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Agriculture wastes conversion for biofertilizer production using beneficial microorganisms for sustainable agriculture applications


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Aims: The emphasis of this study is to generate new valuable bioproducts from non-toxic cleaning waste for environmental healing technology. Methodology and Results: Comparisons between different types of biofertilizer formulations and the field trial effectiveness were done. Results indicated that biofertilizer C contained the highest N value (1.8%) when compared with biofertilizers B and A, which only contained 1.7% and 1.4%, respectively. Biofertilizer A showed significant difference in the total count of yeast, mould, ammonia oxidizing bacteria and nitrate oxidizing bacteria compared to biofertilizer B and C. Meanwhile, biofertilizer C was found to be significantly different from others in Lactobacillus sp. and nitrogen-fixing bacteria count. Photosynthetic total count and Actinomycetes sp. were not noticed in all formulations tested. Conclusion, significance and impact of study: The findings of this study suggest that biofertilizer A is suitable to be used as a promotional biofertilizer in flower and fruit production, biofertilizer B can be used for a leafy crop, while biofertilizer C is good for the growth of roots and stem of plants.
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Malaysian Journal of Microbiology, Vol 9(1) 2013, pp. 60-67
Malaysian Journal of Microbiology
Published by Malaysian Society for Microbiology
(In since 2011)
60 ISSN (print): 1823-8262, ISSN (online): 2231-7538
Agriculture wastes conversion for biofertilizer production using beneficial
microorganisms for sustainable agriculture applications
Siti Zulaiha Hanapi1, Hassan M. Awad1,2*, Sheikh Imranudin Sheikh Ali1, Siti Hajar Mat Sarip1, Mohamad Roji
Sarmidi1, Ramlan Aziz1
1Institute of Bioproduct Development, Universiti Teknologi Malaysia (UTM), 81310, UTM, Johor, Malaysia.
2Chemistry of Natural and Microbial Products Department, National Research Centre (NRC), Dokki, Cairo, Egypt.
Received 27 April 2012; Received revised form 15 August 2012; Acceptance 23 August 2012
Aims: The emphasis of this study is to generate new valuable bioproducts from non-toxic cleaning waste for
environmental healing technology.
Methodology and Results: Comparisons between different types of biofertilizer formulations and the field trial
effectiveness were done. Results indicated that biofertilizer C contained the highest N value (1.8%) when compared with
biofertilizers B and A, which only contained 1.7% and 1.4%, respectively. Biofertilizer A showed significant difference in
the total count of yeast, mould, ammonia oxidizing bacteria and nitrate oxidizing bacteria compared to biofertilizer B and
C. Meanwhile, biofertilizer C was found to be significantly different from others in Lactobacillus sp. and nitrogen-fixing
bacteria count. Photosynthetic total count and Actinomycetes sp. were not noticed in all formulations tested.
Conclusion, significance and impact of study: The findings of this study suggest that biofertilizer A is suitable to be
used as a promotional biofertilizer in flower and fruit production, biofertilizer B can be used for a leafy crop, while
biofertilizer C is good for the growth of roots and stem of plants.
Key words: Biofertilizer, Beneficial microorganisms, Microbiological analysis, Chemical analysis, Field trials.
Currently, there are more than 4.3 million hectares of oil
palm plantation in Malaysia, which is equivalent to
approximately 67 percent of total agricultural land in the
country (DOA, 2010). Malaysia had generated in excess
of 15,000 tons of solid waste per day in the form of
biomass that consists of forest and mill residues, wood
wastes, agricultural wastes, and municipal waste.
Agricultural wastes from agro-based industries are also on
the increase. State of Johore, Selangor, and Perak
collectively accounted for 65.7% of the overall identified
pollution sources in the agro-based and manufacturing
sector (DOE, 2001).
These biomasses bear a huge potential to be
applied as an alternative and beneficial invention for
various applications such as in sustainable agriculture and
etc. because they are high in moisture, organic matter and
other minerals. Thus, they can actually be reproduced into
more useful and value-added products with safety and
profitability. Recently, many countries have made an effort
to recycle 15 50% of the wastes they generated (Diza et
al., 1993). Weeds, stalks, stems, fallen leaves, pruning,
and dead branches (Boraste et al., 2009); animal manure
(Bheki et al., 2010); vermicompost (Warman and
AngLopez, 2010); and agriculture wastes such as
cornstalks, sugarcane bagasse, drops and culls from fruits
and vegetables (Weber et al., 2007) have long been used
as the soil conditioner to fertilize the soil and plant with the
cooperation of beneficial microbes.
Biofertilizers are environmental friendly fertilizers that
not only prevent damages to natural sources but help, to
some extent, in cleaning the nature from precipitated
chemical fertilizers (Food and Agricultural Organization,
2008). The use of organic matter such as sawdust, rice
bran, rice husk and shredded paper in producing
biofertilizer is economical. They also act as the carrier
material for nutrient and microorganisms.
The role of plant nutrients in crop production is well-
established and 16 essential plant nutrients have to be
available to the crops in required quantities to achieve the
yield target. Many studies have also emphasized on the
importance of N, P and K in enhancing the natural ability
of plants to resist stress from drought and cold, pests and
diseases (Debosz et al., 2002; Tsai et al., 2007). Essential
plant nutrients such as N, P, K, Ca, Mg and S are called
macronutrients, while Fe, Zn, Cu, Mo, Mn, B and Cl are
called micronutrients. It is necessary to assess the
capacity of a soil to supply the lacking amounts of needed
plant nutrients (total crop requirement-soil supply) (Food
*Corresponding author
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61 ISSN (print): 1823-8262, ISSN (online): 2231-7538
and Agricultural Organization 2008). This is also important
to produce a good biofertilizer formulation and to supply
nutrients that can improve soil health and fertility of plants.
