Presentation Outline

•Introduction

•Food Borne Pathogens and Diseases

•Risk Factors and Control Measures

•Need for preservatives

•Biopreservatives

•Conclusion
 Introduction (Contd)

Food commodities are the source of nutrients not only for human
beings but also for micro-organisms.

Most of the food products are perishable, it requires some

special care during their preparation, storage and distribution.

Food products are often sold in areas of the world far distant
from their production sites and there is a need for extended safe
shelf-life for products.

Particularly, meat products are highly perishable due to its

biological composition.
 Introduction (Contd)
Meat products also provides a suitable environment for the
proliferation of common food-borne pathogens and disease
causing micro organism.

In addition many interrelated factors influence the shelf life and

freshness of meat such as
holding temperature,
atmospheric oxygen (O2 )
endogenous enzymes,
moisture,
light and
micro-organisms
There is a need for adequate preservation technologies in order
to preserve its safety and quality
 Food Borne Pathogens and Diseases

Food borne diseases due to food contamination by pathogenic

microorganisms is a major concern around the world. (White et
al., 2002).

In 2002, the World Health Organization reported that one out of

every three people is suffering from food borne illness every
year.

Due to high susceptibility of human population to diseases,

changing life styles, and the emergence of newly recognized
human pathogens epidemiology of microbial food-borne
diseases has changed in the last decade (Altekruse and
Swerdlow, 1996; Berkelman, 1994).
 Food Borne Pathogens and Diseases (Contd)
In the United States, it was estimated 76 million cases of illness,
323,000 hospitalizations per year, and 5000 deaths were due to food
borne diseases (Mead, Tauxe et al., 1999).

http://www.cdc.gov/ncidod/eid/vol5no5/meadG.htm
 Food Borne Pathogens and Diseases (Contd)

Food borne illness is a serious and life threatening problem

especially for children and elderly people and to those with
weak immune systems. (FDA, 2009).

Food borne infection by salmonellosis, Campylobacter, or E.

coli O157:H7 appears to be higher in the elderly people (Jouve
et al., 2001).

In US, the annual medical cost due to food borne illness is

estimated to be $10 - $83 billion (FDA,2009).

Though, many advanced technologies have been developed to

eradicate food borne illness new forms of pathogens have been
identified in recent years.
 Pathogens (Contd)
The pathogens that are associated with food and cause
infection are

E. coli O157:H7 and other pathogenic E. coli,

Vibrio vulnificus,

Vibrio cholera,

Cyclospora cayetanensis,

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 Pathogens (Contd)
Listeria monocytogenes,

Clostridum botulinum,

Clostridium perfringens,

Staphylococcus aureus,

http://www.ehagroup.com/
 Pathogens (Contd)

Campylobacter jejuni,
 

Yersinia enterocolitica,

Salmonella enteritidis,

Salmonella typhimurium DT 104,

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salmonellablog.com
 Pathogens (Contd)
Hepatitis A viruses,

Norwalk-like viruses,

Astrovirus,

Rotavirus,

Parasites such as Toxoplasma gondii, Trichinella, Taenia saginata,

Taenia solium, etc..
 Risk Factors and Control Measures

Consumer awareness about the pathogenic microorganisms that

are responsible for food borne diseases are mixed.

The US Department of Health and Human Services, identified

five major risk factors contributing food borne illnesses
 Improper holding temperatures,
Contaminated equipment,
Inadequate cooking such as undercooking
Food from unsafe sources, and
Poor personal hygiene
 Risk Factors and Control Measures

Food Code (2009):

US Public Health Services, Food Code addresses five public
health interventions such as
Demonstration of knowledge,
Employee health controls,
Controlling hands as a vehicle of contamination,
Time and temperature parameters for controlling pathogens,
Consumer advisory
 Risk Factors and Control Measures
Food Safety Management
Hazard Analysis Critical Control Points (HACCP)
Good Manufacturing Practice (GMP)
Good Hygiene Practice (GHP)
Microbiological risk assessment

Biopreservation
 Need for Biopreservatives
Preservative agents are required to ensure that manufactured
foods remain safe and unspoiled.

Some of the oldest methods for the food preservation are the
addition of salts, dehydration & fermentation.

