SEMINAR REVIEW

ON
THE ROLES OF MICRO ORGANISMS IN FOOD SPOILAGE

BY
ONUORAH OBIORAH
NSU/NAS/558/11/12

ZOOLOGY UNIT
DEPARTMENT OF ZOOLOGY
FACULTY OF NATURAL AND APPLIED SCIENCES,
NASARAWA STATE UNIVERSITY, KEFFI

SUBMITTED IN PARTIAL FULFILLMENT OF THE

REQUIREMENTS FOR THE AWARD OF BACHELOR OF
(HONS) DEGREE IN ZOOLOGY,
DEPARTMENT OF ZOOLOGY, NASARAWA STATE
UNIVERSITY, KEFFI, NASARAWA STATE UNIVERSITY

CERTIFICATION
This is to certify that this seminar review was prepared by
Onuorah Obiorah, NSU/NAS/558/11/12 under the supervision of
__________________________ in the department of Zoology (Zoology
Unit), Faculty of Natural and applied sciences, Nasarawa State
University, Keffi.

_____________________________
_____________________________
SUPERVISOR

DATE

_____________________________
_____________________________
H.O.D

DATE

CHAPTER ONE
INTRODUCTION
1.1 BACKGROUND REVIEW
Food spoilage is a complex process and expensive amounts of
foods are lost, due to microbial spoilage, even with modern day
preservation

techniques.

Despite

the

heterogeneity

in

raw

materials and processing conditions, the microflora that develops

during storage and in spoiling foods, can be predicted, based on
knowledge of the origin of the food, the substrate base and a few
central

preservation

parameters

such

as

temperature

atmosphere, and PH. Based on such knowledge more detailed

sensory, chemical, and microbiological analysis can be carried out
on the individual products to determine factors for selection of
spoilage microorganisms, a level of refinement may be found in
some

products

in

which

the

interactive

behavior

of

microorganisms may contribute to their growth and or spoilage

activity. The review gives three of such examples; we describe the
competitive

advantage

of

pseudomonas

spp.,

due

to

the

production of Iron-chelating siderophores the generation of

substrates for spoilage reaction by one organisms from another
microorganisms (so called metabiosis) and the up-regulation of
phenotypes potentially involved in spoilages through cell-to-cell
communication. In particular, we report

for the first time, the

widespread occurrence of N-Acyl homoserine lactones (AHL) in

storey and spoiling fresh foods and we discuss the potential

implications of spoilages and food preservation.

1.2 CONCEPT AND DEFINITION OF TERMS

Food spoilage means the original nutritional value, texture, flavor
of the food are damages, the food becomes harmful to people and
unsuitable to eat. It can also be defined as a disagreeable change
in a foods normal state. Such changes can be detected by smell,
taste, touch or sight. These changes are due to a number of
reasons; air and oxygen, moisture, light, microbial growth and
temperature. Each is briefly explained below

Air and Oxygen

One important cause if food spoilage is air and oxygen. Because
air is colorless, odorless and tasteless, it is often taken for granted
and at times forgotten as a means to cause either condition and
are called facultative anaerobes. Molds and most yeasts that
cause food to spoil require oxygen to grow. They can often be
found growing on the surface of foods when air is present.

Enzymes
Certain enzymes that are naturally present in food are known as
oxidizing enzymes. These enzymes catalyze (speed up) chemical

reactions between oxygen and food components and leads to

food spoilage. Although there are many oxidizing enzymes, two
that cause darkening in diced and sliced vegetables are catalyse
and peroxidase. The browning of vegetables caused by these
enzymes is often accompanied by off-flavors and odors. A simple
heat treatment (blanching) is used to inactivate enzymes.
Oxygen
Oxygen can cause spoilage of foods spontaneously by itself.
Oxidatane spoilage, the chief cause if quality loss in fats and fatty
portions of foods, when lipids oxidized, short chain carbon
compounds are formed; these compounds have very strong odor
and flavor and are very undesirable and unacceptable. The off
odors resulting from this type of spoilage are sharp and acrid and
have been described as linseed oil-like, fishy or perfume like.

Moisture
Water is one of the most common substances on earth. It is an
essential component of all foods. The amount of water in a food
(know and percent water) influences the appearance, texture, and
flavor of the food. All living organism as well as food contain
water. Water makes up about 70% or more of the weight of most
fresh (unprocessed) foods. Even dry foods like beans, flour and
cereals contain some most water between 90% and 95% water.
The amount of water in some common foods is shown below;

Food

Food

Apples

84

lettuce

Bread
Butter

32-37
16

Cornflakes

mayonnaise
milk

peaches

Green beans

92

peanut butter

Flour

12

raisins

Jam jellies

29

strawberries

Sugar

0.5

Although the water content of a food is expressed as a percent,

this number does not reflect how the water exists in the food.
Water in food is classified according to its availability or biological
activity and is either free or bound. Free water is not bound to
any component in a food; it can be used for microbial growth and
also available for chemical reactions. Bound water is physically
bound to large (molecules) components in the food. It is not
available to micro-organisms for their growth and it cannot
participate in chemical reactions.

Water or moisture affects the keeping qualities of food. Excessive

moisture pick up can result in product spoilage and spoilage by:
Microorganisms; microorganisms need water to dissolve the
food the use. Water allow the food to get into bacterial, yeast
and mold cells where it is used for energy and growth. Water
also allows waste products to escape from the cells.
Chemical reactions; The moisture in food also functions to
allow chemical reactions to occur between components in
the product.
Food spoilage & spoilage can occur when there are slight changes
in relative humidity. Moisture can condense on the surface of a
product and this can result in many common food defects.
The molding of grain, soggy cereals and the caking and lumping
of dry products like powder and cake mixes can result in
excessive moisture. Other defects such as motting, crystallization
and stickiness have also been observed. Moisture condensing on
the surface of a food can also provide an environment for bacteria
and molds to grow and multiply. Physical defects such as
cracking, splitting and crumbling occur in excess moisture in lost
from foods.
Water are controlled in foods by
Drying (Dehydration) concentration and evaporation;- by
removing water to a certain level, deteriorative reactions can
be reduced on prevented. Examples of products preserved

by these drying techniques include dry milk, potato flakes,

drink mixes, evaporated milk, and orange juice concentrate.
Freezing:- The freezing of foods changes water from liquid to
solid form and renders it unavailable to microorganisms and
chemical reactions.
Food Additives:- salt and sugar are used in many products to
bind water and thereby making it less available for microbial
growth and biochemical reactions, Jams, Jellies and cured
hams are examples.

