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Relationship Between Temperature, PH and Population of Selected Microbial Indicators During Anaerobic Digestion of Guinea Grass (Panicum Maximum)
Relationship Between Temperature, PH and Population of Selected Microbial Indicators During Anaerobic Digestion of Guinea Grass (Panicum Maximum)
Relationship Between Temperature, PH and Population of Selected Microbial Indicators During Anaerobic Digestion of Guinea Grass (Panicum Maximum)
1, 14-24
Available online at http://pubs.sciepub.com/ajmr/3/1/3
© Science and Education Publishing
DOI:10.12691/ajmr-3-1-3
Department of Microbiology, Faculty of Biological Science, College of Natural and Applied Sciences, University of Port Harcourt,
P.M.B. 5323 Port Harcourt, Nigeria
*Corresponding author: chukwukaogbonna@gmail.com
Received December 01, 2014; Revised December 31, 2014; Accepted January 08, 2015
Abstract In this study, the relationship between process temperature, process pH and population of selected
microbial indicators during anaerobic digestion of guinea grass (Panicum maximum) at ambient condition was
investigated. A one stage batch-type mesophilic anaerobic digestion system was configured using rumen fluid (RF)
as inoculums (ADRF) and a low solid loading of approximately 7.0% total solid (TS). Physicochemical parameters
such as process temperature (PTMRF), process pHRF and volatile fatty acid (VFARF) were monitored with time.
Selected indicator microbial populations were monitored by standard cultural techniques based on metabolic
capacity and oxygen sensitivity with respect to time. Result showed that average PTMRF increased from 27.5°C to
35.2°C, average process pHRF ranged from 6.5 to 7.9 and VFARF ranged from 1,080.00 mg/L to 4,800.33 mg/L with
time. In terms of metabolic capacity and oxygen sensitivity, the populations of cellulolytic bacteria (CBRF), lactose
and glucose fermenting (acidogenic) bacteria (LFBRF and GFBRF), propionate and ethanol oxidizing (acetogenic)
bacteria (POBRF and EOBRF), acetate oxidizing methanogens (AOMRF), obligate anaerobic bacteria (OABRF) and
total facultative bacteria (FAABRF) increased (about 10-fold) respectively with time. Correlation analysis showed
positive relationships between the process temperature (PTMRF) and the population of selected microbial indicators
with time. However, there were negative relationships between the process pHRF and the population of selected
microbial indicators with time. Furthermore, there were positive relationships between the populations of selected
microbial indicators with time. Rumen fluid significantly (P < 0.05) affected the dynamics of the process
temperature (PTMRF) and process pHRF inside the ADRF system with time respectively. These kinds of relationships
between biotic factors and between biotic and abiotic factors could be used to monitor the state of anaerobic
digestion process with respect to time.
Keywords: relationship, microbial populations, temperature, pH, anaerobic digestion, guinea grass
Cite This Article: Ogbonna. C. B., Berebon. D. P., and Onwuegbu. E. K., “Relationship between
Temperature, Ph and Population of Selected Microbial Indicators during Anaerobic Digestion of Guinea Grass
(Panicum Maximum).” American Journal of Microbiological Research, vol. 3, no. 1 (2015): 14-24. doi:
10.12691/ajmr-3-1-3.
temperature on microbial communities during anaerobic populations that were selected included obligate anaerobic
digestion (AD) process (Dela-Rubia et al., 2002; bacteria (OAB) and total facultative anaerobic bacteria
Bouallagui et al., 2009b; Riau et al., 2010; Kim et al., (FAAB) respectively.
