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Pesticide is a generic term that covers a wide range of natural and synthetic chemicals over 700 in total that are used to protect crops from attack from pests, both before and after har

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Sun Z-T et al (1999) Increased risk of hepatocellular carcinoma

in male hepatitis B surface antigen carriers with chronic

hepa-titis who have detectable urinary aflatoxin metabolite M1.

Hepatology 30: 379–383.

Wang L-Y, Hatch M, Chen CJ et al (1996) Aflatoxin exposure

and the risk of hepatocellular carcinoma in Taiwan

Interna-tional Journal of Cancer 67: 620–625.

Wild CP and Turner PC (2002) The toxicology of aflatoxins as a

basis for public health decisions Mutagenesis 17: 471–481.

Yu MW, Lien JP, Chiu YH et al (1997) Effect of aflatoxin

metabolism and DNA adduct formation on hepatocellular

carcinoma among chronic hepatitis B carriers in Taiwan.

Journal of Hepatology 27: 320–330.

Pesticides

M Saltmarsh, Alton, UK

ª 1998 Elsevier Ltd All rights reserved.

This article is reproduced from the previous edition,

pp 869–874, ª 1998, Elsevier Ltd.

What are Pesticides?

Pesticide is a generic term that covers a wide range

of natural and synthetic chemicals (over 700 in

total) that are used to protect crops from attack

from pests, both before and after harvest There

are many different sorts of pests The term includes

insects, slugs and snails, nematode worms, mites,

rodents, weeds, molds, bacteria and viruses The

chemicals can be applied before and during growth

of the plant or on to the stored crop as, for example,

fumigants, which are used to kill pests that have

infested stored cocoa or grain Chemicals used to

treat pests on animals are not included; they are

considered as veterinary medicines

The pesticide formulation used by the farmer will

include the pesticide chemical itself and a number of

other chemicals that enable it to be applied and to

work as effectively as possible These will include

solvents, adhesives, and surface-active agents such as

emulsifiers In some cases other chemicals, known as

‘safeners,’ are applied to minimize the damage done

to the crop while maintaining the effectiveness of the

spray on the target

It is estimated that worldwide usage of pesticides

is around 2.5 million tons with a cost in 1997 of

US$21 billion

Why Do We Need Pesticides?

Food crops are subject to attack by a multitude of

pests and diseases and pesticides are applied to

mini-mize the damage to the crop It has been estimated

that without protection world cereal crop yields would fall by between 46 and 83% History is lit-tered with records of crop failures and famine caused primarily by rodent, insect or fungus Some

of these events have had a wide-ranging and long-lasting effect, like the 1845–1846 Irish potato fam-ine and the 1917–1918 German ‘turnip winter,’ the latter so called because the potatoes rotted and turn-ips were the only stored root crop that was available

to feed the population through the winter Both these events, in which 1.5 million and 700 000 peo-ple died, respectively, were caused by potato blight, infection by the fungus Phytophthora infestans Famine caused by massive swarms of locust is still all too common in Northern Africa and Arabia Less spectacular but as disastrous is the loss of an esti-mated 30% of harvested crops in India to rodents

In addition to the loss of the crop, pesticides are used to control agents which make the crop toxic rather than healthy Two examples are the toxins caused by fungi When an insect bores into a peanut

it allows spores of the fungus Aspergillus flavus to enter and grow, producing the aflatoxins, a series of carcinogens When rye (Secale cereale) grows in damp conditions a fungus, Claviceps purpurea, can grow on the seed If this seed is subsequently ground into flour and made into bread it can cause consu-mers to suffer hallucinations, gangrene, and death Outbreaks amounting to epidemics were common in the Middle Ages in Europe and one occurred as recently as 1951 in France

A second reason relates not so much to quantity

as to quality Supermarkets in the developed nations offer a wide range of fresh produce at competitive prices Consumers do not like holes made by slugs and snails in their fresh lettuce They do not expect scab marks on their apples, or holes made by small maggots in their carrots Flour millers do not expect

to have to clean the grain from weed seeds before milling Even small defects can dramatically reduce the value of the crop, or indeed make it unsaleable, and the need for a competitive price requires mini-mal labor input so that application of pesticide is essential