CHAPTER TWO
LITERATURE REVIEW AND DISCUSSION
2.1 LITERATURE REVIEW
Conservation of food has been characterized for nutrition and
microbiologically stable foods and it has been achieved by
controlling the growth of spoiling and pathogenic food related
microorganisms. Microbial control in foods could be assured by
suppressing one or more essential factors for microbial survival
(Horace, 1982)
There has been increasing concern of the consumers about foods
free or with lower level of chemical preservatives, because these
could be toxic for humans (Bedin et al;1999). Consumers have
been also demanded for food with long shelf and absence of risk
of causing food borne diseases. The perspective has put pressure
on the food industry for progressive removal of chemical
preservatives and adoption of natural alternatives to obtain its
goals concerning microbial safety. This resulted in increasing
serach for new technologies for use in food conservation systems,
which include modified atmosphere packaging combined effects
of underlethal procedures, alternantives antimicrobial compounds
(ecstatic or cidal effects), combination of conventional (used in
low levels) and alternatives antimicrobial. (B rull and coote, 1999)
Recently, there has been increasing interest in discovering new
natural antimicrobial (sagdic et. al., 2003). This is also true in food

microbiology.

Plant

products

with

antimicrobials

properties

notably have obtained emphasis for a possible application in food

production in order to prevent bacterial and fungal growth
(lanciotti et., at., 2003).
Plant

products

are

characterized

for

range

of

volatile

compounds, some of which are important flavor quality factors

(utama et al., 2002). Moreover plant volatiles have been generally
recognized as safe (GRAFS) (Newberne et al, 2000). Systematic
screening for biological interaction between microorganisms and
plant products have been valuable source of new and effective
antimicrobial substances, which could have different action ways
on/in the microbial cell, when compared to other conventional
antimicrobial plant synthesis by a secondary metabolism. Many
compounds with complex molecular structure and some of them
have been related with antimicrobial properties, found in the
plants and their derivatives. Among these secondary metabolic
are found alkaloids, flavonoids, isrflavoids , tannins, cumarins,
glycosides, terpens and phenobiic compounds (simoes et al,
1999)

still

little

information

is

available

emphasizing

the

preservative and antimicrobial role of spices in the prevention of

food against the microbial action(arora and kaur, 1999)
Spices are recognized to stabilize the foods front and microbial
deterioration. This could be observed when spices show initially
high microbial charge and as time progresses , the microbial
growth

becomes

progressively

slower

or

eventually

totally

suppressed (kizil and sogut, 2003). Antimicrobial activity of spices

depend on several factors which includes;
i.
ii.
iii.
iv.
v.

Kind of spices;
Composition and concentration of spices,
Microbial specle and its occurrence level,
Substrate composition,
Processing condition and storage (shelef, 1983; faraj et al.,
1989).

vi.
Spices have been defined as plant substances from indigenous or
exotic origin, aromatic or with strong taste, used to enhance the
taste of foods (Germano and Germano, 1998). Spices include
leaves (bay, mint, rosemary, coriander, laurel, oregano), flowers
(clove), bulbs (garlic, onion), fruits (cumin, red chilli, black
pepper), stems (coriander, cinnamon), rhizomes (ginger) and
other plant parts (Shelef, 1983). Although, spices have been well
known

for

their

medicinal,

preservative

and

antioxidant

properties, they have been currently used with primary purpose of

enhancing the flavor of foods rather than extending shelf-life
(Aktug and Karapinar 1986, Ristori et al., 2002).
Spices active compounds have been included in class of naturally
occurring food preservatives and have their inclusion in foods
allowed by food production regulator offices (Brull and Coote,
1999). Several scientific reports describe the inhibitory effect of
spices on a variety of microorganisms, although considerable
variation for resistance of different microorganisms to a given

spice and of the same microorganisms to different spices has

been observed (Akgul and Kivan, 1988).
. Grohs and Kunz. (2000) observed that spices mixtures were able
to inhibit the growth of various meat-spoiling microorganisms
(Bacillus subtillis, Enterococcus spp., Staphylococcus spp., E. coli
K12 and Pseudomonas fluorescens) providing stabilizing effect on
colour and smell of fresh portioned pork meat.
Study carried out by Al-Jedah et al. (2000) analyzed the action of
combined spices, including cumin (Cuminum cyminum), coriander
(Coriandrum sativum), mustard (Brassica juncea), black pepper
(Pipper

nigrum)

and

lemon

(Citrus

aurantifolia)

on V.

parahaemolyticus, S. aureus, S. tiphy and E. coli count in fish

sauce, which showed that the spices mixtures were able to exert
static effect on all assayed bacteria when in interaction with an
initial inoculum of 1.0 x 104 CFU/mL, except on S. typhy.
Leuchner and Zamparini (2002) studied the growth and survival
of E.coli O157

andSalmonella

enterica serovar enteridis in

mayonnaise in presence of garlic (Allium cepa), ginger, mustard

and ground clove. Garlic (1% w/v) and clove (1% w/v) which
showed

bacteriostatic

towards S.

and

enterica and E.

bactericidal
coli O157,

effect,

and E.

respectively,

coli was

more

sensitive. These results are significant regarding the emergence

of E.

coli 0157

and S.

enterica serovar enteridis as

foodborne

pathogens that have significant impact on the food industry.

Moreover, these bacteria present various undesirable attributes of

virulence that in combination make them some of the most

serious threats for food safety (Proctor and Davis, 2000).
Outtara et al. (1997) analyzed the antibacterial activity of
selected fatty acids and spices essential oils on meat spoilage
bacteria

and

no

fatty

acid

presented

antibacterial

activity. Brochothrix thermosphacta was inhibited by cinnamon,

clove, garlic and rosemary essential oil (1/100 v/v); Serratia
liquefaciens by cinnamon, clove, garlic, pimento and rosemary
essential oil (1/100 v/v); Carnobacterium piscicola by cinnamon,
clove,

pimento

and

rosemary

essential

oil

(1/100

v/v);

and Lactobacillus sake by cinnamon, clove, black pepper, pimento

and rosemary essential oil (1/100 v/v); L. curvatus was inhibited
by black pepper and pimento essential oil (1/100 v/v).
Arora and Kaur (1999) analyzed the antimicrobial activity of
garlic, ginger, clove, black pepper and ground green chilli and
their

aqueous

extracts

including Bacillus
epidermidis,
Pseudomonas

on

human

sphaericus,

pathogenic

Staphylococcus

bacteria
aureus, S.