Several authors such as Debosz et al. (2002); and
Chen et al. (2007) are concentrating on the potential
usage of nitrogen from animal manures. Nonetheless, the
effort to find another source instead of animal manure
needs further study. Granite powder has also been
studied as a good source of slow-release K fertilizer
(Chen et al., 2007). Generally, the addition of nitrogen to
high C: N ratio residues is capable of accelerating the
microorganism activity during the fermentation process
(Saratchandran et al., 2001).
The number of microorganisms and the level of
macro- and micronutrient obviously affect the growth of
plants (Coroneos et al., 1995). One of the benefits of
fertilizers is that they contribute to the availability of
microorganism population (Marrs, 1993). Having a higher
initial count of appropriate microbes in ready biofertilizer
right after the fermentation is essential. One of the ways
to increase the number of selected microorganisms is by
using the concept of an effective microorganism (EM) as
introduced by Higa and Wididana (1991).
Field experiments are needed to determine the
nutrient availability and efficacy of most organic fertilizers.
Such an experiment is important because the nutrient
content of organic fertilizers varies widely (Parr et al.,
1998). The quality is directly governed by the number of
selected microorganisms in the active form per gram and
their capability to promote plant growth and soil fertility.
The aim of the work is to investigate the
conversation and different formulation of the agriculture
wastes for biofertilizer production by beneficial
Raw materials
The raw materials used for the bioorganic fertilizer
production were obtained from a local manufacturer in
Kulai, Johor. The waste was in the form of granules with
chemical characteristics as shown in Table 1.
Table 1: Biofertilizer type combination
Ingredients (%)
Type of biofertilizers
1- Burned soil
2-Nitrogen source
3- Saw dust
4- Burned rice husk.
5- EM
6-Gibberelic Acid
10 ppm
10 ppm
10 ppm
(-) = without addition of burn rice husk, EM= Effective
Biofertilizer preparation
Generally, the ingredients of each biofertilizers differed in
or without following the addition of burned soil, nitrogen
source meal, saw dust and burned rice husk. The
formulation of these biofertilizers types is shown in Table
2. Each of biofertilizers was inoculated with 3% of
effective microorganisms (EM) before the fermentation
Table 2: Microorganisms and specific media used for
isolation and identification.
Lactobacillus sp.
Yeast and Mold
(Leuschner et
al., 2003)
N2 fixing bacteria
(Ashby, 1907,
Harunor et al.,
(Prasertsan et
al., 1993)
(Bhuiya and
Walker, 1977)
(Awad et al.,
2009, Shirling
and Gottlieb,
CGYE = Glucose Yeast Extract Agar, AOB: Ammonia-
oxidizing broth and Nitrogen-oxidizing broth (NOB)
Fermentation and temperature monitoring
The starting fermentation was performed for biofertilizer A,
B and C in different proportions of ingredients, which
differed in or without the addition of burned soil, nitrogen
source meal, saw dust and burned rice husk. The
formulations of these biofertilizer types are shown in Table
1. All biofertilizers were inoculated with 3% of effective
microorganisms (EM) before the fermentation proceeds.
The substrate temperature was measured daily from Day
1 until Day 7 at a depth of 50 cm with a thermometer.
Isolation and enumeration of microorganisms
The total microbial population of the sample was
determined using the following methods and the specific
media for each strain were according to the literatures as
shown in Table 2. The media composition and the
preparation methods which were used in this study are
also listed in Table 2.
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Isolation and enumeration of Lactobacillus sp. by
dilution plate technique
Man Rogosa Sharpe (MRS) medium was used to
encourage the growth of lactic acid bacteria such as
Lactobacilli, Enterococci and Pediococci. Selection of
Lactobacilli was carried out using the pH selection method
(pH 5.5 to 6.2) with Enterocci and Pediococci growing
best in this range. For this purpose, acidified MRS agar
medium (Merk, Darmastdt, Germany) was used (Institute,
Standard method for determining number of yeasts
and molds
A pour plate method following (International Standards
Organization) ISO 7954 (ISO, 1987) using
chloramphenicol glucose yeast extract (CGYE) was used.
The CGYE agar medium contained (g/L): dextrose, 20;
yeast extract, 5.0; chloramphenicol, 0.1; and agar, 15.0.
The medium was adjusted to pH 6.6. ± 0.2 prior
autoclaving. Each substrate of 10 g was suspended in 90
ml sterile saline, shaken thoroughly and 0.1 mL of each
inoculum was inoculated with AMRS and incubated at
25 C ± 1 C for 48 hours. At the same time, the CGYE
medium was incubated at 25 C ± 1 C for five days
(Leuschner et al., 2003).
Determination of nitrogen-fixing bacteria by the
spread plate method
This method is based on the ability of nitrogen-fixing
bacteria to grow in a nitrogen-free medium. The total N2-
Fixing bacteria were counted using Ashby’s medium after
an incubation period (Ashby, 1907). Ashby’s medium
composed of (g/L): mannitol, 20; K2HPO4, 0.2;
MgSO4.7H2O, 0.2; NaCl, 0.2; K2SO3, 0.1; CaCO3, 5.0; and
agar, 15.0. One gram of sample was transferred into 50
ml Ashby’s medium and incubated at 30 C ± 1 C for 2-5
days. Then, the broth surface was examined and using
serial dilution, it was streaked to nitrogen-free medium
agar for enumeration. The colonies that grew on the
medium appeared as white, off white, gray and gray to
white. They were circular, flat, raised, serrate in elevation,
and small and pinpoint in size (Ashby, 1907; Harunor et
al., 2008).
Isolation and enumeration of photosynthetic bacteria
The current method is based on the ability of
photosynthetic bacteria to assimilate CO2 and use light as
their energy source during incubation under bright and
dark conditions. Determination of photosynthetic bacteria
was carried out by incubating 5 g of sample in succinate
broth. It consisted of three media as follows: Mineral
Salts-Succinate Broth medium (1) made up with (g/L):
K2HPO4, 0.33; MgSO4.7H2O, 0.33; NaCl, 0.33; NH4Cl,
0.50; CaCl2.2H2O, 0.05; sodium succinate, 1.0; yeast
extract, 0.02; and agar, 15 g, at pH 6.8-7.2 using 5M
Trace element's medium (2) with (mg/L):
ZnSO4.7H2O 10; MnCl2.7H2O, 3; H3BO3, 30; CoCl2.6H2O,
20; CuCl2.2H2O, 1; NiCl2.6H2O, 2; and Na2MoO4, 3 mg,
the solution was adjusted to pH 3-4 using 5M HCl.
Medium (3) composed of 0.02% FeSO4.7H2O. The
isolates were incubated for four to seven days at 30 °C ±
1 until the appearance of red pigment (bloom) which
indicated the presence of photosynthetic microorganisms.
Positive control (Rhodopseudomonas palustris NRRL B-
4267) was incubated under the same conditions
(Prasertsan et al., 1993).
Isolation and detecting of nitrifying bacteria (Multiple
Five Tube method)
Multiple Five Tube method (Bhuiya and Walker, 1977)
was used in detecting nitrifying bacteria using ammonia-
oxidizing broth (AOB) and nitrogen-oxidizing broth (NOB).
These media were composed of the following
constituents: the AOB-medium (g/L): MgSO4.7H2O, 0.04;
(NH4)2SO4, 0.50; KH2PO4, 0.20; CaCl2.2H2O, 0.04; and
phenol red, 0.001 and the NOB medium (g/L): KNO3,
0.30; MgSO4.7H2O, 0.1875; KHCO3, 1.5; K2HPO4, 0.5;
KH2PO4, 0.5; NaCl, 0.1875; CaCl2.2H2O, 0.0125; and
FeSO4.7H2O, 0.01. Each set of AOB and NOB tubes was
inoculated with 1 ml of sample suspended and incubated
at 25-30 C for 23-28 days in case of (AOB) and for 23
days or more for (NOB).
After the end of incubation, one drop of sulfanilic
acid and N, N-dimethyl-1-naphthylamine were added into
AOB and NOB media in tubes. Red color indicates the
presence of active AOB while an absence of any color
changes is a positive result for NOB. A confirmation test
for nitrite/nitrate was carried out by added one drop of
diphenylamine to a drop of sample on a clean spot plate.
Positive tubes or wells were identified by the development
of a blue color and the absence of color is scored
negatively. All results were computed into the MPN table.
Isolation and enumeration of actinomycete's colonies
by dilution plate technique
Isolation and enumeration of actinomycetes colonies were
performed by a soil dilution plate technique using two
different media: the first medium is an actinomycete's
isolation agar medium (Difco, NJ, USA) at pH 7.0. The
second medium, Streptomyces medium, consisted of
(g/L): glucose, 5; L-glutamic, 4; KH2PO4, 1.0;
MgSO4.7H2O, 0.7; NaCl, 1; FeSO4.7H2O, 3 mg; and agar,
25. This medium was supplemented with 50 µg/L
cycloheximide (Sigma-Aldrich Corp., MO, USA) (Awad et
al., 2009). The isolates were incubated at 28 ± 0.5 ºC for
7-10 days. The results obtained were expressed as the
colony forming unit (CFU).
Actinomycete colonies were characterized
morphologically and physiologically following the
directions given by the International Streptomyces project
(ISP) (Shirling and Gottlieb, 1966).
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Chemical analysis
The total nitrogen, phosphorus and potassium (NPK)
content of samples were analyzed according to the
following methods. Nitrogen was determined by the acid
combustion elemental analysis method using the macro
kjeldahl system (Gerhadt, German) (Tandon 1993). The
phosphorus, potassium and other micronutrients were
digested using the acid digestion method and analyzed
spectrophotometrically (Spectroquant NOVA 60, Merck,
USA) using EPA method 3050B (Tandon, 1993). Moisture
content of the samples was determined using the moisture
analyzer (MX-50, A&D Company Ltd, Japan) to a constant
weight. The pH value was measured in a 5-fold dilution of
distilled water equilibrated with the sample for an hour
with a pH meter (Delta 320, Mettler Toledo, Germany).
Ash content in a dried sample was determined at 550°C
for 24 hours using (CWF 110, Carbolite, England). C %
was determined by APHA 5310 B method according to
APHA (2005). N % was determined by APHA 4500-N org
B (Mod) according to APHA (2005).
Efficacy of biofertilizers
Efficacies for the biofertilizers were carried out for six
months. Soil for this experiment was natural silt loam with
a pH of 7.3 and moisture content of 14.4%. A local variety
of ladyfingers was used as test plant. The experimental
design was the completely randomized design with three
replicates. Four different soil beds at a size of 15 feet x 4
feet (LxW) were prepared for each treatment as followed:
Plot 1 (Biofertilizer A), Plot 2 (Biofertilizer B), Plot 3
(Biofertilizer C) and Plot 4 Controls (without Biofertilizer).
Before the planting started, each plot was treated by
spreading a total of 200 g of the respective biofertilizers
into the loose soil. The plots were then watered regularly
for 14 days. After the soil treatment, a seeding inoculation
was performed. The seeds of ladyfingers were soaked in
water for 10 minutes, and good qualities seeds were taken
out for seeding; good seeds will sink underneath the water
and vice versa. For seeding, about 3-4 seeds were
pressed 1-2 cm into the soil bed. One tablespoon of the
respective biofertilizer (about 14 g) was then dispersed on
the soil surface surrounding the planted seed and water
was applied. This was done weekly and continued to twice
a month until the day of harvesting. During harvesting, the
plants were carefully uprooted from each plot and the
plant height, length of roots, diameter of leaves, fruits and
fruit weight were recorded.
Statistical Analysis
All experiments were carried out in triplicate. All data are
reported as means ± SD (standard deviation). The results
were analyzed statistically by a one-way ANOVA using
(SPSS Inc. 2006) (Levesque, 2007) with the results:
microbiological, chemical and field trial with biofertilizers
as the main factors. The mean of each measurement
parameter was separated statistically using Tukey’s and
Dunnet’s multiple range tests with the plants grown
without the aid of biofertilizer set as control. Significance
was defined as P<0.05, unless otherwise indicated.
Fermentation and temperature monitoring
The temperature was recorded from the first day of
production until the eighth day of biofertilizer fermentation.
The temperature increased rapidly during fermentation,
peaking at 71 C on Day 4 and then decreased gradually
until Day 8 when the biofertilizers achieved maturity. The
result is as shown in Figure 1.
0 2 4 6 8 10
Biofertilizer A
Biofertilizer B
Biofertilizer C
Temperature oC
Time [day]
Figure 1: Temperature profile of different types of
biofertilizers during fermentation period
Isolation and enumeration of microorganisms
During fermentation, the results of total microbial
population in different biofertilizers of A, B and C
measured using the CFU/g biofertilizer are as shown in
Table 3.
In all biofertilizers tested, Lactobacillus sp. was the
major population while the nitrogen-fixing bacteria were
the minority population. The results showed that the total
count of Lactobacillus sp was 3.3 x 105 CFU g/L, 4.9 x 105
CFU g/L, and 2.3 x 104 CFU g/L in biofertilizer A, B and C,
respectively. Meanwhile, biofertilizer B and C showed the
greatest growth of yeast total count, which were 3.0 x 107
CFU g/L and 3.5 x 107 CFU g/L and the lowest total count
of yeast, 2.4 x 105 CFU g/L, was recorded in biofertilizer
On the other hand, the population of nitrifying
bacteria in terms of biofertilizer A was significantly
different from biofertilizer B and C and ranged between
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3.20 x 102 to 3.60 x 102 CFU g/L for AOB and NOB,
respectively. Concurrently, the results showed that no
significant difference had been observed for all types of
biofertilizer formulation in the case of total nitrogen-fixing
bacteria count. Also, the results indicated that no growth
was detected in both photosynthetic bacteria and
Actinomycetes in all biofertilizers examined.
Chemical analysis
The results in Table 4 show that the pH value of all
formulated biofertilizers was slightly alkaline and ranged
between 8.2 - 8.5 due to the degradation of nitrogen-
containing materials to soluble organic nitrogen. The
moisture content for each formulated biofertilizer differed
from one another based on the formulation composition
and ranged from 16.60 to 22.30%.
In addition, the ash content of all biofertilizers ranged
between 0.42-0.61 percent, depending on the formulation
composition. The stability of ash content can be used as
the parameter of compost maturity. The total organic
carbon and nitrogen content was determined and the
results showed that biofertilizer B possessed the highest
content (19.2) followed by biofertilizer C (12.0) and lastly
biofertilizer A (6.0). The results also indicated that the
macro- and micronutrients content of biofertilizer B was
the highest followed by biofertilizer A and C.
Efficacy of biofertilizers
The field trail results in Table 5 show that the plants
treated with biofertilizer C grew more vigorously than the
other plants grown in different treatments. On the other
hand, the plants treated with biofertilizer A and contained
burned rice husk exhibited the largest fruit diameter as
well as the fruit weight. The plants treated with biofertilizer
B grew better than the plants treated with biofertilizer A
and C for the diameter of leaves recorded. Overall, it was
observed that the plants treated with biofertilizer A, B and
C were growing well and had better yields than the plants
which grew without any treatment (control).
During biofertilizer preparation, microbes decompose the
organic matter and release the fermentation heat (Yang,
2003). Temperature changes have to be recorded during
fermentation to monitor the activity of the microbes. The
results showed that the temperature increased from 41 C
to 71 C at Day 4 and gradually decreased to 50 C at
Day 8, indicating that the biofertilizers had achieved
maturity. The increasing temperature during fermentation
occurs due to the active microbial growth. The
temperature changing patterns in this work are similar to
the commercial composting process made by Pai et al.
Proper fermentation also will effectively destroy
pathogens and weeds through the metabolic heat
generated by the microorganisms (Yang 2000; Nakasaki
et al., 1996). These results are in accordance to those
obtained by Tsai et al. (2007) who found that the
inoculation of appropriate microbes during fermentation
will shorten the period of maturity and thus improve the
quality of biofertilizers. Nevertheless, there is a lack of
reported studies in the number of actinomycetes and
photosynthetic bacteria present in biofertilizer samples.
Many of the literatures only showed that the isolation of
these microorganisms from environmental samples such
as oil was noticeable (Fuentes et al., 2010). In order to
prepare a multi-functional biofertilizer, thermo-tolerant
phosphate-solubilizing microbes, including bacteria,
actinomycetes and fungi have to be isolated from different
compost plants and biofertilizers (Chang and Yang, 2009).
Biofertilizers of three different formulations were
analyzed for their microbiological, chemical and physical
components. The presence of certain microorganisms and
the nutrient mineralization are favorable to support plant
growth and yields (Parthasarathi and Ranganathan,
1999). Another study was done by Edward and Fletcher
(1988); they stated that the increase of microbial
populations increases the performance of biofertilizer
microbiologically, chemically, and physically.
The large number of Lactobacillus sp. and yeast
isolated from the final product of biofertilizer indicated the
success of fermentation. The total number of Lactobacillus
sp. and yeast were in between 5.00 x 105-8 CFU g/L.
These results are consistent with the microbial analysis
results from liquid biofertilizers produced by several
authors such as Ngampinol and Kunathigan (2008); and
Department of Agriculture (2004).
Microorganism Total Count
Lactobacillus sp.
3.30 x 105 a
4.90 x 105 a
2.30 x 104 b
2.40 x 105 b
3.00 x 107 a
3.50 x 107 a
>1.60 x 103 b
3.3 x 102 a
3.5 x 102 a
6.20 x 102 b
3.2 x 102 a
3.6 x 102 a
Photosynthetic bacteria
Nitrogen-fixing bacteria
4.5 x 101 b
5.2 x 101 b
1.4 x 101 b
Table 3: The populations of Effective Microorganisms (EM) examined in Biofertilizer A, B and C in CFU/g.
Significance difference (P<0.05) NG= no growth, AOB=Ammonia-oxidizing bacteria, NOB= Nitrite-oxidizing bacteria
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Table 4: Macro- and micronutrients and other chemical analysis for Biofertilizer A, B and C.
Percentage (%)
Concentration (mg/L)
Significance difference (P<0.05)
Table 5: Physical analysis during field trial for ladyfingers fertilized with Biofertilizer A, B and C.
Physical analysis
Biofertilizer A
Biofertilizer B
Biofertilizer C
Plant height (cm)
Root length (cm)
Leaves diameter (cm)
Fruits diameter (cm)
Fruits weigh (g)
Significance difference (P<0.05)
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The nitrifying and nitrogen-fixing bacteria total counts
were low (< 1.00x 103 CFU/g) in all formulations prepared.
The chemical analysis results derived from the
composting process showed that the inoculated
biofertilizers with tested microbes had a significantly
higher temperature, ash content, pH, total nitrogen, and
soluble phosphorus content. Adding these microbes can
shorten the period of maturity, improve the quality,
increase the soluble phosphorus content, and enhance
the populations of phosphate-solubilizing and proteolytic
bacteria in the biofertilizers (Chang and Yang, 2009). The
pH value of each biofertilizer in these experiments was in
the range of 8.20 - 8.50 which is slightly alkaline than
other solid biofertilizers as reported by many authors such
as Debosz et al. (2002); and Tsai et al. (2007). The
slightly alkaline pH is beneficial because this will
contribute to the neutralization of acidic agricultural soil
(Fageria and Baligar, 2001).
The moisture content of compost decreased during
the incubation period because the inoculation of the
biofertilizer with EM increased the temperature and
decreased the moisture content of biofertilizer. The same
phenomena has also observed in open field composting
(Pai et al., 2003).
Total ash content in the biofertilizer samples was
determined. The stability of ash content can be used as a
parameter of compost maturity. The ash content
significantly increased during preparation since the
organic materials were decomposed to form the metabolic
gases (Yang, 2003; Chang and Yang, 2009). Total
organic carbon content (C:N ratio) decreased from 19.2 in
case of biofertilizer B to 12.0 and 6.0 for biofertilizer A and
C, respectively. Total organic carbon content significantly
decreased during composting due to the degradation of
organic matter. These results are in accordance with
those results obtained by Chang and Yang (2009). It has
been noted that the properties of the initial material, in
particular is affecting the C:N ratio of the biofertilizers.
Higher C:N ratio (>30%) contributed longer composting
process to occur (Tiquiaa and Tam, 2000). Other factors
such as aeration condition, moisture content, and
temperature are also affected by the degree of N loss.
The field trial study was conducted to monitor and
observe the differences in the biofertilizers’ effectiveness
regarding their abilities to encourage plant growth.
Significant reduction of all physical properties in the case
of non-treatment plant can be explained by lack of or low
soil fertility. The plant height, length of roots, diameter of
leaves and fruits as well as the ripe fruit weight increased
when the plants were treated with biofertilizers.
It has been noted that the addition of burned rice
husk in biofertilizer B provided a higher percentage of
potassium (6.6%), which contributed to extra growth in
fruit diameter and weight compared to other biofertilizers
(without burned rice husk). These results are in
agreement with Seripong (1989) who mentioned that the
dry weight of shoots and fruits significantly increased as
the burned rice husk was added. Similarly, with the
addition of more than 3% nitrogen source meal from a
total of 7% in biofertilizer A and C gave a good yield of
leaves diameter for plant treated with biofertilizer B.
On the other hand, the plants treated with
biofertilizer C recorded the highest root and stem lengths
(237.6 ± 4.96 cm) in comparison to plants treated with
biofertilizer A and B, which were 185.0 ± 7.00 cm and
217.5 ± 6.93 cm, respectively. Total nitrogen content for
all biofertilizers (> 1%) had no effect on populations of
total bacteria, yeast, as well as ammonia and nitrite
utilizing bacteria. These results are in accordance with
those results obtained by Sarathchandran et al. (2001)
who reported that the nitrogen content in biofertilizer
around 048 - 0.69% did not give any significant difference
to the total count of microbes studied.
In conclusion, the microbiological, chemical and physical
properties of biofertilizer A, B, and C were determined.
Based on these properties, we suggest that biofertilizer A
is the best in encouraging flower and fruit growth, while
biofertilizer B is superior in leaf production and biofertilizer
C is good for the strength development of roots and
stems. Furthermore, the formulated biofertilizers in this
study was prepared from an economical and low-cost raw
material with the inoculation of special microorganisms,
which is a feasible and potential market for the
commercialization as well as to promote environmental
friendly technology. This will reduce the country’s
reliability on chemical fertilizers in a way to produce,
increase and sustain food production. Therefore, the
utilization of agricultural waste converted to biofertilizer
can be one of the successful alternative ways of
optimizing the use of resources and to generate income.
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... A sample of the biofertilizer was taken to the Regional Soils Laboratory of the Department of Agriculture located in Parola, Iloilo City, Philippines. Soil test data (Table 1) from the laboratory analysis reflected that the biofertilizer produced from the bioreactor composting technology has pH level of 8.23, 1.51 (2013) [16] which obtained a biofertilizer pH range of 8.20 -8.50. Though higher than the neutral pH, the slightly alkaline pH is beneficial because it will contribute to neutralizing acidic agricultural soil [17] that is dominant in the locality. ...
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Establishing reliable technological information on the safety of biofertilizers produced from a bioreactor composting technique is a must prior to its commercialization. A phytotoxicity study of biofertilizer made from the bioreactor composting technology at Aklan State University – Banga, Aklan, Philippines was conducted for fourteen (14) days using commercially available lettuce seeds (Lactuca sativa L.). Standard phytotoxicity attributes such as hypocotyl length, radicle length, relative germination percentage, and relative radicle growth observed during the germination stage were evaluated. Results revealed no significant difference in the radicle lengths of the germinated lettuce seeds as affected by the varying levels of biofertilizer dilution at H(3)=10.567, p=0.061>0.05. On the other hand, the hypocotyl length of the lettuce showed significant differences in response to varying levels of biofertilizer dilution with Welch’s F(5,5.163)= 8.175, p=0.017<.05. Also, the different levels of biofertilizer affected significantly the germination percentage of lettuce seeds F(5,12)=5.822, p=0.006<0.05. All levels of biofertilizer treatments indicated a decrease in relative germination percentage. However, those seeds applied with 10% biofertilizer have the highest reduction of germination percentage, equivalent to 86.9%(RGP=13.10%). All levels of biofertilizer showed an increase in radicle growth in contrast to negative control except for the 10% level of biofertilizer. Seeds that received the 10% biofertilizer showed an extremely high reduction in radicle growth, equivalent to 72.22% (RRG=27.78%). The study shows that applying low levels of the bioreactor-produced biofertilizer will observably reduce the measure of the germination characteristics of lettuce seeds, but not necessarily low enough to be considered phytotoxic. However, the application of at least 10% bioreactor-produced biofertilizer can presumptively lead to phytotoxicity.
... The world now has tended to reduce the use of chemical fertilizers for their negative effects on the environment, and tended to use microorganisms as bio-fertilizers, that some of which fix nitrogen and some work to dissolve the important elements necessary for plants (Hanapi et al., 2013). Among the microorganisms which work to fix atmospheric nitrogen is the root nodule bacteria (Rhizobium sp). ...
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تمت دراسة المىكبات الكيمياءية في العقد الجذرية لصنفين من نبات الفاصوليا المعاملة ببكتيريا الرايزوبيا وعنصري الحديد والموليبدنيوم النانويين
... The population count of microbes; bacteria, actinomycetes, fungi and two functional groups viz., nitrifying bacteria and phosphorus solubilising microbes (PSM) were taken to examine the effect of pesticides on their respective populations. Microbial counts were carried out using the plate count agar (PCA) for bacteria [29], Rose Bengal chloramphenicol agar for fungi [30], starch casein agar for actinomycetes [31], Pikovskaya's medium for phosphate solubilizing bacteria [32] and Ashby medium for nitrifying bacteria [33]. Serial dilution plate count method was used for enumeration of colony forming units (cfu). ...
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Introduction: The presence of pesticides in soils could result in alterations in microbial activities (enzyme activities, microbial populations), soil physical and chemical properties. Research gap/Challenges: Insufficient literatures on extensive monitoring of soil quality through enzyme activity, during pesticides application. Existing literatures concerning analysis of effect of pesticide application on soil enzyme activity are not comprehensive with respect to number of soil enzymes analysed. Aim/Objective: The study was conducted to investigate the effect of carbofuran and paraquat on soil biochemical characteristics on certain soils in the Niger Delta region of Nigeria. Methodology: These pesticides were applied at recommended doses, their effects on soil organic carbon, enzymes activity and microbial populations were assessed using standard methods. The enzymes monitored were amylase, invertase, protease, urease, phosphatase and dehydrogenase. Microbial counts were carried out for total heterotrophic bacteria, fungi, actinomycetes, nitrifying bacteria and phosphate solubilizers using the spread plate method. Original Research Article Ataikiru et al.; SAJRM, 4(2): 1-16, 2019; Article no.SAJRM.50500 2 Results: There were variations in the different enzyme activities in carbofuran-and paraquat-treated soil during this research. Dehydrogenase activity increased in treated soils. Also, urease activity was lower compared to other enzyme activities. As the study progressed, variations in values of soil organic carbon were observed. There was a gradual increase in microbial counts and can be traceable to their ability to temporarily mineralize and use the pesticides as carbon and energy source. The soil organic carbon, enzymes and microbial counts values were significantly different at P=0.05. Conclusion: This research revealed that the pesticides cause temporal impact on microbial populations and enzyme activities, associated with the pesticide type at recommended field application rates. A change in numbers, activity and diversity of soil microorganisms may act as indicators of soil fertility and reflect the soil quality.
... Actinomycetes were enumerated using starchcasein agar [29] and Pikovskaya's medium for phosphate solubilizing microbes [30]. Ashby culture medium was used to enumerate nitrogen fixers [31] and individual colonies were recorded as colony forming units (cfu). ...
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This study aimed at determining the impact of Carbofuran and Paraquat use on soil microbial biomass and microbial population as soil health index. Pot experiment, set-up as a randomized block design with replicates was done, with both pesticides applied at recommended rates for eight weeks. Twenty-four (24) soil samples were taken from the pesticides polluted soil as well as the unpolluted soil. These samples were used to assess the effect of pesticides on microbial biomass carbon (MB-C), nitrogen (MB-N) and phosphorus (MB-P). Also, microbial population (determined by aerobic spread plate count) of the pesticide-polluted soils was used as health index. The assessments were done weekly. The microbial biomass values increased from 273.48 µg/g to 293.15 µg/g (MB-C), 17.275 µg/g to 18.52 µg/g (MB-N) and 10.605 µg/g to 11.37 µg/g (MB-P) in carbofuran treated soil while increases from 277.26 µg/g to 288.365 µg/g (MB-C), 17.515 µg/g to 18.22 µg/g (MB-N) and 10.745 µg/g to 11.18 µg/g (MB-P) were observed in paraquat treated soil. The microbial counts in treated soils were within the ranges of 1.95 x 106 cfu/g to 1.03 x 107 cfu/g, 8.83 x 104 to 1.90 x 105 cfu/g, 1.08x 104 to 2.43 x 104, 1.15 x105 to 2.17 x 105 cfu/g, 1.38 x 105 to 2.22 x 105 cfu/g for total heterotrophic bacterial, fungal, actinomycetes, phosphate solubilizers, nitrifiers counts, respectively. The pesticides had no negative effects on the MB-C, MB-N, MB-P and soil microorganisms at recommended field rates, hence their use must be strictly based on these rates. These findings indicate that the relationship between soil nutrients and microbial biomass is significant in facilitating the use of microbial biomass as an important soil quality indicator.
... Nilai manfaat penggunaan pupuk hayati terhadap perbaikan kualitas tanah dan peningkatan hasil pertanian sudah banyak dilaporkan pada berbagai tanaman, termasuk padi (Kantachote et al. 2016, Wartono et al. 2015, jagung (Obidiebube et al. 2010, Soleimanzadeh dan Farshad 2013, Beyranvand et al. 2013, Hipi et al. 2013, Asih et al. 2016, Tao et al. 2017, kedelai (Noor 2003, Zarei et al. 2012, stroberi (Pesakovic et al. 2013), kapas (Egamberdiyeva et al. 2006), kelapa sawit (Hanapi et al. 2013). Pupuk hayati mampu menekan berbagai penyakit tanaman seperti fusarium pada pisang (Shen et al. 2015), hawar daun bakteri pada padi hingga 21,7% (Wartono et al. 2015), dan meningkatkan sifat antibakteri (Abdel-Aziez et al. 2014). ...
Industrial waste contains many organic and inorganic contaminants which pollute the land and cause health and environmental hazards. The circular economy (CE) phosphorus approach is the optimization of P fertilization, collection and recycling of P-rich wastes, improvement of household sewer systems and the application of biogenic wastewater treatment. Nitrogen fertilizers make plants produce proteins and nucleic acids and can be used to feed farm animals. The bacterial consortia can metabolize proteins and other inorganic nutrients from waste and improve crop yield. Biomass valorisation composting with an increased concentration of nutrients (N, P, K) can be directly applied to fields. In composting, the transformation of complex organic matter is achieved by earthworms and microbes. The mixture of waste with the addition of fungal strains serves as a multi-component fertilizer and the production is designed as waste-free. Further, physical and biological treatment of liquid and solid wastes can be converted into bio-fertilizers and organic manure. In biological treatment, the microorganisms such as bacteria, fungi, Actinomycetes, algae, cyanobacteria etc. could be the main factors. Application of industrial treated waste is possible to make soil fertile, good climatic factors and thereby improvement of biodiversity. Bio-based wastes can be delivered as fertilizers such as basal, foliar and also irrigate the plants to get more yield. The use of bio-fertigations may be a practical solution to recover valuable fertilizer components from various types of industrial liquid and solid waste using low-cost technology. Finally, the problems of contaminants in industrial wastes which could be converted into different types of economically viable bio-products in the possible directions also will be discussed.KeywordsIndustrial wasteBiological treatmentBio-fertigationDegradationVermicompostingMicroorganisms
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This study aimed to investigate the influence of the microbial biohumus obtained from organic non-toxic wastes on the yield of three varieties of peppers, identification of the effect of biohumus on soil physicochemical parameters, to determine the ecological significance and economic feasibility of biohumus application. The photosynthetic activity of plants and accumulation of ascorbic acid in the tissues of peppers were investigated. The experiments in the vegetation cabin condition have been carried out according to the following variants: 1. Control, 2. Biohumus, 3. N60P60K60, 4. Biohumus + N60P60K60. The yield of a variety of Jermatnayin hska pepper in the vegetation cabin conditions in the Biohumus (237 c/ ha) variant was lower than in the “Biohumus + N60P60K60” variant and was higher from “N60P60K60” and “Control” variants (277c/ha, 229 c/ha, and 191c/ha respectively). The yield of a variety of Arajnek pepper in the Biohumus (286 c/ha) variant was lower than in the “Biohumus + N60P60K60” variant and was higher “N60P60K60” and “Control” variants (320c/ha, 265c/ha, and 228c/ha respectively). The yield of a variety of Loshtak pepper in the “Biohumus” (335.7 c/ha) variant was lower than in the “Biohumus + N60P60K60” variant and was higher in “N60P60K60” and “Control” variants (391.9c/ha, 314c/ha, and 239.8c/ha respectively).
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The intensive amount of chemical usage in agricultural practices could contribute to a significant impact on food safety issues and environmental health. Over-usage of chemical fertilisers may alter soil characteristics and contaminate water sources, leading to several human and animal health issues. Recently, there have been efforts to use microbial biofertilisers as a more sustainable and environmentally friendly agricultural practice in the common household of Southeast Asia. Traditionally, this method tends to utilise leftover food materials and readily available bacterial cultures, such as yoghurt drinks, and ferment them under a specific period in either solid or liquid form. So far, most of the testimonial-based feedbacks from local communities have been positive, but only limited information is available in the literature regarding the usage of biofertiliser fermented food (BFF). Previously, raw food waste has been used in the agriculture system to promote plant growth, however, the functional role of fermented food in enhancing plant growth have yet to be discovered. An understanding of the symbiotic relationship between fermented food and plants could be exploited to improve agricultural plant production more sustainably. Fermented food is known to be rich in good microbial flora (especially lactic acid bacteria (LAB)). LAB exist in different sources of fermented food and can act as a plant growth-promoting agent, improving the nutrient availability of food waste and other organic materials. Therefore, in this review, the potential use of seafood-based, plant-based, and animal-based fermented food as biofertiliser, especially from Southeast Asia, will be discussed based on their types and microbial and nutritional contents. The different types of fermented food provide a wide range of microbial flora for the enrichment of proteins, amino acids, vitamins, and minerals content in enhancing plant growth and overall development of the plant. The current advances of biofertiliser and practices of BFF will also be discussed in this review.
