University of Pretoria etd – Chale-Matsau, J R B (2005)

Chapter 2
Literature Review

2.1 Introduction

The use of human excreta for fertilizer, ranging from night soil application to
irrigation with sewage has been a world-wide practice for many years, especially
in highly populated countries such as China and India (Rudolfs et al., 1950). It is
especially advantageous because it recycles nutrients back to the land and can
be economically attractive (Zenz et al., 1976).

In South Africa sludge production is increasing rapidly, and at the same time the
soil condition has deteriorated markedly. As sludge contains high levels of
organic matter and nutrients (Hu et al., 1996), use of this product in agricultural
land could provide an alternative means of disposal, and also benefits the poor
soil quality of most of South Africa’s agricultural land. It is believed that when
treated properly, and provided certain industrial contaminants are restricted from
entering the sewage, the resultant sewage sludge can become a relatively
innocuous organic fertilizer and soil conditioner of significant value for growing
trees, grass and certain crops (WRC, 1997).

The beneficial use of sludge for soil amendment in South Africa was also recently
shown by Snyman and colleagues (1998). At present, sewage sludge is used for
crop growing but limited only to fenced areas to restrict access to unauthorized
persons as well as milk-, meat- and egg producing animals (WRC, 1997). Other
recommendations suggest that application may only be done with planting and
during the period subsequent to harvesting and prior to the next growing season.
Snyman and Van der Waals (2003) reported that South African farmers, noting
the increased crop production as a result of enhanced soil properties from
sewage sludge use, are in favour of using sewage sludge and show adherence
to the recommended dosage of 8 ton/ha as stipulated in the guidelines (WRC,
1997).
5
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Elsewhere, application of sewage sludge to deteriorated soil, resulted in

increased yields (Tester and Parr, 1983). Consequently, municipal sewage
sludges are routinely utilized on agricultural lands in various parts of the world. In
Canada, it is becoming a common practice such that as much as 43% of the
produced sewage sludge is applied to land. By comparison, the United States
and Europe apply approximately 60% and 34% respectively of their sewage
sludge to agricultural land (EPA, 1999; Apedaile, 2001).

2.2 Metals and Toxic Organic Pollutants in Sludge

The composition of wastewater sludge may be highly variable depending on the

quantity and quality of contributions from industrial and domestic sources. The
types of constituents include among others, chlorinated hydrocarbons,
polynuclear aromatic hydrocarbons and metals (Brown et al., 1991). Hyde
(1976) pointed out that heavy metals are retained in soils following sludge
application and can accumulate to the point at which they are toxic to plants.
Thus, due to their uptake by crops, they may also be toxic to humans and
animals. This observation was confirmed by Rost and colleagues (2001) who
recently reported that heavy metals have long lasting adverse effects on
biological functions in soil. The heavy metal of major concern, because of its
possible phytotoxicity and danger to the human food chain, is cadmium (Cd).
Other heavy metals of importance are copper (Cu), nickel (Ni) and zinc (Zn), and
they are also known to be phytotoxic (Hyde, 1976; Purves, 1990).

Organic compounds such as pesticides, polychlorinated biphenyls, halogenated

aliphatics, ethers and aromatic hydrocarbons are the products of industrial
wastewater which could land up in wastewater sludges (Korentajer, 1991;
Vorobieva et al., 1996; Kouloumbis et al., 2000). The concentration of these
compounds needs to be monitored and limited by implementing source
reduction.

6
 University of Pretoria etd – Chale-Matsau, J R B (2005)

2.3 Socio-economic Issues Regarding Sludge Use

Farmers and the food industry have expressed their concern that agricultural use
of untreated sludge may affect the safety of food products and the sustainability
of agricultural land, and may carry potential economic and liability risks (NRC,
1996). There is also concern that the use of contaminated sewage sludge for
crop production could negatively affect the export market. For fear of foodborne
illness, some countries may refuse importation of vegetables and foods produced
under such agricultural practices (Sobsey, 1996; Doyle, 2000).

There has been increased public scrutiny of the potential health and
environmental consequences of land spreading of sewage sludge. It appears
that once fear of pathogens, odours, nuisances and possible environmental
deterioration have been generated in a community, people have great difficulty in
accepting the risks, even if there aren’t any, of applying sewage sludge to
agricultural land (Hyde, 1976; Tauxe, 1997). Thus, it is essential that aesthetic
characteristics and matters affecting both long-term quality of the land and the
public health must be thoroughly understood before using sewage sludge on
farmland.

In spite of the increasing concerns, in their recent report to the United States
Environmental Protection Agency, the National Research Council pointed out
that there is no evidence that proper use of wastewater treatment sludge on land
has any detrimental effect on either the people working at the site, on the
population surrounding the land application site, or on people eating the crops
grown in the sludge-amended soil (NRC, 2002). Vesilind (2003) is of the opinion
that the aversion to sludge use emanates from the knowledge of its origin and
not necessarily from diseases linked to sludge use.

Kirby (2001) pointed out that exposure to potentially lethal pathogens is linked to
social factors such as class, education and income. Carneiro and his colleagues
(2002) have observed that less intense Ascaris infection came from affluent

7
 University of Pretoria etd – Chale-Matsau, J R B (2005)

households with higher socio-economic profile. In many African countries

including South Africa, a large percentage of the population live in poverty
(Parliamentary Bulletin, 1996), thus it can be expected that these households
would be intensely affected by contaminated crops. The high incidence of
HIV/AIDS (Human Immunodeficiency Virus/ Acquired Immune Deficiency
Syndrome) infection in the country could translate into more pathogenic
infections due to their depressed immune systems, if such communities are
exposed to contaminated crops. South Africa is a comparatively large country,
covering 1,221,042 square kilometers and with an estimated population of about
40 million. It has been estimated that 14.2% of people in South Africa have been
infected with HIV/AIDS (Dorrington et al., 2002). The routine surveillance
conducted by the Department of Health has shown that among pregnant women
attending public health clinics for antenatal care, the prevalence has increased
from less than 1% in 1990 to 26.5% in 2002 (Dorrington et al., 2002). Overall, it
is estimated that 23.3% of men and 23.5% of women are infected whereas the
prevalence amongst the male and female youth is 5.8% and 21.6% respectively
(Dorrington et al., 2002).

2.4 Microorganisms Encountered in Sewage Sludge

Infectious diseases are transmitted primarily through human and animal excreta,
particularly faeces. If there are active cases or carriers in the community, then
faecal contamination of water sources will result in the causative organisms
being present in water. Pathogens in domestic sewage are primarily associated
with insoluble solids. Many of these organisms become bound to solids following
wastewater treatment and are transferred to wastewater sludge (Bitton, 1994).
As the wastewater treatment processes concentrate these solids into sewage
sludge, the sewage sludge has higher quantities of pathogens than incoming
wastewater (EPA, 1999). However, the transmission of pathogens can be
minimized by reducing the infectivity of sludges through effective treatment
processes (Smith, 1996).

8
 University of Pretoria etd – Chale-Matsau, J R B (2005)

The actual species and quantity of pathogens present in sewage sludge from a
particular municipality depend on the health status of the local community and
may vary substantially at different times (EPA, 1999). The four major types of
human pathogenic organisms, namely bacteria, viruses, protozoa and helminths
may all be present in sludge. These organisms can cause infection or disease if
humans or even animals are exposed to sufficient levels. The infective dose,
that is, the number of pathogenic organism to which a human must be exposed
to become infected, varies depending on the organism and on the health status
of the exposed individual (EPA, 1999). While some pathogens may cause
infections in a susceptible host by a single organism, others may require several
hundreds to be present before an infection can be initiated. Symptoms may vary
in severity from mild gastroenteritis to severe and sometimes fatal diarrhoea,
dysentery, hepatitis or typhoid depending on the type of pathogen and pathogen
load. Thus, when reclaimed water or sludge is used on fields producing food
crops, it is critical to protect public health.

In the sections that follow, the major bacterial, viral and parasitic organisms
found in wastewater sludge are described.

2.4.1 Bacteria

Wastewater normally contains many bacterial species, and strains (Vilanova et

al., 2002) that may end up in the wastewater sludge. If such sludge is not
adequately treated and used in agricultural land, crop contamination may be
imminent. As Bubert and colleagues (1999) have pointed out, contamination of
food material does not only occur during food processing, but may also begin
with the production of raw food materials in the environment.