2006; Trzcinski and Stuckey, 2010; Kashyap et al., 2003;
Briski et al., 2007; Fezzani and Cheikh, 2010; Ward et al., 2.3. Statistical Analyses
2008). Generally, the AD process is carried out at
mesophilic temperatures (El-Mashad et al., 2003) because Using SPSS software (version 20), multiple correlation
operation in the mesophilic range is more stable and analysis was performed to determine the relationships
requires a smaller energy expense (Fernandez et al., 2008; among microbial populations, process temperature (PTM)
Ward et al., 2008). Overall, a temperature range between and process pH inside the anaerobic digester with rumen
35 and 37°C is considered suitable for the production of fluid inoculation (ADRF) and the anaerobic digester
methane (Azeem et al., 2011). A number of researchers without rumen fluid inoculation (ADNRF) with respect to
such as Agdag and Sponza (2007),Ward et al., (2008), Lee time. Furthermore, the effect of rumen fluid on the
et al., (2009b), Kim et al., (2003) and Liu et al., (2008) dynamics of process temperature and process pH with
have also reported a range of pH values suitable for time was determined using one – way ANOVA. Finally,
anaerobic digestion but the optimal pH for Chi – square (Goodness of fit) analysis was used to
methanogenesis has been found to be around 7.0 (Huber et determine the stability of the process temperature and
al., 1982; Yang and Okos, 1987). process pH with respect to time.
The objective of this study was to determine the
relationship between process temperature, process pH and
population of selected microbial indicators during
3. Result and Discussion
anaerobic digestion of guinea grass (Panicum maximum)
at ambient condition. 3.1. Microbial and Physicochemical Analysis
The population dynamics of selected indicator
microbial groups observed during 105 days retention time
2. Materials and Methods of anaerobic digestion of Panicum maximum have been
presented in Ogbonna et al., (2014). Generally, these
2.1. Anaerobic Digestion Set-Up microbial populations increased about 10-fold with respect
One-stage anaerobic digestion (AD) systems were to time inside the anaerobic digester with and without
conFigured for batch-type mesophilic reactors as rumen fluid inoculation (ADRF and ADNRF) respectively.
described by Ogbonna et al., (2014). The anaerobic Average ambient temperature (ATM) around the
digestion system with rumen fluid inoculation (ADRF) was anaerobic digesters ranged from 27.5°C to 29.5°C with
the experimental set-up, while the anaerobic digestion time (Figure 1a). In the anaerobic digester with rumen
system without rumen fluid inoculation (ADNRF) was the fluid inoculation (ADRF), average process temperature
control set-up. Anaerobic digestion (AD) of the feed was (PTMRF) increased from 27.5°C (at day 0) to 35.2°C (at
performed (at ambient conditions) inside the mesophilic day 70) and then dropped to 33.3°C at day 105. In the
fermentors with a retention time of 105 days. The feed for anaerobic digester without rumen fluid inoculation
anaerobic digestion was also prepared as described by (ADNRF), average process temperature (PTMNRF) increased
Ogbonna et al., (2014). from 27.5°C (at day 0) to 34.4°C at day 105 (Figure 1a).
This result showed that the anaerobic digester with rumen
fluid inoculation (ADRF) was slightly more heated than the
2.2. Sample Collection and Determination of anaerobic digester without rumen fluid inoculation
Physicochemical/Microbial Parameters (ADNRF) with time. Nevertheless, the temperatures
To monitor the AD process, digester slurry samples observed in both anaerobic digesters (ADRF and ADNRF)
were collected with respect to time. Daily ambient lie within the operational mesophilic temperature
temperature (ATM), process temperature (PTM) and requirement (i.e., 20°C to 45°C) for biogas production
weekly process pH were measured using thermometer during anaerobic digestion of organic matter with the
(SCT-lilliput, Scichem Tech.) and a general purpose pH optimum at around 35°C to 37°C (Schnurer and Jarvis,
meter (SCT-lilliput, Scichem Tech.), respectively. Process 2010; Azeem et al., 2011). Average process pH (pHRF and
temperature and pH were respectively determined by pHNRF) of the feed inside the anaerobic digest er with
dipping the thermometer and the pH meter into the rumen fluid inoculation (ADRF) and anaerobic digester
digester slurry samples immediately after collection in without rumen fluid inoculation (ADNRF) dropped from
beakers. Volatile fatty acid (VFA) was determined using 7.9 and 8.1 (at day 0) to 6.5 and 6.8 (at day 42 and day 56)
the titrimetric method described by Buchauer (1998). and then increased again to 7.4 and 7.3 at day 105
Bacteria populations were monitored by enumerating respectively (Figure 1b). These pH ranges that were
selected indicator groups based on metabolic capacity and observed in both anaerobic digesters (ADRF and ADNRF)
oxygen sensitivity with respect to time (Ogbonna et al., lie within the operational pH requirement (i.e., 6.0 to 8.5)
2014). According to metabolic capability, the microbial for biogas production during anaerobic digestion of
populations that were selected included cellulolytic organic matter with the optimum at around 7.0 (Huber et
bacteria (ACB), lactose fermenting (acidogenic) bacteria al., 1982; Yang and Okos, 1987; Ward et al., 2008; Lee et
(LFB), glucose fermenting (acidogenic) bacteria (GFB), al., 2009b; Kim et al., 2003; Liu et al., 2008).
propionate oxidizing (acetogenic) bacteria (POB), ethanol In the anaerobic digester with rumen fluid inoculation
oxidizing (acetogenic) bacteria (EOB) and acetate (ADRF), the concentration of volatile fatty acid (VFARF)
oxidizing methanogens (AOM). The O2-sensitive increased from 1,080.00 mg/L (at day 0), peaked at
16 American Journal of Microbiological Research
4,800.33 mg/L (at day 42) and then dropped to 1,630.53 day 56) and then dropped to 2,694.70 mg/L at day 105
mg/L at day 105. Likewise, in the anaerobic digester (Figure 2). This result correlates with findinds of Claudia
without rumen fluid inoculation (ADNRF), the et al., (2009) who also reported significant increase (and
concentration of volatile fatty acid (VFANRF) increased decrease) in the concentration of volatile fatty acid during
from 729.34 mg/L (at day 0), peaked at 4,632.04 mg/L (at anaerobic digestion of municipal soild waste (MSW).
Figure 1. (a) Ambient (ATM) and process temperature dynamics (PTMRF and PTMNRF) and (b) Process pH dynamics (pHRF and pHNRF) inside the
anaerobic digester with and without rumen fluid inoculation (ADRF and ADNRF)
Figure 2. Volatile fatty acid dynamics (VFARF and VFANRF) inside the anaerobic digester with and without fluid inoculation (ADRF and ADNRF)
populations of selected indicator microbial groups inside and ADNRF) increased to some degree with time, the
the anaerobic digester with rumen fluid inoculation (ADRF) microbial populations and consequently degree of
and the anaerobic digester without rumen fluid inoculation digestion of the substrate also increased and “vice versa”
(ADNRF) with time (Figure 3). This may be further (Schnurer and Jarvis, 2010; Geradi, 2003; Azeem et al.,
underscored by the near sigmoid pattern exhibited by the 2011). This may have led to a corresponding rise in the
process temperatures (PTMRF and PTMNRF) with time concentration of volatile fatty acids (VFARF and VFANRF)
(Ogbonna et al., 2014). This was also predicted because to some degree and a consequent fall in the process pH
anaerobic degradation of organic matter by microbes leads (see Figure 3 and Figure 4) inside both anaerobic digesters
to the production of little amount of heat energy which (ADRF and ADNRF) with time and “vice versa” (Schnurer
may accumulate with time and thus increase the and Jarvis, 2010; Kim et al., 2003).