Types of Pesticides

There are currently around 600 pesticides, both natural and synthetic Natural pesticides include both chemicals derived from plant sources and biological agents such as parasitic wasps, mites, bacteria, and chemicals contained within or exuded by plants or bacteria While there is no inherent reason why natural products should be any safer than synthetic ones (after all, insect

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venoms and toxins and poisonous plants are

nat-ural), it appears that the risks do lie in their

potential impact on the environment rather than

on their effect in food There are also increasing

numbers of cases where plants have been given a

gene which expresses a natural pesticide (see

Bacil-lus thuringiensis, below)

At the time of writing, naturally derived pesticides

make up less than 5% of the world pesticide market,

but a great deal of work is being devoted to the

screening of natural sources and this proportion

will certainly increase The most successful natural

product development so far has been that of the

pyrethrin insecticides, of which 33 are currently

available

The largest classes of pesticides are pyrethrins,

organochlorines, organophosphates, and

carba-mates, although there are many smaller classes

with only one or two members The chemical

struc-tures of the key members of the major groups are

given inTable 1

Important Pesticide Groups

This list covers the important pesticide groups and

some individual pesticides but does not attempt to

be comprehensive

Pyrethrins

Pyrethrins are chemically related to pyrethrin,

which is a secondary metabolite found in the

flow-ers of the pyrethrum plant (Chrysanthemum

ciner-ariaefolium) Dried pyrethrum flowers were used as

an insecticide in ancient China and in the middle

ages in Persia The dried flowers are still used

Current production is around 20 000 tons per

an-num centered in Kenya and Tanzania The

pyre-thrins are effective insecticides, having very low

dose rates and rapid knockdown of insects but

being harmless to mammals under all normal

con-ditions Natural pyrethrins break down rapidly

under the influence of oxygen and UV light This

limits their use in agriculture, but recently synthetic

analogs have been developed to overcome these

problems Starting from the structure of the natural

product a large number of synthetic compounds

have been made It is worth noting how they differ

in effectiveness: deltamethrin is a broad range

insec-ticide; allethrin is particularly toxic to house flies

(Musca domestica) but much less effective with

other insects; flumethrin is active against cattle

ticks; while others are acaricides or miticides with

little or no insecticidal activity

Bacillus thuringiensis

Bacillus thuringiensis is a widely distributed bacter-ium that during sporulation produces a crystal inclusion which is insecticidal when ingested by the larvae of a number of insect orders Susceptible orders include Lepidoptera, Diptera, and Colcop-tera The action of B thuringiensis was first observed in 1901 as the cause of a disease of silk-worms Several strains of the bacterium have been identified with activity against a range of insects including cabbage looper, tobacco budworm, mos-quito, black fly, and more recently nematodes, ants and fruit flies While the bacterium appears an ideal insecticide (having a toxicity 300 times greater than synthetic pyrethroids), it requires careful use It is most effective against neonates and early larval instars so that spraying must be timed for egg hatch It also has no contact activity and must be ingested so the plant must be well covered to ensure the insect receives a lethal dose Furthermore it has

a half-life in the field as short as 4 h, so careful timing is essential for it to be effective Despite these limitations, it has been shown to be an impor-tant component of crop management programes One way of overcoming the problems of applica-tion of B thuringiensis is to incorporate the gene responsible for expression of the protein into the crop plant This has been achieved with maize (Zea mays) to protect against the European corn borer, with cotton (Gossypium hirsutum) to protect against

a range of budworms and bollworms, and with potato (Solanum tuberosum) against Colorado bee-tle (Cotton may seem irrelevant in a text on food but cottonseed oil is used extensively in cooking oils, margarines, and industrial fats.) This genetic mod-ification has great benefits but care has to be taken that the food product has not changed in some unpredicted way All genetically modified foods have to be extensively tested and cleared by regula-tory agencies before release