Faecal coliforms and enterococci have been used widely as faecal pollution
indicators (Vilanova et al., 2002). Both bacterial groups include several species.
For example, the genus Enterococcus contains 19 recognized species (Manero
and Blanch, 1999). Salmonella spp, Shigella spp, Campylobacter spp, Yersinia

9
 University of Pretoria etd – Chale-Matsau, J R B (2005)

spp, Leptosporia spp and Escherichia coli are bacterial pathogens of primary
concern in sludge. Escherichia coli is particularly abundant in human and animal
faeces, where numbers may reach 109 g-1 of faeces (Bitton, 1994). The major
bacterial groups or species are tabulated in Table 2.1 and some of these (*) are
discussed in sections i to viii. Several case studies have been cited in these
sections. These case studies do not necessarily detail outbreaks due to
wastewater sludge use, but are indicators of what the effects could be if the
pathogens manage to survive and infect a receptor, as a worst-case scenario.

Table 2.1 Bacterial pathogens to be expected in sewage sludge (Source:

EPA, 1999; Strauch, 1991)
Pathogen Disease

Salmonella spp * Salmonellosis (gastroenteritis)

Shigella spp * Bacillary dysentery
Escherichia coli * Urinary infection; diarrhoea
Yersinia enterocolitica * Yersniosis (gastroenteritis)
Clostridium spp * Gas gangrene
Leptospira spp Leptospirosis
Mycobacterium spp Tuberculosis and leprosy
Vibrio cholerae spp Cholera
Staphylococcus spp Osteomyelitis
Streptococcus spp Rheumatic fever; glomerulonephritis
Klebsiella spp Pneumonia; urinary tract infection
Enterobacter spp Urinary tract infection
Serratia spp Meningitis; endocarditis
Citrobacter spp Neonatal meningitis
Proteus spp Urinary tract infection
Providencia spp * Urinary tract infection
Listeria monocytogenes * Listeriosis

10
 University of Pretoria etd – Chale-Matsau, J R B (2005)

i Escherichia coli

Escherichia coli is normally found in the gastrointestinal tract of humans and

other warm-blooded animals (Brooks et al., 1991) and is the most common
cause of foodborne illness. Foods that have been implicated with E. coli include
cheese, beef, fish, poultry, apple cider and lettuce (Reis et al., 1980; Kornacki
and Marth, 1982; Ackers et al., 1998).

Escherichia coli, depending on the infective strain, can cause a variety of

illnesses that include infantile diarrhoea, traveler’s diarrhoea, hemorrhagic colitis
(HC), hemolytic uremic syndrome (HUS) and thrombocytopenic purpura (TP)
(Pell, 1997; Penner, 1998). Hemorrhagic colitis is a severe illness and is
characterized by bloody diarrhoea and severe abdominal cramps while HUS is
characterized by bloody diarrhoea followed by renal failure. Thrombocytopenic
purpura yield symptoms similar to those of HUS but the central nervous system
is also affected. Death often occurs in patients with HUS and TP (Pell, 1997).
Hemolytic uremic syndrome can be a serious complication in children and is a
leading cause of acute kidney failure (Penner, 1998).

ii Salmonella spp

Salmonellosis was normally associated with contamination of food of animal

origin, but in recent years, it has been indicated that Salmonella spp
contamination may also occur in foods of plant origin. For instance Salmonella
spp outbreaks have been associated with consumption of celery, watercress,
watermelon, lettuce, cabbage, tomatoes, potatoes and carrots (Wells and
Butterfield, 1997; Guo et al., 2000). These organisms have also been implicated
in infections due to wastewater spreading (Melloul and Hassani, 1999).
Salmonella spp are capable of surviving and multiplying in fruits and vegetables.
Asplund and Nurmi (1991) have demonstrated that tomatoes can provide a
favourable environment for growth of S. enteitidis, S. infantis and S. typhimurium
in spite of their low pH value, showing that the high acidity is not necessarily

11
 University of Pretoria etd – Chale-Matsau, J R B (2005)

effective enough to inhibit Salmonella spp growth. Salmonella spp can grow and
multiply at temperatures of 22oC (Asplund and Nurmi, 1991) suggesting that
once produce has been contaminated, microorganisms may continue to grow on
the shelf in retail stores increasing the risk of infection.

Enteric fever is caused by the microorganism S. typhosa, in which the organism,

ingested along with food finds its way into the bloodstream. Another organism,
S. cholera-suis causes septicemia resulting in blood poisoning. The S.
typhimurium and S. enteritidis cause gastroenteritis, an infection very commonly
associated with contaminated food. Symptoms of Salmonellosis include nausea,
vomiting, headache, chills, diarrhoea, fever and can even lead to reactive
arthritis. In most cases the disease is short-lived, but salmonellosis can be fatal
(Penner, 1998). Infants, once infected, frequently become long-term carriers and
cause family outbreaks (Burge and Marsh, 1978).

iii Listeria spp

Listeriosis is rare in non-pregnant healthy adults, however, adults with conditions

such as type 1 diabetes, cardiovascular disease, renal transplant, neoplasm,
alcoholism and AIDS are more susceptible (Penner, 1998). Due to its ability to
survive for long periods and its capability to grow at refrigerator temperature
(Penner, 1998) this organism poses a serious threat in regard to foodborne
illness. Healthy animals can be asymptomatic carriers of L. monocytogenes
(Pell, 1997).

Listeria monocytogenes is a human and animal pathogen capable of causing

nausea, vomiting, headache, fever, and severe infections like septicemia,
encephalitis and meningitis, especially in immunocompromised individuals,
newborns and pregnant women where it can result in stillbirths. About 100 cells
of L. monocytogenes are sufficient to cause illness (Brooks et al., 1991). In the
USA this organism has a fatality rate of 20 – 40% (Penner, 1998).

12
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Several outbreaks have been associated with contaminated commercial

foodstuffs, such as vegetables, milk and meat products on which these bacteria
can multiply even at low temperatures (Bubert et al., 1999). Both L.
monocytogenes and L. innocua have been isolated from various environmental
samples such as soil, vegetation and human and animal faeces (Bubert et al.,
1999).

iv Yersinia enterocolitica

Yersinia enterocolitica causes yersiniosis and is found in a variety of animals,

particularly pigs. It will grow at refrigerator temperatures, but grows best at room
temperature. Infection with this organism yields symptoms that range from a mild
gastroenteritis to sever conditions of polyarthritis and meningitis (Prescott et al.,
2002).

v Shigella spp

Shigellosis is caused by bacteria of the genus Shigella (Brooks et al., 1991).

This disease is characterized by diarrhoea, abdominal pain, vomiting and fever.
As few as 10 to 100 microorganisms are sufficient to cause an illness (Penner,
1998). Shigella are readily killed by heat and do not survive well in acidic
environments (Prescott et al., 2002).

vi Clostridium spp

Clostridium botulinum produces a neurotoxin that cause botulism. After the toxin
is absorbed, it binds to nerve endings and causes vomiting and diarrhoea,
fatigue, dizziness and headache. Later there may follow constipation, double
vision, difficulty speaking and swallowing, involuntary muscles may become
paralyzed leading to cardiac and respiratory failure and eventually death.
(Penner, 1998). The C. botulinum spores are heat resistant (Brooks et al., 1991).

13
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Clostridium perfringens produces toxins that cause diarrhoea and severe

abdominal pain. However, death is uncommon. Although spores of this organism
are common in raw foods and they are heat resistant, large numbers of
vegetative cells of C. perfringes are necessary for an illness to occur (Penner,
1998).

vii Campylobacter jejuni

Campylobacter jejuni causes camphylobacteriosis characterized by cramps,

nausea, diarrhoea, headache and fever. Onset of the disease following
consumption of contaminated food is within two to five days. Prolonged illness
may lead to complications such as meningitis, urinary tract infection and reactive
arthritis, but death occurs rarely (Penner, 1998). The high incidence of C. jejuni
infections in persons infected with the human immunodeficiency virus points to
the widespread transmission of low levels of this organism (Blaser, 1996).
Campylobacter cells survive for several weeks at temperatures even as low as 4
o
C (Waage et al., 1999). The infective dose of C. jejuni is very small, it has been
estimated that about 500 cells of this organism can cause human illness (Black
et al., 1988). Also, Campylobacter cells may enter a viable but non-culturable
state due to starvation and physical stress, making them even more difficult to
detect (Brooks et al., 1991).

viii Providencia spp

Providencia spp are members of the normal intestinal flora. They cause urinary
tract infection and are often resistant to antimicrobial therapy (Brooks et al.,
1991).

14
 University of Pretoria etd – Chale-Matsau, J R B (2005)

2.4.2 Persistence of Bacteria in Soil

The survival of microorganisms added to soil is influenced by a number of factors

that include, water-holding capacity of soil, temperature, rainfall, sunlight, organic
material in soil and the hydrogen ion (Rudolfs et al., 1950; deRopp, 1999).

