INTRODUCTION

Composting is more safe and natural process for reducing biomass and volume of organic waste
and therefore also help in recycling of tough contents present in MSW that are not easily
degradable. (Juan F. Saldarriaga · Jorge L. Gallego 2018) In devolping countires the main source
of MSW comes from food waste with average percentage of 55. (Alejandra Cerda, Adriana
Artola 2017). It is observed that MSW is being produced between 1.3 an 1.9 billion tonnes per
year and it was thought that it will rise upto 2.2 billion tonnes per year by 2025. (J. Jara-
Samaniego, M.D. Pérez-Murcia 2017)

Succesive Microbial population carried out on organic substrate that leads to biological
decomposition under controlled condition, as a result the final product, more-stable humus like
compound is formed having less environmental effect on agriculture field, this simple process
is known as composting (J.A. Lopez, F.Suarez 2014). It is carried out under aerobic conditions
and kills pathogenic organism. Microorganism inculding fungi bacteria , worms etc convert
organic waste into more stable organic coumpound that can be used as a resourse for mankind
called as compost (M. Pilar Bernal, Sven G. Sommer2017)

There are different key parameters that are interlinked and support stability of the final product
of composting. Microbial succesion community are affected by evaluating some factors such as
pH, temperature etc when extensive study of heterotophic and their metabolic activities is
observed. Furthermore these communities are also affected by different raw materials and
objects that will be used in comosting. (Awasthi 2017)
On the bright side microbes plays an important role in the decomposition of organic waste
therefore it is necessary to understand changes and structures of different microbial
community.(M.M Jurado, F suarez-Estrella 2015).Agriculture wastes are decomposed rapidly
with the help of lignocellulolytic microorganism (MAM Ayesha Ameen intro)

Composting comprise of different stages .Among them the most effective stage of composting is
thermophillic phase. (Sutripta Sarkar,Rajdeep Benerjee 2009) The second phase, Thermophillic
phase(35-65 C) starts by completely replacing the microphillic flora, here mesophilic organism
died and thermophilic organisms starts their actions. This is the fast process and continues until
62 C temperature is reached. Thermophllic bacteria can tolerate high temperature but
thermophilic fungi have growth maxima of (33-55 C).The last phase is cooling where final
product is obtained by lower activity of microbes and temperature . (H Insam and M de Bertoldi
2007)(M. Pilar Bernal, Sven G. Sommer 2017 In Biodegration phase thermophiles play an
important role, the thermophilic phase of composting was found to be dominate by Bacillus spp.,
; Geobacillus. (Sutripta Sarkar, Rajdeep benerjee 2009) According to temperature ranges
thermophiles are divided into three grouping catogeries: Thermophiles having temperature
ranges from 30-60C with optimum temperature of 50C, extreme thermophiles with temperature
rangs from 40-70 and an optimum temperature of 65C and the last Ultrathermophiles or
Hyperthermophiles, their mimimun temperature are not defined with am optimum temperature of
100C having maximum range od 115-120C. Most important bacteria among thermophiles that
are extensively studied are Bacillus (B. stearothermophillus) and Thermus (T. aquaticus).
(Edward 1993)

The thermophilic phase can be made more effective by increasing microbial population or
treating them with desired enzymes or by improving microbiological quality, that can be done
by one of the process called microbial inoculation. Inoculation can be made more affected by
selecting more efficient bacteria (Shabnam Ghaffari1, Abbas Akhavan Sepahi 2011). Some of
the effective microorganism is originated from Japan, lactic acid bacteria, yeast and
photosynthetic bacteria are few commercially used microbial inoculants, they prove hugh
nutrient content in compost with less odour and high stability (Yee Van Fan, Chew Tin Lee
2017)

Some of the efficient isolates in compost belongs to the genus of Geobacillus, its an all round
group of thermophillic facultative anaerobic bacteria. These rod shaped, gram positive bacteria
have ability to sporulate. Some strains of these bacteria can degrade cellulose or xylan (Martinus
J. A. Daas , Antonius H. P. van de Weijer) Another bacteria cellulolytic bacteria Anoxybacillus
sp. MGA110 was served to be more efficient in promoting biodegradation of waste products.
(Shabnam Ghaffari1, Abbas Akhavan Sepahi 2011)

There are many reports published for the isolation of efficient degrading bacteria from
thermophillic stage. One of the study (B Trello et al., 1996) show isolation of different strains
during thermophillic stage at different temperature. The temperature range of thermophillic stage
is (65-80C), only strain that are related to Bacillus stearothermophillus were identified at this
temperature range of (65-69C). On the other hand some of the facultative autotrophic (Bacillus
schlegelii) and obligately autotrophic (Hydrogenobacter spp.) were observed in the hot composts
at temperature rang above 70C but these bacteria does not efficiently contribute to the
degradation. This study shows a great number of gram negative, aerobic, heterotrophic, non-
spore forming, thermophillic bacteria related to Thermus strains at a temperature range of 65-
82C) were isolated from a thermogenic compost. By a thorough procedure of isolation including
sample collection, protein profiling, DNA-DNA hybridization, 16S Ribotyping shows that these
strains were closely related to Thermus thermophillus HB8.(B Trello et al., 1996) One study
shows that there were low diversity of obligately heterotrophic thermophiles that were closely
related to Bacillus stearothermophillus, there great number of oxygen and suphur hydrogen
bacteria contributes its important role in mineralization.( B Trello et al., 1995)

