




































BANGLADESH JOURNAL OF MULTIDISCIPLINARY SCIENTIFIC RESEARCH 9(1) (2024), 15-24 

 

15 

-  

     MULTIDISCIPLINARY SCIENTIFIC RESEARCH 

 
        BJMSR VOL 9 NO 1 (2024)  P-ISSN 2687-850X  E-ISSN 2687-8518 

 
        Available online at https://www.cribfb.com 

     Journal homepage: https://www.cribfb.com/journal/index.php/BJMSR 

                                                                                                                                                                                                    Published by CRIBFB, USA 
                                                                                                                                            

A STUDY ON EFFICIENT MICROORGANISMS ISOLATED FOR 

DEGRADATION FROM MUNICIPAL SOLID WASTE OF 

CHHATRAPATI SAMBHAJINAGAR, MAHARASHTRA, INDIA

                   
 Abhijit Thorat  (a)  Rakshanda Ingale (b)1    Vijay Rakte (c)    Ketki Sangle (d)    Sanjaykumar Thorat (e)    

 
(a) Research Scholar, School of Environment and Earth Sciences, KBCNMU Jalgaon Maharashtra, India; E-mail: thoratabhi782@gmail.com 
(b) Research Scholar, School of Environment and Earth Sciences, KBCNMU Jalgaon Maharashtra, India; E-mail: rmingale01@gmail.com 
(c) Research Scholar, School of Environment and Earth Sciences, KBCNMU Jalgaon Maharashtra, India; E-mail: raktevijay777@gmail.com 
(d) Research Scholar, School of Environment and Earth Sciences, KBCNMU Jalgaon Maharashtra, India; E-mail: ketkisangle310@gmail.com 
(e) Professor and Director, School of Environment and Earth Sciences, KBCNMU Jalgaon Maharashtra, India; E-mail: drst118@gmail.com 

 

 
A R T I C L E I N F O 

 
 

Article History: 
 

Received: 11th December  2023 
Reviewed & Revised: 12th December 2023 

to 9th March 2024 

Accepted: 10th March  2024 

Published: 16th March 2024 

 
Keywords: 

 

Characteristics, Solid Waste Dumping Site, 

Chhatrapati Sambhaji Nagar (Aurangabad) 

 
JEL Classification Codes: 

 

      G32, F65, L66, L25, M41    

  

      Peer-Review Model:  

 
      External peer-review was done through  

      double-blind method.        

 
A B S T R A C T 

 

With rapid industrial development and the progress of civilization, the problem of increased waste 

generation has become more complex in urban areas. This research evaluates the composition and 

characteristics of municipal solid Waste produced in a representative residential neighbourhood. Ten 

samples were taken from the disposal site of the Chhatrapati—Sambhajinagar (Aurangabad) city area. 

Gathered from garbage, nine bacterial isolates were made using a nutrient agar medium. Investigations 

were conducted into the best culture conditions, microbiological traits, biochemical traits within the 

strains, tolerance to five heavy metals (Cadmium, zinc, Arsenic, lead and mercury), sensitivity to four 

different antibiotics (penicillin, streptomycin, oxytetracycline, and gentamycin), and extracellular 

enzyme production of the microbial strains. All six strains that could produce protease were used for the 

waste degradation efficiency test. Due to these findings, there is now a greater chance of identifying 

bacteria of scientific significance from municipal waste disposal sites, and these isolates may be a key 

source of compounds with practical applications in industry. Based on the research, it is possible to 

extract beneficial bacteria for the environmentally friendly bioconversion of solid Waste from the 

(Chhatrapati. Sambhajinagar (Aurangabad) city area.                         

 
 

© 2024 by the authors. Licensee CRIBFB, U.S.A. This open-access article is distributed under the 
terms and conditions of the Creative Commons Attribution (CC BY) license 
(http://creativecommons.org/licenses/by/4.0/).  

            

 

INTRODUCTION     

Solid Waste is defined as waste type that includes principally household waste/domestic Waste, with sometimes the addition 

of commercial Waste collected by a municipality within a given area. They are generally in either solid or semisolid form. 

Exclude industrial hazardous wastes. Solid Waste Microflora is the collective name for microorganisms living in solid Waste 

(SWM). Fungi and bacteria are the most prevalent organisms typically found in solid Waste. The waste materials serve as 

a growth substrate for these microorganisms. They develop and increase this Waste using the many solid waste elements. 

Furthermore, it has been found that these organic wastes contain a wide range of harmful microbes. The study of microflora 

has yet to be conducted in this field. So, we are interested in investigating the Bacteria present in solid wastes and their 

applications. Since the beginning of humankind, Waste has been generated in the form of bones and other parts of animals 

they slaughter for their food or the wood they cut to make their carts; with the progress of civilization, waste generation has 

become more complex.  