Faecal coliforms can survive for several years under optimum conditions, and the
Salmonella spp may survive for a year in rich, moist organic soil (deRopp, 1999).
The survival period of Salmonella spp has been reported to be as long as
between 15 – 117 weeks in contaminated soil (Rudolfs et al., 1950; Jones,1980;
Strauch, 1991; Sidhu et al., 1999; Baloda et al., 2001). The L. monocytogenes
grows well in sewage and survives for long periods in soil (Strauch, 1991). Other
bacteria such as Streptococcus jaecelis, Clostridium botulinium, Clostridium
tetani, Clostridium perfringes and butyl-butyric Clostridia spp were found in small
numbers 7 months after sludge application (Hyde, 1976).

Campylobacters spp are not capable of proliferating in the environment, but may
remain dormant and survive in the environment for several weeks at low
temperatures (Waage et al., 1999). However, the infective dose is very small
which increases the risk of infection (Black et al., 1988).

One of the most important factors influencing the survival of pathogenic bacteria
in soil is competition with the existing soil microflora. In soils with low microbial
activity, the newly added microorganisms may persist for much longer (Bitton,
1994). Thus the application of large quantities of sludge to soil with low existing
microbial activity will increase the ability of the pathogens to persist in soil
environment and hence increase the potential risk for transfer of pathogens to
crops grown in the soil. On the other hand, in biologically active soils,
microorganism numbers are rapidly reduced due to competition (Penner, 1998).
The soils in South Africa are typically biologically active, which could be
advantageous due to the fact that introduced microorganisms are rapidly out

15
 University of Pretoria etd – Chale-Matsau, J R B (2005)

competed. However, as a result of high microbial activity, the organic material in

agricultural soil is low (Korentajer, 1991).

Microorganisms may move through the contaminated soil as a result of rainfall or

irrigation (Gerba et al., 1975). Gagliardi and Karns (2000) have indicated that if
soil pores do not become clogged, E. coli can travel below the top layers of soil
for more than two months. They also indicated that E.coli from manure applied
to soil could survive, replicate and move vertically in the soil (Gagliardi and Karns
2000). While soil contaminated with sewage sludge could lead to crop
contamination, it has been indicated that water bodies such as groundwater,
storm-water and rivers could be contaminated following rainfall or irrigation as a
result of runoff from contaminated agricultural land (Lee and Jones-Lee, 1993;
Bilgrami and Kumar, 1998).

2.4.3 Viruses

Sludge from wastewater treatment may contain demonstrable numbers of viruses

even after anaerobic digestion (Damgaard-Larsen et al., 1977). Some of the
viruses that can be expected in sewage sludge are tabulated in Table 2.2.

Human enteric viruses are excreted in faeces, and can be shed in high numbers
(108 to 1010 particles per gram of faeces) by infected individuals (Abbaszadegan
et al., 1999). The persistence of enteroviruses in sludge and sludge-amended
soil was demonstrated by Damgaard-Larsen et al. (1977) and by Straub et al.
(1994). The virus of greatest potential concern appears to be Hepatitis A, a
disease with potential for long-term liver damage (Pahren et al., 1979).

16
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Table 2.2 Viruses that can be expected in sewage sludge (Sources: EPA,
1999; Strauch, 1991; Bofill-Mas et al., 2000)
Pathogen Disease

Enteroviruses
Coxsackievirus A Acute hemorrhagic conjunctivitis
Coxsackievirus B Meningoencephalitis
Echovirus Acute hemorrhagic conjunctivitis
Poliovirus Poliomyelitis
Adenovirus Respiratory and systemic infections
Reovirus Acute respiratory infections
Hepatitis A virus Infectious hepatitis
Rotavirus Acute gastroenteritis
Astrovirus Gastroenteritis
Calicivirus Acute gastroenteritis
Coronavirus Gastroenteritis
BK virus Uretal stenosis and hemorrahgic colitis
JC virus Multifocal leukoencephalopathy
Norwalk and Norwalk-like viruses Acute gastroenteritis

Human polyomaviruses JC virus and BK virus were also indicated as being

present in urban sewage obtained from widely divergent geographical areas in
Europe and Africa (Bofill-Mas et al., 2000). The JC virus is aetiologically
associated with a fatal demyelinating disease known as progressive multifocal
leukoencephalopathy, which has emerged as a frequent complication of AIDS in
HIV infected individuals. Infection with BK virus has been associated with
diseases of the urinary tract including hemorrhagic cystitis and ureteral stenosis
(Bofill-Mas et al., 2000).

Virus inactivation under natural conditions is a slow process (Damgaard-Larsen

et al., 1977). Viruses may become eluted and travel through the soil (Damgaard-
Larsen et al., 1977) which includes both vertical and lateral migration (Straub,

17
 University of Pretoria etd – Chale-Matsau, J R B (2005)

1995). For instance, other enteroviruses such as the coxsackie B3 virus have
been isolated 18m below the soil surface after wastewater recharge (Straub et
al., 1995). Rainfall and irrigation events may contribute to viral transport (Straub
et al., 1995). Viruses readily adsorp to soil particles, and this has been reported
to prolong their survival (WHO, 1979). However these viruses remain as
infectious to humans as free viruses.

Viruses can survive for up to six months in cold weather and for three months in
warm weather. Enteric viruses can survive up to 170 days in loamy and sandy
soil. Poliomyelitis virus has been detected in soil irrigated with infected sewage
sludge and effluent after 96 days in winter and 11 days in summer in the UK, and
on the surface of mature vegetables 23 days after irrigation had ceased (Tierney
et al., 1977; WHO, 1979). Viral survival on crops may be shorter than in the soil
if viruses on crops surfaces are directly exposed to detrimental environmental
factors such as sunlight and desiccation (Pahren et al., 1979; WHO, 1979). The
warm climate in some regions of South Africa may reduce the survival of these
viruses. However, more prolonged survival can be expected in the moist or more
protected parts of plants, such as within the folds of leafy vegetables, in deep
stem areas and on rough cracked surfaces of edible roots. It is also likely that
viruses can penetrate damaged roots and under certain conditions enter the
stem and leafy parts of edible plants (Pahren et al., 1979).

Once crops are harvested, enteric viruses can survive for prolonged periods
during commercial and household storage at low temperature. The risk of
human infection associated with virus-contaminated crops is greatest in the case
of fruits and vegetables consumed raw (WHO, 1979).

2.4.4 Parasites

Parasites are a group of foodborne pathogens that have received relatively little
attention. Parasites that are usually encountered in sludge are indicated in Table
2.3, and some of these (*) are briefly discussed.

18
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Table 2.3 Parasites that can be expected in sewage sludge (EPA, 1999;
Strauch, 1991)
Pathogen Disease

Entamoeba histolytica Amebiasis

Giardia lamblia * Giardiasis
Toxoplasma gondii Toxoplasmosis
Sarcocystis spp Intestinal infection
Taenia spp * Taeniasis
Diphyllobothrium latum Pernicious anaemia
Echinococcus granulosus Echinococcosis
Ascaris spp * Ascariasis
Toxocara spp Pneumonic symptoms
Trichuris trichiura Trichuriasis
Toxoplasma gondii Toxoplasmosis
Cryptosporidium * Cryptosporidiosis

Some of the common types of parasites that have been detected in fresh fruits
and vegetables include Giardia lamblia, Entamoeba histolytica and Ascaris spp.
(Brackett, 1987). As little as 10 or fewer Giardia cysts are sufficient to cause
illness (Brooks et al., 1991). Ayres and colleagues (1992) recovered viable
Ascaris eggs from lettuce irrigated with raw sewage, while Gaspard and
Scwartzbrod (1993) recovered viable Ascaris from both tomatoes and lettuce
following raw sewage irrigation. It has also been demonstrated that farm workers
may be infected with enteric parasites as a result of occupational exposure (Clark
et al.,1984). It should be noted that these incidents were associated with the
irrigation of raw sewage and not wastewater sludge. However, it does give an
indication of potential risk.

The parasites most often found in sludge are Ascaris species such as A.
lumbricoides (human intestinal roundworm) and A. suum (pig’s roundworm) as

19
 University of Pretoria etd – Chale-Matsau, J R B (2005)

well as some Toxocara and Trichuris species (Bitton, 1994; Gaspard et al.,
1995).