temperature of the anaerobic digester environment Chi-square (goodness of fit) analysis suggested that
(Schnurer and Jarvis, 2010). Our result also suggested that there were no significant (p > 0.05) highs or lows in the
the process temperatures (PTMRF and PTMNRF) inside process temperature inside the anaerobic digester with and
both anaerobic digesters (ADRF and ADNRF) were likely to without rumen fluid inoculation (ADRF and ADNRF) with
be more favourable to the other microbial populations than time. However, one-way ANOVA suggested that there
the population of the methanogens (Geradi, 2003; Azeem was a significant difference between the temperature
et al., 2011). This is because the positive relationship dynamics (PTMRF) inside the anaerobic digester with
between the process temperature (PTMRF and PTMNRF) rumen fluid inoculation (ADRF) and the temperature
and the population of the methanogens (AOMRF and dynamics (PTMNRF) inside the anaerobic digester without
AOMNRF) inside the anaerobic digesters (ADRF and ADNRF) rumen fluid inoculation (ADNRF) with time. The process
was the weakest (Figure 3). temperature (PTMRF) of the ADRF system was always
There was a moderate positive relationship(r = 0.67 and higher than the process temperature (PTMNRF) of the
r = 0.60) between the process temperature (PTMRF and ADNRF system during the anaerobic digestion process
PTMNRF) and the concentration of volatile fatty acids (Figure 1a). This may be attributed to the influence of
(VFARF and VFANRF) inside the anaerobic digester with rumen fluid inoculation inside the ADRF system. Rumen
and without rumen fluid inoculation (ADRF and ADNRF) fluid contains various microbial groups that are
with respect to time (Figure 3). An increase in the specialized in anaerobic digestion of cellulosic substrates
concentration of volatile fatty acids to some degree have for biogas production and was added for the purpose of
been associated with a decrease in process pH (see Figure increasing the microbial populations inside the ADRF
4) especially when alkalinity of the AD process was system (Aurora, 1983; Allison and Leek, 1993; Budiyono
significantly consumed by accumulation of these acids et al., 2009). As discussed earlier, there were positive
(Schnurer and Jarvis, 2010; Claudia et al., 2009; Azeem et correlations between the process temperature and the
al., 2011). Figure 3 underscores this phenomenon because microbial populations inside ADRF and ADNRF systems
it showed that there was a strong negative relationship (r = with time (Figure 3). Therefore, the anaerobic digester
0.80 and r = 0.79) between the process temperature (ADRF) with higher microbial populations should exhibit
(PTMRF and PTMNRF) and the process pH (pHRF and higher (or more efficient) degree of digestion of the
pHNRF) inside the anaerobic digester with and without substrate. This should lead to the production of higher
rumen fluid inoculation (ADRF and ADNRF) with time. In amount of heat energy and thus, a higher process
other words, as process temperature inside the anaerobic temperature with time (Schnurer and Jarvis, 2010; Levén
digester with and without rumen fluid inoculation (ADRF et al., 2007).
Figure 3. Relationship between process temperature (PTMRF and PTMNRF) and ambient temperature (ATM), process pH, volatile fatty acid (VFA),
cellulolytic bacteria (ACB), lactose fermenting bacteria (LFB), glucose fermenting bacteria (GFB), propionate oxidizing bacteria (POB), ethanol
oxidizing bacteria (EOB), acetate oxidizing methanogens (AOM), obligate anaerobic bacteria (OAB) and total facultative bacteria (FAAB) inside the
anaerobic digester with and without rumen fluid inoculation (ADRF and ADNRF) with respect to time
18 American Journal of Microbiological Research
metabolites such as CO2 and NH3 may have also lignocellulose content. Because of this, the rate of organic
contributed to the rise in process pH with time (Schnurer acid production during anaerobic biodegradation of the
and Jarvis, 2010; Zinder, 1993; Liu and Whitman, 2008; grass may have been slow or moderate (Susan, 1995).
