Microbial Risks Associated with Biogas and Biodigestor Sludge
B Vinnerås, Swedish University of Agricultural Sciences, Uppsala, Sweden, and National Veterinary Institute,
Uppsala, Sweden
C Schönning, Swedish Institute for Infectious Disease Control, Water and Environmental Microbiology, Solna,
Sweden
& 2011 Elsevier B.V. All rights reserved.
Abbreviations
ABP animal by products
HRT hydraulic retention time
MRT minimal retention time
VTEC vero toxin-producing Escherichia coli
Introduction
Anaerobic digestion is an attractive alternative for
treatment of biodegradable waste, farm waste, and was-
tewater. In the industrialized world, sewage sludge, ani-
mal manure and biodegradable household and industrial
waste, and wastewater sludge are treated anaerobically in
medium- to large-scale systems. The majority of the
small-scale anaerobic reactors that exist are found in the
developing world. Most of these reactors are local, on-
farm reactors for on-site treatment of manure and farm
waste together with human-derived waste (e.g., excreta
and biodegradable household waste).
In most of the cases in the industrialized countries, the
biodigestor sludge (biodigestate) is used as a fertilizer and
its standard of hygiene is important when used on food
crops. For example, sewage sludge is mainly used for
other purposes (e.g., soil production), among other fac-
tors, due to the uncertainty regarding the hygiene quality.
The biogas produced is used as an energy source, elec-
tricity, and car fuel; but in some cases biogas can also be
found on the national gas grid alone or in combination
with natural gas. The latter has raised concerns about the
risk of spreading pathogens into household gas systems.
In developing countries most of the treatment is
performed on small scale, with different semiunder-
ground systems such as fixed dome, floating drum, or bag
reactors. The gas from these systems is mainly used for
household purposes such as cooking and heating, and the
remaining biodigestate is either used in agriculture or
released to a nearby water recipient.
Biodigestion
The degradation of organic matter during anaerobic di-
gestion occurs in four different steps: hydrolysis, acid-
ogenesis, acetogenesis, and methanogenesis. In contrast to
aerobic treatment, where oxygen is the electron
acceptor and the organic material can be degraded all
the way to carbon dioxide, every step of the anaerobic
degradation is performed separately using different
chemical compounds as electron acceptor and each of the
steps is performed by separate organisms. During the
initial steps of hydrolysis, acidogenesis, and acetogenesis,
where the redox potential is more than 300 mV, intes-
tinal organisms such as Escherichia coli and Salmonella can
aid in the process and grow. The methane formation step
maintains a redox potential less than 300 mV and is
performed by strictly anaerobic methane-forming bacteria.
Most reactors run in the mesophilic temperature
range (30–40 1C), with a few exceptions of psychrophilic
(o 30 1C) and thermophilic (445 1C) reactors. No ex-
ternal heating is needed to create a mesophilic process in
tropical regions. In the different temperature intervals
within the range 20–90 1C, different organisms dominate
methane formation depending on their optimum tem-
perature, for example, Methanogenum spp. have an opti-
mum of 20–40 1C and Methanothermum spp. an optimum
of 80–90 1C. Moreover, most reactors run at neutral pH
and produce low concentrations of chemicals that can act
as toxins (e.g., ammonia). However, after a long adap-
tation period most processes can be adapted to highly
specific local conditions due to the substrate used (e.g.,
ammonia concentrations of o5 g l 1 due to digestion of
slaughterhouse waste).
A wide variety of material originating from house-
holds, industry, and agriculture can be treated by an-
aerobic digestion. The main requirement is that it is
biodegradable in the absence of oxygen. Solid household
waste and industrial waste, including that from shops,
markets, and restaurants, can contain pathogens de-
pending on how the material is produced and processed
(Table 1). The main sources of pathogens are con-
taminated food and food products but in general the level
of contamination is low in these fractions. Most medium-
and large-scale sewage plants treat their dewatered
sludge anaerobically. The content of enteric pathogens in
sewage sludge is high as it reflects a large population, and
when using the treated sludge as a fertilizer even for
energy production it is important that no zoonotic
pathogens (e.g., Salmonella and VTEC (verotoxin-pro-
ducing E. coli )) are transmitted from the population to
the surrounding wild and domestic animals.
