Population Dynamics of Azotobacter sp., Azospirillum sp., Bacillus

subtilis, Talaromyces pinophilus, Trichoderma asperellum, and

Syncephalastrum racemosum in the Medium with Various Earthworm

Applications

Nur Ulina Warnisyah Sebayang

, Tengku Sabrina

, Abdul Rauf

, Mariani Sembiring

, Asmarlaili

Sahar

, and Erman Munir

Department of Agriculture, Universitas Sumatera Utara Padang BulanMedan, Indonesia 20155

Department of Math and Science, Universitas Sumatera Utara Padang BulanMedan, Indonesia 20155

Keywords: Earthworm, microorganism, population, dynamics.

Abstract: The quality of the biological fertilizer was based on the population of inoculated microorganism. The

important efforts made to increase the population and maintain the population of microorganisms during

storage was using the best carrier. The aim of this study was to measure the population dynamics of some

microbes due to the application of various types of earthworms in producing biofertilizer. The study was

conducted in Biology and Biotechnology Soil Laboratory, Department of Agriculture, Universitas Sumatera

Utara. The design used was a Factorial Randomized Block Design consisting of 2 factors. The first factor

was a type of earthworm with 4 treatments viz; without earthworm, anecic earthworms, endogeic

earthworms, and epigeic earthworms. The second factor was a type of microorganism inoculants with

6treatments, viz; Azotobacter sp., Azospirillum sp., Bacillussubtilis, Trichodermapinophilus, Talaromyces

asperellum, and Syncephalastrum racemosum. The results showed that there was a significant interaction

between the types of earthworms in influencing the population of each microorganisms in the medium. The

highest alteration of microbial population was found in Azotobacter sp. and Trichodermaasperellum. The

best type of earthworm that could be used on the inoculum medium was the epigeic earthworm.

1 INTRODUCTION

The use of organic fertilizers has been used to

reduce the use of chemical fertilizers. Development

of biofertilizer technology is being actively

conducted to resolve the limitations of biofertilizer

that is a relatively short storability thus inhibiting its

commercialization. Many studies have been done to

get the best carrier. Fungus phosphate-solvent

growth of 2.0x10

after 7 days of storage can be

maintained from wheat skin mixed with 20% (w / w)

perlite when compared to peat carrier, corn cobs +

20% (weight / weight) perlite and cow manure

compost with 20% (weight / weight) perlite(Wang,

2015).Increased proportion of carrier material

vermicast in improving the survival of microbes.

The results also showed that vermicasts can be used

as alternative carriers for Azotobacter chroococcum,

Bacillus megaterium and Rhizobium leguminosarum

(Sekar and Karmegam, 2015).

Earthworms have the ability as a vector of

microorganisms and the vermicast have

highavailability nutrient and population

ofmicroorganisms. Earthworm is an

organismcapable of selecting and increasing

thepopulation of microorganisms through

peptidesderived from its excretion. Digestive fluid of

earthworms released amino acids, sugars and

organic molecules from organic residues. Cellulotic

bacteria are present in the digestive canal and casts

of earthworms indicating that they live in the

digestive canal of earthworms and actively degrade

food containing cellulose (Suhartanti, 2013).

By using the role of the earthworm, a new

innovation can be found in biofertilizerproduction

(enriched product of vermicompost and some

microbial inoculant). The biovermi to be produced

must be investigated in advance to identified the

quality through the parameters of microbial

population observed for some time.

Sebayang, N., Sabrina, T., Rauf, A., Sembiring, M., Sahar, A. and Munir, E.

Population Dynamics of Azotobacter sp., Azospirillum sp., Bacillus subtilis, Talaromyces pinophilus, Trichoderma asperellum, and Syncephalastrum racemosum in the Medium with Various

Earthworm Applications.

DOI: 10.5220/0010073900260030

In Proceedings of the International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches (ICOSTEERR 2018) - Research in Industry 4.0, pages

26-30

ISBN: 978-989-758-449-7

The observed microbial population dynamics

indicate that the quality of the biofertilizer to be

developed in terms of both variation of earthworm

and microorganism and to know the fertilizer's

storage capacity. Therefore, it is necessary to know

the effect of calibrationvariation of earthworms and

microorganisms in organic compost carrier to know

the dynamics of the microbial population in order to

produce qualified biofertilizers that can be utilized

as an innovation and environmentally technology.