Waste disposal threatens cooperation between man, animals and the soil. Like chemical hazards, aetiologic agents 

might be dispersed in the environment through water and wind. Poisonous plants, insects, animals and indigenous pathogens 

                                                      
1Corresponding author: ORCID ID: 0000-0003-2172-6030 

© 2024 by the authors. Hosting by CRIBFB. Peer review under the responsibility of CRIBFB, U.S.A.  
https://doi.org/10.46281/bjmsr.v9i1.2196 

  

To cite this article: Thorat, A., Ingale, R., Rakte, V., Sangle, K., & Thorat, S. (2024). A STUDY ON EFFICIENT MICROORGANISMS ISOLATED 
FOR DEGRADATION FROM MUNICIPAL SOLID WASTE OF CHHATRAPATI SAMBHAJINAGAR, MAHARASHTRA, INDIA. Bangladesh 

Journal of Multidisciplinary Scientific Research, 9(1), 15-24. https://doi.org/10.46281/bjmsr.v9i1.2196 

 

https://orcid.org/0000-0001-5429-4970
http://creativecommons.org/licenses/by/4.0/)
http://creativecommons.org/licenses/by/4.0/)
https://www.openaccess.nl/en
https://doi.org/10.46281/bjmsr.v9i1.2196
https://orcid.org/0000-0003-2172-6030
https://orcid.org/0009-0009-4765-3571
https://orcid.org/0009-0009-9841-874X
https://orcid.org/0000-0002-6499-4986


Thorat et al., Bangladesh Journal of Multidisciplinary Scientific Research 9(1) (2024), 15-24 

 

16 

are biological hazards that might be encountered at the waste site (Khupe, 1996; Chetan et al., 2017). Solid Waste Microflora 

is the collective name for microorganisms living in solid Waste (SWM). Fungi and bacteria are the most prevalent organisms 

typically found in solid Waste. The waste materials serve as a growth substrate for these microorganisms. Using the many 

elements that comprise solid Waste, they develop and increase this Waste. 

Furthermore, it has been found that these organic wastes contain a wide range of harmful microbes (Amalra et al., 

2006). There was no significant study of microflora. So, we are interested in investigating the microorganisms present in 

solid wastes and their applications. The solid waste disposal industry divides solid Waste into four major categories for 

disposal depending on the state in which they are disposed of. The Waste produced in urban areas is generally known as 

municipal solid Waste (M.S.W.). 28.8% of India's population resides in urban areas (census 2011). It is predicted that 41% 

of the population will reside in cities by 2011. In India, waste quantity increased from 46 million tons in 2001 to 65 million 

tons in 2011 (Kumar & Gaikwad, 2011)—solid Waste in Chhatrapati. Sambhajinagar (Aurangabad) generates large amounts 

of poorly disposed and untreated Waste, so there is an urgent need to design scientifically unified solid waste management 

for the town. Earlier studies showed that people in urban areas produce half a kilogram, 10% of being burned. The average 

solid waste generation rate in low-income states or cities is only 0.4 to 0.6 kg/person/day, compared to 0.7 to 1.8 

kg/person/day in fully commercial states (Cointreau, 1982; Blight & Mbande, 1996). Every day, massive amounts of 

waste materials are generated in all cities and municipal areas of India. Solid Waste in urban areas has a very high 

organic content, ranging from 70% to 85%. 

India's urban solid trash generation is increasing with the country's population growth and per capita G.D.P.—

the only municipality in Chhatrapati. Sambhajinagar (Aurangabad), which has a population of 10.07 million and an 

area of 325 square kilometres, believes that 3000 tons of Waste are generated daily by the A.M.C. In the Chhatrapati. 

Sambhajinagar (Aurangabad) city area, the daily rubbish generation per capita ranges from 0.35 kg to 0.4 kg. The 

formal system efficiently collects all garbage generated. A formal system recycles about 10%–15% of Waste, but 

illegal or self-disposable discarded uncollected Waste makes up 35–50% (Joerger et al., 2009). Therefore, this 

biodegradable fraction could be combined or co-composted as biofertilizers and soil conditioners (Parr & Hornick, 

1992). The five main microorganisms in soils are actinomycetes, bacteria, fungi, algae, and protozoa. Bacteria make up 

the majority and are crucial for the breakdown of Waste (Sultana, 1997). Bacteria utilize Waste for their metabolism, 

and as a result, they create a few straightforward yet helpful mixes crucial for plant growth, soil health, and maintaining 

the overall equilibrium of a natural ecosystem. Composting involves the regulated breakdown or conversion of organic 

matter, typically in the presence of oxygen, into a stable, soil-like substance known as manure. The quantity of 

microorganisms, in addition to rodents and insects, is essential for the decomposition of solid Waste. Since bacteria 

are the most significant, it is possible to use efficient microorganisms to aid in solid organic Waste's breakdown. This 

study was conducted considering the critical importance of garbage decomposition to create a repeatable procedure 

and search for more active decomposer bacteria that could efficiently and effectively break down organic wastes and 

provide valuable components for plant nutrition (Sultana, 1997; Zaved et al., 2008; Chetan et al., 2017). 

 

MATERIALS AND METHODS 

Study Area: Chhatrapati. Sambhajinagar (Aurangabad) is an A-grade municipal corporation and the capital of Marathwada 

located at N19°53'47"–E75°23'54" of Maharashtra state. The city is bounded by mountains in all directions, titled "The City 

of Gates", and the strong presence of these can be felt as one drive through the city. Chhatrapati. Sambhajinagar 

(Aurangabad) is Maharashtra's tourism capital and the fifth largest city in Maharashtra. It has an average rainfall of 756.6 mm, 

and the maximum and minimum temperatures of the town are 42.60C and 9.10C, respectively. Temperature and wetness 

during the rainy season lead to a higher humidity content in municipal solid Waste, which raises the weight of the trash. 