Enterobacter

aerogenes,

Escherichia

coli,

aeruginosa,

Salmonella

tiphy and Shiguella

flexneri and found that all tested bacteria were sensitive to

ground garlic and its extract. Moreover, garlic extract showed
considerable cidal effect on S. epidermidis, S. tiphy andE.
aerogenes.
Kivan et al. (1991) studied the effect of spices on starter cultures
(Lactobacillus

plantarum and Leuconostoc

mesenteroides)

considering that lactic acid bacteria are relatively resistant to

toxic effect of spices and derivatives (Jansen et al., 1987) and that
some

spices

have

exerted

stimulatory

effect

on

these

microorganisms resulting in enhanced acid production (Tiwari and

Pandey, 1981). Cumin (0.5, 1.0 and 1.0% w/w) and its essential oil
(150,

300

and

600ppm)

stimulated

the

growth

of L.

plantarum and L. mesenteroides and acid production. Oregano

was able to stimulate the growth of L. plantarum and acid
production, however this behavior was not observed in L.
mesenteroides.
Exact mechanism of antibacterial action of spices and derivatives
is not yet clear (Lanciotti et al., 2004). Although some hypothesis
habe been given, which involve: i) hydrophobic and hydrogen
bonding of phenolic compounds to membrane proteins, followed
by partition in the lipid bilayer (Juven et al., 1994); ii) perturbation
of membrane permeability consequent to its expansion and
increased fluidity causing the inhibition of membrane embedded
enzymes (Cox et al., 2000); iii) membrane disruption (Caccioni et
al., 2000); iv) destruction of electrons transport systems (Tassou
et al., 2000); v) cell wall perturbation (Odhav et al., 2002).
Generally, gram-negative bacteria have been reported to be more
resistant than Gram-positive to essential oils antimicrobial effect
because of their cell wall lipopolyssaccharide (Russel, 1991). Cell
wall lipopolyssacaride may prevent that essential oils active
compounds reach the cytoplasmic membrane of Gram-negative
bacteria (Chanegriha et al., 1994).

Antifungal activity of spices and derivatives has been studied

regarding viable cells count, mycelial growth and mycotoxins
synthesis. Juglal et al. (2002) studied the effectiveness of nine
essential oils to control the growth of mycotoxins-producing
moulds and noted that clove, cinnamon and oregano were able to
prevent

the

growth

ofAspergillus

parasiticus and Fusarium

moniliforme, while clove (ground and essential oil) markedly

reduced the aflatoxin synthesis in infected grains. These findings
could be useful for rural communities to prevent the synthesis of
fungal toxins in contaminated grains by simple measures.
Thyagaraja and Hosono (1996) assayed the ability of chilli,
coriander, pepper, cumin and asafoetida to inhibit food spoilage
moulds

(Rhizopus

azygosporus,

Mucor

dimorphosphorous,

Penicillium commune, Fusarium solani) and asafoetida showed

promising results in inhibiting the fungal growth. Among the
ethanolic and aqueous asafoetida extract, only ethanolic fraction
showed

antifungal

property. Little

information

is

available

emphasizing the preservative and antimicrobial role of spices in

the prevention of foods of the microbial action (Arora and Kaur,
1999).
Little information on spices and derivatives action on/in the fungal
cell in order to promote fungistatic or fungicide effect. In general,
inhibitory action of natural products on mould involves cytoplasm
granulation, cytoplasmic membrane rupture and inactivation
and/or inhibition of intercellular and extracellular enzymes. These
biological events could take place separately or concomitantly

culminating with mycelium germination inhibition (Cowan, 1999).

Also, it is reported that plant lytic enzymes act in the fungal cell
wall causing breakage of b-1,3 glycan, b-1,6 glycan and chitin
polymers (Brull and Coote, 1999).
Presence of pathogenic and spoiling microorganisms in spices
could act as vehicles for microorganisms to enter in foods.
Frequently, spices are grown and harvested in warm and humid
areas where the growth of wide variety of microorganisms is
readily supported (Mousuymi and Sarkat, 2003). As many other
agricultural commodities, spices are exposed to a wide range of
environmental

microbial

contamination

during

harvest,

processing, and in retail markets by dust, waste water, and

animal and even human excreta (Freire and Offord, 2003). The
International Commission on Microbiological Specifications for
Foods (1974) has set up maximum limit of 106, 104 and 103 CFU of
total aerobic mesophilic bacteria (TAMB), fungi, coliforms and E.
coli, respectively, per gram spice (Zamboni et al., 1991). Brazilian
Microbiological Standard for Foods (ANVISA, 2001) has set
maximum limit of 5 x 102 e 102 CFU/g for faecal coliforms and
positive coagulaseStaphhylococcus, respectively, and absence in
25g of spices for Salmonella. In German legislation, standard limit
value for TAMB, Bacillus cereus and S. aureus is 105, 104 and
102 CFU per gram of spices, respectively (Mousuymi and Sarkat,
2003). The microbiological quality, the load of heterotrophus
or Enterobacteriaceae in particular, often acts as indicator of the

hygienic situation of the region where the spices are produced

and processed (Schwab et al., 1982).
Mousuymi and Sarkat (2003) reported the presence of various
microorganisms

including

total

heterotrophus,Bacillus

cereus, Clostridium perfringens, Escherichia coli, Salmonella and

toxigenic moulds in spices. Thus, there is strong need to evaluate
and control the microbial quality of spices including bacterial and
mycological analyses and presence of microbial toxic metabolites
(Fernndez et al., 1984; Zamboni et al., 1991; Oliveira et al.,
1992; Toro Santa Maria et al., 1993; Hofman et al., 1994; AbdelHafez and Al-Said, 1997; Pereira et al., 1999; Freire and Offord,
2003; Benezet et al., 2003).
Antimicrobial activity of spices could be recognized as important
factor for providing their inclusion in food conservation systems
when pertinent measures are taken to assure their satisfactory
microbiological quality. These measures must include action to
control the moisture, good sanitary conditions in the processing,
workers training, satisfactory transport conditions, proper storage,
microbiological quality monitoring and actions of sanitary mark
applied since harvest until insertion in foods.
It is established that spices and their derivatives could be suitable
alternatives for inclusion in food conservation systems and could
act sometimes as main or adjuvant antimicrobial compounds.
Before

including

spices

and/or

their

derivatives

in

food

conservation systems, some evaluations about microbiological

quality, economic feasibility, antimicrobial effect for a long time

and toxicity should be carried out.
Cinnamon (Cinnamon Zeylancium): Cinnamon is used to To treat
upset stomach and diarrhea (Skidmore- Roth, 2003) and For the
treatment of gastric ulcers (Tanaka et al., 1989). Cinnamon bark
may possess a potentiating effect on insulin (Khan et al., 1990),
and can be useful in the treatment of type 2 diabetes; as well as
lowering triglyceride levels and serum cholesterol (Khan et al.,
2003; Broadhurst et al., 2000; Onderoglu et al., 1999) For the
treatment of bronchitis, coughs and other respiratory ailments
(Martinez,
1989). It is also Against nervous disorders and For loss of appetite
and

dyspepsia

(Blumenthal,1998).

To

promote

conception,

cinnamon is sometimes used alternately with damiana (Turnera

diffusa Wild) (Adame and Adame, 2000). For the treatment of
hypertension (high blood pressure). It is used as an invigorating
tonic, externally as a poultice to treat minor bacterial and fungal
infections

of

the

skin.