Emerging new methods of preservation include

Non-thermal processes like High Pressure Processing (HPP),
Pulsed Electric Fields (PEF), Irradiation,
New packaging systems such as Modified Atmosphere
Packaging (MAP) and active packaging,
 Natural antimicrobial compounds and

Consumers are concerned about the synthetic chemicals used as

preservative in food, and there is a resulting trend towards less
 Biopreservatives

The use of non-pathogenic microorganisms and/or their

metabolites or the extract of plant parts to improve
microbiological safety and to extend the shelf life of foods is
defined as biopreservation (De Martinis et al, 2001).

The demand for high quality, natural, nutritious, fresh

appearance and convenient meat products with natural flavor and
taste with extended shelf-life leads to the development of natural
or biopreservatives..
 Biopreservatives- Lactic Acid Bacteria & its
Metabolites
Lactic acid bacteria or (LAB) have been employed in the
preservation of food products in many countries. Widely
used as starter cultures in the meat industry for sausage
fermentation

LAB control the growth of pathogens such as

Listeria,
Clostridium,
Staphylococcus
Bacillus spp.
Enterococcus spp.
 Biopreservatives- Lactic Acid Bacteria & its
Metabolites (Contd)
LAB include the genera Lactococcus, Streptococcus,
Lactobacillus, Pediococcus, Leuconostoc, Enterococcus,
Carnobacterium, Aerococcus, Oenococcus, Tetragenococcus,
Vagococcus, and Weisella.

LAB produce acid (low pH),hydrogen peroxide and

bacteriocins contributes the preservative activity
(ogunbanwo,2005).

Bacteriocins is a kind of proteinaceous substance (protein or

protein complexes) that inhibits the growth of the related
species by exhibiting bactericidal activity and it is toxic to
the other bacterial strains.
 Biopreservatives- Lactic Acid Bacteria & its
Metabolites (Contd)
K. Intarapichet & S. Gosaarak found that bacteriocin from
lactobacillus lactis inhibited the growth of “Brochothrix
thermosphacta” on surface of pork meat balls kept at 4oC for
12 days while untreated meat balls showed higher bacterial
count of 2 log cycles than the treated one.

Sakhare and Narasimha Rao (2003) determined that the use

of a mixture of LAB could extend the shelf life of minced
meat at high temperature and inhibited the growth of
pathogenic microorganisms in the pH range of 4.0-4.2.

For stabilization of meat the desirable low pH (4.0-4.2) was

achieved faster at ambient temperature using a LAB culture.
 Biopreservatives- Lactic Acid Bacteria & its
Metabolites (Contd)

The combinations of bacteriocins and other antimicrobial

agents or processes enhance the antimicrobial activity.

Meets to the requirement of less processed food and by

extending the shelf life of food.
 Biopreservatives- Enterocins

Enterocin is a pediocin family of bacteriocins produced by

Enterococcus faecium strains.

Type depends on the bacterial strain.

Enterocins A and B exhibited antimicrobial effect in meat

and meat product which includes deboned chicken breasts,
pâté, and slightly fermented sausages, cooked ham, minced
pork,etc.

Enterococcus faecium P13 isolated from dry-fermented

sausages produces Enterocin P which exhibits strong
inhibitory activity against Listeria monocytogenes.
 Biopreservatives- Enterocins

Aymerich (et al.,2000) investigated the effect of Enterocin A

and B isolated from Enterococcus faecium CTC492 by
injecting into the meat at different concentrations which was
already inoculated with L. innocua.

Enterocins at 4800 AU/g reduced the Listeria count by 7.98

and 9 log cycles in cooked ham and pate stored at 7 ºC for 37
days and the Listeria count was decreased to below 50 CFU/g
in which there was no reduction found in control samples.
 Biopreservatives- Pediocin

Pediocin is a bacteriocin produced by Pediococcus

acidilactici & generally recognized as safe(GRAS) organism,
commonly found and used in fermented sausage production.

Pediocin AcH has been proven to be effective against

pathogenic & spoilage organisms, including Listeria
monocytogenes, Enterococcus faecalis, Staphylococcus
aureus and Clostridium perfringens.

Active at a wide range of pH and most of them are thermo

stable proteins
 Biopreservatives- Nisin

Nisin is a bacteriocin produced by the bacterial starter culture

Lactococcus lactis that possesses a natural antimicrobial
activity against the gram positive bacteria including the food
borne pathogen L. monocytogenes.

Nisin was awarded (GRAS) status in the United States in 1988

Nisin has low molecular weight compound can withstand

temperature up to 100°C for 100 min (Mahadeo, 1995).