Ascaris eggs and certain larval stages of trichostrongylids can survive for over a
year in soil that has been irrigated with sewage sludge (Strauch, 1991), and the
eggs of Cryptosporidium parvum and Taenia saginata are known to survive in
sewage for more than 12 months (NRC, 1996). Cryptosporidium species and
Giardia species pose a serious threat to human health as these organisms are
difficult to inactivate with disinfectants and their infective doses in humans are
very low (Finch and Belosevic, 2001).

Protozoan parasites, such as Giardia spp have been found in sludge in Western
Australia where they remain the most common cause of enteric disease (Hu et
al., 1996). The most noxious are the Ascaris eggs and coccidial oocysts as they
have high resistance (Pahren et al., 1979; Gaspard and Schwartzbrod, 1993).
Helminths larvae are usually killed by composting, but often remain viable in
slurry during storage (Shuval et al., 1984).

Also encountered in sludge are the organisms of the genus Cryptosporidium

(Kuckzynska and Shelton, 1999; EPA, 1999). Of the Cryptosporidium species,
C. parvum is the agent of clinical cryptosporidiosis in humans and livestock. The
C.parvum oocysts are shed by infected mammals and are known to be resistant
to standard disinfectants (Champliaud et al., 1998). Waterborne C. parvum
oocysts may remain viable for several months (Kuczynska and Shelton, 1999).

Table 2.4 indicates the concentrations of pathogens as indicated by other

countries.

20
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Table 2.4 Concentrations of pathogens in sludge from other countries

(Jimenez et al., 2002)
Pathogen Concentration Country

Fecal coliforms (MPN/gTS) 3.6 X 104 – 1.4 X 106 United Kingdom

2.3 X 107 – 9.3 X 1010 Mexico
2.0 X 107 United States
E.coli (PFU/gTS) 1.0 X 106 – 1.9 X 106 Mexico
1.3 X 105 United States
Ascaris/gTS 2.40 – 8.98 United Kingdom
66 – 136 Mexico
1.4 – 9.7 United States
0.60 – 2.4 France

2.5 Disinfecting Treatment Processes

Previous sections provided detailed discussions on the occurrence of

microorganisms in sludge and their potential presence in crops if inadequately
treated sludge is used for land application. However, the transmission of
pathogens can be minimized by reducing the infectivity of sludges through
effective treatment processes (Smith, 1996). Various techniques are used to
eliminate or reduce the number of microorganisms to levels that do not threaten
human health (EPA, 1999).

Many of these treatment processes are applied either to stabilize the sludge, i.e
reduce its vector attraction potential and odour or render the sludge easier to
handle, store and transport by reducing the volume or drying the wastewater
sludge. Additional treatment technologies need to be employed to reduce the
viable content. Some of these techniques recommended in the US Part 503 rule
are indicated in Table 2.5.

If effective treatment is not available, long term storage could be used to

accelerate inactivation and thus reduce the number of infective species before

21
 University of Pretoria etd – Chale-Matsau, J R B (2005)

sludge is spread onto soil (Jenkins et al., 1999). Jenkins and colleagues (1999)
warned that although storing prior to spreading could be an effective
management practice for reducing infective oocyst load, spreading of sludge
during the cold season may have the opposite effect by sustaining the survival of
C. parvum oocysts and positioning them for transport in surface runoff (Jenkins
et al.,1999).

22
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Table 2.5 Techniques listed in the 40 CFR Part 503 and their effectiveness
in removing pathogens (EPA, 1999)
R = Reduction, E = Elimination, 3 = effective in pathogen reduction/elimination
and 2 = not effective in pathogen reduction /elimination
Effectiveness in Eliminating
Technique Description Pathogens

Viruses Bacteria Parasites

R E R E R E
Aerobic Sewage sludge is agitated
3 2 3 2 2 2
Digestion with air or oxygen to maintain
aerobic conditions
Air Drying Sewage sludge is dried on
3 2 3 2 2 2
sand beds or on paved or
unpaved basins. The sewage
sludge dries for a minimum
duration of 3 months
Anaerobic Sewage sludge is treated in
3 2 3 2 2 2
digestion the absence of air at a specific
temperature. The values of
the temperature shall be
o
between 15 days at 35 C and
o
60 days at 20 C
Composting Using either the within-vessel,
3 2 3 2 3 2
static aerated pile, or widow
composting methods. The
temperature of sewage sludge
o
is raised to 40 C or higher
and remains at 40 oC or higher
for 5 days. Fours in the 5 day
period, the temperature in the
o
compost pile exceeds 55 C
Lime Sufficient lime is added to the
3 2 3 2 2 2
Stabilization sewage sludge to raise the pH
of the sewage sludge to 12 for
2 hrs.
Thermal Sewage sludge is heated to a
o
3 2 3 3 3 2
Treatment temperature of 180 C or
higher for 30 minutes

22
 University of Pretoria etd – Chale-Matsau, J R B (2005)

2.6 Treatment and Sewage Sludge Classification in South Africa

Snyman and colleagues (2003) documented the treatment technologies

employed by South African wastewater treatment plants. According to this study,
57% of the sludge that is produced employs anaerobic digestion of primary and
humus sludge (Snyman et al., 2003). The sludge types generated from these
plants are presented in figure 2.1.

Activated Sludge
20%

Anaerobic Digestion Blended sludge Petro sludge

57% 12% 2%

Aerobic digestion
1%
Oxidation Dams
0.3%

Figure 2.1 Sludge types produced by the wastewater treatment plants

surveyed in South Africa on a mass percent basis. The blended sludge
represents primary and activated sludge blended before or after digestion
(Snyman et al., 2003).

Figure 2.2 illustrates the tertiary and additional stabilisation technologies

employed by the wastewater treatment plants surveyed in South Africa. The
majority (74% mass) of the sludge producing treatment plants surveyed do not
treat the sludge further than the traditional anaerobic digestion and activated
sludge treatment. Composting is used by both metropolitan city councils and
plants in smaller town councils while pelletisation is only employed by large
metropolitan councils (Snyman et al., 2003). Aerobic digestion is employed as an

23
 University of Pretoria etd – Chale-Matsau, J R B (2005)

additional treatment method after anaerobic digestion in one major site (Snyman
et al., 2003).

Aerobic digestion
2%

Composting
19%

None
74%

Pellets
4%

Figure 2.2 The tertiary and additional stabilisation technologies employed

by the wastewater treatment plants surveyed in South Africa on a mass
percent basis (Snyman et al., 2003).

The sewage sludge produced from treatment plants in South Africa is used for a
number of activities, including application onto golf courses and use by
municipalities for lawn cultivation, while some is collected by farmers for
agricultural use. The disposal and beneficial use of sewage sludge in South
Africa are summarized in figure 2.3.

25
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Contractor
Sludge Dams
10%
0.4%
Golf course
Landfill 3%
2% Compost
2%
Municipal Gardens
Stockpile 21%
11%
Farmers
10%
Instant Lawn
12%
Land Application
12% Sold
17%
Other
75%

Figure 2.3 the major disposal methods employed by the wastewater

treatment plants surveyed in South Africa on a mass percent basis
(Snyman et al., 2003).

Table 2.6 summarizes the classification of sewage sludge indicated in the South
African sludge guidelines. The South African guidelines classify sludge at three
levels (Types A, B and C) and a fourth category (Type D) that stipulates ceiling
limits for pollutants is added. Although the hygienic quality of Type D is similar to
Type C, the Type D sludge is produced for unrestricted use on land at maximum
application of 8 tonnes per hectare per year, the levels of metals and inorganic
content are limited to acceptable low levels (WRC, 1997).

26
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Table 2.6 Classification of Sewage Sludge to be used or disposed off on

Land (WRC, 1997)
Sewage Sludge Treatment Characteristics-Quality of Sewage sludge

Type A Sludge Cold digested sludge Usually unstable and can cause odour nuisances and
Septic tank sludge fly-breeding
Oxidation tank sludge Contains pathogenic organisms
Variable metal and inorganic content
Type B Sludge Anaerobic digested Fully or partially stabilized – should not cause significant
sludge odour nuisance or fly-breeding
Surplus activated Contains pathogenic organisms
sludge Variable metal inorganic content
Humus tank sludge
Type C Sludge Pasteurised sludge Certified to comply with the following quality
Heat treated sludge requirement:
Lime-stabilised Stabilized – should not cause odour nuisances or fly-
sludge breeding
Composted sludge Contains no viable Ascaris ova per 10 gram of dry
Irradiated sludge sludge
Maximum 0 Salmonella organisms per 10 gram dry
sludge
Maximum 1000 Faecal coliform per 10 gram dry sludge,
immedialtely after treatment (disinfection/sterilization)
Variable metal and inorganic content
Type D Sludge Pasteurised sludge Certified to comply with the following quality
Heat-treated sludge requirement:
Lime-stabilised Stabilized – should not cause odour nuisances or fly-
sludge breeding
Composted sludge Contains no viable Ascaris ova per 10 gram of dry
Irradiated sludge sludge
Maximum 0 Salmonella organisms per 10 gram dry
sludge
Maximum 1000 Faecal coliform per 10 gram dry sludge,
immedialtely after treatment (disinfection/sterilization)
Has specific limits for metal and inorganic content
(summarized in WRC, 1997)
Product must be registered in terms of Act 36 of 1947 if
used for agricultural activities

27
 University of Pretoria etd – Chale-Matsau, J R B (2005)

2.7 Resistance of Microorganisms to Disinfection

The previous section discussed disinfecting techniques employed by wastewater

treatment plants to reduce or eliminate the numbers of infective species. If the
sewage sludge used is not adequately treated, there is potential for crop
contamination.