The

essential

oil

is

employed

in

aromatherapy as a rub to promote blood circulation (Tisserand,

1995).
Some of the plant constituents have proven value against
bacteria and fungi,
including the molds that produce the carcinogenic aflatoxins
(Gruenwald, 2004; McCann, 2003; Juglal et al., 2002).
Cinnamon constituents possess antioxidant action and may prove
beneficial against free radical damage to cell membranes

(Dragland et al., 2003; Jayaprakasha et al., 2003; Lee and

Shibamoto, 2002) and Its essential oil contains both antifungal
and antibacterial principles that can be used to prevent food
spoilage due to bacterial contamination (Fabio et al., 2003;
Guynot et al., 2003; Kalemba and Kunicka, 2003; Suhr and
Nielsen, 2003; Valero and Salmeron, 2003; Friedman et al., 2002;
Mejlholm and Dalrgaard, 2002; Ranasinghe et al., 2002; SmithPalmer et al., 2002; Yuste and Fung, 2002; Mau et al., 2001).
Cinnamon oil has proven to be particularly effective against some
species of
toxicogenic fungi (Juglal et al., 2002; Soliman and Badeaa, 2002;
Montes-Belmont

and

Carvajal,

1998),

as

well

as

against

respiratory tract pathogens, including species belonging to the

genera Aspergillus, Candida, Cryptococcus, and Histoplasma
(Viollon and Chaumont, 1994; Inouye et al., 2002). Even though
cinnamon has antibacterial effects, clinical trials against the
species Helicobacter pylori, associated with gastric ulcers, have
shown contradictory results (Martin and Ernst, 2003; Tabek,
1999).
Gram negative bacteria are more resistant to the antibacterial
activity

of

essential

oils

than

gram-positive

bacterial

P.aeurogeinosa and e.coli were the most resistamt strains to spice

samples and that they were susceptible to cinnamon only as per
sema agaogiu, nursel dustbin, and suleyman alemdar (2007)_.
They also found that cinnamon was detected to exhibit a similar
inhibitory effect against p.aeurginosa and e.facecalis and the

weakest activity was against e.coli and M.luteus. the anti

microbial activity og cinnamon may be explained by its volatile oil
components. The most important active substances found in
cinnamon oils are cinnamon aldehyde ad eugenol. A number o
finvestigations (Azzouz M.A, Bullerman L.R 1982) reported that
cinnamon inhibited the growth and toxin production of some
mould species, with an activity emerging from cinnamic alheyde
and eugenol.
In another study (outtara B. Simmard D.E, holley R.A, Pitte G.J.P,
Begin A. 1999) eugenol was shown to have a stronger bacterial
activity against E.coli and K.pneumonia thatn some antibiotics.
The sults of the present study concur to the results of the study
mentioned before D.Afyue Atep I., Z lem Turgag Erdourul in 2003
suggested that the extractes of cinnamon species showed
antibacterial activity to the microorganisms tested. The alcohol
extracts showed inhibition zone against B. megaterium and
E.faecalis. a number of investigations reported that cinnamon
inhibited the growth of some mould species with an activity
emerging from cinnamic aldehyde and eugenol
Colve (syzygium aromaticum); most workers have investigated
the properties of spices inhibiting agaents of mycelid toxin
production. Syzigium aromaticum (colves) have been found to be
particulary; effective often completely inhibiting both fungal
growth and toxin production (

et al 1980; Mubarak and

elshayeb 1980). Growth was completely inhibited in three

toxigenic strains of aspergilla (a.flavns ATCC 15548, A. flavns

) cultured on ground clove (1.5g) and sterile

water for 30 days. Cloves have been shown to be significantly
efficient in preventing oxidation of lipids in food model systems
(madhari et al 1996; chizault et al 1952; pruth, 1980). Aqeous
clove infussor germinated spores of baccilus subtillis (al-khayat
and blank 1985). It also inhibits pathogens, campylobacter jejuni,
E.coli and s.aureus at levels of 0.03-0.05 percent (smith et al
1998). The ethanolic extract of clove showed the broadest
antibacterial activity by inhibition mostly the growth of all
bacterial strains tested, S.Typhimerins was the most susceptible
sterotype to the extracts of clove. Sema ogaoglu, nursel dosbile
and suleyman alemdar in 2006 found that cloves exhibited similar
results as cumin. This spice was effective against s.aureus,
e.faecalis, m.smegatis, m.luteus and c.albleeans; however in
contrast

to

cumin

it

was

found

to

be

effective

against

K.pneumooniae. arora and kaur (1999) assayed the sensitivity of

yeast spices aqsous extracts and found that colves extract was
able to inhibit candida acutus c.albicans (
Rhodotorula,

rubra,

sacharomyces

cervisae

c.catenulata,
and

trignopsis

cariabilis and in some cases, strong cidal effects was observed.

Essential oils of clove possess anti microbial properties (kalemba
and kunicka 2003) clove oils was effective against e.coli,
l.mon________________s. enteric (friedman et al, 2002). The
antibacterial activity of cloves against two garm negative
bacterial such as pseudomonas floureus and serratia
liquefaciemsc and four gram positive bacteria, such as

bronchothrx thermosphacta, carnobacterium piscicola

lactobacillus curvatus and lactobacillus involved in eat spoilage
was found effective. The 1/100 dilution of cloves oil inhibited the
bacterial growth of five of the six tested bacteria mentioned
above.
Garlic (Allium sativum) decocted garlic extracts that are left to
set overnight are very effective in healing wounds. In 1858, Louis
Pasteur observed garlics antibacterial activity and it was used as
an antiseptic to prevent gangrene during world war I and world
war II. Two percent (w/v) ground garlic in potato dextrose agar
completely inhibits the growth of seven mycotoxin producing
mouls for up to 21 days. (Azzouz and bullerman, 1982). Whole
plant plant contain an antbiotic essential oils including allicine and
diallyl sulphide (

and paris 1977). There are many

organisms inhibited by garlic among which the important are viz,

coliforsms garlic extracts) chung et al, 1997).
L. monocytogens (garlic oil). Firozi et al, 1998. Salmonella
lypimurium (49 micrograms /ml allian) (feldberg 1988) and also
other organisms (both gram positive and gram negative ) as an
anti oxidant agent, it was found to have an excellent antioxidant
capacity

among 22 common vegetables (cao et al. 1996).