Furthermore, heat and high humidity accelerate the breakdown of the organic waste component, creating challenges for 

processing and disposal that have an immediate negative impact on the residents' and garbage workers' environmental health. 

 

Sample Collection: Ten waste samples were collected; five came from the Chhatrapati. Sambhajinagar (Aurangabad) 

Municipality's garbage transfer facility and the other five came from the city's industrial zone and landfill in Naregaon. 

Blended soil and garbage were evaluated, collected aseptically, stored at 40°C, and stamped as per their source and location 

as needed. Microbe samples were brought to the laboratory for soil isolation, and pH and moisture content were noted. In 

nutrient agar, all the microorganism cultures were kept at 40C. Every culture was subcultured every fifteen days. S1: Solid 

Waste collected from Housing colony A.M.C. Area; S2: Solid Waste from dumping (Garbage, behind Ghati Hospital) area 

at Chhatrapati. Sambhajinagar (Aurangabad) City Area Dump; S3: Soil surface, Salim Ali lakeside of Delhi Gate, 

Chhatrapati. Sambhajinagar (Aurangabad); S4: Transportation of Solid Waste in AMC Area Wet soil, drain side of 

Naregaon Area Chhatrapati. Sambhajinagar (Aurangabad); S5: Solid Waste Dump Sites in A.M.C. Area drain side 

garbage, Chhatrapati. Sambhajinagar (Aurangabad); S6: Soil surface, Segregation of Waste and Recycled items in A.M.C. 

Area, Chhatrapati. Sambhajinagar (Aurangabad); S7: Waste Encroached Road in Chikalthana Industrial Area wet soil of 

drain.  S8:   Waluj Industrial Area Housing Colony Waste Dumping Sites in wet soil of drain. S9: Garbage, Chitegaon area, 

Chhatrapati. Sambhajinagar (Aurangabad) and S10: Refuse, slaughterhouse, Padegaon Area, Chhatrapati. 

Sambhajinagar (Aurangabad). Three bacterial samples, S1 through S3, had superior growth on their ideal Medium. 

These three samples were labelled as S3, S2, and S1, respectively, since I.U.L. was observed on Nutrient Agar media 

(N.A.), Dump was observed on Basic Czapek-Dox-Agar medium (BCDA), and S1 was observed on Basic Czapek-

Dox-Agar medium (BCDA). Previous research teams have noted this unappealing pattern in the literature (Bundela et 

al., 2010; Chatterjee, 2010; Thorat & Chavan, 2021). Residential areas should not be too close to waste disposal sites, and 

covered rubbish bins are a good idea. Waste disposal plants are sporadically located near the A.M.C. area's residential areas 



Thorat et al., Bangladesh Journal of Multidisciplinary Scientific Research 9(1) (2024), 15-24 

 

17 

and public buildings. A typical view of these types of trash disposal sites can be found at the Naregaon site, which is in one 

of the government-reserved zones of Chhatrapati. Sambhajinagar (Aurangabad) Municipal Council area (AMC). 

 

Characteristics of Waste: The following parameters of the synthetic normal for the test were broken down: organic matter 

(%), absolute N (%), P(%), and K(%). Following a quick titration technique, natural carbon was resolved Agarwal 2005. 

The trash was processed using a combination of acids (HClO4, HNO3, and H2SO4) to determine the aggregate nitrogen 

content. This was done using the Kjeldahl methodology, as demonstrated by the method presented by Agarwal 2005. A 

colourimetric approach was used to evaluate all the phosphorus using stannous chloride and ammonium molybdate. The 

Flame photometric approach was used to measure absolute potassium. 

 

Pure Culture: A solitary colony was detected and subsequently re-streaked onto the surface of a nutrient agar plate 

along with Basic Czapek-dox agar (BCDA) medium as the major inoculant. After that, the plates were incubated at 

either room temperature or 30°C. Pure cultures were examined using nutrient agar plates and Basic Czapek-dox agar 

(BCDA). Gramme dye was chosen to confirm that the cell morphology and gram reaction were identical to the original 

colony's. At this stage, pure cultures were again examined using the previously described procedures, and a new 

nutrient agar plate and Basic Czapek-dox agar (BCDA) were re-streaked with the appropriate colony. Once a pure 

culture was obtained, the same colony was streaked onto a BCDA slant and nutrient agar. These cultures were 

refrigerated for a full day of incubation. The isolates from the soil were used for further experiments. 

 

Biochemical Tests: The catalase test detects soil bacteria; 2-3 ml of the hydrogen peroxide solution was poured into a 

test tube. Using a sterile wooden stick, several colonies of the test organisms were removed from the nutrient  agar 

plate and Basic Czapek-dox agar (BCDA) medium and immersed in the hydrogen peroxide solution. Bubble formation 

was then observed. Lactose and mannitol fermentation differentiate the microorganisms fermenting carbohydrates such 

as lactose and mannitol. Voges Proskauer test was carried out to detect the production of acetylmethylcarbinol acetoin, 

a natural product formed from pyruvic acid during glucose fermentation. The buffered glucose broth and the organism 

were inoculated and incubated at 370C for three days. Approximately 3 ml of alpha naphthol was added, followed by 

1 ml of 40 % K.O.H. and mixed well for 30 minutes. For the result, the pink solution means V.P. (+) and no change 

means V.P. (-). 