Studies have shown that once fruits and vegetables have been contaminated, it
may be difficult to disinfect them (Maxy, 1982; Takeuchi et al., 2000; Wachtel et
al., 2002a; Wachtel et al., 2002b). Some microorganism such as E. coli show
preferential attachment to the interior of damaged fruits and vegetables than on
the surface (Takeuchi et al., 2000) as the juice within the vegetable provides
good growth medium (Maxy, 1982). Itoh and coworkers (1998) found that E. coli
was internalized when radish sprouts were produced from contaminated seeds
and therefore would be protected from surface decontamination treatment. E.
coli is capable of attachment to the interior of stomatal pores (Seo and Frank,
1999; Takeuchi and Frank, 2000; Takeuchi and Frank, 2001) and has a tendency
to form aggregate associations (Wachtel et al., 2002a). These attachment sites
and aggregation tendencies may cause bacterial resistance to physical methods
of surface disinfection as well as chemical treatment such as chlorination
(Wachtel et al., 2002b).

2.8 Protecting the Public and Environment through Regulatory Management

Most countries adopt a similar approach to protect the public from infection due
to pathogens originating from wastewater sludge. The use of wastewater sludge
is regulated and these regulations stipulate how the sludge should be disinfected
and/or how to minimize the chance of infection through prescribed management
practices. In the United States, the use and disposal of treated sewage sludge is
regulated under CFR Part 503 (EPA, 1999).

The regulation protects public health and the environment through requirements
designed to reduce the potential for contact with disease-bearing pathogens in
28
 University of Pretoria etd – Chale-Matsau, J R B (2005)

sewage sludge applied to the land or placed. These requirements are divided
into:
• Requirements designed to control and reduce pathogens in treated
sewage sludge and
• Requirements designed to reduce the ability of the treated sewage sludge
to attract vectors (insects and other living organisms that can transport
sewage sludge pathogens)

It includes both performance and technology-based requirements. Wastewater

plants have the freedom to modify conditions and combine processes with each
other to meet the requirements.

At present in South Africa humans and the environment are protected under the
National Water Act 36 of 1998 (NWA), National Environmental Management Act
107 of 1998 (NEMA), Water Services Act 108 of 1997(WSA), the Constitution of
the Republic of South Africa (Act 108 of 1996) and the Health Act 63 of 1977.

The Department of Water Affairs and Forestry is the custodian of water

resources in South Africa. The guidelines for sewage sludge classification and
application are summarized in a document on permissible utilization of sewage
sludge (WRC, 1997; WRC, 2002). If the sludge reuse or disposal method does
not comply with the requirements detailed for the applicable classification its
reuse or disposal requires permission, which could be in a form of a licence or
permit (WRC, 2002).

In South Africa, there aren’t any specified restricted techniques for sludge
treatment, but the chosen technologies need to yield the sludge quality as
required in the guidelines (WRC, 1997).

29
 University of Pretoria etd – Chale-Matsau, J R B (2005)

2.9 Public Perception

The benefits of sewage sludge are well understood by the scientific community,
and through consultation, most governments around the world recognize the
benefits of using sludge in crop production. It is for this reason that a number of
countries have since engaged in utilizing sewage for land application purposes.

Despite the advancements in sludge use in agriculture, the main recipients of

these services have often been neglected. This often led to fear and rejection of
sewage sludge among some members of the public as a result of misinformation
due to media coverage (Sunday Times, 2003). Due to lack of scientific
knowledge, the public will generally reject any association with a product or
service if it is linked to odour or discolouration (Small Wright, 2002). Tyson
(2002) reported that if sewage sludge did not smell, the public probably would not
complain.

In a small preliminary survey done in South Africa, it has emerged that only a
small percentage (39%) of low income earners were aware of what sewage
sludge was (Snyman and Van der Waals, 2003). Snyman and Van der Waals
(2003) also noted that the respondents did not understand the risks associated
with using sewage sludge for agricultural soil amendment. Of the respondents
from a higher income bracket, 79% were found to have knowledge of sewage
sludge and its potential benefits. The majority of the respondents from this group
also expressed their willingness to purchase vegetables from a sewage sludge
fertilized farm, with 45% prepared to consume vegetables grown on sewage
sludge (Snyman and Van der Waals, 2003). It appears from this survey that if
members of the general public are informed of the benefits of sludge, reception
of the use of sewage sludge might increase in the future. It is thus the
responsibility of the sludge producers together with the governments to introduce
mechanisms of educating the public of sewage sludge and its use in agriculture.

30
 University of Pretoria etd – Chale-Matsau, J R B (2005)

2.10 Assessing Human Risk Exposure

The sections preceding indicate that many wastewater plants generate sludges
that still contain pathogens. However, these sludges are still used in agricultural
practices. The question to address therefore is “What is the risk associated with
this practice?” If the risk of using such sludge is unacceptable, what
management practices should be adapted to reduce this risk to an acceptable
level?

While complete elimination of pathogens from sludge is ideal, it has been

indicated that if the numbers of pathogens in sludge are reduced to an
acceptable level, the use of such sludge in agricultural land does not appear to
result in unacceptable risk to human health (Apedaile, 2001; Tanner et al., 2003).
According to Vesilind (2003), coming into contact with small doses of pathogens
is the “sufficient challenge” our bodies need to stay healthy as our enhanced
health comes not from zero exposure, but from a sufficient exposure to
pathogens. Although this is true for healthy individuals, this could be different for
the South African population, as a large percentage of the population is HIV
positive and therefore immunocompromised (Dorrington et al., 2002).

One of the concerns often raised regarding sludge application is the emission of
pathogenic aerosols during land application (Pillai et al., 1996). The risk of
release rises as the pathogenic content in sludge increases. Raw sludge from
municipal sewage would be more likely to release airborne pathogens than those
that have been treated to reduce the pathogens (Straub et al., 1993). Tanner
and colleagues (2003) evaluated the potential for bio-aerosols from sludge
application, and concluded that the risk of adverse public health effects from bio-
aerosols following land applied sludge is low. Forcier (2002) indicated that
although quantities of bio-aerosols could be released during storage, loading and
land application, they are diluted and scattered through atmospheric dispersion in
ambient air. The survival of and the potential for infection from these organisms
are lessened by the natural processes of attenuation such as ultra-violet radiation

31
 University of Pretoria etd – Chale-Matsau, J R B (2005)

and desiccation (Forcier, 2002). Bio-aerosol emissions are also lessened when
applied sludge is subsequently incorporated into the soil (Straub et al., 1993). It
appears that the methods used for sludge land application do not result in
airborne release of biological agents to the same extent as in wastewater
treatment facilities (Apedaile, 2001).

Tools exist to measure the risk to human health associated with the use of
sewage sludge that contains pathogens in agricultural practices. The following
section details one of the tools used in this thesis.

2.10 .1 Health Risk Assessment

The health risk assessment provides a means to estimate the probability of

adverse effects associated with measured or estimated levels of the hazardous
agents, and a tool for predicting the extent of potential or probable health effects.
The protocol was originally developed for carcinogen assessments. However,
current trends favour the application of similar procedure to establish the risk of
microbiological hazards. The process as defined by the US EPA, is comprised of
four distinguishable but interacting phases, namely:
- Hazard identification;
- Exposure assessment;
- Dose-response assessment and
- Risk characterisation (Zwietering and van Gerwen, 2001)
The interrelation of these phases is depicted in Figure 2.4.