Garcia et al., 2000). Therefore, the effect on the process pH may not have been
Chi-square analysis suggested that there were no as significant as it would if the substrate was easily
significant highs or lows in the process pH (pHRF and degradable (Schnurer and Jarvis, 2010). An easily
pHNRF) inside the anaerobic digester with and without degradable substrate degrades faster and may rapidly
rumen fluid (ADRF and ADNRF) with time respectively. produce higher concentration of VFAs which may lead to
This has been predicted for most substrates (or feedstock) a significant drop in the process pH and thus, negatively
with low or moderate degradability (Schnurer and Jarvis, affecting the anaerobic digestion process by slowing or
2010). Matured guinea grass (used as substrate in this inhibiting methanogenesis (Ogbonna et al., 2014; Kim et
study) is not readily biodegradable due to its al., 2008; Azeem et al., 2011; Schnurer and Jarvis, 2010).
Figure 4. Relationship between process pH (pHRF and pHNRF) and volatile fatty acid (VFA), cellulolytic bacteria (ACB), lactose fermenting bacteria
(LFB), glucose fermenting bacteria (GFB), propionate oxidizing bacteria (POB), ethanol oxidizing bacteria (EOB), acetate oxidizing methanogens
(AOM), obligate anaerobic bacteria (OAB) and total facultative bacteria (FAAB) inside the anaerobic digester with and without rumen fluid inoculation
(ADRF and ADNRF) with respect to time
Furthermore, it is highly likely that oxygen was present Likewise, the population of acetate oxidizing
inside the biodigesters (ADRF and ADNRF) because it may methanogens (AOMRF and AOMNRF) was very strongly
not have been possible to evacuate all the air (oxygen) and positively related to (or associated with) the
from the 500L-capacity anaerobic digestion tanks before population of obligate anaerobic (OABRF and ADNRF) than
loading. with the population of facultative aerobes and anaerobes
The population of propionate oxidizing acetogens (FAABRF and FAABNRF) inside the anaerobic digesters
(POBRF and POBNRF) was very strongly and positively with and without rumen fluid inoculation (ADRF and
related to (or associated with) the populations of ethanol ADNRF) respectively with time (Figure 7b).
oxidizing acetogens (EOBRF and EOBNRF) and acetate Methanogenesis can only occur in an obligate anaerobic
oxidizing methanogens (AOMRF and AOMNRF) with time environment (Schnurer and Jarvis, 2010). This may
inside both AD systems (ADRF and ADNRF), respectively explain (to some degree) why their population was
(Figure 6b). The population of these acetogens (POBRF strongly positively related to the population of obligate
and EOBRF and POBNRF and EOBNRF) may have increased anaerobic bacteria (Figure 7b). Their population may have
or decreased together because they may have been also increased together with the population of total
positively influenced by the population of the acidogens facultative bacteria because they may have needed the
(LFBRF and GFBRF and LFBNRF and GFBNRF) during population of the facultative groups to create the anaerobic
acidogenesis as indicated in Figure 5b and Figure 6a environment that enabled them increased with respect to
which showed that there were very strong positive time (Ogbonna et al., 2014; Schnurer and Jarvis, 2010).
relationships between the acetogens and the acidogens Facultative bacteria are capable of consuming any oxygen
inside the anaerobic digester with rumen fluid inoculum which may have been present inside the anaerobic
(ADRF) and the anaerobic digester without rumen fluid digesters (ADRF and ADNRF) at the start of the AD process
inoculum (ADNRF) with time(Labat and Garcia, 1986; thus, eliminating all the oxygen inside the anaerobic
Claudia et al., 2009; Schnurer and Jarvis, 2010; Yassar, digesters with time (Schnurer and Jarvis, 2010). This
2011). The population of propionate oxidizing acetogens would have promoted the growth of obligate (or strict)
(POBRF and POBNRF) may have correlated strongly and anaerobic bacteria population such as the methanogens to
positively with the population of acetate oxidizing some degree with time (Schnurer and Jarvis, 2010).
