ORIGINAL RESEARCH
The effectiveness of aerobes used as manure additives for
swine manure odor control
Jun Zhu, PhD, EIT
Zhu J. The effectiveness of aerobes used as manure additives
for swine manure odor control. Swine Health Prod. 2000;8(1):5-9.
This article is available in PDF format
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University of Minnesota, Southern Research and Outreach Center,
35838 120th Street, Waseca, Minnesota 56093; zhuxx034@tc.umn.edu
Summary
Objective: To examine the growth of aerobic bacteria
in swine manure when used as manure additives for odor control.
Methods: Hydrogen peroxide was used as an oxygen provider
added to the test manure at fixed time intervals to enhance the
dissolved oxygen level in manure, simulating intermittent aeration.
Results: Adding hydrogen peroxide failed to establish
an aerobic environment in the top liquid. The added aerobes could
not outgrow the indigenous anaerobes even when the dissolved oxygen
levels in the manure were raised for short intervals on a regular
basis.
Implications: The frequency of running intermittent
aeration in order to maintain an active aerobic flora requires
further study. Without enough aeration, the effectiveness of microbial-based
manure additives for odor control purposes in actual manure storage
systems is questionable.
Keywords: swine,
manure, odor, microbes
Received: July 8, 1999
Accepted: October 6, 1999
The idea of using biological
manure additives to control odors was proposed about 20 years
ago and a considerable amount of research effort has been expended
in this field. Past researchers rarely found pit additive products
to be effective in reducing odor levels of swine manure,1-4
perhaps due to the complexity of odorous components in swine manure.
The key hindrance to developing effective manure additive products,
however, is our lack of understanding of the biological activities
that occur in stored swine manure. Trial-and-error methods to
evaluate manure additive products, although common, are both time
consuming and fail to measure the growth of the added bacteria.
To develop effective additive products, it is important to examine
the growth kinetics of microbes added to manure as well as characteristics
of the manure environment that may affect the chemical, physiological,
and biological processes of additives. The objective of our study
was to measure the growth of aerobic bacteria added to test manure
under partial aeration. Factors that affect bacterial growth are
also discussed.
Materials and methods
Experimental design
Our experiment was conducted over a period of 30 days in an
environmentally controlled room with the temperature maintained
between 18.3-23.9 degrees C (65-75 degrees F).
The experimental design was created to simulate a surface aerated
layer on animal waste storage facilities (Figure 1). The system was composed of a permeable
membrane of woven, nondegradable material, which was held at a
depth of approximately 30 cm (12 inches) below the liquid surface.
The liquid portion above the membrane served both as a substrate
for growth of aerobic microorganisms and a semi-permeable barrier
for migration of volatile materials from the subsurface layer.
The experimental apparatus consisted of PVC columns 38.2 cm
(15 inches) in diameter and 183.2 cm (72 inches) high. Three test
columns were assigned to each treatment to obtain three replications.
The columns were filled with manure to a depth of 137.4 cm (54
inches) at the beginning of the test and were maintained at that
level throughout the entire testing period. Air circulated continuously
through the headspace of the column (top 45.8 cm [18 inches])
at a rate of 14 L per minute (0.5 cfm) to simulate a typical ventilation
situation in either a deep pit or an outside storage basin.4
The manure was a mixture of 25% fresh manure with 75% lagoon water
to make up to 1% total solids content to mimic the top portion
of liquid manure in the storage facilities. Hydrogen peroxide
(H2O2) was used as oxidizing agent for enhancing
the level of dissolved oxygen in the top liquid to simulate aeration.
There were a total of five treatments (four treatments and
one control) in this study:
- 50 ppm of H2O2 (twice a day: 8:30 AM
and 4:30 PM) + microbial additive (ADD),
- 50 ppm of H2O2 (twice a day: 8:30 AM
and 4:30 PM) + daily catalyst (1:450 dilution) + microbial additive
(CATADD),
- 50 ppm of H2O2 (twice a day: 8:30 AM
and 4:30 PM) (50H2O2),
- 10 ppm of H2O2 (twice a day: 8:30 AM
and 4:30 PM) (10H2O2), and
- control manure.
The biocatalyst and the mixed aerobic microbial strains used
in this study were provided by Olympic Environmental Company (Denver,
Colorado). The biocatalyst is a water-like substance containing
grass extracts with neutral pH and salts of natural groundwater.
According to the results of experiments conducted by the manufacturer
(personal communication) the biocatalyst is capable of enhancing
the availability of oxygen to both aerobic and facultative microbes
in a minimal oxygen environment, thereby increasing the growth
of these microbes.