 

Isolation of Microorganisms: Municipal sludge and sewage were the sources of the soil samples. These germs were 

isolated using serial dilution and streaking methods until a single colony was obtained. Twenty agar plates were prepared 

aseptically. After the agar plate was ready, the fast-growing bacteria plate was taken. The bacteria on the plate are chosen 

based on the different shapes. The selective bacteria on the agar plate are streaked using an aseptic technique. The plate is 

divided into four parts. The streak agar plate is sealed and kept in a 30°C incubator for growth. After 24 hours, the bacteria 

was purified by another spread plate technique and incubation. Besides that, gram staining was done on the previous plate. 

Samples were observed under a microscope and characterized based on Chetan et al. (2017). 

 Biochemical tests were performed to identify microorganisms using a standard procedure based on Bergey’s 

manual. Numerous biochemical identification techniques were employed to identify these bacteria from municipal sludge, 

including the Gramme stain, spore formation, strict anaerobes, starch hydrolysis, Voges-Proskauer, and swollen cell test. 

Municipal sludge can be identified biochemically using various techniques, including the Gram stain, starch hydrolysis, 

citrate test, and Voges-Proskauer. A urease test was used to detect soil bacteria; a dense "milky" suspension of the test 

organism was prepared in 0.25ml   physiological saline in a small tube. A urease tablet was added into the tube and 

incubated at 35- 370C for up to 4h or overnight. The color change was observed in the test organism. An indole test was 

done to detect the soil bacteria. The culture contained tryptophan for the development of test organisms. This Medium was 

prepared in a bijou bottle with 3 ml of sterile tryptone water. The Medium was then added with 0.5 ml Kovac’s reagent 

(4p-dimethylamino-benzaldehyde) with gentle shaking, and the colour was observed. The ability of an organism to use 

citrate as a carbon source and ammonia as a nitrogen source is known as the Citrate utilization test. The media will 

turn green to blue if the citrate utilization test is positive. 

 

RESULTS AND DISCUSSIONS 

This study involved isolating and characterizing bacterial strains from various locations within the Chhatrapati—

Sambhajinagar (Aurangabad) Municipality area and from industrial zones. Numerous physiochemical factors affect 

bacterial growth, including Medium, pH, temperature, incubation time, carbon source, etc. Bacteria can grow in a wide 

range of moisture levels. The current investigation began with the discovery that the moisture content of the samples 

obtained ranged from 25.09 to 78.19%. The samples from S3 and S2 had the highest moisture content (78.19%), while 

those from S1 had the lowest moisture content (25.09%). The relationship between a soil's bacterial population and 

moisture content is well-established. The maximum bacterial density is found in regions of high moisture content, and 

the optimum level for the activities of aerobic bacteria is often 60 to 70 % of the soil's moisture-holding capacity 

(Sonawane et al., 2010). Most aerobic soils include large numbers of Achromobacter, Pseudomonas, and Bacillus 

species; anaerobic and damp environments favor the growth of Clostridium. Under such circumstances, actinomycetes 

are proven to a comparable reckonable growth (Sonawane et al., 2010) observed using a variety of growth media, 

including potato dextrose agar (P.D.A.), nutrient agar (N.A.), and Czapek-Dox agar (both acidic and basic), the 

investigation's isolated strains were seen to develop. Nutrient agar (N.A.) medium was found to be fitting for the 

massive growth of the S3 strain in Table 2, whereas basic Czapek-Dox-agar (BCDA) was found to be suitable for the 



Thorat et al., Bangladesh Journal of Multidisciplinary Scientific Research 9(1) (2024), 15-24 

 

18 

massive growth of the S1 and S2 strains in Table 1. 

 

Table 1. Effect of different pH on the growth of isolated strains in BCDAa medium 

 
pH of 

Medium 

Strains Incubation Period 

6 h 12 h 24 h 36 h 48 h 72 h 

 

7.1 

S1 NG PG GG GG GG GG 

S2 NG PG    GG    GG   GG     GG 

S3 NG PG    MG    GG   GG     GG 

S1 NG PG   GG    GG GG     GG 

7.6 S2 NG PG    GG    GG GG     GG 

S3 NG NG PG     MG GG     GG 

S1 NG NG PG       PG  MG    MG 

9.1 S2 NG NG PG PG PG     PG 

S3 NG NG NG NG PG    MG 

S1 NG NG NG NG NG NG 

10.6 S2 NG NG NG NG NG NG 

S3 NG NG NG NG NG NG 

S1 NG NG NG NG NG NG 

12.10 S2 NG NG NG NG NG NG 

S3 NG NG NG NG NG NG 
aBCDA = Basic Czapek-Dox-Agar; NG = No growth; PG  = Poor growth; MG = Moderate   growth;     GG = Good growth. 