757
758 Microbial Risks Associated with Biogas and Biodigestor Sludge
Table 1 Materials entering biodigestors and their sources of
pathogens
Fraction to digestor Main source of pathogens
Food waste from Contaminated food and foodstuffs
households and
groceries
Other household waste Feces-contaminated material,
e.g., disposable nappies, toilet
paper, and feces from humans
and animals
Sewage sludge, including Infected humans
latrine waste
Manure Infected livestock
Slaughterhouse waste Infected livestock
Waste from the food Contaminated food and foodstuffs
industry
Park and garden waste Excreta from humans, pets, and
wild animals
In most places where anaerobic digestion is practiced,
manure serves as the main source of incoming material.
Several large-scale municipal biodigestors in western
Europe use some manure in their treatment, the major
exception being reactors for treatment of sewage sludge.
However, at least in northern Europe, this is decreas-
ing as most of the reactors are used for household-
and industrial waste and these wastes are associated
with a treatment fee, whereas manure is not. The
second treatment alternative found is farm-based diges-
tors where the material treated is mainly manure
and the main purpose is energy production – in most
cases electricity. In some cases, for example, in farms
located close to urban areas, reduction of smell is also
an important factor. In developing countries, a number
of farm-based treatment systems are also used, and
here the material treated is often a mixture of animal
manure and human excreta and agricultural waste
(e.g., harvest residues), and the gas is used mainly for
cooking purposes. A reactor type that is rapidly
increasing in interest is the bag reactor, where a plastic
bag is used for biodigestion purposes.
Slaughterhouse waste (animal by-products (ABP)
category 3) can have high pathogen content depending
on the health status of the animals. In Europe, ABP must
be treated for 1 h at 70 1C or similar according to EC
regulations 1774/2002 and 208/2006. In many places in
developing countries, slaughterhouse waste is not treated
at all but just left as a pollutant in the closest water
recipient. By collection and some sort of treatment, the
pollution and risk of transmission of diseases could be
managed. One attractive alternative in this case is bio-
digestion treatment, where in addition to biogas pro-
duction, the material can be stabilized and, with
additional sanitization treatment, the risks to the en-
vironment can be minimized.
Management of Incoming Material
In highly advanced treatment systems, all incoming ma-
terial is heat-treated, especially in the case of slaugh-
terhouse- and food waste. The temperature and duration
of pretreatment are regulated by the type of material;
where manure is treated in central systems, food waste
and ABP category 3 must be heated at 70 1C for 1 h or
subjected to a treatment giving a similar effect. The
majority of intestinal pathogens are easily managed with
this kind of treatment, the exceptions being some heat-
resistant viruses that may survive or be only slightly
reduced during the treatment. For example, Parvoviruses
are only reduced by 2–3 log10 by pasteurization. Sewage
sludge, which in many cases holds a higher risk of
pathogen content, is not treated with regard to its
pathogen content before biodigestion. In biogas digestors,
where the outgoing gas is used for electricity production,
a large amount of excess heat is produced from the gas
engine and this can be used for both heating the reactor
and for pasteurization of the incoming material. If the
biogas is produced for other purposes, for example,
cooking or car fuel, external heat is required as the in-
ternal heat production within the process is too small to
achieve a significant rise in temperature, since most of the
energy produced is in the form of chemical energy, as
methane.
The most critical parameter for pathogen inactivation
is the temperature in combination with time, but in some
cases effects from chemicals present can also be included
(e.g., high concentration of ammonia).
Monitoring of pathogen reduction in different stages
of treatment of biodegradable waste shows that con-
ventional treatment by mesophilic digestion gives a
limited reduction in the content of pathogens (Table 2).
Most studies show that conventional mesophilic treat-
ment can reduce the incoming organisms by up to 1 log
(90%) during the treatment, but it may be possible to
improve this, as discussed later. Using thermophilic di-
gestion or extending treatment by composting or storage
of the treated sewage sludge improves the final hygiene
quality. However, long-term storage or composting de-
creases the fertilizer value of the final product and in-
creases the risk of pollution of the environment. Table 2
reviews a number of studies, but these were generally
based on the analysis of a few microorganisms. When
considering the final reduction in indicator bacteria or
bacterial pathogens, it may be difficult to differentiate
between actual inactivation and regrowth. The data given
for bacteria are therefore uncertain. One important
conclusion is that high temperature treatment (pasteur-
ization) is the most effective treatment, but even during
pasteurization it is possible for some viruses to survive.