32
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Hazard Identification

Dose-Response Assessment Exposure Assessment

Risk Characterisation

Figure 2.4 The interrelation of the risk assessment phases (Genthe 1998).

i Hazard Identification

This involves the identification of biological, chemical and physical agents

capable of causing adverse health effects and that may be present in a particular
food or group of foods (Rocourt et al., 2001). Once the health hazard has been
identified, the remainder of the process encompasses the description of the
properties of the hazardous agent and the identification of both acute and chronic
health effects (Genthe, 1998).

ii Hazard Characterisation

This involves the qualitative and or quantitative evaluation of the nature of the
adverse health effects associated with the hazard present in food. It provides
description of the severity and duration of adverse effects that may result from
ingestion of a microorganism in food. This involves a dose response assessment
by establishment of a relationship between the dose of an agent and the rate of
infection. Dose response assessment is considered a key ingredient of
quantitative risk assessment as it is supposed to provide the link between
exposure to a hazardous agent and the probability of ensuing health effects
(Teunis and Havelaar, 2000).

Some microorganisms when present at sufficient levels are capable of causing

disease, while others may produce toxins that contribute to the development of a

33
 University of Pretoria etd – Chale-Matsau, J R B (2005)

disease (Brooks et al., 1991). Toxins produced by bacteria are generally

classified into two groups, exotoxins and endotoxins. Exotoxins are excreted by
living cells, while endotoxins are released on bacterial death (Brooks et al.,
1991).

iii Exposure Assessment

This involves the qualitative and or quantitative evaluation of the likely intake of
biological, chemical and physical agents via food, as well as exposure from other
sources if relevant (Rocourt et al., 2001). It is usually defined as a process of
measuring or estimating the intensity, frequency and duration of human exposure
to a contaminant. The task of exposure assessment is to provide the actual
exposure conditions required to predict risk, and to identify and predict the effects
of the proposed control options (Genthe, 1998).

iv Risk characterisation:

This involves the qualitative and or quantitative estimation, including attendant

uncertainties of the probability of occurrence and severity of known or potential
adverse health effects in a given population based on hazard identification,
hazard characterization and exposure assessment (Rocourt et al., 2001). Risk
characterisation has been defined as the process of calculating the incidence of
the health effect under the conditions of exposure described in exposure
assessment. A major component of risk assessment is an evaluation of all
assumptions used and all sources of uncertainty (Genthe, 1998). In risk
characterisation all results of the former steps are integrated, bringing together all
inaccuracies from the former steps (Zwietering and van Gerwen, 2000). Thus
risk characterization is defined as the process of estimating the likelihood or
probability of experiencing the adverse effects of an identified hazard, the impact
or consequences of those effects and describing the attendant uncertainty of the
estimates.

34
 University of Pretoria etd – Chale-Matsau, J R B (2005)

2.11 Factors Affecting Management of Sewage Sludge Use in South Africa

South Africa has unique factors that could influence management of land
application of sewage sludge. These factors include population density, high
incidence of HIV/AIDS, unique climatic conditions and soil quality, amongst
others. A detailed description of these factors will be provided in later sections to
indicate how they influence management of sludge use in South Africa.

2.12 Conclusion

Sewage sludge could be used beneficially in agricultural practices, especially in

South Africa’s carbon depleted soils. It appears there are vast agronomic and
economic benefits to sludge use, particularly as the cost of fertilizers are on the
increase.

However, pathogens do occur in a large percentage in what is regarded as

sewage sludge ready for agricultural use. In South Africa, little information is
available on the risks associated with using sewage sludge that has not been
disinfected.

International authors have investigated and quantified these risks. As a result of

the factors that are unique to South Africa, it would not be appropriate to adopt
work from other countries. These factors justify an investigation to assess the
risks associated with the use of pathogen rich sewage sludge in agricultural
practices.

A high risk crop was chosen to illustrate a worst case scenario. It was therefore
decided to investigate the prevalence of microorganisms in a crop grown in
sewage sludge amended soil. A risk assessment will provide a means of
estimating the probability of adverse effects associated with measured or
estimated levels of hazardous agents, and a tool for predicting the extent of
potential health effects.

35
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Based on our understanding and findings a functional management plan for

sewage sludge application to agricultural land can be formulated.

36
 University of Pretoria etd – Chale-Matsau, J R B (2005)

2.13 References

Abbaszadegan, M., Stewart, P. and Lechevallier, M. 1999. A Strategy for

detection of viruses in groundwater by PCR. Applied and Environmental
Microbiology. 65(2). 444 - 449

Ackers, M-L., Mahon, B.E., Leahy, E., Goode, B., Damrow, T., Hayes, P.S., Bibb,
W.F., Rice, D.H., Barett, T.J., Hutwagner, L., Griffin, P.M. and Slutsker, L. 1998.
An Outbreak of Escherichia coli O157:H7 Infections Associated with Leaf Lettuce
Consumption. The Journal of Infectious Diseases. 177. 1588-1593

Apedaile, E. 2001. A Perspective on Biosolids Management. The Canadian

Journal of Infectious Diseases. 12(4)
http//www.pulsus.com/Infdis/12_04/aped_ed.htm

Asplund, K. and Nurmi, E. 1991. The Growth of Salmonellae in Tomatoes.

International Journal of Food Microbiology. 13. 177-182

Ayres, R.M., Stott, R., Lee, D.L., Mara, D.D. and Silva, S.A. 1992. Contamination
of Lettuce with Nematode Eggs by Spray Irrigation with Treated and Untreated
Wastewater. Water Science and Technology. 26(7-8) 1615-1623

Baloda, S.B., Christensen, L. and Trajcevska, S. 2001. Persistence of a

Salmonella enterica Serovar Typhimurium DT12 Clone in a piggery and in
Agricultural Soil Amended with Salmonella-Contaminated Slurry. Applied and
Environmental Microbiology. 67(6). 2859-2862

Bilgrami, K.S. and Kumar, S. 1998. Bacterial Contamination in Water of the River
Ganga and its Risk to Human Health. International Journal of Environmental
Health Research. 8. 5-13

Bitton, G. 1994. Wastewater Microbiology. Wiley-Liss, New York. 478pp

37
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Bofill-Mas, S., Pina, S. and Girones, R. 2000. Documenting the Epidemiologic

Patterns of Polyomaviruses in Human Populations by Studying their Presence in
Urban Sewage. Applied and Environmental Microbiology. 66(1). 238 - 245

Black, R.E., Levine, M.M., Clement, M.L., Hughes, T.P. and Blaser, M.J. 1988.
Experimental Campylobacter jejuni Infection in Humans. Journal of Infectious
Diseases. 157. 472-479

Blaser, M.J. 1996. How Safe is Our Food? The New England Journal of
Medicine. 334(20). 1324-1325

Brackett, R.E. 1987. Microbiological Consequences of Minimally Processed

Fruits and vegetables. Journal of Food Quality. 10. 195-206

Brooks, G.F., Butel, J.S. and Ornston, L.N. 1991. Medical Microbiology. 9th
Edition. New York. 488 pp

Brown, K.W., Thomas, J.C. and Donnelley, K.C. 1991. Bacterial Mutagenicity of
Municipal Sewage Sludge. Journal of Environmental Science and Health A26(3).
395-413

Bubert, A., Hein, I., Rauch, M., Lehner, A., Yoon, B., Goebel, W. and Wagner, M.
1999. Detection and Differentiation of Listeria spp. By a Single Reaction Based
on Multiplex PCR. Applied and Environmental Microbiology. 65(10). 4688 - 4692

Burge, W.D. and Marsh, P.B. 1978. Infectious Disease Hazards on Land
spreading Sewage Wastes. Journal of Environmental Quality. 7(1) 1-9

Carneiro, F.F., Cifuentes, E., Tellez-Rojo, M.M. and Romieu, I. 2002. The Risk of
Ascaris lumbricoides Infection in Children as an Environmental Health Indicator
to Guide Preventive Activities in Caparao and Alto Caparao, Brazil. Bulletin of the
World Health Organization. 80(1). 40-46

38
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Champliaud, D., Gobet, P., Naciri, M., Vagner, O., Lopez, J., Buisson, J.C.,
Varga, I., Harly, G., Mancassola, R. and Bonnin, A. 1998. Failure to Differentiate
Cryptosporidium parvum from C. meleangridis Based on PCR Amplification of
Eight DNA Sequences. Applied and Environmental Microbiology. 64(4). 1454 -
1458

Clark, C.S., Linnemann, C.C., Clark, J.G. and Gartside, P.S. 1984. Enteric
Parasites in Workers Occupationally Exposed to Sewage. Journal of
Occupational Medicine. 26(4). 273 - 275