 

Table 2. Effect of different pH on the growth of isolated strains in NAa medium 

 
pH of 

Medium 

 

Strains 

Incubation period 

6 h 12 h 24 h 36 h 48 h 72 h 

4.2 S1 NG NG NG NG NG NG 

S2 NG NG NG NG NG NG 

S3 NG NG NG NG NG NG 

5.7 S1 NG NG NG NG PG PG 

S2 NG NG NG NG PG PG 

S3 NG NG NG NG PG PG 

7.2 S1 NG PG MG MG GG GG 

S2 NG PG MG MG GG GG 

S3 NG PG MG GG GG GG 

8.7 S1 NG NG MG MG      MG MG 

S2 NG NG PG PG      MG MG 

S3 NG NG PG MG      MG GG 

10.2 S1 NG NG NG PG PG PG 

S2 NG NG NG PG PG PG 

S3 NG NG NG PG PG PG 
aNA = Nutrient Agar; NG = No growth; PG = Poor growth; MG = Moderate growth; GG = Good growth. 

 

It was discovered that the pH range of the two media was 7-8. Bacterial strains grew most readily in N.A. 

and BCDA at pH 7.2 and 7.6, respectively. Based on the previously mentioned data, the pH values of the S2, S1, and 

S3 strain samples were 7.79, 7.95, and 7.86. This could be the cause of the bacteria's successful in vitro growth at pH 

7-8 in BCDA and N.A. Although bacteria can react with soil at pH values ranging from 4 to 10, most prefer a pH 

somewhat on the alkaline side of neutrality. Certain Bacillus spp. can thrive at pH 11, but Thiobacillus thiooxidans 

and Acetobacter spp. can only grow at pH levels 0 and 2. Agarwal (2005). Thermo-actinomycetes can only develop 

at temperatures between 50°C and 65°C. Their optimal growth occurs at pH levels 8 or 9, while reactions at 

approximately five significantly inhibit their growth. (Amalraj, 2006). Mycobacterium tuberculosis var. grows well 

but slowly (2–6 weeks) at 37°C on glycerin agar or a solid medium such as coagulated egg, serum, or blood. 

Escherichia coli, Vibrio, and Streptococcus faecalis can also withstand an alkaline reaction (pH 8–9) (Amalraj, 2006). 

Three strains of bacteria were used in this experiment, and their cultures were cultured at various temperatures—25, 

29, 34, 37, and 40°C. 37°C was the temperature at which all strains grew massively. For bacteria, the ideal temperature 

range is between 25 and 36°C. The temperature range of 10-40°C is suitable for the growth of many microorganisms. 

Sultana (1997) noted that the optimal temperature range for bacterial growth was 33–40°C. Some bacteria grow most 

quickly at temperatures lower than 200 degrees Celsius. Some thermophiles can multiply below 40°C, and 

thermophiles often grow at temperatures between 45 and 65°C 2010 saw (Sonawane et al., 2010). While M. Chelonei 

and M. Marinum grow well at lower temperatures (18–30°C), Mycobacterium avium thrives best at 40°C. On tomato 

juice agar, Lactobacillus sp. grows best at 25 to 39°C. On standard laboratory media, Agrobacterium sp. grows well at 

pH of 6.8 to 25 and 39°C (Sultana, 1997). At around 25°C, streptococcus lactis can be grown on agar plates with milk, 

whey, tomato juice, or sterile milk. It grows most readily when lactose or glucose is present. Around 35°C is better for 

growing than 25°C (Amalraj, 2006). The strains used in this investigation underwent varying lengths of incubation (6, 

12, 24, 36, 48, and 72 hours). The S1 and S2 strains grew well with a 24-hour incubation period. However, the S3 strain 

required a 36-hour incubation period. Coliform bacteria grow in the incubation period of 24±2 h and at 32°C, and they 

show good growth at 370C for 48h of incubation. Three methods were used to characterize the selected strains: non-

microscopic or visual observation, microscopic observation, and biochemical tests. Upon visual inspection, it was 



Thorat et al., Bangladesh Journal of Multidisciplinary Scientific Research 9(1) (2024), 15-24 

 

19 

discovered that following a 24-hour incubation period, S1 had turned pale orange, S2 had turned white, and S3 had 

turned light brown in their chosen media (BCDA and N.A.). Following a 48–72-hour incubation period, S1 turned 

orange, S2 turned yellow, and S3 turned brown. While S3 had a creamy colony type, the S1 and S2 strains had wet 

colonies. Staphylococci and Micrococci develop golden brown, yellow, or white colonies on regular media. Certain 

enterococci, coryneform, and enterobacteria can form black colonies on a regular medium. According to Chatterjee 

(2010), Staphylococcus aureus produces glossy, convex, black colonies on the Baird-Parker medium. An established 

and trustworthy technique for observing microorganisms is gram staining—alcohol-decolored gram-negative 

bacteria, causing them to lose their crystal violet-purple hue. According to Uwadiegu and Iyi (2014), gram-positive 

bacteria did not decolourize and continued to be purple. Every isolated strain found in the current study tested positive 

for gram-positive bacteria. Because the isolated strains (S1, S2, and S3) produced oxygen gas from hydrogen peroxide 

(H2O2) through enzymatic degradation, they all showed positive catalase test results. A similar result was observed in 

Zurbrugg (2002). Several biochemical tests have been performed to study the characteristics of bacterial strains, as 

shown in Table 3. A fermentation test differentiates the microorganisms that ferment carbohydrates, such as lactose and 

mannitol. The isolated strains exhibited positive lactose and mannitol fermentation tests because open and sealed tubes 

produced a yellow color. 