One alternative hygiene treatment method presented
in recent years is using ammonia as a sanitation agent
 Microbial Risks Associated with Biogas and Biodigestor Sludge
Table 2 Potential percentage reduction in pathogenic microorganisms during treatment of biodegradable waste and wastewater,
including animal manure
Percentage reduction
Enteric bacterial
pathogensa
Mesophilic digestion (35 1C) 0–90
Thermophilic digestion (55 1C) 90–100
Long-term storage (41 year) 90–100
Pasteurization (70 1C, 1 h) 100
a
The organisms included here are mainly pathogens from the Enterobacteriaceae group (e.g., Salmonella spp., E. coli, Campylobacter jejuni ).
b
Ascaris spp. used as model parasite.
c
Adenovirus used as model for viruses. When data are not available, other pathogenic viruses are used (e.g., Hepatitis A, Rotavirus, and for
Parvovirus pasteurization).
within the reactor. By increasing the input of proteins to
the reactor, the ammonia content of the biodigestate is
increased. If this is done slowly, the methane-producing
organisms are able to adapt, whereas newly introduced
organisms such as pathogens and organisms that cannot
grow within the reactor (i.e., parasites and viruses) are
reduced faster than with conventional anaerobic treat-
ment. Mesophilic digestion in high ammonia reactors has
an efficient reduction of bacterial pathogens and the
ammonia present assure no regrowth of these bacteria.
However, the potential regrowth of pathogenic clostridia
needs to be resolved. Spore-forming bacteria are not
reduced during pasteurization or digestion and are thus
not included in regulation EC 208/2006. However, po-
tential pathogenic clostridia (e.g.,Clostridium chauvoei ) can
regrow during anaerobic fermentation at nonacidic pH,
constituting a risk if the residue is used as a fertilizing
agent.
Thermophilic treatment has in some cases been pre-
sented as an alternative treatment for highly pathogenic
material such as sewage sludge, as it has an efficient
treatment process that inactivates the pathogens if a
practical hydraulic retention time (HRT) and minimal
retention time (MRT) are maintained (see later). The
disadvantage with this kind of treatment is that external
heating is always needed to run the process at these high
temperatures and that the degradation of organic pollu-
tants is much slower in this temperature range.
Hygiene Risk of Outgoing Material
Biodigestate
Treatment for reduction of potential pathogens in bio-
digestate is only one of the several barriers that should be
used to prevent disease transmission. If the risks of
pathogen survival are high, selective use of the treated
biodigestate can be a complement. Selective use can in-
volve, for example, application of sewage sludge-based
biodigestate to energy crops. However, regardless of end
759
Parasitesb Virusesc
0–50 0–90
90–100 90–100
0–90 90–100
100 99–100
use, zoonotic diseases first have to be managed, otherwise
there is a risk of transmission of diseases to surrounding
wild- and domestic animals. The use of manure-based
biodigestate for fertilization of food products is a similar
case, as zoonotic diseases can be transmitted with the
biodigestate.
The composition of the biodigestate regarding the
pathogen components is closely correlated to the sources
of incoming material (Table 1) and the treatment process
used (Table 2). When excreta (human and animal) are
treated, either directly in local reactors or as sewage from
sewage treatment plants, there is a high risk of pathogens
being present in the material. Studies of the pathogen
content in mesophilically digested sewage sludge in
Sweden show a low number of helminths but higher
numbers of viruses and pathogenic bacteria such as
Salmonella spp. Corresponding studies of the pathogen
content in sewage sludge in the United States and Spain
show higher occurrence of helminths and also high
numbers of pathogenic viruses and bacteria. Studies of
the pathogen content in anaerobic digestors for animal-
and human excreta in Vietnam show even higher counts
of helminths and bacterial pathogens.