Damgaard-Larsen, S., Jensen, K.O., Lund, E. and Nissen, B. 1977. Survival and
Movement of Enterovirus in Connection with Land Disposal of Sludges. Water
Research. 11. 503 – 508

DeRopp, 1999. Worms and Disease. In: Humanure Handbook. Jenkins, J (Ed.)
Jenkins Publishing, PA. www.weblife.org/humanure/chapter7.html

Dorrington, R., Bradshaw, D. and Budlender, D. 2002. HIV/AIDS Profile in the

Provinces of South Africa. Indicators for 2002. Medical Research Council, South
Africa. 31pp

Doyle, M.P. 2000. Reducing Foodborne Disease: What Are the Priorities?
Nutrition. 16(7/8). 647-649

Easton, J.S. 1983. Utilisation and Effects of Anaerobically Digested Sludge on a

Red Sandy Soil of Natal. Water SA. 9(2). 71 - 78

EPA, 1999. Environmental Regulations and Technology. Control of pathogens

and vector attraction in sewage sludge. U.S. Environmental Protection Agency.
EPA/625/R-92-013. 111pp

39
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Finch, G.R. and Belosevic, M. 2001 Controlling Giardia spp. and Cryptosporidium
spp. in drinking water by microbial reduction processes. Canadian Journal of Civil
Engineering. 28(Suppl. 1). 67-80

Forcier, F. 2002. Biosolids and Bioaerosols: The Current Situation. Biosolids and
Bioaerosols Solinov. Quebec Ministry of Environment. 22pp

Gagliardi, J.V. and Karns, J.S. 2000. Leaching of Escherichia coli O157:H7 in
Diverse Soils Under Various Agricultural Management Practices. Applied and
Environmental Microbiology. 66(3). 877-883

Gaspard, P. and Schwartzbrod, J. 1993. Determination of the Parasitic

Contamination of Irrigated Vegetables. Water Science and Technology. 27(7-8).
295-302

Gaspard, P. G., Wiart, J. & Schwartzbrod, J. 1995. Urban sludge reuse in

agriculture: Waste treatment and parasitological risk. Bioresearch and
Technology. 52: 37-40

Genthe, B. 1998. Specialist Study on the Potential Health Impacts of the

Proposed wastewater treatment facility on the West Bank of East London.
Environmentek, CSIR. 20pp

Gerba, C.P., Wallis, C. and Melnick, J.L. 1975. Fate of Wastewater Bacteria and
Viruses in Soil. Journal of Irrigation and Drainage Engineering. 101. 157-174

Guo, X., Chen, J., Beuchat, L.R. and Brackett, R.E. 2000. PCR Detection of
Salmonella enterica Serotype Montevideo in and on Raw Tomatoes Using
Primers Derived from hilA. Applied and Environmental Microbiology. 66(12).
5248 – 5252

Hyde, H.C. 1976. Utilization of Wastewater Sludge for Agricultural Soil

Enrichment. Journal of Water Pollution Control Federation. 48(1). 77 - 90
40
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Hu, C.J., Gibbs, R.A., Mort, N.R., Hofstede, H.T., Ho, G.E. and Unkovich, I.
1996. Giardia and Its Implications for Sludge Disposal. Water Science and
Technology. 34(7-8). 179-186

Itoh, Y., Sugita-Konoshi, Y., Kasuga, F., Lwaki, M., Hara-Kudo, Y., Saito, N.,
Noguchi, Y., Konuma, H. and Kumagai, S. 1998. Enterohemorrhagic Escherichia
coli O157:H7 Present in Radish Sprouts. Applied and Environmental
Microbiology. 64(4). 1532-1535

Jimenez, B., Maya, C., Sanchez, E., Romero, A., Lira, L. and Barrios J.A. 2002.
Comparison of the Quantity and Quality of the Microbiological Content of Sludge
in Countries with Low and High Content of Pathogens. Water Science and
Technology 46(10) 17-24

Jenkins, M.B., Walker, M.J., Bowman, D.D., Anthony, L.C. and Ghiorse, W.C.
1999. Use of a Sentinel system for field Measurements of Cryptosporadium
parvum oocyst inactivation in soil and animal waste. Applied and Environmental
Microbiology. 65(5). 1998-2005

Jones, P.W. 1980. Health hazards associated with the handling of animal
wastes. Veterinary Record. 106. 4-7

Kirby, J.B. 2001. Exposure, resistance, and recovery: a three-dimensional

framework for the study of mortality from infectious disease. Social Science and
Medicine. 53. 1205-1215

Korentajer, L. 1991. A review of the agricultural use of sewage sludge: Benefits

and potential hazards. Water SA. 17(3). 189-196

Kornacki, J.L. and Marth, E.H. 1982. Foodborne Illness Caused by Escherichia
coli: A Review. Journal of Food Protection. 45(11). 1051-1067

41
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Kouloumbis, P., Rigas, F. and Mavridou, A. 2000. Environmental Problems from

the Disposal of Sewage Sludge in Greece. International Journal of Environmental
Health Research. 10. 77-83

Kuckzynska, E. and Shelton, D.R. 1999. Method for Detection and Enumeration
of Cryptosporidium parvum Oocysts in Feces, Manures and Soils. Applied and
Environmental Microbiology. 65(7). 2820 - 2826

Lee, G.F. and Jones-Lee, A. 1993. Public Health Significance of Waterborne

Pathogens in Domestic Water Supplies and Reclaimed Water. Report to state
California Environmental Protection Agency Comparative Risk Project, Berkley,
CA, 27pp

Manero, A. and Blanch, A.R. 1999. Identification of Enterococcus spp. with a

biochemical key. Applied and Environmental Microbiology. 65. 4425-4430

Maxy, R.B. 1982. Fate of Microbial Contaminants in Lettuce Juice. Journal of

Food Protection. 45(4). 335-339

Melloul, A.A and Hassani, L. 1999. Salmonella Infection in Children from the
Wastewater-spreading Zone of Marrakesh City (Morocco). Journal of Applied
Microbiology. 87. 536-539

NRC, 1996. Use of Reclaimed Water and Sludge in Food Crop Production.
Committee on the Use of Treated Wastewater Effluents and Sludge in the
Production of Crops for Human Consumption. Water Science and Technology
Board. Commission on Geosciences, Environment and Resources. National
Research Council. National Academy of Press, Washington, DC

NRC, 2002. Biosolids Applied to land. National Research Council. National

Academy of Press, Washington, DC

42
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Pahren, H.R., Lucas, J.B., Ryan, J.A. and Dotson, G.K. 1979. Health risks
associated with land application of municipal sludge. Journal of Water Pollution
Control Federation. 51(11). 2588 - 2601

Pell, A.N. 1997. Manure and Microbes: Public and Animal Health Problem?
Journal of Dairy Science. 80. 2673-2681

Penner, K. 1998. Microorganisms and Foodborne Illness. Food Safety.

http://www.oznet.ksu.edu

Pillai, S.D., Widmer, K.W., Dowd, S.E. and Ricke, S.C. 1996. Occurrence of
Airborne Bacteria and Pathogen Indicators during Land Application of Sewage
Sludge. Applied and Environmental Microbiology. 62(1). 296 - 299

Parliamentary Bulletin, 1996. Poverty in South Africa – Poverty Week Debate. 21

October 1996. Parliamentary Bulletin No. 7. Republic of South Africa

Prescott, L.M., Harley, J.P. and Klein, D.A. 2002. Microbiology 5th ed. McGraw-
Hill. New York. 1026pp

Purves, D. 1990. Toxic Sludge. Nature. 346. 617 - 618

Recke, H., Schnier, H.F., Nabwile, S. and Qureshi, J.N. 1997. Responses of Irish
Potatoes (Solanum Tuberosum L.) to Mineral and Organic Fertilizer in Various
Agro-Ecological Environments in Kenya. Experimental Agriculture. 33(1). 91-102

Reis, M.H., Vasconcelos, J.C. and Trabulsi, L.R. 1980. Prevalence of

Enterotoxigenic Escherichia coli in Some Processed Raw Food from Animal
Origin. Applied and Environmental Microbiology. 39(1). 270-271

43
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Rocourt, J., Hogue, A., Toyofuku, H., Jacquet, C. and Schlundt, J. 2001. Listeria
and listeriosis: Risk assessment as a new tool to Unravel a multifaceted problem.
American Journal of Infection Control.29(4). 225-227

Rost, U., Joergensen, R.G. and Chander, K. 2001. Effects of Zn enriched

sewage sludge on microbial activities and biomass in soil. Soil Biology and
Biochemistry. 33. 633-638