In the urease test, due to red and pink color formation through the media, the S2 strain showed a positive 

urease test, which could decompose urea to ammonia. S1 and S3 showed negative tests because they did not produce a 

red-pink color in this Medium. The indole test demonstrates the ability of certain bacteria to split the amino acid 

tryptophan into indole, which accumulates in the Medium. The S3 strain showed a positive test and produced a red color 

in the media. S2 and S1 strains showed negative tests because they did not produce a red color in the media (Butu & 

Mshelia, 2014). They also observed the same result. S2 and S3 strains showed positive results in the hydrogen sulphite 

production test because they produced black color in the media. However, S1 displayed a negative test as this strain 

did not produce a black color in the media. Sultana (1997) observed the same result. The citrate utilization test was based 

on an organism's ability to use citrate as its only carbon source and ammonia as its only nitrogen source. The S3 strain 

showed a positive citrate utilization test because the Medium turned green to blue. S1 and S2 did not turn the media blue; 

therefore, they showed negative tests, as shown in Table 4. 

Numerous researchers conducted studies demonstrating how domestic waste decomposition accumulated and 

separated lower volatile fatty acids and volatile organic compounds (V.O.C.s) (Sonawane et al., 2010). The weight 

and volume of the stuff diminish as bacteria break down the Waste. Because of the bacteria that broke down the Waste 

and transformed it into simple molecules, we also saw a decrease in the weight and volume of the treated rubbish in 

the current breakdown study. After 30 days of treatment with S2 strain, the maximum weight loss in suspension 

treatment for the breakdown of solid Waste without additives was observed at 50.70%. Chatterjee (2010) initiated the 

highest weight loss (26.06 %) using the Trichoderma strain after 30 days in a similar study. 

 

Table 3. Biochemical tests of some waste-decomposing bacteria 

 
 

Strains 

 

Catalase 

Lactose 

Fermentat

ion 

Test 

Mannitol 

Fermenta

tion Test 

Voges- 

Proskauer 

Test 

 

Urease 

Test 

Indole 

Test 

Citrate 

Utilization 

Test 

 

Identified Bacteria 

S3 +ve +ve +ve -ve -ve /+ve -ve /+ve +ve Xanthomonas spp. 

S2 +ve +ve +ve +ve +ve -ve -ve Bacillus spp. 

S1 +ve +ve +ve +ve -ve -ve -ve Pseudomonas spp. 

 

Table 4. Changes in temperature during garbage decomposition by bacterial suspension and different 

concentrations of molasse solution at 3-day intervals. 

 

a M.S., Molasses solution, b C.T., Control. 

After 30 days, the highest volume loss for the S2 strain in suspension treatment was 38.67%, and in pellet 

treatment, it was 36.73%. 

Additives such as sucrose can be used with garbage for decomposition, increasing the growth of bacteria 

(Mohapatra, 2006). It was observed that by treating with sucrose solution, the growth of bacteria was high, and the 

decomposition rate was also high. It added sucrose solution to the garbage and enhanced the decomposition rate through 

 

 

Days 

Temperature (°C) of decomposition 

garbage by treating B.C. Dump strain 

and molasses solution. 

Temperature (°C) of 

Decomposition garbage by treating 

S1 and molasses solution 

Temperature (°C) of decomposition 

garbage by treating S3 strain and 

molasses solution 

5%

MSa 

10%

MSa 

15%

MSa 

C.T. 
b 

5%

MSa 

10%

MSa 

15%

MSa 

C.T. 
b 

5%

MSa 

10%

MSa 

15%

MSa 

CT b 

3 29 29 29 29 29 29 29 29 29 29 29 29 

6 31 31 32 29 31 31 31 29 31 31 31 29 

9 33 33 34 30 33 33 33 30 32 32 33 30 

  12 35 35 36 30 34 35 35 30 33 33 34 30 

  15 37 37 39 30 36 37 37 31 35 35 36 31 

  18 37 38 40 31 36 37 38 31 35 36 37 31 

  21 35 38 40 31 34 35 37 31 33 34 36 31 

  24 33 36 37 31 32 34 35 31 31 32 34 31 

  27 32 34 34 30 31 32 33 30 30 31 31 30 



Thorat et al., Bangladesh Journal of Multidisciplinary Scientific Research 9(1) (2024), 15-24 

 