Liquid sample collection and analysis
Liquid samples were collected only from the top portion of
manure in the columns (above the permeable membrane) and analyzed,
at each sampling time, for total anaerobic and aerobic bacterial
counts. Methods presented by Zhu, et al.,5 were used
to incubate and calculate bacterial counts.
Concentrations of dissolved oxygen in the top liquid layer
were monitored using oxygen probes (OxyGuard Mk I(R),
Point Four Systems Inc.; British Columbia, Canada). The oxygen
probes were calibrated using Zero Oxygen Solution (Cole Parmer
Company; Chicago, Illinois). Each column was assigned one probe
and the probes were connected with an onsite computer so the concentrations
of dissolved oxygen in the liquid were recorded automatically
at 1-minute intervals. Recording started immediately after hydrogen
peroxide was added and lasted approximately 2.5 hours. This time
frame was determined by several trials prior to the test in which
a typical descending feature of the concentration of dissolved
oxygen was observed; i.e., the H2O2-induced
increase in dissolved oxygen concentration in the test liquid
phased out after approximately 2-3 hours.
Statistical analysis
A complete randomized design with three replications per trial
was employed. Student's t test at significance level of
.05 was used to compare the 10-, 20-, and 30-day samples.
Results
Dissolved oxygen concentrations
Dissolved oxygen (DO) concentrations for all the treatments
(except control) increased rapidly after addition of H2O2
(Figure 2). The CATADD, ADD, and 50H2O2
treatments reached a maximum mean concentration of approximately
20 mg per L in about 12 minutes and then decreased gradually.
As compared to these, the DO increase for the 10H2O2
treatment was much lower. At the end of the recording time (2.5
hours), the 50H2O2 treatment
had DO concentrations of approximately 5 mg per L, while the
other four treatments each had approximately 2 mg per L. The DO
levels for all treatments with H2O2 dropped
to a range from 0.1-0.3 mg per L before the next addition of H2O2.
Bacterial counts
After 10 days, both ADD and 50H2O2 had
significantly higher aerobic bacterial counts than the control
(Figure 3). Aerobes did not grow better
in the CATADD than in the control. Adding aerobic microorganisms
to the liquid manure did not necessarily result in an increase
in the populations of aerobes (e.g., in the CATADD treatment).
In no test columns did aerobic bacteria outgrow anaerobic bacteria
after 10 days.
After day 20, the number of anaerobes in all H2O2-treated
columns was significantly decreased and the aerobic bacterial
counts were not significantly different among the treatments containing
H2O2 (the control had concentrations of
330 for aerobes and 424 for anaerobes) (Figure
3). At the end of the test, the aerobic bacterial counts for
the 50H2O2 treatment and the 10H2O2
treatment rebounded significantly as compared to those in the
CATADD and ADD treatments (Figure 3).
Discussion
Adding H2O2 to liquid manure can dramatically
raise DO levels within a short time but can not maintain the raised
concentration. The ability of liquid to hold oxygen is temperature
dependent;6 under the test environment (approximately
20 degrees C), the saturation concentration of oxygen in manure
liquid is around 8-9 mg per L. Thus, it can be assumed that excess
oxygen in the manure liquid escaped into the air shortly after
being added.
Our observation that aerobic bacteria failed to outgrow anaerobic
bacteria after 10 days is consistent with the results obtained
by Bourque, et al.,7 who concluded that inoculated
microbes could not become dominant in nonsterilized swine manure
samples and that indigenous flora of the manure always grew better
than inoculated microorganisms. Goldstein, et al.,8
have offered this as an explanation for the possible failure of
inoculation to enhance biodegradation. There have been numerous
studies of the use of microbial additives for swine manure odor
control,1-4 but the success has been relatively limited,
as indicated by Ritter.9 Results from our test showed
that externally adding microbes failed to establish an active
aerobic flora in liquid manure.
The significantly higher anaerobic bacterial counts we observed
for most of the treatments after the first 10 days suggests that
a fully aerobic environment may not be established by adding H2O2
into the liquid. Since most indigenous bacterial genera in swine
manure are strict anaerobes, they can always outcompete the added
aerobes for nutrients and maintain their growth under anaerobic
conditions. It appeared that H2O2 had some
inhibitory effect on the growth of anaerobes, but this effect
was not apparent until after the first 10 days.