Nisin is mostly active against gram positive bacteria and it can

be made to be effective against Gram-negative bacteria by
providing some chelating agents to induce damage to the outer
membrane.
 Biopreservatives- Nisin

Nisin was found to have a synergistic effect when it combines

with other antimicrobial agent.

Synergistic effect of nisin and lysozyme in retarding the growth

of L. monocytogenes by the pre-surface application before in-
package pasteurization at 65°C for 32 s effectively reduced the
microbial count of L. monocytogenes to below detectable levels
in 2-3 weeks of storage (Sunil, 2006)
 Biopreservatives- Lactoferrin

Lactoferrin is a natural bioactive glycoprotein that is

commonly found in milk, saliva and tears and in trace
quantities in meat tissue.

Lactoferrin disrupts cell membranes by binding to

lipopolysaccharide of bacterial walls and its iron part affects
(oxidizes) the membrane permeability by forming peroxides.

ALF an activated form of lactoferrin is effective against variety

of foodborne pathogens such as E. coli O157:H7, Listeria
monocytogenes and Salmonella, and also spoilage bacteria.
 Biopreservatives- Natamycin

Natamycin also known as primaricin is a polyene macrolide

antibiotic produced by streptomyces natalensis and related
species.

Natamycin is used as a food additive to control the growth of

yeasts and moulds on the surface of non sterile products such
as meat and sausages.

FAO/WHO recommended an acceptable daily intake of (ADI)

0-0.3 mg/kg body weight.
 Biopreservatives- Antimicrobial enzymes

Due to health concerns of the consumer the producers has been

interested to produce biopreservatives in antimicrobial
packaging.

Such as
Lysozyme,
Lactoperoxidase,
Chitinase and
Glucose oxidase
 Biopreservatives- Antimicrobial enzymes
Lysozyme
Lysozyme is an antimicrobial enzyme that inhibits the growth of
bacteria and fungi secreted by polymorphonuclear leukocytes
(Jolles and Jolles, 1984).

It is mainly active against Gram-positive bacteria as they lyse the

cell walls by splitting the bond between N-acetylmuramic acid and
N-acetylglucosamine of the peptidoglycan in the bacterial cell wall.

It is widely distributed in various biological fluids and tissues,

including avian egg, plant and animal secretions, tears, saliva,
respiratory and cervical secretions.

It was awarded safe (GRAS) status by WHO/FDA

 Biopreservatives- Antimicrobial enzymes
Lactoperoxidase system
Lactoperoxidase system (LPS) can inhibit the growth of E. coli
O157:H7, L. monocytogenes, S. aureus, and naturally present
microbial populations in ground beef.

Kennedy(et al.,2000) explored the effect of LPS on the pathogen

control(inhibition) in ground beef stored at 0, 6 and 12°C .
It was observed that the growth of the microbial populations
was strongly inhibited by the presence of LPS while number of
bacteria has been increased without LPS in 4 hr incubation
time.

It was also reported that LPS was a temperature dependent and its
effect was maximum at a temperature permissive and the condition
did not favor the growth of the test strain.
 Biopreservatives- Antimicrobial enzymes
Chelators

Chelators (Citrate and EDTA) which bind metal ions, are not by
themselves restrict the growth of L. monocytogenes in foods.

However, these compounds interact with other preservatives and

sometimes aid in suppressing the growth of L. monocytogenes in
meats.
 Biopreservatives- Anti-oxidants
Tocopherols
Tocopherols are considered as effective natural antioxidants for the
foods rich in lipids.

As an antioxidant it competes with the substrate for the chain

carrying peroxyl radicals.

Researches has been carried out for improving the stability of

lipids & myoglobin in lamb, beef and turkey. α-tocopherol was
found to retard the decomposition of hydroperoxides.
 Biopreservatives- Anti-oxidants
Rosemary extract
Rosemary extracts have a potent antioxidant activity and are
widely used in the food industry.

A number of authors have reported the effectiveness of rosemary

for reaching higher sensory scores and for retarding oxidation in
foods rich in lipids.

In addition to inhibition of lipid oxidation, several authors have

reported that some of the compounds present in rosemary extracts
possess antibacterial properties

It has been demonstrated that rosemary extracts effectively

inhibited hydroperoxide formation
 Biopreservatives- Chitosan

Chitosan as Antioxidant & biopreservative: Chitosan exhibits in

vitro antimicrobial activity against a range of food borne
microorganisms and it is considered as a potential natural food
preservative.