Especially the action of garlic (bacteria cause of ulcers) is a major

therapeutic benefit obtained from foods rich in garlic (or
thiosulphate compounds)
Fennel (foeniculum vulgare)

the antimicrobial activity of

fennel has been reported to be effective against

E.coli,

salmonella typhimurium, staph aeureus, candida albicans at

levels of 0.25 1.0 percent (v/v) of essential oils (hammer 1999).
Fennel has also been reported to have some antioxidant activity
at o.2% level in the prime stream lard (chipault 1952). Fennel was
effective only against s.aeureus as per sema agaolu, nursel
dostbill and suleyman alemdar (2007)
Ginger (Zinigiber Officinale). The presence of gingerol is
responsible for its microbial activity. Ginger has a potent
antioxidant property Kikuzaki and NAkatani (1993) have reported
to have a higher antioxidant activity than alpha tocopherol. This is
mainly because of the presence of gingerol besides Zingerone.
Also ginger effectively acts as an antioxidant in prime. Steam lard,
piercuss. Although the antibacterial activity is limited, but there
are reports of its being effective against yeast and molds (4%
extract) in mango juice (Ejechi 1998). The minimum inhibitory
concentration (MIC) of ginger extracts on and bacillus subtillis
showed that ginger was more effective and produced remarkable
inhibitory effective on them.
Traditionally, ginger has been used to treat intestinal infections,
especially related with digestive probles. Equally its antibacterial
power is effective against preveting numerous intestinal
problems that take place as a result of the alteration of intestinal
flora. Ginger has the capacity to eliminate harmful bacteria such
as eschericha coli; responsible for most of the diarrhea especially
in children. Ginger

both diarrhea and constipation; hence it

should have impact on the growth of bacillus cereus which mainly

causes diarrhea and nausea. Bacillus subtillis has been implicated

in various foods spoilage including ropiness in bread, production
of Co2 in canned meats, sliminess and coagulation in milk, etc. it
was clear from this work that the solvent of extraction affected
the degree of antibacterial activity of the xtracts. It was observed
that ethanolic extract of ginger gave the widest zone of inhibition.
It ws also observed that raw ginger extracts has activity only on
salmonella typh; and no effect of e.schema coli and bacillus
subtillis. It has been reported that ginger extracts and pngent
compound demonstrated greater antibacterial activity against a
variety

of

bacteria

species

including

helicobacter

pylori,

staphylococci aureus and pseudomona aeruginosa, althoush

mixed result is attributed to different ginger preparation and
varying strength.
Zinziber officinale (Z. officinale) has been shown to have
antimicrobial activity (HAbsah 2000; srinivasan;2001). Ethanolic
extracts of the rhizomes of z. officianales

showed significant

inhibition of growth of both certain gram positive and gram

negative bacteria methanol extracts of the dried powdered ginger
zhizome and the isolated constitutnts , 6-,8-,10- gingerol and 6shogaal were tested against 19 strains of a 19 strain in vitro with
a minimum inhibitory concentration range of 6.25-so ug/ml. the
crude extract, containing gingerols, ing=habited the growth of all
strains of H.pylor with a MIC range of 0.78 to 12.5 ug/ml and with
significant activity against the coa+ strain (mahady et al: 2003).

The extracts of ginger exhibited antibacterial activity against the

pathogens s.aureus, s. pyrogens, s. pneumonia and H.influenza.
Coriander (coriandrum sativum):- its antimicrobial activity is
shown to be against enterococus faecalis, candida albican at
levels of 0.25-1.0 percent (v/v) of essentialoils (hammer 1999).
The antioxidant activity of coriander, has been reported to help
stabilize prime steam lard at 0.2% level (Chipault 1952)
Black pepper (piper nigrum): the antimicrobial activity of
black pepper has been shown to be effective against enterococus
faecalis (hammer 1999) and clostridis species (Huhtanen 1980).
Black pepper is reportedly more potent than the alpha tocopherol
(shahidi and warasundra 1992). Also (chipault 1952) has been
reported them to effective in prime lard and piecrusts at 0.2%
levels. It has been shown to have antimicrobial activity (perez and
anesini) 1994: Dorman and deans, 2000). Both aqueous and
ethanol extracts of black pepper screened for antibacterial
activity against a penicillin G resistant strain of staphylococcus
aureus, showed antibacterial activity which was determined by
the agar well diffusion method, using cephalolin as a standard
antibiotic

(perez

and

anesini,

1994).

Pipering

[1-,51,1,3-

benzodioxcol -5 yl] -1-ox-2-4, pentadienyl, piperdine a pungent

alkaloid present in P. Nigrum =, enhanced the bioavailability of
various structurally and therapeutically diverse drugs. A concise
mechanism of its bioavailability enhancing action is poorly
understood.

However data suggested that piperine is absorbed very fast

across the intestinal barrier, it may form non-polar complexes
with drugs and solutes increasing permeability across the barrier
(Khajurra, 1998).
Mustard (Brassica Juncea): Kirk, 1964 amd Kharcheneko 1964
reports allyl isothiocyanate as the main component of mustard oil
(szakacs dobozi, 1988) demonstrated that addition of 0.2%
triochdema reeser cellulose increases the yield 50% higher than
without it. Its antifungal activity has been reported by Oliver,
1999 Nielsen and Rios 2000.
The chemical composition of Brassica Juncea essential oil has
studied by (jimm, 2003). Allyl isothiocynate showed strong
andtimicrobial activity against a wide spectrum if bacterium as
observed by (furuya and ishikl, and chan 1995). From the above
literature it is clear that herbs and spices have great potential and
could be utilized as antimicrobial or preservative agents. The
literature is not equally available for all, some are more worked
out and some are little. No systematic work with full information
about the components and their antimicrobial activity has been
done.
Due to isolated and insufficient information. These material have
not

so

far

been

widely

accepted

in

pharmaceutical

and

preservative industries. Therefore this work has been planned

that materials could have a wider application.
DISCUSSION

Often we talk about the negative issues cause by microorganisms

in food, such as food spoilage. But they can be used as efficient
live factories to produce health beneficial food. Without some
microorganisms production if certain food items may not be
possible or may be costly. Microbes are chest resurces that
consists of numerous enzymes which can convert complex
chemical, structures into simple digestible molecules with a high
efficiency. These microorganisms use the nutrients in the food and
other required precursors for their growth in which a fermented
food results.
People realize long time ago that micro organisms could improve
nutritional value of the food they are eating. Some types of food
that is comsumed today have a hundreds or thousands year long
tradition. Evidence (like excavated jars) suggest that some
fermented beverages were produced in Babylon 5000 years BC
while milk fermentation started a bit later around 3000 BC.
Fermentation is one of the oldest methods used for food
preservation. List of food that could be processed and improved
by microorganisms is long as well as rthe list of beneficial effects
fermented food has on our health.
Fermentation process increases nutitional value of food by
increasing the level of proteins, vitamins, amino acids, essential
fatty acids. It also increases food digestibility. Typical example if
lactose intolerance that prevents many people from drinking milk.
Microorganisms used in milk fermentation digest lactose and
decrease it to the level that vcould be safely consumed even with

lactose intolerant people. Bacterial enzymatic hydrolysis can

enhance

intestinal

bioavailability

of

the

fat

and

proteins.