The higher aerobic counts we observed in the control compared
to the other treatments could be due to a treatment effect on
the growth rates of aerobes. For example, it is possible that
the growth of aerobes in the treatments with H2O2
was accelerated by the H2O2 . The nutrients
(such as N, P, K, S, and C) in the top layer of liquid are depleted
more rapidly than in the control.10 In the control,
the aerobes grew slowly but steadily, reaching their maximum around
day 20. The decrease in aerobe counts in the control after day
20 could also be due to the depletion of nutrients. Perhaps only
a small quantity of nutrients could diffuse up from underneath
the permeable membrane. However, the overall concentrations of
aerobes were low for all the treatments, which implied that the
available nutrients were exhausted in the top layer of liquid.
This nutrient deficiency probably constricted the growth of both
aerobic and anaerobic bacteria, resulting in a reduction in bacterial
population. Because nutrients were not analyzed in the manure
liquid, we were not able to verify this hypothesis in our study.
According to Grubbs,11 the key to using bacterial
cultures for deodorizing manure is to enable the added bacteria
to become the predominant strains. Results from our study showed
that merely adding aerobes to manure did not result in a flourishing
population of the added bacteria. There are many factors that
may affect bacterial growth,12 few of which are controllable
in a real environment. Although attempts have been made to study
mechanisms of odorous compound decomposition by aerobic bacteria,
past work was mainly focused on determining bacterial functions
in digesting odorous compounds under optimum conditions.13
This usually does not guarantee that bacteria growing well
under optimum conditions will also grow well in the field, suggesting
that the types of aerobes may also be important in terms of growth
and function. In general, microbial species with the fastest growth
rate and the ability to utilize most of the available organic
matter will be the predominant species.12 Because the
ability of different bacterial genera or species to tolerate the
living environment varies, as does their ability to effectively
digest odorous organic compounds, it will be critical to identify
and select aerobic bacteria specifically for treating swine manure
stored in typical storage facilities to develop effective odor
controlling additive products. Unfortunately, there is little
information to inform these decisions.
The performance of aerobic bacteria in this study raises the
question of the feasibility of using microbial additives for odor
control. The majority of the bacterial genera used in commercial
additives are obligate aerobes, while most storage lagoons, earthen
basins, and deep pits are anaerobic (despite claims that some
of the outside ponds are aerobic). Deficiency of oxygen in such
manure storage systems will kill bacteria in the supplement culture
shortly after inoculation, and dominant concentrations of aerobic
bacteria will never be achieved. This may explain the limited
success of using microbial additives to control odor.
In a number of studies, researchers have tried to forcibly
increase the concentrations of aerobic bacteria in the manure
by massive inoculation. Ohta and Ikeda13 reported that
odorous compounds in swine manure could be greatly decomposed
by added bacteria (2 g of bacterial culture per 10 g of swine
feces). Another study showed that a dose of about 4.5 kg of bacterial
material was needed for each pig marketed to control odor.14
Obviously, such massive inoculation can be achieved only in the
laboratory, not on farms where the volumes of manure to be treated
are considerable.
Under the experimental design used in this study, the effect
of temporarily raising dissolved oxygen levels in liquid manure
on the growth of aerobic bacteria is negligible. To date, no data
have been reported regarding the minimal concentration of oxygen
that should be maintained in liquid manure to assist aerobic bacterial
growth. It appears that, without aeration, the possibility of
controlling odor by any of the microbial-based manure additives
that have been developed so far is questionable.
Implications
- Adding H2O2 at the concentrations used
in this study can temporarily raise dissolved oxygen concentration
in the manure liquid; however, it contributes little to establishing
an aerated layer that can function as a suitable environment
for growth of aerobic bacteria. This implies that intermittent
aeration, widely recommended to save energy for odor control,
may not be able to maintain aerobic activities for a prolonged
period.
- Data from this study show that adding aerobic microorganisms
alone into swine manure may not necessarily increase the population
of aerobic microbial flora, possibly because they were unable
to compete with indigenous anaerobes for nutrients. Therefore,
it could be inferred that supplemental bacterial culture in an
actual manure handling system might not be able to achieve a
dominant population. The effectiveness of microbial-based manure
additives currently in the market for odor control purpose may
be questionable.
- Due to the complex nature of bacterial involvement in swine
manure odor reduction and production, research regarding how
to control odors using microbial manure additives is still in
its infancy. Appropriate aerobic bacterial genera or species
that can survive the swine manure storage environment and establish
their dominance without aeration to decompose odorous compounds
have yet to be found. More research is needed to study the biology
of aerobes, to search for suitable aerobic bacterial species,
and, if possible, to develop new species using contemporary recombinant
DNA techniques.
- Further research is needed to discover how aeration influences
nutrient consumption by aerobes as well as the growth kinetics
associated with the depletion of nutrients in liquid manure.
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