20 

increasing fermentation. After 30 days of inoculation, the S2 strain showed the most significant percentages of weight 

loss (52.73%, 55.32%, and 65.12%, respectively) in the 5%, 10%, and 15% sucrose solution treatment (together with 

the bacterial strain). Of all those treatments, the S2 strain with a 15% sucrose solution had the most significant weight 

loss at 65.12%. A similar pattern for volume loss was discovered. With a 30% sucrose solution added and 30 days of 

inoculation, the S2 strain showed the most significant volume loss (51.34%). Thus, regarding weight reduction and 

volume loss, S2 and a 15% sucrose solution were the most effective strains for sucrose treatment . Since sucrose 

treatment successfully facilitated trash decomposition, molasses was eventually employed as a less expensive and 

alternative source of carbohydrates to aid in the breakdown of solid Waste. Following a 30-day inoculation, the S2 

strain showed the highest weight loss percentages of 56.70%, 62.70%, and 80.24% in the 5%, 10%, and 15% molasses 

solution treatments, respectively. Of all those treatments, the S2 strain's 15% molasses solution resulted in the most 

significant weight loss, at 80.24%. A similar pattern for volume losses was discovered. After 30 days of inoculation  

with the S2 strain and 15% molasses solution, the most significant volume loss was discovered to be 64.27% (both in 

terms of weight loss and volume loss). Again, comparing the effect between different concentrations of sucrose and 

molasses treatment for degradation of solid Waste with bacterial suspension, the highest weight loss was 80.24 % in 

the 15 % molasses solution in the S2 strain. 

In contrast, the 15% sucrose solution in S2 resulted in the most significant weight loss, 65.12%. A similar 

pattern for volume losses was discovered. Thus, the waste breakdown method using 15% molasses was the most 

successful regarding volume losses, followed by a 15% sucrose solution. On the other hand, bacterial suspension 

without any additives reduced volume change more than treatments including additives. The maximum volume loss 

(%) in the 15% sucrose treatment was 51.34% after 30 days of inoculation in the S2 strain; the highest volume loss 

(%) in the 15% molasses solution was 64.27%. A related investigation (Ramesh & Mathivanan, 2009) discovered that 

Trichoderma strains with 4% glucose concentrations lost 66% of their body weight. After two weeks of composting, 

the ultimate weight of the dead and after-birth piglets in the composting pile was just 3.1 kg (6.9 lb), according to (Zaved et 

al., 2008). The remaining tissue easily crumbled in the sawdust medium. Over 6% of the initial animal mass was lost on 

average per day in this experiment. The current study found that molasses treatment outperformed sucrose treatment 

regarding volume loss (%) and weight loss (%) when breaking down organic solid Waste with bacterial suspension. It can 

be explained in this way that in molasses, some extra components (which were absent in sucrose) could give extra 

nutrients to the rapid growth of bacteria and increase the number of bacterium cells. So, the degradation process with 

molasses solution was higher with more bacteria than with sucrose solution treatment. So, the growth of bacterial strains 

was the main factor in decomposing garbage. 

The relationship between volume loss and weight loss was linear, and no interaction was found between the 

two parameters. Both volume loss (%) and weight loss (%) increased gradually with the progression of the 

decomposition process, and the result was the same for each strain treatment. The weight loss (%) was higher than 

volume loss (%) in garbage decomposition in each strain treatment, as shown in Figure 1. 

 

 
(A)                                                                                     (B) 

                                            
(C)  

 

Figure 1. Weight loss (%) and volume loss (%) of decomposed garbage at 7-day intervals using S2 suspension and 

different concentrations of molasses solution. (A) S2 + 5% molasses solution; (B) S2 +10% molasses solution; (C) S2 

+15% molasses solution. 

 

 

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Thorat et al., Bangladesh Journal of Multidisciplinary Scientific Research 9(1) (2024), 15-24 

 

21 

According to Ezeah and Roberts (2012) and Bundela et al. (2010), heat is produced, and the temperature rises 

when bacteria break down garbage. In this investigation, we found that the temperature grew steadily in all cases after 

4-6 days, peaked after 15–24 days, gradually decreased after 28–30 days, and finally decreased to its starting point. 

Designed for decomposition in the composting method of solid Waste in vitro conditions by bacterial suspension and 

culture pellet, the highest temperature was 37°C after 15-18 days in suspension treatment of S2 and S1 (Figures 2 and 

3). 

 

Figure 1(A)x 

 

 Figure 1(A) 

 
Figure 1(B) 

 
Figure 1(C) 

Figure 2. Changes of temperature (°C) during garbage decomposition at 3-day intervals using suspension and culture 

pellet of the three strains: (A), S2 strain; (B), S1 strain; (C), S3 strain. 

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3 6 9 1 2 1 5 1 8 2 1 2 4 2 7 3 0

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Thorat et al., Bangladesh Journal of Multidisciplinary Scientific Research 9(1) (2024), 15-24 

 

22 

 
Figure 2(A)  

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Figure 3. Changes of temperature (°C) during garbage decomposition at three-day intervals using the three strains 

separately, along with the most effective concentration of additives. (A), three strains + 15% sucrose solution; (B) Three 

strains + 15% molasses solution. 

 

The maximum temperature reached by the S2 strain with 15% sucrose solution after 18 days was 39°C. After 

18 days, the maximum temperature for the molasses solution, which included 15% molasses solution in the S2 strain, 

was 40°C. Thus, it may be concluded that a high decomposition rate may be related to temperature. This study's 

treatment with the highest temperature had the most significant weight and volume loss. The S2 strain had the most 

weight and volume loss in all instances, along with a high temperature. The original pH of the fresh rubbish used in 

this experiment was 4.11, which is acidic. The pH of degraded Waste in suspension and pellet treatments and 

treatments with sucrose and molasses changed from an acidic to an alkaline condition in all cases. The pH ranged from 

7.31 to 11.06. The S2 strain-decomposed 15% molasses solution yielded the highest pH of 11.06. 