Disorders like constipation, irritable bowels syndrome, yeast

infection and some kind of allergies, could be prevented or
decreased with well balanced gut microfloral
Here is the list of some less known fermented products and their
effects on the health.
Keffir is popular in eastern Europe and middle eastern
countries. This is easily digested alcoholic milk beverage that
could improve immune response and is often given to
immune compromised patients like those suffering from AIDS
or cancer
Koumiss is Caucasus derived fermented dairy product used
in the treatment of pulmonary tuberculosis
Chinese douche is another soybean product that is effective in
lowering high blood pressure.
Apart from benefits of bacteria in food industries, some of them
are causing food poisioning. For this reasons the purity of culture
f important for any fermentation process. The most important part
of microbial inoculums preparation is maintenance of purity,
which helps in avoiding unwanted microorganisms in food
processing. Therefore, the knowledge of bacteria which may
cause or damage food or equally important.
Today many bacterial has toxins and also possess the capability to
cause disorders like food poisioning in human and animals. The
consumption of these foods may show symptoms like headache,
fevers, cold, vomiting etc. one of such bacteria is bacillus cereus.

These bacterial groups consists of six members. These are

B.anthatcis,

B.cereus,

B.mycoides,

B.pseudomycoides,

B.welhenstephanensis and B.Thuringglensis. these specre are

closely related and therefore should be placed within one specres.
B.Cereus is many at times isolated from variety of foods. These
bacterium is soil inhabitant. It causes diarrhea, vomiting and
illness simultaneously. It is many times found in deserts, meat
dishes, dairy products and are associated with diarrhea illness.
These toxin has been isolated from these bacterium

and is

characterized.
It

is small

peptide

ring

synthesized

by

enzymes

peptide

synthetase . in many outbreaks, it has been observed that three

typical B.Cereus enterotoxins are involved. Two out of three are
enterotoxins which are three component proteins and they are
related. The third protein iis CYTK which is one component protein
in nature. This species has unique characteristics to adhere to
many surfaces and thus they are able to survive normal washing
and disinfection.
In such scenarios, one needs to have knowledge of such bacteria
their metabolism, habitat, prevention, etc., so as to maintain the
quality of certain products.
All food should be safe and free from contamination and spoilage
at all points in its journey from its source, until it reaches the
consumer. However, food contamination is a serious public health
problem in many countries such as Ethiopia resulting in food
borne diseases that affect many people every year.

Hence, awareness of potential sources of food contamination is an

important component of good nutrition and good health. Food can
be contaminated by microorganisms, chemicals and physical
factors. Food may be contaminated by different microorganism or
by chemicals that can cause health problems for anyone who eats
it.
Food spoilage is a metabolic process that causes foods to be
undesirable or unacceptable for human consumption due to
changes in sensory characteristics. Spoiled foods may be safe to
eat, i.e. they may not cause illness because there are no
pathogens or a toxin present, but changes in texture, smell, taste,
or appearance cause them to be rejected. Some ecologists have
suggested these noxious smells are produced by microbes to
repulse large animals, thereby keeping the food resource for
themselves [1]. Food loss, from farm to fork, causes considerable
environmental

and

economic

effects.

The

USDA

Economic

Research Service estimated that more than ninety-six billion

pounds of food in the U.S. were lost by retailers, foodservice and
consumers in 1995. Fresh produce and fluid milk each accounted
for nearly 20% of this loss while lower percentages were
accounted for by grain piroducts (15.2%), caloric sweeteners
(12.4%), processed fruits and vegetables (8.6%), meat, poultry
and fish (8.5%), and fat and oils (7.1%) [2]. Some of this food
would have been considered still edible but was discarded
because it was perishable, past its sell-by date, or in excess of

needs.

There

are

also

environmental

and

resource

costs

associated with food spoilage and loss. If 20% of a crop is lost,

then 20% of the fertilizer and irrigation water used to grow that
crop was also lost. Shelf life of a food is the time during which it
remains stable and retains its desired qualities. The wide array of
available dairy foods challenges the microbiologist, engineer, and
technologist to find the best ways to prevent the entry of
microorganisms, destroy those that do get in along with their
enzymes, and prevent the growth and activities of those that
escape

processing

treatments.

Troublesome

spoilage

microorganisms include aerobic psychrotrophic Gram-negative

bacteria, yeasts, molds, heterofermentative lactobacilli, and
spore-forming bacteria. Psychrotrophic bacteria can produce large
amounts of extracellular hydrolytic enzymes, and the extent of
recontamination of pasteurized fluid milk products with these
bacteria is a major determinant of their shelf life. Fungal spoilage
of dairy foods is manifested by the presence of a wide variety of
metabolic by-products, causing off-odors and flavors, in addition
to visible changes in color or texture.
Coliforms, yeasts, heterofermentative lactic acid bacteria, and
spore-forming bacteria can all cause gassing defects in cheeses.
The rate of spoilage of many dairy foods is slowed by the
application of one or more of the following treatments: reducing
the pH by fermenting the lactose to lactic acid; adding acids or
other approved preservatives; introducing desirable microflora
that restricts the growth of undesirable microorganisms; adding

sugar or salt to reduce the water activity (aw); removing water;

packaging to limit available oxygen; and freezing. The type of
spoilage

microorganisms

because

of

production,

the

differs

selective

formulation,

widely

effects

of

processing,

among
practices

dairy

foods

followed

packaging,

in

storage,

distribution, and handling [3].

Scenario of food spoilage worldwide

The issue of food losses is of high importance in the efforts to
combat hunger, raise income and improve food security in the
worlds poorest countries. Food losses have an impact on food
security for poor people, on food quality and safety, on economic
development and on the environment. The exact causes of food
losses vary throughout the world and are very much dependent
on the specific conditions and local situation in a given country. In
broad terms, food losses will be influenced by crop production
choices

and

patterns,

internal

infrastructure

and

capacity,

marketing chains and channels for distribution, and consumer

purchasing and food use practices.
Irrespective of the level of economic development and maturity of
systems in a country, food losses should be kept to a minimum.
Food losses represent a waste of resources used in production
such as land, water, energy and inputs. Producing food that will
not be consumed leads to unnecessary CO2 emissions in addition
to loss of economic value of the food produced. Economically