Neem oil

This is an oil obtained from the neem tree, Azadirachta indica A Juss It has been used as an insecticide in India and Africa but is increasingly being developed as a significant commercial pro-duct It contains a number of compounds, one of the most active being azardirachtin, which is an insect antifeedant but also shows growth inhibitory and endocrine disrupting effects This product and its individual components is at the beginning of its commercial development, which is likely to result in

a series of products as significant as those from pyrethrum

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Microbial phytotoxins

These are herbicides and include the highly

com-mercially successful glufosinate, a synthetic form of

phosphinothricin, first isolated from Streptomyces

hygroscopicus, a soil-borne microbe This com-pound is a potent, irreversible inhibitor of gluta-mine synthetase which is used in plants for photorespiration Many attempts have been made

to make synthetic variants of phosphinothricin

Table 1 Chemical structure and acceptable daily intake (ADI) of some pesticides

body weight) Deltamethrin Pyrethrin

O C

CN

H

CO2 C

Br

Br CH

CH3

CH3

0.01

CH

CCl3

0.02

Lindane (HCH) Organochlorine

Cl Cl

Cl Cl

Chlorfenvinphos Organophosphate

(mixture of two isomers)

Cl ,

C C H

Cl O

(CH3CH2O)2P (CH3CH2O)2P

O

Cl

C C Cl

H O

Malathion Organophosphate

CHCH2CO2CH2CH3

CO2CH2CH3 PS

S (CH3O)2

0.02

OCH(CH3)2

0.02

Simazine Triazine

N N

N

Cl NHCH 2 CH 3

NHCH 2 CH 3

0.005

CH3PCH2CH2CHCO2H

OH NH2

0.02

HO2CCH2NHCH2P(OH)2

0.3

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without success Other members of this group

include anisomycin and herboxidiene, derived

from other Streptomyces strains The veterinary

insecticide, avermectin is derived from Streptomyces

avermitilis

Organochlorines

The organochlorines were the first group of

syn-thetic insecticides and without them the dramatic

decrease in malaria observed in the 1950s would

have been impossible The best known of this

class is DDT (dichlorodiphenyltrichloroethane) but

others include 2,4 DD, hexachlorbenzene, and

lin-dane Of these only lindane

(-hexachlorocyclohex-ane, see Table 1) is still in use in the developed

world

These compounds are very slow to break down in

the environment and one result of this persistance

was the decline in bird numbers graphically

described by Rachel Carson in the book Silent

Spring The problem was that DDT was

concen-trated through the food chain and predator birds in

particular were failing to raise chicks Since the

organochlorine pesticides and other sources of

orga-nochlorines in the environment have been largely

phased out, numbers of many species of birds are

rising again It is recognized that pesticides are still

having an adverse influence on numbers of some

birds that inhabit farmland However, this is not a

straightforward effect In the case of the grey

par-tridge, for example, it is because herbicides have

reduced the number of weeds, which in turn has

reduced the number of insects that feed on the

weeds, resulting in fewer insects for the chicks

to eat

The mechanism of action of the organochlorines

is not known in detail although they appear to act

on the central nervous system In humans the

orga-nochlorine compounds tend to accumulate in the

body fat and in mothers’ milk While there is no

direct evidence that they cause mutations or cancers,

there is concern that lindane may be a carcinogen

and its role in breast cancer is still under review

However, in contrast, DDT and -HCH have both

been shown to inhibit tumors in mice initiated by

aflatoxin B1

Although organochlorine pesticides have largely

been phased out in Europe, analysis for them

con-tinues and low levels of lindane are still being

detected in milk in the UK (typically at

0.005 mg kg 1compared with the maximum residue

limit (see below) of 0.008 mg kg 1and an acceptable

daily intake (see below) of 0.05 mg per kg body

weight)

Organophosphorus compounds

Organophosphorus compounds generally contain both sulfur and phosphorus linked to carbon atoms Their discovery was a by-product of the development of nerve gases The group includes parathion, malathion, dimethoate, diazinon, and chlorfenvinphos They are used as herbicides, insec-ticides, and fungicides They break down quickly in the environment and do not concentrate in body fats, although they may be stored for some time However, their mode of action – inhibition of acet-ylcholine esterase – means that they affect both insects and mammals and their use depends on the effective dose in the target species being below the sensitivity of other species

Acute effects of sublethal doses of organopho-sphates in man include sweating, salivation, abdom-inal cramps, vomiting, muscular weakness, and breathing difficulties Concern has also been expressed about long-term effects following acute exposure Research suggests that some victims may show reductions in some neurobehavioral tests when tested some months after exposure There are also concerns that people who do not appear to have suffered acute poisoning have subsequently devel-oped debilitating illnesses Symptoms include extreme exhaustion, mood changes, memory loss, depression, and severe muscle weakness