Rudolfs, W., Falk, L.L. and Ragotzkie, R.A. 1950. Literature review on the
occurrence and survival of enteric, pathogenic, and relative organisms in soil,
water, sewage, sludges, and on vegetation. Sewage and Industrial Wastes. 22.
1261-1281

Seo, K.H. and Frank, J.F. 1999. Attachment of Escherichia coli O157:H7 to
Lettuce Leaf Surface and Bacterial Viability in Response to Chlorine Treatment
as Demonstrated by Using Confocal Scanning Microscopy. Journal of Food
Protection. 62(1). 3-9

Shuval, H.I., Yekutiel, P. and Fatal, B. 1984. Epidemiological evidence for

Helminth and Cholera transmission by Vegetables Irrigated with Wastewater:
Jerusalem – a case study. Water Science and technology. 17. 442-443

Sidhu, J., Gibbs, R.A., Ho, G.E. and Unkovinch, I. 1999. Selection of Salmonella
Typhimurium as an Indicator for Pathogen Regrowth Potential in Composted
Biosolids. Letters in Applied Microbiology. 29. 303-307

Small Wright, M. 2002. Sewage Sludge/Biosolids. A Health and Environmental

Crisis and Scandal. http://www.pereladra-ltd.com/AB1473/webpage.cfm

Smith S. R. 1996. Agricultural recycling of sewage sludge and the environment.

Biddles Ltd., Guildford. 446pp

44
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Snyman, H.G., De Jong, J.M. and Aveling, T.A.S. 1998. The Stabilization of
Sewage Sludge Applied to Agricultural Land and the Effects on Maize Seedlings.
Water Science and Technology. 38(2). 87-95

Snyman, H.G., Terblanche, J.S. and Van der Westhuizen J.L.J. 1999.
Management of Land Disposal and Agricultural Reuse of Sewage Sludge Within
the Framework of the Current South African Guidelines. Procedures of
Specialised Conference on Disposal and Utilization of Sewage Sludge:
Treatment Methods and Application Modalities. Athens, Greece

Snyman, H.G. and Van der Waals, J. 2003. Laboratory and field scale evaluation
of agricultural use of sewage sludge. Final Report of WRC Project Number
K5/1210

Snyman, H.G., Herselman, J.E. and Kasselman, G. 2003. A Metal Content of

South African Sewage Sludge and an Evaluation of Analytical Methods for their
Determination in Sludge. WRC Report

Sobsey, M.D. 1996. Microbial Aspects of Water Quality and Health in the
Developed and Developed World. Stockholm Water Symposium 1996.
Safeguarding Water Resources for Tomorrow: New Solutions to Old Problems.
Stockholm. 195-197
Straub, T.M., Pepper, I.L. and Gerba, G.L. 1993. Hazards from Pathogenic
Microorganisms in Land-disposed Sewage Sludge. Rev of Environmental
Contamination and Toxicology. 132. 55-91

Straub, T.M., Pepper, I.L., Abbaszadegan, M. and Gerba, C.P. 1994. A Method
to Detect Enteroviruses in Sewage Sludge-Amended Soil Using the PCR.
Applied and Environmental Microbiology. 60(3). 1014 - 1017

Straub, T.M., Pepper, I.L. and Gerba, C.P. 1995. Comparison of PCR and Cell

45
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Culture for Detection of Enterovirises in Sludge-Amended Field Soils and

Determination of their Transport. Applied and Environmental Microbiology. 61(5).
2066 - 2068

Strauch, D. 1991. Survival of Pathogenic Microorganisms and Parasites in

Excreta, Manure and Sewage Sludge. Revue Scientifique et Technique office
International des Epizooties. 10(3). 813 - 846

Sunday Times, 2003. Putting Our Pooh to Good Use. Location: Sunday 24
August 2003. Gauteng News

Tanner, B., Brooks, J., Josephson, K., Gerba, C. and Pepper I. 2003. Evaluation
of the Potential for Bioaerosols from Land Applied Biosolids. Proceedings of the
IWA Biosolids 2003 Conference, wastewater Sludge as a Resource. 23-25 June
2003. Trondheim. Norway

Takeuchi, K., Matute, C.M., Hassan, A.N. and Frank, J.F. 2000. Comparison of
the Attachment of Escherichia coli O157:H7. Listeria mococytogenes, Salmonella
typhimurium and Pseudomonas fluorescens to Lettuce Leaf. Journal of Food
Protection. 63(10). 1433-1437

Takeuchi, K. and Frank, J.F. 2000. Penetration of Escherichia coli O157:H7 into
Lettuce Tissue as Affected by Inoculum Size and Temperature and the Effect of
Chlorine Treatment on Cell Viability. Journal of Food Protection. 63(4). 434-440

Takeuchi, K. and Frank, J.F. 2001. Quantitative Determination of the Role of

Lettuce Leaf Structures in Protecting Escherichia coli O157:H7 from Chlorine
Disinfection. Journal of Food Protection. 64(2). 147-151

Tauxe, R.V. 1997. Emerging foodborne diseases: an evolving public health

challenge. Emerging Infectious Diseases. 3. 425-434

46
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Tester, C.F. and Parr, J.F. 1983. Intensive Vegetable Production Using Compost.
Biocycle. 24(1). 34-36

Teunis, P.F.M. and Havelaar, A.H. 2000. The Beta Poisson Dose-Response
Model is not a Single-Hit Model. Risk Analysis. 20(4). 513- 520

Tierney, J.T., Sullivan, R. and Larkin, E.P. 1977. Persistence of Poliovirus 1 in

Soil and on Vegetables Grown in Soil Previously Flooded with Inoculated
Sewage Sludge or Effluent. Applied and Environmental Microbiology. 33(1). 109-
113

Tyson, J.M. 2002. Perceptions of Sewage Sludge. Water Science and

Technology. 46:373-380

Vesilind, P.A. 2003. Pathogens in Sludge: A Case of Sufficient Challenge.

Proceedings of the IWA Biosolids 2003 Conference, wastewater Sludge as a
Resource. 23-25 June 2003. Trondheim. Norway

Vilanova, X., Manero, A., Cerdà-Cuéllar, M. and Blanch, A.R. 2002. The effect of
a sewage treatment plant effluent on the faecal coliforms and enterococci
populations of the reception river waters. Journal of Applied Microbiology. 92.
210-214

Vorobieva, L.V., Selyuzhitskii, G.V. and Chernova, G.I. 1996. Ecologic and
Hygienic Evaluation of Lake Ladoga as a Source of Drinking Water.
Hydrobiologia. 322. 137-141

Waage, A.S., Vardund, T., Lund, V. and Kapperud, G. 1999. Detection of Small
Numbers of Campylobacter jejuni and Campylobacter coli Cells in Environmental
Water, Sewage and Food Samples by a Seminested PCR Assay. Applied and
Environmental Microbiology. 65(4). 1636-1643

47
 University of Pretoria etd – Chale-Matsau, J R B (2005)

Wachtel, M.R., Whitehead, L.C. and Mandrell, R.E. 2002a. Association of

Escherichia coli O157:H7 with Preharvest Leaf Lettuce Upon Exposure to
Contaminated Irrigation Water. Journal of Food Protection. 65(1). 18-25

Wachtel, M.R., Whitehand, L.C. and Mandrell, R.E. 2002b. Prevalence of

Escherichia coli Associated with a Cabbage Crop Inadvertently Irrigated with
Partially Treated Sewage Wastewater. Journal of Food Protection. 65(3). 471-
475

Wells, J.M. and Butterfield, J.E. 1997. Salmonella Contamination Associated with
Bacterial Soft Rot of Fresh Fruits and Vegetables in the Marketplace. Plant
Disease. 81. 867-872

WHO, 1979. Human Viruses in Water, Wastewater and soil. Report of a WHO
Scientific Group. World Health Organization Technical Report Series 639. World
Health organization, Geneva. 19 -20

WRC, 1997. Permissible Utilisation and Disposal of Sewage Sludge. Water

Research Commission. TT 85/97

WRC, 2002. Addendum to the Guideline on Permissible Utilisation and Disposal

of Sewage Sludge. Edition 1 Water Research Commission. TT 154/01

Zenz, D.R., Peterson, J.R., Brooman, D.L. and Lue-Hing, C. 1976. Environmental
Impacts of Land Application of Sludge. Water Pollution Control Federation.
48(10) 2332-2342

Zwietering, M.H and Gerwen, S.J.C. 2000. Sensitivity Analysis in Quantitative

Microbial Risk Assessment. International Journal of Food Microbiology. 58. 213-
221

48