For better understanding, a compared study showing number of thermophillic strains at variable
temperature in young and old compost samples were identified. Different strain Thermus
thermophillus, Hydrogenobacter spp., and Bacillus spp were isolated by the same techinique
involving molecular characterization of bacterial coommunity by restriction enzyme
analysis.This study investigated 200 clones out of which 69 clones were identified as
Thermophillic Bacillus and Thermus thermophillus and thus showed a gradual change is both
phsyichemical evolution and temperature in young (13-18 days) and old samples (39-41 days).
(Blanc M et al., 1999)

In 1989, a study shows thermophillic methanogenic bacteria isolation and characterization from
the mushroom compost. Methogens are said to be isolated from those places where oxygen is not
available usually from thermophllic volcanic environment, sediments, rumens and sewage sludge
digesters and this study shows presence of thermophillic methagonic bacteria although oxygen is
available in mushroom compost. Those isolated bacteria show morpjological similarity,
moreover their similarities were obtained from the cofactor content and mol% G + C. According
to these similarities the strains of M. thermoautotrophicum were indicated as M.
thermoautotrophicum strain CNC-1 and M. thermoautotrophicum strain CNC-2. (Derikx P.J.L et
al., 1989)

17 thermophillic bacteria were isolated from a solid compost they were incubated at 60C for 2
day under anaerobic condition on different agar media out of which only single colony were
identified and isolated by repeated streaking on BHI agar plates that were incubated under
anaerobic condition at 60C. The isolates are isolate M5EXG and M10EXG that are identified
from compost comprised with 1% (w/v xylose), 1% (w/v glucose) and 5 or 10% (v/v ethanol) the
other 2 strains that were isolated from compost supernatant were B10EXG and isolates C1
however there observation remain unpublished in this study. Their 16s rDNA sequences and
phylogenetic analysis shows that both isolates resembles with the Bacillus subgroup 5 and were
closely related to G. thermoglucosidasius with 99.5% and 98.9% 16S rDNA sequence
similarities. (Fong J.C.N et al., 2006)

The isolation and characterization of thermophillic bacteria from a sewage sludge was also
observed, on the basis of size reappearance margin and colour characterstic colonies on the agar
plate were preselected, they were streaked repeatedly on the agar plates to confirm those
characterstic that they are of ineterest. 12 bacilliform strain with 11 aerobes and 1 anaerobes
were isolated as a pure cultures, when the isolated strains are characterized 9 strains were
identified as endospore-forming, gram positive bacilli and 2 strains as non-endosporing bacilli.
Those 9 strains of genus Bacillus on the basis of their characterstics were identified as B
stearothermophillus growth inhibition with 3% NaCl. Anaerobic isolates were not tested and on
the other hand the 2 left strains 1T and 2T resembles with the genus Thermus. (Hakozaki et al.,
1991)

One study showed different bacterial population during thermophillic phase composting of
mixture of raw material (cattle manure, rice bran and plant ash ). The samples were identified
and inoculated on suitable culture media suitable for bacteria or actinomycetes. Those strains
were further identified with their morphological and physichemical characterstic followed by
16S rDNA sequence analysis.The diversity of thermophillic bacteria at the initial stage is much
higher total strain of 243 x106 that belongs to 13 genera involving Bacillus, Geobacillus,
Ureibacillus, Pseudoxanthomanas, Brevibacillus, Rhodococcus, Pseudomonas, Chelatococcus,
Proteus, Paenibavillus, Parvibaculum, Luteimonas and Actinotalea. Bacillus were the one's that
was isolated in the peak heating phase of fresh waste and have higher degrading abilities in the
early phase. In rice straw composting the dominant genera observed were Bacillus and
Actinomycetes in their thermophillic phase. However Geobacillus and Ureibacillus were
frequently detected dominant genera during thermophllic composting stage. U.
thermosphaericus, B aerius and B licheniformis were detected in initial and final samples of
thermophillic phase of composting having higher degrading abilities in the final thermophillic
phase of composting. The efficiency of degrading ability in thermophillic phase of composting
depends upon microbial activity of different bacteria. (Li R et al., 2014)

Here, strains were isolated after four repetitions on LB plates by streaking colonies, T1 strains
and T2 strains from sewage sludge. T1 strains had 98% resemblance with both Petrobacter sp.
and Tepidiphilus margaritifer on the other side T2 strains had 99% and 100% similarity with
Pseudoxanthomonas sp. and Pseudoxanthomonas taiwanensis. The study of these strains showed
that they can establish in thermophillic digesters. (Liu S et al., 2011)