methanogens (AOMRF and AOMNRF) inside the anaerobic This fact may be further underscored by Figure 7c
digesters (ADRF and ADNRF) as shown in Figure 6b which showed that as the population of total facultative
because propionate oxidation usually produces CO2, H2 bacteria (FAABRF and FAABNRF) increased, the
and acetic acid which is the substrate for acetotrophic population of obligate anaerobic bacteria (OABRF and
methanogenesis (Schnurer and Jarvis, 2010; Yassar, 2011; OABNRF) also increased to some degree inside the
Chanakya and Sreesha, 2012). anaerobic digesters (ADRF and ADNRF) with time and
In terms of O2-sensitivity, the population of propionate “vice versa”. In fact some authors like Labat and Garcia
oxidizing acetogens (POBRF and POBNRF) was very (1986) and Schnurer and Jarvis (2010) have suggested that
strongly and positively related to (or associated with) the an AD process may not be seriously negatively affected if
population of obligate anaerobic bacteria (OABRF and there was an accidental introduction of air (or oxygen)
OABNRF) than with the population of total facultative inside the bio-digesters because the facultative bacteria
bacteria (FAABRF and FAABNRF) inside the anaerobic groups may quickly consume any oxygen that was
digester with and without rumen fluid inoculation (ADRF introduced before it significantly harms the population of
and ADNRF) respectively with time (Figure 6b). This may obligate (or strict) anaerobes. Of course there may have
be because acetogenesis (via propionate oxidation) may been other factors (such as the process temperature and
only proceed under obligate anaerobic condition (Schnurer pH) which may have also affected the populations of these
and Jarvis, 2010). Furthermore, if propionate oxidation microbial groups in such a way that would have made
(for acetogenesis) may only occur under anaerobic them to increase or decrease together to some degree with
conditions, then the population of facultative bacteria time (Figure 5 to Figure 7).
(FAABRF and FAABNRF) which are capable of consuming It is very important to note that an increase in the
any oxygen that may have been present inside the population of methanogens may also positively affect the
biodigesters (ADRF and ADNRF) at the beginning of the populations of hydrolytic bacteria and acid (and hydrogen)
AD process must increase to some degree with time forming (i.e., acidogenic and acetogenic) bacteria groups
(Claudia et al., 2009; Labat and Garcia, 1986; Schnurer to some degree with time (Figure 5 to Figure 7). This is
and Jarvis, 2010). Therefore, as the concentration of the because when the respective populations of hydrolytic,
oxygen inside the digester reduced with time (because of acidogenic and acetogenic bacteria increased with time,
an increase in the population of total facultative bacteria), hydrolysis, acidogenesis and acetogenesis also increased
the population of obligate anaerobic bacteria like the to some degree (Ogbonna et al., 2014). Some of the
propionate oxidizing acetogens (POB) may also begin to products of acidogenesis and acetogenesis include
increase and become established with time (Schnurer and alcohols, carbon dioxide, hydrogen, organic acids (or
Jarvis, 2010). This may have been one of the reasons why acetic acids in the case of acetogenesis), etc. Therefore, as
the population of propionate oxidizing acetogens (POBRF acidogenesis and acetogenesis increase with respect to
and POBNRF) increased together with the population of time, the concentrations of these by-products may also
obligate anaerobic bacteria (OABRF and OABNRF) and total increase with time. The accumulation of hydrogen and
facultative bacteria(FAABRF and FAABNRF) inside the organic acids (and/or acetic acid) to some degree may be
anaerobic digesters (ADRF and ADNRF) to some degree inhibitive to the respective populations of the hydrolytic,
with time (Figure 6b). acidogenic, acetogenic and even the methanogenic groups
with time (Hickey et al., 1987; Labib et al., 1992). Thus,
American Journal of Microbiological Research 21
sufficient system assimilation capacity for hydrogen and products below inhibitory levels (Parkin and Owen, 1986;
acetate removal must be maintained to ensure continuous Yassar, 2011; Schnurer and Jarvis, 2010). This metabolic
acid production (Yassar, 2011). The methanogens usually relationship between the population of methanogens and
convert the hydrogen (with carbon dioxide) and acetate to the population of other bacteria groups monitored in this
biogas. As their populations increase with time, this study may be one of the reasons why their populations
conversion rate may also increase to some degree and thus, increased together to some degree with time and “vice
continuously keeping the concentration of these by- versa” (Figure 5 to Figure 7).