Carbamates

Carbamates are derived from carbamic acid and are used against both insects and weeds They are also acetylcholine esterase inhibitors They are very reac-tive and are used up rapidly after application

Methyl bromide

Methyl bromide was for many years the fumigant of choice for destroying insects in stored crops, but it is now being withdrawn as part of the general restric-tion on volatile organohalogen compounds because

of their damaging effect on the ozone layer It is being replaced by a number of less environmentally damaging compounds, including phosphine, although none currently available is as effective or

as cheap as methyl bromide

Phosphine

Phosphine has been used as a fumigant for many years It is highly reactive and leaves no residues but great care has to be taken in its application because it is very toxic to humans

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Control of Pesticides

Control over pesticides is exercised in two ways:

stringent testing on new pesticides before they are

permitted and measurement of the residue in the

crop

Testing pesticides

There are a number of national and international

bodies that approve new pesticides within their

areas of responsibility These include Codex

Alimen-tarius, the European Union, and the US Food and

Drug Administration (USFDA) Currently, within

the European Union, registration of pesticides is

being harmonized under Directive 91/414 EEC

Annexe 1 of this directive will identify all active

ingredients permitted in pesticides As yet this

annexe is incomplete and member states are still

acting under their national laws

Within the UK pesticide registration is carried out

under the Control of Pesticide Regulations 1986 and

is the responsibility of the Ministry of Agriculture,

Fisheries, and Food who are advised by the Advisory

Committee on Pesticides

In the USA a new Food Quality Protection Act of

1996 replaced both the Food, Drug, and Cosmetic

Act and the Insecticide, Fungicide, and Rodenticide

Act to provide a comprehensive regulatory scheme

for pesticides

In order to gain approval for use, pesticides are

subjected to an extensive testing program including

toxicity tests on mammals, plants, insects, fungi,

birds, bees, fish, earthworms, and other soil

organ-isms The toxicity studies include effects of

pesti-cides on fetuses and infant animals There are also

environmental tests which include laboratory tests

on the breakdown and movement of the chemical in

plants, soil, water, air, mammals, birds, and fish

These latter tests determine the rate of decay in the

various species Laboratory tests are followed by

prolonged field trials to determine the fate of the

chemical and its breakdown products in the

envir-onment and to estimate how the pesticide is

concen-trated up the food chain On average it takes about

10 years to develop a new pesticide at a cost of

about £50 million The complete dossier of results

has to be submitted to the approval body who

deter-mine whether the tests have been sufficiently

rigor-ous to allow an acceptable daily intake (ADI) of the

pesticide to be set The ADI is defined as the amount

of a pesticide that can be taken in each day

through-out a person’s life with the practical certainty, on

the basis of all known facts, that no harm will result

This is determined on the basis of the highest level at

which the pesticide has no observable effect in

animal tests This is then reduced by a factor of 10

in case humans are more sensitive than the animals used in the tests, and by a further factor of 10 to allow for cases where some humans may be more sensitive than others In some cases, where the data show unusual effects, the safety factor can be increased from 100 to 500 or 1000 In practice the amount of pesticides to which the population is exposed is far below this level

Table 1 includes the ADI for a number of the more common pesticides There is no evidence that there are any cases where the combined effects of two pesticides are greater than the sum of their individual effects, in other words there is no evi-dence of synergy in toxicology between the different pesticides Once maximum residue limits (MRL see below) for foodstuffs have been set on the basis of good agricultural practice, a total dietary intake is determined by considering all commodities in which the pesticide is likely to be used, and assuming the upper range of consumption, all foodstuffs at the MRL and no losses during transport, storage or food preparation This figure is then compared with the ADI For all permitted pesticides in the

UK the figure is below the ADI

Maximum residue limits

Maximum residue limits (MRLs) are statutory limits set on individual active ingredient and foodstuff combinations They are based on residue levels which result when the pesticide is used according

to the instructions on the label and in accordance with good agricultural practice (GAP) MRLs may

be used to ensure that the pesticides are only being used in accordance with GAP Many countries have codes of good operating practice with training for farmers and operators to ensure that pesticides are used at optimal levels Some countries rely on the Codex Alimentarius Committee on Pesticide Resi-dues to establish MRLs, while others set their own (Codex Alimentarius is an international body which has over 120 countries as members and their stand-ards are increasingly being accepted as the basis of world trade in foodstuffs.)

In the USA the FDA used to set tolerances for pesticide/foodstuff combinations but under the

1996 Act it sets a level for each pesticide in all foods based on the principle of a reasonable cer-tainty of no harm This is defined as a lifetime cancer risk of less than 1 in a million There is also

a requirement that residue tolerances must be speci-fically determined as being safe for children Within the EU, individual member states have historically set their own MRLs which differ from

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state to state Directive 76/895 established a

com-mon MRL setting regime and a series of subsequent

directives has fixed the levels for a series of

pesti-cides in fruit, vegetables, cereal products, and

pro-ducts of animal origin There is an ongoing program

to harmonize the levels throughout the Union

Most industrialized countries have pesticide

sur-veillance programs which cover both

home-produced and imported commodities and these report

annually The EU has an annual specific coordinated

program to check compliance in nominated

combi-nations of pesticide and foodstuff MRLs require

sophisticated equipment for their determination

because the levels are so low and the minimum

detectable limit depends on the foodstuff For

exam-ple the tolerance for aldrin and dieldrin (two

orga-nochlorines) in the USA is between 0.05 and

0.1 mg kg 1 (parts per million), depending on the

foodstuff There are over 600 different active

ingre-dients available commercially Because there are so

many, laboratories around the world have

devel-oped sophisticated rapid analytical techniques to

allow them to screen pesticides by class so that

retailers, food manufacturers, and governments can

carry out analyses as a matter of routine

The MAFF 7th Report of the UK Working Party

on Pesticide Residues in 1996 showed 68% of

sam-ples had no detectable residue, 31% had residues

below the MRL, and <1% were over the MRL

Similar results were obtained by the FDA who

report results with relation to the tolerance to the

pesticide/commodity combination In 1995, of over

9000 samples analyzed, 64% had no detectable

resi-dues, 34% had residues below the tolerance, <1%

had residues over the tolerance and <1% had

residues for which there is no tolerance in that

par-ticular pesticide/commodity combination

In all cases where MRLs or tolerances are

exceeded follow-up action is taken For

home-produced materials, this involves investigation of the

grower and prosecution if necessary For imports,

exceeding the level causes the consignment to be

refused entry

Maximum levels of pesticides are also set for

drink-ing water Pesticides get into water from spraydrink-ing,

runoff, percolation or from treatment of fish in

aqua-culture Good practice is increasingly being developed

to minimize the levels in raw water and treatment

works are developing systems to reduce incoming

levels to levels acceptable for drinking water

Endocrine Disruption

The possibility that a number of chemicals

dis-charged into the environment as a result of human

activity may disrupt the endocrine system of a wide range of mammals has recently been given consider-able prominence Among the chemicals cited are the organochlorine pesticides, most of which have now been withdrawn for other reasons While there is no doubt that there are a significant number of cases of endocrine disruption, the evidence to point to any particular chemical as a cause is lacking It is also worth noting that deliberate endocrine disruption is

a mechanism of a number of natural insecticides which act so as to inhibit development of juvenile larvae to adults Fortunately these pesticides are reactive and usually have a short life in the field

It is also true that there are very many naturally occurring endocrine disruptors, including the phytoestrogens present in vegetables, notably soya beans, peas, beans, cabbage, and hops However, since this issue is very serious a considerable amount

of work has now been initiated and its results will have implications for future testing of pesticides

Future Prospects

In many parts of the world it is recognized that there has been too great a reliance on pesticide use and not enough on improving agricultural practices There is increasing pressure to move towards mini-mizing pesticide usage in order to both improve the environment and to reduce cost This is being done

by using newer, more specific pesticides and by adopting improved agricultural practices and inte-grated pest management (a combination of biologi-cal and chemibiologi-cal control)

Biological control is not new In the 1930s Macro-centrus homonae was introduced into Sri Lanka from Indonesia to control the tea small leaf roller (Adoxophyes) with such success that no chemical control measures are needed for this pest even today More recently there have been some impress-ive results from using predator insects, for example

in the control of cassava green mite (Mononchellus tanajoa) in West Africa and white fly in European greenhouses