Chitosan has the capacity of forming complexes with the anionic

charges .The cell surface of the gram negative bacteria contains
anionic ions and so chitosan containing cationic charges binds with
the cell surface of bacteria and it disrupts the barrier properties of
the outer cell membrane.

The restriction on the use of chitosan has been removed and it was
given GRAS status by the US FDA.
 Biopreservatives- Synergistic effect

Shun Wada, Xing Fang (2007) observed a synergistic effect

between a mixture of α-tocopherol and rosemary extract (0.05%+
0.02%) on frozen-crushed fish meat as it exhibited a strongest
antioxidant activity.
They noted that α-tocopherol remained longer for 5 more days
in the sample when it was mixed with rosemary extract.

Dimitrios Georgantelis (et al., 2006) found the synergistic effect of

a mixture of chitosan, rosemary extract and α-tocopherol on fresh
pork sausages stored for 20 days at 4 °C (separately and in
combination).
Combination of chitosan & rosemary provided the best
antimicrobial and antioxidative effects.
 Biopreservatives- Antimicrobial activities of
plants/spice/herb/fruit extracts
Bio-preservatives are mainly derived from plant extracts as
because most of them contains phenolic compounds that acts as
antioxidants by reducing the free hydroxyl groups.

It also inhibits the microbial growth due to their protein binding

capacity.

Plant extracts such as hop extracts , eugenol , cassia, clove, garlic,

sage, oregano, thyme, rosemary, scutellaria and forsythia suspensa
(Thunb), pimento leaf , horseradish distillates, cinnamic acid ,
furanocoumarins , and carvacol have antimicrobial potency .
 Biopreservatives- Antimicrobial activities of
plants/spice/herb/fruit extracts
Antimicrobial potency differ according to their chemical
composition, content of extracts, essential oils and it also depends
upon the type of food and micro organism.

Antimicrobial activity of hop extracts was higher in lower

fat meats

Rosemary was found effective against Listeria spp. in

meat including in ready-to-eat pork liver sausage while
horseradish distillates on roast beef, and eugenol & pimento leaf
on refrigerated cooked beef but their inhibitory effect depends on
the amount of fat content in meat.
 Biopreservatives- Antimicrobial activities of
plants/spice/herb/fruit extracts
Ha et al. (2001) evaluated the antimicrobial activity of grape seed
extract in ground beef by incorporating it in to Polyethylene films
showed inhibitory effect against E. coli, S. aureus, and Bacillus
subtilis.

The mixture of commercial ingredients such as 1.5% blend of

lemon/cherry/vinegar powder, 2.0% buffered vinegar and a 3.0%
cultured sugar/vinegar exhibited antimicrobial activity against L.
monocytogenes present in ham, roast beef and turkey without
affecting the sensory attributes (Glass and Sindlear, 2010)
 Biopreservatives- Antimicrobial activities of
plants/spice/herb/fruit extracts
Lemon, mustard, cumin, and pepper added to a fish sauce Although
lemon and pepper had no effect, mustard was highly effective in
controlling E. coli in the fish sauce, which was stored at 25°C for
28 days (AI-Jedah et al.,2000).

Natural spice extracts serves as a strong antioxidants in cooked

pork products (Kong et al.,2010).
Effect of total phenolic content & DPPH scavenging activity of
six spice extracts namely cassia bark, liquorice, nutmeg, clove,
rosemary and round cardamom and found they were strong
inhibitors of TBARS formation ( Thio-Barbituric Acid
Reactive Substances) and reduced off flavor and a provided a
stable red colour in cooked pork products during storage.
 Conclusion

Due to increase in demand by consumers for less processed food,

the food industries started focusing on producing products with
natural antimicrobials.

Many researches has been carried out for using various natural
antimicrobials in meat and meat products as it is one of the major
food in many countries.

The recent researches has been showing promising results from

foods.
 Conclusion

Consumer acceptance of new processes is likely to be increased

when a direct consumer benefit is recognized.

However, huge information gaps exist among scientists and

particularly between scientists and consumers.

To match the promise offered by technological advances and

optimize nutrition, the overcoming of barriers in psychological and
- as a consequence - political feasibility is required.

There is needs not only research efforts but education in all stages
of the food chain and in all sectors of communication