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Science and Technology Towards Sustainable Development
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Liquid biofertilizer is increasingly available in the market as one of the alternatives to chemical fertilizer and pesticide. One of the benefits from biofertilizer is a contribution from population of microorganisms available. Traditionally, liquid biofertilizer produced from fermentation of effective microorganisms (EM) was recommended to be used within three months. This experiment showed that shelf life of the liquid biofertilizer produced from vegetable waste contains high amount of viable microbial population after four months of storage. The two conditions of storage, with and without light, were tested and it was found that there was no significant difference (p>0.05) upon viable microbial population, chemical and physical characteristics. However, there was significant difference from batch to batch of production due to raw materials.
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Organochlorine pesticides are notorious, due to their high toxicity, persistence in the environment and their tendency to bioaccumulate. Their extensive use in the northwest of Argentina has left residues in the environment.Microbial degradation is an important process for pesticide bioremediation and actinomycetes have a great potential for that.The current study examined organochlorine pesticides in contaminated soil. Indigenous actinomycetes were isolated from contaminated samples to evaluate bacterial growth as well as pesticide removal and release of chloride ions as a result of degradation.Most of the isolated microorganisms belonged to the Streptomyces genus, except one, which belonged to Micromonospora. Bacterial growth depended on the microorganism and the pesticide present (chlordane, lindane or methoxychlor). Highest growth and pesticide removal were observed with chlordane. Twelve out of 18 studied strains released chloride into culture supernatants, and percentages were higher with chlordane as carbon source than with lindane or methoxychlor. These results are supported by principal component analysis.This is the first report about actinomycetes isolated from an illegal storage of organochlorine pesticide in Argentina with capacity to growth, remove and use different organochlorine pesticide.
The effect of Bacillus licheniformis HA1 cell density on the acceleration of organic waste composting was tested in a bench-scale composting system utilizing a process limit temperature of 60°C. Variables measured during composting were CO2 evolution rate, conversion of substrate carbon and pH. When an initial cell density of 2.0×104 cfu/g-dry solid was used, the strain HA1 increased in number and prevented the decrease in pH during the early stage of composting. This resulted in enhanced populations of other thermophiles and increased the rate of organic matter decomposition. By contrast, no effect was observed at a lower cell density of HA1. It was found that the minimum cell density of HA1 to accelerate organic decomposition was around 104-105 cfu/g dry solid of raw material.
Four photosynthetic bacteria, isolated from 14 samples taken from seafood processing plants, were identified as species of Rhodocyclus gelatinosus, belonging to the purple, non-sulphur bacteria of the family Rhodospirillaceae. Cultivation in synthetic medium under four different conditions indicated that all four strains gave maximum carotenoid and bacteriochlorophyll synthesis under anaerobic conditions in the light, with values of 11 to 12.6 and 102 to 108 mg/g dry cell wt, respectively. These values are 87% higher than the pigment content obtained from aerobic cultivation, although the cell biomass of all strains (1.7 to 2.3 g/l) was 22 to 38% higher under aerobic conditions. Protein content was always between 32 and 43%. The specific growth rates of all isolates in aerobic cultivation (0.04 to 0.06 h(-1)) were twice those in anaerobic conditions in the light. No growth occurred in anaerobic conditions in the dark.
Processing procedures described are size reduction, air classification, screening and magnetic separation. Size reduction is the single most used and costliest of the unit processes involved. Non-compostable contaminants must be removed prior to screening. The cost of processing increases as the quality of the compost increases. One potential processing scheme that the authors consider technically reliable and economically practical is described. 6 references.
Some years before the appearance of Hellriegel and Wilfarth's work on the sources of nitrogen of leguminous plants, and while the part played by atmospheric nitrogen in the nutrition of crops was under active discussion, Berthelot was making some exact observations on the behaviour of uncropped soils towards the free element. He found that when 50 kilograms of air-dry arable soil were exposed to the air and rain in a vessel for seven months a great increase in the nitrogen content could be observed; the total nitrogen of the original soil had increased from 50 grams to 63, or a gain of over 25 per cent, after allowing for the small amount of combined nitrogen brought down by rain; in another experiment where the soil had first been washed free from nitrates, a gain of 46 per cent, of nitrogen was proved. In many other cases, however, the gain was only from 10–15 per cent, of the original nitrogen present in the soil.
Food waste is approximately one quarter of the total garbage in Taiwan. To investigate the feasibility of microbial conversion of food waste to multiple functional biofertilizer, food waste was mixed with bulking materials, inoculated with thermophilic and lipolytic microbes and incubated at 50 1C in a mechanical composter. Microbial inoculation enhanced the degradation of food wastes, increased the total nitrogen and the germination rate of alfalfa seed, shortened the maturity period and improved the quality of biofertilizer. In food waste inoculated with thermophilic and lipolytic Brevibacillus borstelensis SH168 for 28 days, total nitrogen increased from 2.01% to 2.10%, ash increased from 24.94% to 29.21%, crude fat decreased from 4.88% to 1.34% and the C/N ratio decreased from 18.02 to 17.65. Each gram of final product had a higher population of thermophilic microbes than mesophilic microbes. Microbial conversion of food waste to biofertilizer is a feasible and potential technology in the future to maintain the natural resources and to reduce the impact on environmental quality.