In terms of agricultural practice, improved crop hygiene, crop rotation, better understanding of opti-mal timing of application, and varying sowing dates, together with the development of more powerful and more discriminating pesticides has brought about a decrease in pesticide inputs This is seen dramatically in the case of oil seed rape (canola) Less than 1% of the weight of herbicide applied to this crop in 1983 was applied in 1993

Unfortunately pests develop resistance to indivi-dual pesticides over time and research is continually

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needed to develop both new pesticides and resistant

varieties of crops to keep the pests in check There

has been some success with new pesticides having

new modes of action such as the antifeedants and

antimolting agents, but this will be a continuing

battle for the foreseeable future

See also: Phytochemicals: Classification and

Occurrence; Epidemiological Factors

Bacterial Contamination

N Noah, London School of Hygiene and Tropical

Medicine, London, UK

ª 2005 Elsevier Ltd All rights reserved.

The burden of gastroenteritis (GE) in the world, in

terms of both morbidity and mortality, is enormous

In the developing world (e.g., Southeast Asia),

diar-rhea vies with acute respiratory tract infection as the

leading cause of death in childhood Even in the

more developed world, infectious GE is a significant

cause of illness and time lost from work, and death

does occur Infectious intestinal disease in England is

estimated to cost the country £743 million p.a (in

1994–95 prices) GE caused by bacteria was far

more costly than that caused by viruses The more

sophisticated surveillance systems become, the more

GE they uncover

Not all GE is caused by food Probably most GE

is caused by poor hygiene leading to direct or

indir-ect transmission of infindir-ection without the assistance

of food Nevertheless, a major cause of infectious

GE throughout the world is contaminated food The

definition of food poisoning (FP) is not

straightfor-ward In essence, FP is an acute gastroenteritis

caused by food Hepatitis A, typhoid, and

brucello-sis, however, are not usually considered as FP,

whereas botulism is, even though it causes paralysis

and not GE, as is listeria, which causes septicemia

and meningitis

Bacteria are the most common known cause of FP

and, with the possible exception of the Norwalk-like

viruses, of GE also As one would expect, bacterial

FP is more common in summer than winter

Bacteria produce their effects on the intestinal tract

either by direct invasion of the mucosa or by the

production of toxin Some of the toxins are produced

outside the intestinal tract—in the food; others are

formed in the intestine Some invasive bacteria also

produce a toxin in the intestine This article provides

an overview of the bacterial causes of FP

Bacterial Toxins

There are three main forms of bacterial enteric toxin:

Enterotoxin producing excess fluid secretion into the gut (cholera and some types of Escherichia coli) Cytotoxin causing inflammation and mucosal damage (shigella and enterohemorrhagic E coli) Neurotoxin affecting the nervous system (botulism and staphylococcal toxin)

Some E coli strains produce toxin; these are dealt with under Invasive Bacteria Red kidney bean, scombrotoxin and other fish toxins, and heavy metal poisoning are dealt with elsewhere in this book

Staphylococcal Food Poisoning

Background Staphylococcal food poisoning (SFP)

is one of the few causes of bacterial FP that can commonly be attributed to a food handler Humans frequently carry staphylococci either in an infected site or asymptomatically Infected sites include wounds and abscesses, which may be the source of large numbers of staphylococci Asymptomatic sites include throat, nostrils, fingernails, or hair In gen-eral, only coagulase-positive staphylococci (Staphy-lococcus aureus), and only certain types, produce enterotoxin Rarely, some coagulase-negative strains may occasionally produce toxin Because the organ-ism is also carried by many animals, outbreaks attributable to inadequate processing of a precon-taminated food can occur also

Growth and survival Staphylococci are killed by normal cooking temperatures Any staphylococci that survive because of inadequate heat penetration

or, more frequently, by postcooking contamination from a food handler will, if it is an enterotoxigenic strain and given the right conditions of warmth, moisture, pH, and time, produce toxin Growth of staphylococci and production of toxin are optimum

at approximately 20–37C, but growth can occur between 8 and 48C This toxin is fairly heat stable; boiling for approximately 30 min is required to destroy it Canning is usually, but not always, suffi-cient The toxin is also resistant to radiation Many foods can cause SFP Because the organism can grow in foods with high salt or sugar content (possibly because there is less competition from other organisms), ham is a common cause of SFP,

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