When bacteria are grown in a medium with a pH of 7, it is quite probable that this pH will alter due to 

substances the organism produces, which could be primary or acidic (Thorat & Chavan, 2021). It has been noted that 

as composting progresses, the pH changes over time. Most bacteria thrive in the pH range of 6.0 to 7.5. For the 

remaining composting, use 7.5 to 8.5. Composting produces organic acids at the first stage. The pH value lowers in 

the first few days for this acidic state. However, after a few days, the temperature rises, and the pH value rises as more 

organic Waste decomposes. After 10, 20, and 30 days, an unpleasant smell was associated with the bacterial culture 

pellet decomposition of organic kitchen waste, suggesting a slow degradation of organic materials. After ten days, 

there was a pungent stench in the solid waste decomposition process caused by bacterial suspension. After 30 days, 

there was no stench, suggesting that the organic kitchen wastes had possibly wholly degraded. In a related study, 

Ahsan (1999) found that composting reduced the odour during the breakdown of solid Waste. According to the study, 

beneficial bacteria may be separated from the surrounding environment to favourably facilitate the bioconversion of 

solid organic Waste. This study demonstrates how valuable and successful the established method of waste 

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Figure 2(B) 



Thorat et al., Bangladesh Journal of Multidisciplinary Scientific Research 9(1) (2024), 15-24 

 

23 

decomposition is in safeguarding human health and the environment from waste-related issues. The study was 

conducted to see the effect of fresh, decomposed garbage and bacterial suspensions on biomass production on a fresh-

weight basis of potato Solanum tuberosum. For the experiment, decomposed garbage using bacterial suspension and 15 

% molasses, fresh garbage, and bacterial suspensions were used separately in the soil pot of potato. 

A control treatment using simple garden soil was carried out for every treatment. The highest biomass output 

(65.61 g) was obtained in the decomposed garbage treatment, broken down by S2 suspension and 15% molasses. In 

contrast, fresh garbage and bacterial suspension showed 50.98 and 39.31 g of biomass production, respectively. The 

maximum biomass output (58.41 g) was shown by the decomposed garbage treatment, which was broken down by S1 

suspension and 15% molasses; fresh garbage and bacterial suspension showed 48.45 and 37.07 g of biomass 

production, respectively. The highest biomass output (52.18 g) was obtained in the decomposed garbage treatment, 

broken down by 15% molasses and S3 suspension. 

In contrast, fresh garbage and bacterial suspension produced 47.69 and 34.19 g of biomass, respectively. In 

all cases, the addition of decomposed garbage to soil enhanced biomass production when compared with fresh garbage 

or bacterial suspension treatment. With decomposed Waste and a 15% molasses solution, S2 suspension produced the 

most biomass in terms of fresh weight, followed by S1 and S3. Fresh trash and bacterial suspension outperformed the 

control: garden soil alone. 

 

CONCLUSIONS  

Solid Waste contains different microorganisms. Furthermore, it is concluded that several drug-resistant organisms are found 

in solid Waste and garbage in Chhatrapati—Sambhajinagar (Aurangabad) city and industrial areas, which spread bacterial 

diseases. Hence, solid Waste should be segregated and purified before being sent for recycling. Because civil solid Waste 

is a blend of many substrates, it is an ideal advancement medium for the growth of different microorganisms. Our current 

research shows that a metropolitan trash dump may serve as a breeding ground for various mechanical and antimicrobial 

microorganisms. 

Furthermore, it can be a valuable tool for bio-searching new or rare species, which may produce important bioactive particles 

essential for environmentally harmful trash contamination. In combination, it can also serve as a respectable adjunct in the 

industrial sector. In contrast to other natural conditions, the metabolically dynamic nature of the microbes in this 

environment produces various catalysts and bioactive mixtures. Therefore, it is crucial to understand the Waste implied by 

tiny creatures from a biological perspective and how they might benefit environmental biotechnology. 
 

 

 

Author Contributions: Conceptualization, A.T. and S.T.; Methodology, A.T.; Software, V.R.; Validation, S.T. and R.I.; Formal Analysis, K.S.; 

Investigation, A.T.; Resources, K.S.; Data Curation, V.R.; Writing – Original Draft Preparation, V.R.; Writing – Review & Editing, V.R.; Visualization, 

R.I.; Supervision, S.T.; Project Administration, S.T.; Funding Acquisition, A.T., R.I., V.R., K.S. and S.T. Authors have read and agreed to the published 
version of the manuscript. 

Institutional Review Board Statement: Ethical review and approval were waived for this study because the research does not involve vulnerable groups 

or sensitive issues. 
Funding: The authors received no funding for this research. 

Acknowledgements: The authors thank the School of Environmental and Earth Sciences, Kavayitri Bahinabai Chaudhari North Maharashtra University, 

for supporting this research. They are also thankful to Chhatrapati. Sambhajinagar (Aurangabad) Municipal Corporation (A.M.C.) for permitting us to 
collect samples and encouraging us during this work.  

Informed Consent Statement: Informed consent was obtained from all subjects involved in the study. 

Data Availability Statement: The data presented in this study are available on request from the corresponding author.  
Conflicts of Interest: The authors declare no conflict of interest.       

                                                                                                                                                                                                                             

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