2 MATERIALS AND METHODS

2.1 Experimental Design

The experiment was conducted in Laboratory of

Biology and Biotechnology Soil,

AgriculturalFaculty, Universitas Sumatera Utara,

Medan, Indonesia.

The experiment used Factorial

Random Block Design with 2 factors with 2

replications, so this study gets 48 treatments

. The

first factor is type of earthworms with 4 stage of

treatments that are; without

earthworm (C

anecic earthworm (C

), endogeic earthworm (C

and epigeic earthworm (C

). The second factor is

type of microorganismwith 6 stage of treatments that

are; Azotobactersp. (M

), Azospirillumsp. (M

Bacillus subtilis(M

), Talaromycespinophilus(M

Trichodermaasperellumstrain G (M

), and

Syncephalastrumracemosumisolate VPCI 1857/11

). Data wereanalyzed statistically by using

Analysis ofVariance (ANOVA) and then

following by Duncan Multiple Range Test

(DMRT) at 5 % level.

2.2 Experimental Pot Preparation

The experimental pot used was made of plastic with

size 19cm x 13cm x 9cm as figure 1.

Figure 1 : The Sketch of Experimental Pot.

2.3 Earthworm Preparation

Identification of earthworm was based on Blakmore

identification (Blakmore, 2002). The same weight of

each type earthworms was chosen for further

experiment. All selected earthworms soaked in

sterile water in order to remove its sewage so the

microbes on the digestive canal. Then they were

acclimatized in the laboratory with culture media

compost that has been sterilized.

2.4 Preparation of Inoculant Microbial

Azotobacter sp. and Azospirillum sp. were obtained

from the AgriculturalFaculty, Universitas Sumatera

Utara, Medan, Universitas Padjadjaran, Bandung.

Bacillus subtiliswas obtained from the Laboratory of

Institut Pertanian Bogor Culture Collection, Bogor,

Indonesia. Talaromycespinophilus was obtained

from the collection of Soil Biology Laboratory of

AgriculturalFaculty, Universitas Sumatera Utara,

Medanfrom potato plant (Sembiring, 2015). T.

Asperellum strain strain G and

Syncephalastrumracemosum isolates VPCI 1857/11

were obtained from a previous study (Sabrina, 2017)

derived from oil palm trunkchips and have been

identified. Multiplication of each isolate was carried

out using specific media such as Jensen medium for

Azotobacter sp., Okonmedium for

Azospirillumsp.,Pikovskayamedium to

Talaromycespinophilusand Bacillus subtilis.Potato

Dextrose Broth medium for

Trichodermaasperellumand

Syncephalastrumracemosum. After each isolate was

inoculated then incubated for 3-5 days, then

microbial population was calculated using colony

counter with the Pour Platedilution technique.

2.5 Preparation of Culture Media

Pot experiment of earthworm culture medium filled

with compost sterile as much as 400 gram and

covered with cloth net. Culture media regulated

moisture by adding water using hand sprayer.

2.6 Application of Microbes and

Earthworm

Each microbial was applied to a sterile culture

medium (compost sterile) of 10 mL and mixed

evenly. After 1 week of microbial application, the

earthworm application was performed with the same

individual weights selected and fed into the

experimental pot in accordance with the treatment

and then covered with a net cloth.

Population Dynamics of Azotobacter sp., Azospirillum sp., Bacillus subtilis, Talaromyces pinophilus, Trichoderma asperellum, and

Syncephalastrum racemosum in the Medium with Various Earthworm Applications

Table 1: The average of microbialpopulation in differenttypes of earthwormapplication on the second observation.