The incoming material is the main source of patho-
gens in the biodigestate. However, several studies indi-
cate an accumulation of some pathogenic bacteria during
the anaerobic treatment process. For example, both
Campylobacter jejuni and Salmonella spp. have shown this
ability. The intestinal pathogenic bacteria can grow
during the initial hydrolyzing and acidifying steps of
anaerobic digestion, as most of the pathogens are facul-
tative anaerobic bacteria (e.g., Salmonella spp.). By intro-
ducing an additional second digestion, with longer
retention time, the bacterial pathogens are reduced
together with the available nutrients and electron ac-
ceptors for energy production, as shown by lower redox
potential. In biodigestate from several commercial
digestors with pasteurization of all incoming material, it
is still possible to find intestinal flora (e.g., Enterococcus
spp.). Close examination of such biodigestors reveals
760 Microbial Risks Associated with Biogas and Biodigestor Sludge
evidence of colonization and growth of Enterococcus spp
within the reactor, particularly in reactors that hold a
high ammonia concentration (44 g l 1). Therefore,
Enterococcus spp. is not a good indicator for the removal of
intestinal pathogens in biogas treatment processes. This
puts the emphasis on the quality of the inoculate used for
the reactor, which initially comes from unpasteurized
material. There are only a few obligate anaerobic
pathogenic bacteria that can infect through the gastro-
intestinal tract. One important example is Clostridium
chauvoei (black leg, a worldwide infectious disease of
ruminants), which conventional treatment, even heat
treatment at 70 1C for 1 h, cannot remove due to its spore
form. The survival and growth of this organism in the
biodigestion process needs to be further investigated, but
it is likely to survive the anaerobic treatment well.
The reduction of undesirable microorganisms during
biodigestion relies on a number of factors relating to the
composition of the material and the temperature. One of
the most important factors is the actual duration of
treatment. Most discussed is the effect of the HRT, the
average length of time that the compound remains in the
reactor. The HRT varies greatly, from 1 to 2 days in small
on-farm bag reactors in Vietnam, 10–30 days in com-
mercial reactors in northwest Europe, to almost 100 days
in farm-based reactors in the same region. The main
value of HRT is in calculating the dilution of incoming
substrate before the next emptying of a continuous-feed
reactor. In a completely mixed reactor, an HRT of 10
days results in a dilution corresponding to approximately
90% within a time span of 24 h. The most important
factor here is the MRT, the shortest time the material
remains in the reactor. Most commercial reactors have an
MRT of only a few hours. The HRT together with the
MRT defines the reduction potential of the process, as a
reasonable HRT only results in a reduction, by dilution,
corresponding to 90%. In plug flow reactors, the HRT is
closer to the MRT; and in a defined plug flow reactor, the
retention time is longer and the reduction in pathogens is
thereby greater.
Reinfection and regrowth of pathogens in treated
(hygienized, sanitized) organic material is a known
problem that has been documented in a number of
studies. Reinfection may occur when treated material,
which is basically free of pathogens, is reinfected from an
external source. In open storage of digested residues this
can occur, for example, via bird droppings, which can
contain pathogens (e.g., Salmonella). It can also occur if the
treated product is not managed properly. In Sweden,
reinfection has been documented from transport vehicles,
for example, when tankers are not cleaned properly after
transporting untreated material. Moreover, if satellite
storage is used, this also needs to be properly cleaned and
managed before initial usage, as it has shown that the risk
of contamination and regrowth in such storage is high.
For safe reuse of biodigestate, posttreatment processes
also need to be free from contaminants.
Bacteria are the only organisms that can grow in or-
ganic material. Regrowth may occur if the conditions are
appropriate for remaining bacteria, or bacteria can enter
the material via reinfection. The possibility of regrowth is
higher if the material is pasteurized after digestion,
compared to pasteurization before. In the latter case, the
remaining bacteria that have been active in the digestion
process may compete with the newly arrived potential
pathogens.
The number of biogas plants is currently increasing
worldwide, both in developing and industrialized coun-
tries. In addition, the increased demand for bioenergy
will probably further increase the number of biogas
plants. In developed countries there is a demand for re-
duced pollution of water recipients and for recycling of
organic material, and the need for stabilization and
sanitization of the material before disposal or reuse is
acknowledged. Within the European Union (EU), organic
waste may not be dumped in landfill; thus alternative
treatments and uses (or disposal) are needed. In de-
veloping countries, biodigestion could have a great po-
tential for treatment of different types of organic wastes,
including manure and human excreta, as mentioned
earlier.
Biogas
The presence of microorganisms in biogas and natural
gas systems has only been looked into in a few studies.
On examining the microbiological content, those studies
found a variety of microorganisms in the gas and in
biofilm growing on the surface of the pipes.
The microflora found in the condensate water from
the gas piping can be used as an indicator of content of
microorganisms in the biogas. In general, the total
number of microorganisms from the condensate water
culture on agar plates was up to 105 cfu ml 1 for bacteria
and 104 cfu ml 1 for fungi, whereas some bacteriophages
can also be found in the condensate. With regard to the
composition of the gas itself, the distribution was similar
to that in the condensate water with mainly bacteria,
some fungi, and no detection of viruses at all in, for
example, 2 m3 gas tested for viral content.
The main proportion of the organisms found com-
prised bacteria. Some fungi were also isolated, and al-
though the number of species was small, the total number
of organisms was high. In condensate water from systems
where wastewater was used for upgrading the gas in a
scrubber system, it was possible to find coliphages, in-
dicating that viruses could be present in the gas. In that
system, endotoxins, which upon inhalation have a general
negative impact on health, were also present.
 Microbial Risks Associated with Biogas and Biodigestor Sludge
It is also possible to find bacteria in the gas collected
from natural gas systems. Studies of both the develop-
ment of biofilm within the pipes and analysis of the
outcoming gas from the system indicate that there is a
growth of microorganisms within the system itself. The
different types of bacteria found in the systems are to a
large extent a reflection of the environment from where
the organisms enter the system, as the ambient microflora
can be found in the pipes. The concentration of micro-
organisms found in natural gas is similar to the concen-
tration in biogas and consists of both bacteria and fungi.
When there is a growing biofilm present within the
system, parts of this biofilm can be removed into the gas,
resulting in point release of organisms.
Bacteria and fungi are thus found in both condensate
water and gas. The diversity of the bacteria is generally
much higher than that of the fungi. In the studies re-
viewed, only three groups of fungi were found; Fusarium
spp., Mucor spp., and yeast. The bacterial diversity was
much higher and anaerobic Clostridia spp., as well as
several facultative anaerobic organisms were found in the
raw gas. The biogas process is generally considered to be
driven by strictly anaerobic organisms such as Clostridia
spp. However, several facultative anaerobic organisms
may contribute to the initial degradation of the organic
matter during the acidification process. A factor to con-
sider is the relatively short retention time in the reactor
combined with batch feed as the reduction of pathogens,
in most cases of mesophilic anaerobic digestion, is in
relation to the retention time.
In analysis of biogas and the condensate from biogas
systems, no actual pathogens have been found. However,
several opportunistic pathogens have been found in
the system, together with microorganisms from groups
in which pathogens are included, for example, the
Enterobacteriaceae.
Looking at the microbial community in different gas
systems, it is possible to detect 1–100 cfu m 3, mainly
bacteria but also fungi. The concentration of micro-
organisms can to some extent be correlated to the
moisture content, with fewer microorganisms at lower
moisture contents. The microorganisms detected in the
gas are to some extent reflected in the microorganisms
detected in the condensate water.
Upgrading the biogas by removal of carbon dioxide,
either for fuel or for introduction to the natural gas grid
can be achieved with several different methods. Those
most commonly used are a scrubber technique or an
absorption technique. When using the scrubber techni-
que, the source of the waste has a great effect on what is
found in the treated gas. For example, if human sewage is
the treatment substrate, there is a high risk of finding
intestinal pathogens in the treated gas. In addition, the
growth of bacteria and fungi in the carrier material is
high in the scrubber, which can lead to enrichment of
761
some of the organisms. In the absorption technique,
few external organisms are introduced, but there is still
growth of biofilm, and thereby a risk of growth of un-
wanted organisms. The microorganisms found in the
upgraded gas are probably a combination of the micro-
organisms in the water used for upgrading and the or-
ganisms growing in the packing material in the treatment
reactor.
Legionnaires’ disease is an acute respiratory infection
caused by the bacterium Legionella pneumophila, which can
cause a broad spectrum of disease from mild cough and
fever to serious pneumonia. Bacteria thrive at tempera-
tures between 20 and 50 1C and can be found in water
supply, central heating, and air-conditioning systems.
Low temperature scrubbers can be a potential source of
Legionella bacteria. Specialist cleaning and disinfection
precautions should always be taken when the tempera-
ture in the scrubber is in the ‘Legionella range.’
Regardless of gas system (natural or biogas) and
treatment of the gas, no significant difference has been
detected between the concentrations of microorganisms.
This indicates that if no active treatment is used for re-
moval of microorganisms, one can expect gas systems to
contain a microbial load of approximately 10–100
cfu m 3. The main factor to take into account is the risk
of introducing pathogens into the system. Pathogens can
enter the biogas system in scrubbers, especially the type
that use wastewater for upgrading the gas. The second
source of pathogens is biogas systems where no sani-
tization of the sludge is carried out before digestion, for
example, sewage sludge digestors or small-scale house-
hold/farm-based reactors where the gas is used for
cooking. The final source of pathogens for the biogas
system is the inoculum used when the digestor is initially
started. The easiest method for removal of unwanted
organisms is drying of the treated gas and, as in gas fuel
pumps, using a 1 mm particle filter to remove any po-
tentially pathogenic bacteria present in the gas.
Risk of Exposure
In practice, the different exposure routes can be classified
according to the handling chain of incoming material and
treated material (Figure 1). At the treatment plant the
products are handled in different steps that may result in
human exposure. The treated product is either used
directly or stored before use in, for example, agriculture.
The density of pathogens in the treated material, and
thereby the risk, is reduced over time, depending on the
survival of pathogens in the material. The risk from
direct contact with the finished product is sometimes
regarded as the highest. Transmission of disease may then
occur through unintentional ingestion of the material,
either through splashes or through contaminated hands
or food produce.
762 Microbial Risks Associated with Biogas and Biodigestor Sludge
Biogas
Manure
biowaste
Sanitati
on
Transpo
rt Diges
handlin tion
g Tran
sport
hand
ling Trans
p Vector
o rt
hand
ling
Figure 1 Potential pathogen transmission routes from anaerobic digestion of biodegradable waste and wastewater and biogas
production to humans and animals.
The risk from contaminated crops is largely
dependent on the application method used. A close-to-
ground application decreases the exposure from splashes
or aerosols and from crops if the material is placed
only on the soil when applied in a growing crop, rather
than on the plant foliage. Immediate incorporation by
tillage or direct injection further decreases the risk of
exposure of humans and animals. If pathogens remain in
the soil they can be transported to groundwater or sur-
face water. The risk is lower if a drier product is applied
(also involves less risk of splashing and aerosol for-
mation). Heavy rain can increase pathogen transport and
potentially lead to infections by ingestion of drinking
water or recreational water. The interaction between
microorganisms and soil particles is complex and the
risks are difficult to estimate. However, the survival of
microorganisms in the soil has in several cases been
shown to be long, for example, over 3 months survival
after application of Salmonella-contaminated manure to
Swedish soils.
The most obvious risk of transmission of infections is
through contaminated crops (food and feed crops).
However, it is difficult to establish direct connections
between fertilization and crop contamination and, to our
knowledge, no actual cases have been reported. However,
there are reports of higher incidence of enteric infections
when vegetables are irrigated with inappropriately trea-
ted wastewater. In developing countries (e.g., in China),
the prevalence of intestinal parasites is high in areas
where human excreta are used as fertilizer. These risks
can be reduced by restrictions on the crops fertilized.
Exposure may also occur from use on other types of land,
for example, grassland and in soil production. Possible
exposure routes are summarized in Table 3.
In comparison with other risks related to biogas sys-
tems, the risk when using the upgraded biogas is probably
insignificant. The risks of coming in contact with raw gas
or condensate water in the plant are higher, since at that
Food/fo
d
Storag Surface der
e runoff
an
Fertili
z Environ imals
Transp diges ing ment
o tate
handli rt
ng
time these substances have been subjected to a lower
degree of treatment than the upgraded gas. However, the
system is generally closed and, according to professional
assessments, workers are seldom exposed. The majority
of the microorganisms in the digestor remain in the
biodigestate rather than being transported with the gas.
One study concluded that the wastewater used in the
scrubber and the biogas produced from sewage sludge
constituted a greater risk of contributing enteric patho-
gens than the biogas produced from biodegradable solid
waste. However, further treatment of the gas when waste-
water is not used in the scrubber would probably
transform the microbial community toward more en-
vironmental microorganisms and fewer fecal indicators,
as new microorganisms would enter from the sur-
rounding environment. Upgrading the gas to car fuel
standard would significantly reduce any microorganisms
present as the gas is compressed to 200 bar and filtered
through a 1 mm particle filter. Possible exposure routes
are summarized in Table 4.
The suggestion of introducing biogas into piping
systems constructed for natural gas has been questioned,
partly due to hygiene issues. However, natural gas has
also been found to contain spore-forming bacteria such as
Bacillus spp. and the densities of microorganisms found do
not differ greatly from those found in the upgraded
biogas. Theoretically, the biogas system could contribute
pathogens to the system. However, from the results
reviewed in the present study and from a systematic
risk assessment, the indications are that this is quite
unlikely.
Recommended Management to Minimize Risk
of Transmission of Diseases
There are defined regulations and recommendations re-
garding the treatment and use of biodigestate, for ex-
ample, at EU and national level. These recommendations
 Microbial Risks Associated with Biogas and Biodigestor Sludge 763

Table 3 Possible exposure points and evaluation of exposure scenarios in a biogas system for digestion
Exposure point Possible exposure Individuals exposed Pathogens identified a Comment on risk identified
(exposure scenario)

Handling of incoming Accidental ingestion by Workers at the plant Enteric pathogens,

material splashes or similar zoonotic agents
The digestion process Accidental ingestion by Workers at the plant Enteric pathogens,
plant splashes or similar, zoonotic agents
e.g., during
maintenance
Management of Accidental ingestion by Workers at the plant Enteric pathogens, The risk is negatively
outgoing material splashes or similar, and contractors or zoonotic agents that correlated to the
e.g., during transport farmers. Animals survived treatment efficiency of the
and storage that come in contact and bacteria treatment as efficient
with the material regrowing or treatment removes the
during storage contaminating majority of unwanted
during storage organisms
Fertilization of Accidental ingestion by Contractors or farmers Enteric human The risk is negatively
agricultural fields or splashes or similar pathogens, zoonotic correlated to the
other types of land pathogens that efficiency of the
survived treatment treatment, as efficient
treatment removes the
majority of unwanted
organisms
Consumption of Ingestion Consumers Enteric human Restrictions on the use of
fertilized crops pathogens, zoonotic fertilizer can improve the
pathogens that quality, e.g., waiting
survived treatment periods and selection of
crop
Grazing animals on Ingestion Grazing animals Enteric animal Restrictions in the use of
fertilized field pathogens, zoonotic fertilizer can improve the
pathogens that quality, e.g., quarantine
survived treatment periods and selection of
crop
Wild animals on Ingestion Wildlife Enteric animal
fertilized fields pathogens, zoonotic
pathogens that
survived treatment
a
Pathogens identified as constituting a potential risk, either in theory or in studies.

take into consideration the incoming material, the
treatment process used, and reuse practices. By adapting
the treatment to the possible pathogen load from the raw
material, it is possible to significantly decrease risks.
However, in line with, for example, WHO guidelines on
wastewater reuse, it is important to include several bar-
riers to protect various groups of humans and animals.
These barriers may include restrictions on the crops
fertilized (food, feed, energy), a minimum quarantine
period between fertilization and grazing or sowing/har-
vesting, and type of application method.
In developing countries it is less likely that stan-
dardized procedures are followed, especially on the
small-scale level. The prevalence of enteric infections is
generally higher in developing countries, which results
in higher loads of pathogens in human excreta and
manure. It is also likely that foodstuffs have higher rates
of contamination due to the use of untreated organic
fertilizers and irrigation waters (e.g., wastewater) and
less hygienic handling. In tropical climates it is also
more likely that bacteria will grow on various types of
food and feed crops. Since most processes are run at
mesophilic temperatures, a significant inactivation of
pathogens is not likely because a reduction corres-
ponding to 1–2 log10 is expected. However, containment
and some degree of treatment may still be more ad-
vantageous than other alternatives. Generally, risks from
small-scale systems are lower than those from large-
scale systems. On a household level, in particular, the
risks of transmission of disease may be outweighed by
other routes.