avoidable food losses have a direct and negative impact on the

income of both farmers and consumers. Given that many
smallholders live on the margins of food insecurity, a reduction in
food losses could have an immediate and significant impact on
their livelihoods. For poor consumers (food insecure or at-risk
households), the priority is clearly to have access to food products
that are nutritious, safe and affordable. It is important to note that
food insecurity is often more a question of access than a supply
problem. Improving the efficiency of the food supply chain could
help to bring down the cost of food to the consumer and thus
increase access. Given the magnitude of food losses, making
profitable investments in reducing losses could be one way of
reducing the cost of food. But that would, of course, require that
financial gains from reduced losses are not outweighed by their
costs. How much food is lost and wasted in the world today and
how can we prevent food losses? Those are questions impossible
to give precise answers to, and there is not much ongoing
research in the area. This is quite surprising as forecasts suggest
that food production must increase significantly to meet future
global demand [4].
Worldwide postharvest fruit and vegetables losses are as high as
30 to 40% and even much higher in some developing countries.
Reducing postharvest losses is very important; ensuring that
sufficient food, both in quantity and in quality is available to every
inhabitant in our planet. The prospects are also that the world
population will grow from 5.7 billion inhabitants in 1995 to 8.3

billion in 2025. World production of vegetables amounted to 486

million ton, while that of fruits reached 392 million ton. Reduction
of post-harvest losses reduces cost of production, trade and
distribution, lowers the price for the consumer and increases the
farmer's income. Fruits and vegetables are very important food
commodities not only in India but all over the world. India, which
is the second most populated country of the world, is still
struggling to achieve self-sufficiency to feed about 800 million
people. For this purpose, fruits and vegetables have got their
specific importance to provide a balance and healthy diet to the
people. India is the second largest producer of vegetables and
fourth largest producer of fruits in the world. Though India is
producing adequate quantities of fruits and vegetables, yet on
account of losses in the field as well as in storage, they become
inadequate. Generally, about 30 % fruits and vegetables are
rendered unfit for consumption due to spoilage after harvesting.
India annually produces fruits and vegetables of the value of
about Rs. 7000 crores and wastage may be of the order of Rs.
2100 crores. This is a huge loss of valuable food even when the
mini- mum food requirement of the population is not met.
Therefore, it is important not only to grow more, but also to save
what is grown at high cost.
Post harvest loss of fruits and vegetables has been defined as
"that weight of wholesome edible product (exclusive of moisture
content) that is normally consumed by human and that has been
separated from the medium and sites of its immediate growth and

production by deliberate human action with the intention of using

it for human feeding but which for any reasons fails to be
consumed by human." Not only quantity and quality but even the
appearance of fruits and vegetables are affected and their market
value is reduced. Most 'skin deep' injuries such as 'fly speck' in
apples do not affect the edible part. Some infections are not
harmful as they occur on inedible parts and can be trimmed off
before.
Fresh fruits and vegetables are perishable and highly prone to
these losses because they are composed of living tissues. These
tissues must be kept alive and health throughout the process of
marketing. These are composed of thousands of living cells which
require care and maintenance.Therefore, the reduction of postharvest loss of fruit and vegetables is a complementary means for
increasing production. It may not be necessary to considerably
step up the production of fruits and vegetables with the growing
demand if the post-harvest loss is reduced to a great extent. The
cost of preventing losses after harvest in general is less than
preventing a similar additional amount of fruit and vegetable crop
of the same quality. Attention to the concept of post-harvest food
loss reduction, as a significant means to increase food availability,
was drawn by the World Food Conference held in Rome in 1974.
The global dairy industry is impressive by large. In 2005, world
milk production was estimated at 644 million tons, of which 541
million tons was cows milk. The leading producers of milk were
the European Union at 142 million tons, India at 88 million tons,

the United States at 80 million tons (20.9 billion gallons), and

Russia at 31 million tons. Cheese production amounted to 8.6
million tons in Western Europe and 4.8 million tons in the United
States [5]. The vast array of products made from milk worldwide
leads to an equally impressive array of spoilage microorganisms.
A survey of dairy product consumption revealed that 6% of US
consumers would eat more dairy products if they stayed fresher
longer [6]. Products range from those that are readily spoiled by
microorganisms to those that are shelf stable for many months,
and the spoilage rate can be influenced by factors such as
moisture content, pH, processing parameters, and temperature of
storage.
Scenario of food spoilage in India
India is the second major producer of fruits and vegetables and
ranks next to Brazil and China respectively, in the world. It
contributes 10 percent of world fruit production and 14 per cent of
world vegetable production. Fruits and vegetables are more prone
to spoilage than cereals due to their nature and composition, and
this

spoilage

occurs

at

the

time

of

harvesting,

handling

transportation, storage, marketing and processing resulting in

waste. Efficient management of these wastes can help in
preserving vital nutrients of our foods and feeds, and bringing
down the cost of production of processed foods, besides
minimizing pollution hazards. According to India Agricultural
Research Data Book 2004, the losses in fruits and vegetables are
to the tune of 30 per cent. Taking estimated production of fruits

and vegetables in India at 150 million tones, the total waste

generated comes to 50 million tones per annum. The post-harvest
technologies for perishable horticultural produce serve as an
effective tool for getting better return to the produce and also
help in avoiding wastage both at production site and distribution
centers, which will help in regulating the market infrastructure.
Like any other food, fruits and vegetables are also prone to
microbial spoilage caused by fungi, bacteria, yeast and moulds. A
significant portion of losses of fruits and vegetables during postharvest period is attributed to diseases caused by fungi and
bacteria. The succulent nature of fruits and vegetables makes
them easily invaded by these organisms. Besides attacking fresh
fruits and vegetables, these organisms also cause damage to
canned and processed products. Many serious post-harvest
diseases occur rapidly and cause extensive break down of the
commodity,

sometimes

spoiling

the

entire

package.

It

is

estimated that 36 % of the vegetable decay is caused by soft rot

bacteria. Similarly fruit rot in aonla and other soft fruits caused by
fungi is also very destructive. As far as
vegetables are concerned, naturally the source of infection is from
the field, water used for cleaning the surface, contact with
equipment

and

storage

environment.

The

most

common

pathogens causing rots in vegetables and fruits are fungi such as

Alternaria, Botrytis, Diplodia, Monilinia, Phomopsis, Rhizopus,
Pencillium, Fusarium, etc. Among bacteria Ervinia, Pseudomonas,
etc. cause extensive damage.