Figure 5. (a) Relationship between the population of cellulolytic bacteria (ACBRF& ACBNRF) and the populations of lactose fermenting acidogens
(LFBRF& LFBNRF), glucose fermenting acidogens (GFBRF& GFBNRF), propionate oxidizing acetogens (POBRF& POBNRF), ethanol oxidizing acetogens
(EOBRF& EOBNRF), acetate oxidizing methanogens (AOMRF& AOMNRF), obligate anaerobic bacteria (OABRF& OABNRF) and total facultative bacteria
(FAABRF& FAABNRF) inside the anaerobic digester with and without rumen fluid inoculation (ADRF& ADNRF) respectively with time at ambient
condition. (b) Relationship between the population of lactose fermenting acidogens (LFBRF& LFBNRF) and the populations of glucose fermenting
acidogens (GFBRF& GFBNRF), propionate oxidizing acetogens (POBRF& POBNRF), ethanol oxidizing acetogens (EOBRF& EOBNRF), acetate oxidizing
methanogens (AOMRF& AOMNRF), obligate anaerobic bacteria (OABRF& OABNRF) and total facultative bacteria (FAABRF& FAABNRF) inside the
anaerobic digester with and without rumen fluid inoculation (ADRF& ADNRF) respectively with time at ambient condition
Figure 6. (a) Relationship between the population of glucose fermenting acidogens (GFBRF& GFBNRF) and the populations ofpropionate oxidizing
acetogens (POBRF& POBNRF), ethanol oxidizing acetogens (EOBRF& EOBNRF), acetate oxidizing methanogens (AOMRF& AOMNRF), obligate anaerobic
bacteria (OABRF& OABNRF) and total facultative bacteria (FAABRF& FAABNRF) inside the anaerobic digester with and without rumen fluid inoculation
(ADRF& ADNRF) respectively with time at ambient condition.(b) Relationship between the population of propionate oxidizing acetogens (POBRF&
POBNRF) and the populations of ethanol oxidizing acetogens (EOBRF& EOBNRF), acetate oxidizing methanogens (AOMRF& AOMNRF), obligate
anaerobic bacteria (OABRF& OABNRF) and total facultative bacteria (FAABRF& FAABNRF) inside the anaerobic digester with and without rumen fluid
inoculation (ADRF& ADNRF) respectively with time at ambient condition
22 American Journal of Microbiological Research
Figure 7. (a) Relationship between the population of ethanol oxidizing acetogens (EOBRF& EOBNRF) and the populations of acetate oxidizing
methanogens (AOMRF& AOMNRF), obligate anaerobic bacteria (OABRF& OABNRF) and total facultative bacteria (FAABRF& FAABNRF) inside the
anaerobic digester with and without rumen fluid inoculation (ADRF& ADNRF) with time at ambient condition.(b) Relationship between the population of
acetate oxidizing methanogens (AOMRF& AOMNRF) and the populations of obligate anaerobic bacteria (OABRF& OABNRF) and total facultative bacteria
(FAABRF& FAABNRF) inside the anaerobic digester with and without rumen fluid inoculation (ADRF& ADNRF) respectively with time at ambient
condition.(c) Relationship between the populations of obligate anaerobic bacteria (OABRF& OABNRF) and the populations of total facultative anaerobic
bacteria (FAABRF& FAABNRF) inside the anaerobic digester with and without rumen fluid inoculation (ADRF& ADNRF) respectively with time at
ambient condition
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