Both in the digestion process and during the pro-
duction of gas, exposure of workers should be minimized
by supplying them with adequate information and per-
sonal protection equipment, if needed. When mainten-
ance work is being performed, it is particularly important
to bear in mind that there is a risk of disease transmission.
Upgrading the gas to car fuel standard would signifi-
cantly reduce any microorganisms present because the
gas is compressed to 200 bar and filtered through a 1 mm
764 Microbial Risks Associated with Biogas and Biodigestor Sludge

Table 4 Possible exposure points and evaluation of exposure scenarios in a biogas system for upgrading biogas
Exposure point Possible exposure Individuals exposed Pathogens identified a Comment on risk identified
(exposure scenario)

Biogas production Inhalation by leakage
of gas or during
maintenance
Biogas production – Accidental ingestion by
scrubber splashes or similar,
e.g., during
maintenance of the
scrubber
Usage of the gas as Inhalation of the gas at
fuel final use
Gas introduced to the Inhalation of the gas at
grid system for final use
natural gas (city gas)
Inhalation during
service of the piping
system
Other usage of the gas
a
Pathogens identified as constituting a potential risk, either in theory or in studies.
Workers at the plant
Workers at the plant
Personnel/car owners
filling vehicles
Household inhabitants
(the public)
Workers on the piping
system
Others
Opportunistic, and Insignificant gas leakages.
groups possibly During maintenance the
including pathogens system is shut off
Opportunistic, and Closed system.
groups possibly Condensate led to
including pathogens. WWTP. Possible
Enteric pathogens if exposure via water from
wastewater used in the scrubber and seals
scrubber during maintenance
– Requirement for particle
filter means that only
viruses and spores can
be released. Back-
suction – minimal outlet
of gas
Low volumes of gas
released. Particle filters
could further minimize
release of
microorganisms

particle filter. By having a general 1 mm particle filter,
when using the biogas or introducing the biogas into
larger gas systems, it would be possible to prevent pas-
sage of the majority of fungi and nonspore forming
bacteria. From a general risk assessment of biogas use, it
can be concluded that the risk of explosion probably
outweighs the risk of disease transmission, even without
final filtration.
Further Reading
Bagge E, Sahlström L, and Albihn A (2005) The effect of hygienic
treatment on the microbial flora of biowaste at biogas plants. Water
Research 39: 4879--4886.
Eisenhardt A, Lund E, and Nissen B (1977) The effect of sludge
digestion on virus infectivity. Water Research 11: 579--581.
Gantzer C, Gaspard P, Galvez L, Huyard A, Dumouthier N, and
Schwartzbrod J (2001) Monitoring of bacterial and parasitological
contamination during various treatment of sludge. Water Research
35: 3763--3770.
Gerardi MH (2003) The Microbiology of Anaerobic Digesters. Hoboken,
NJ: Wiley. Wastewater Microbiological Series.
Guzmán C, Jofre J, Montemayor M, and Lucena F (2007) Occurence
and levels of indicators and selected pathogens in different sludges
and biosolids. Journal of Applied Microbiology 103: 2420--2429.
Horan NJ, Fletcher L, Betmal SM, Wilks SA, and Keevil CW (2004)
Die-off of enteric bacterial pathogens during mesophilic anaerobic
digestion. Water Research 38: 1113--1120.
Ottoson J, Schnürer A, and Vinnerås B (2007) In situ ammonia
production as a sanitation agent during anaerobic digestion at
mesophilic temperature. Letters in Applied Microbiology 46:
325--330.
Pepper IL, Grooks JP, and Gerba CP (2006) Pathogens in biosolids.
Advances in Agronomy 90: 1--41.
Vinnerås B, Schönning C, and Nordin A (2006) Identification of the
microbiological community in biogas systems and evaluation of
microbial risks from gas usage. Science of the Total Environment
267: 606--615.
WHO (2006) Volume 4: The Use of Excreta and Greywater in
Agriculture. Geneva: WHO.
Zhu X, Lubeck Y, and Kilbane Z (2001) Characterization of microbial
communities in gas industry pipelines. Applied and Environmental
Microbiology 69: 5354--5363.