High temperature and relative humidity favour the development

of post-harvest decay organisms. More acidic tissue is generally
attacked by fungi, while fruits and vegetables having pH above
4.5 are more commonly attacked by bacteria, ego bacterial soft
rot of potato caused by Ceratocystis, fimbriata, water soft rot of
carrot by Sclerotinia
sclerotiorum etc.
In India, there is a vast scope for growing fruit and vegetable
throughout the yearin one or other part of the country because
the climatic conditions are highlysuitable for growing various
types of fruits and vegetables. Fruit and vegetable are highly
perishable but most important commodity for human diet due to
their high nutritional value. They are the cheapest and other
source of protective food supplied in fresh or processed or
preserved form throughout the year for human consumption.
Hence the national picture will improve significantly. Fruit and
vegetable are available in surplus only in certain seasons and
availability in different regions. In peak season due to improper
handling practices, marketing, storage problems around 20-25%
fruit and vegetable are spoilt in various stages. Fruit and
vegetable are living commodities as they respire. Hence, proper
post harvest management handling and processing is required in
horticulture crops. A variety of fresh fruit and vegetable in India
can be made available in plenty due to favourable agro-climatic
situations. Hence there is no derth for raw material for processing.
Product profile being developed in India at present is limited to

few fruit and vegetable like mango, pineapple, grapes etc. But
there is a wider potentiality for processing of papaya, banana,
jack, guava, aonla, carambola and other minor fruits. Similarly
there is a greater scope for processing cauliflower, carrot, bittergourd onion, garlic, watermelon, muskmelon etc. Proper handling,
packaging, transportation and storage reduce the post-harvest
losses of fruit and vegetables. For every one percent reduction in
loss will save 5 million tons of fruit and vegetable per year.
Processing and preservation technology helps. There are about
4000 small and large scale processing units in the country which
process only about 2.5% of the total fruit and vegetable as
against 40-85% in developed countries.

REFERENCES
[1] D.E. Burkepile, J.D. Parker, C.B. Woodson, H.J. Mills, J. Kubanek,
P.A. Sobecky and M.E. Hay. Ecol., 2006, 87: 28212831.
[2] L.S. Kantor, K. Upton, A. Manchester and V. Oliveira. Food Rev.,
1997, 20: 212.
[3] P. Martorell, M.T. Fernandez-Espinar and A. Querol. Int. J. Food
Microbiol., 2005, 101: 293302.
[4] V.B. Fonnesbech, K. Venkateswaran, M. Satomi and L. Gram.
Appl. Environ. Microbiol., 2005, 71: 66896697.
[5] T. Kutzemeier. Europ. Dairy Magzin., 2006, 7: 3436.
[6] P. Lempert. Progressive Grocer, 2004, 83: 18p.
[7] L. Gram, L. Ravn, M. Rasch, J.B. Bruhn, A.B. Christensen and M.
Givskov. Int. J. Food Microbiol., 2002, 78:7997.
[8] R.E. Mandrell, L. Gorski and M.T. Brandl. In: Microbiology of
fresh fruits and vegetables (Eds. G.M. Sapers,J.R. Gorney, and A.E.
Yousef ), New York: Taylor and Francis Group, 2006, pp. 33-73 .
[9] P. Elez-Martinez, J. Escola-Hernandez, R.C. Soliva-Fortuny and
O. Martin-Belloso. J. Food Prot., 2004, 67:25962602.
[10]G.D. Casey and A.D.W. Dobson. J. Appl. Microbiol., 2003, 95:
1322.
[11] J.A. Couto, F. Neves, F. Campos and T. Hogg. Int. J. Food
Microbiol., 2005, 104: 337344.
[12] J.I. Pitt and A.D. Hocking. Fungi and Food Spoilage. Blackie
Academic and Professional, New York, 1977.
[13]K. Boor and H. Fromm. In: Food Spoilage Microorganisms (Ed.
C. Blackburn), CRC Press LLC, Boca RatonFL, 2006, pp. 171193.

[14]M. Rasch, J.B. Andersen, K.F. Nielsen, L.R. Flodgaard, H.

Christensen, M. Givskov and L. Gram. Appl.Environ. Microbiol.,
2005, 71: 33213330.
[15] J.B. Bruhn, A.B. Christensen, L.R. Flodgaard, K.F. Nielsen, T.O.
Larsen, M. Givskov and L. Gram. Appl.Environ. Microbiol., 2004,
70: 42934302.
[16]F. Russo, D. Ercolini, G. Mauriello and F. Villani. Food
Microbiol., 2006, 23: 797802.
[17]M.C. Hozbor, A.I. Saiz, M.I. Yeannes and R. Fritz. Food Sci.
Technol., 2006, 39: 99104.
[18]V.H. Tournas. Int. J. Food Microbiol., 2005, 99: 7177.
[19] J. Calvo, V. Calvente, M.E. de Orellano, D. Benuzzi and M.S. de
Tosett. Food Microbiol., 2007, 21:703714.
[20]D.S. Thomas and R.R. Davenport. Food Microbiol., 1985, 2:
157-169.
[21]D.J. Fitzgerald, M. Stratford, M.J. Gasson and A. Narbad. J.
Food Prot., 2004, 67: 391395.
[22]L. Cocolin, N. Innocente, M. Biasutti and G. Comi. Int. J. Food
Microbiol., 2004, 90: 8391.
[23]M. Walker and C.A. Phillips. Food Microbiol., 2007, 24: 313
318.
[24]H.C. Deeth and C.H. Fitz-Gerald. In: Developments in dairy
chemistry (Ed. P.F. Fox) Applied Science,London, 1983, pp. 195239.
[25]B. Dogan and K.J. Boor. Appl. Environ. Microbiol., 2003, 69:
130138.

[26]R.G. Stevenson, M.T. Rowe, G.B. Wisdom and D. Kilpatrick. J.

Dairy Res., 2003, 70: 293296.
[27]R. Mayr, K. Gutser, M. Busse and H. Seiler. Milchwissenschaft,
2004, 59: 262266.
[28] J.D. Meyer, J.G. Cerveny and J.B. Luchansky. Int. J. Food
Microbiol., 2003, 103: 3745.
[29]M.B. Mayoral, R. Martin, A. Sanz, P.E. Hernandez, I. Gonzalez
and T. Garcia. Int. J. Food Microbiol., 2005,105: 2734.
[30]N. Arneborg, L. Jespersen and M. Jakobsen. Arch. Microbiol.,
2000,174: 125-128.
[31]S.E. Lindow and M.T. Brandl. Appl. Environ. Microbiol., 2003,
69: 18751883.
[32]P. Ludovico, M.J. Sousa, M.T. Silva, O.C. Lea and M. Corte-Real.
Microbiol., 2001, 147: 2409-2415.
[33]C. Restuccia, C. Randazzo and C. Caggia. Int. J. Food
Microbiol., 2006, 109: 146150.
[34]L. Fernandes, M. Corte-Real and O.C. Lea. Lett. Appl.
Microbiol., 1999, 28: 345-349.
[35]O. Pepe, G. Blaiotta, G. Moschetti, T. Greco and F. Villani. Appl.
Environ. Microbiol., 2003, 69: 23212329.
[36]G. Fleet. Crit. Rev. Biotechnol., 1992, 12: 1- 44.
[37]P. Munsch, V.A. Geoffroy, T. Alatossava and J.M. Meyer. Appl.
Environ. Microbiol., 2000, 66: 48344841.