in the oceans, soil, the water and food
we consume. They can be used efficiently in modern biotechnology, as well as alternatives
to antibiotics for many antibiotic resistant bacterial strains. Phages can be used as vehicles
for vaccines both DNA and protein, for the detection of pathogenic bacterial strain, as biocontrol
agents in agriculture and food industry. This review outlines the properties as well
as the influence of different external physical and chemical factors like temperature and
acidity on phage persistence. A better understanding of the complex problem of phage
sensitivity to external factors may be useful for other researchers working with phages.
Furthermore, the applications of bacteriophages were described in this paper as well.
Bacteriophages are the viruses of bacteria and are widely distributed in the biosphere, exhibiting
dramatic manifestations both in liquid cultures and on solid media. In this study, bacteriophages
were isolated from different types of food (beef, chicken meats, cucumber, lettuce, clam,
cockles and shrimp) and sewage samples using 6 reference pathogen strains (Salmonella
Enteritidis, Salmonella Typhimurium, Campylobacter jejuni, Vibrio parahaemolyticus, Listeria
monocytogenes and Escherichia coli). A total of 29 bacteriophage isolates were obtained and
further examined for titer via agar overlay assay. The titers were determined within the range
of 108
to 1011 PFU/mL. Our results showed that diverse of bacteriophages are naturally present
in a variety of foods.
Listeriosis and salmonellosis are the major foodborne illnesses worldwide. Over the last decade,
increasing reports about the antibiotic resistance of Listeria monocytogenes and Salmonella from diverse sources have prompted public health concerns, especially in developing countries with over reliance or misuse of antibiotic drugs in the treatment of humans and animals. In this study, antibiotic susceptibility profiles of 58 L. monocytogenes and 12 Salmonella Enteritidis strains from vegetable farms and retail markets in Malaysia were testedby the standard disk diffusion method. Listeria monocytogenes isolates were found to exhibit 100% resistance to penicillin G. Also, high resistance patterns were observed for meropenem (70.7%) and rifampicin (41.4%). The multiple antibiotic resistance (MAR) index of L. monocytogenes isolates ranged from 0.11 to 0.56. Besides, the antibiogram results revealed that multidrugresistant (MDR) S. Enteritidis were detected and all the S. Enteritidis isolates demonstrated resistance to at least four antibiotics. Ampicillin, amoxicillin, and trimethoprim failed to inhibit all the S. Enteritidis strains. Salmonella Enteritidis isolates also displayed high resistance to nalidixic acid (75.0%), trimethoprim-sulfamethoxazole (75.0%), and chloramphenicol (66.7%). Findings in this study indicated that vegetables could be potential sources of multidrug resistance of L. monocytogenes and S. Enteritidis, which can be a serious issue and a major concern for public health. Thus, there is a great need for surveillance programs in Malaysia to continuously monitor the antibiotic resistance profiles of important pathogens.