2.7 Maintenance and Observation of

Microorganism Population

Maintenance of various types of earthworms by the

addition of sufficient water using hand sprayer into

each culture medium. Observations were made on

the number of microorganisms in the culture

medium. Observations and sampling were

performed at the start of the study, a week after

application the microbial and a week after

application the earthworm (it means that 2 weeks

after microbial application). Ten (10) g of media

using a spatula and dissolved in 90 mL of sterile

water and shaker until homogeneous (dilution 10

-1

3 RESULT

3.1 Microbial Population (CFU/ml)

The results showed thatapplication oftypes of

earthworms, various microbes and theinteractions of

both in compost have a significantly effect

onmicrobial population. The average population of

microbes after a week earthworm application

available in table 1.

The population of Azospirillum sp. on boxtreated

with endogeic earthworms was the (27.5 x 10

-8

CFU/mL), however was not significant different

with its population in treatment anecic earthworms

(26 x 10

-8

CFU/mL). The lowest microbial

population was at treatment anecic earthworms and

Trichodermaasperellum(0.2 x 10

-8

CFU/mL) which

is a fungal decomposers and has a value of the

average population of microbes similar to

earthworms anecic treatment with the microbial

Syncephalastrumracemosum (table 1).

3.2 The Microbial Population

Dynamics

The dynamics of microbial populations was

determined from the difference between early

microbial population (a week after the application of

microbial) with microbial populations after

application of earthworm (figure 2).

Treatments

Azotobacte

Azospirillu

acillus

subtilis

Talaro

mycespinophilu

Trichoder

maasperellum

Syncephalast

rumracemosu

Mean

-8

CFU/mL

Without

Earthwor

12,1cdefg 11,5defg

2,4cde

0,4defg 0,9defg 0,4efg 6,3

Anecic

Earthwor

23,7b 26,0a 22,5bc 0,3fg 0,2g 0,2g 12,1

Endogeic

Earthwor

22,5c

27,5a 21,4c

0,3fg 0,7defg 1,1defg 12,2

Epigeic

Earthwor

20,2c

10,7defg 12,7cde 0,3fg 4,9defg 0,5defg 8,2

Mean 19,6 18,9 17,2 0,3 1,7 0,6



ICOSTEERR 2018 - International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches

FU/

Figure 2 : Dynamics of Microbial Population

The dynamics of microbial populations in biovermi

compost suistainthe increased and decreased. The

bacterial population increased from the first

observation to the second observation, but the

average of fungus population decreased from the

first observation to the second observation (figure

2). However, the treatment without earthworms was

the lowest population in the first observation,

whereas in the second observation the lowest

population was treated with anecic earthworms. In

this study, the application of earthworms in the

second week after the application of earthworms had

the percentage of live earthworms continued to

decline, indicating that earthworms are not able to

survive on the medium that used in this study. On

the second day after the application of earthworms,

the percentage of live anecic earthworms dropped

dramatically, so that all anecic earthworm were died.

Anecic earthworms did not survive on carrier media,

it may be due to anecic type earthworms usually live

in the soil and not live in waste habitats as

decomposer worms. The anecics earthworm (length

13-17 cm; diameter 10-15 mm) is larger than other

types and usually move vertically in soil. The same

thing happens withendogeic earthworms prefer live

on the soil with medium clay content for their

habitat. While the comparison of culture media used

may not be comparable, so that this endogeic cannot

survive. In this case the endogeic worm used is

Pontocolexcorethrurus. However, it is inversely

proportional to the epigeic earthworm that used

Eisenia Andrei that survive from application to

completion of observation. This happens because the

epigeic earthworm is an active earthworm as a

decomposer of organic matter in soil. Furthermore,

epigeic earthworms are also more relative tolerant to

pH and can tolerate pH levels and are relatively

tolerant of environmental conditions of organic

waste (

Dominguez and Edwards, 2011),

(

Neuhauser, 1988).

4 CONCLUSION

The interaction between the types of earthworms and

microorganisms influenced the population of

microorganisms. The type of earthworm that could

be used on the medium was the epigeic earthworm

(Eisenia andrei). The highest microbe population

were Azospirillum sp. (bactery) and Trichoderma

asperellum (fungus).

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Population Dynamics of Azotobacter sp., Azospirillum sp., Bacillus subtilis, Talaromyces pinophilus, Trichoderma asperellum, and

Syncephalastrum racemosum in the Medium with Various Earthworm Applications

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ICOSTEERR 2018 - International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches