Bacillus Cohnii Research Articles

IMPACT OF FREEZE-THAW CYCLES ON BACTERIAL VIABILITY IN CONCRETE

Calcium carbonate precipitating bacteria can improve mechanical and physical properties of building materials. Most commonly, bacteria are used for the crack repair, production of self-healing concrete, or as a cementitious agent for low-strength materials. The favorable effect of bacterial metabolism can only be achieved if the sufficient number of cells remain viable in the structural material. Besides the limited amount of oxygen, extremely high pH level and mechanical pressure on cell walls, bacteria must survive the repetitive freezing and thawing in concrete-type materials. Freezing water may generate additional internal pressure damaging the bacterial cells and suppressing the bacterial activity. In present study, the viability two of bacteria strains (Bacillus pseudofirmus and Bacillus cohnii), was studied under repeated freezing and thawing conditions. It was shown that concrete-embedded bacteria may survive fluctuations of low temperatures and freeze-thaw cycles, compromising approximately 50% of viable spores. This survivability may only be achieved if bacterial spores are protected from the concrete environment itself, for example, spores could be injected to the expanded clay particles and coated with the styrene-acrylate emulsion. These findings confirm that Bacillus pseudofirmus and Bacillus cohnii bacteria could be used in cold climatic regions, retaining viable bacteria spores within the concrete.

IMPACT OF FREEZE-THAW CYCLES ON BACTERIAL VIABILITY IN CONCRETE

Calcium carbonate precipitating bacteria can improve mechanical and physical properties of building materials. Most commonly, bacteria are used for the crack repair, production of self-healing concrete, or as a cementitious agent for low-strength materials. The favorable effect of bacterial metabolism can only be achieved if the sufficient number of cells remain viable in the structural material. Besides the limited amount of oxygen, extremely high pH level and mechanical pressure on cell walls, bacteria must survive the repetitive freezing and thawing in concrete-type materials. Freezing water may generate additional internal pressure damaging the bacterial cells and suppressing the bacterial activity. In present study, the viability two of bacteria strains (Bacillus pseudofirmus and Bacillus cohnii), was studied under repeated freezing and thawing conditions. It was shown that concrete-embedded bacteria may survive fluctuations of low temperatures and freeze-thaw cycles, compromising approximately 50% of viable spores. This survivability may only be achieved if bacterial spores are protected from the concrete environment itself, for example, spores could be injected to the expanded clay particles and coated with the styrene-acrylate emulsion. These findings confirm that Bacillus pseudofirmus and Bacillus cohnii bacteria could be used in cold climatic regions, retaining viable bacteria spores within the concrete.

Molecular diagnosis for camel raw milk microbiota.

The existence of diverse microbes in unprocessed camel milk poses a significant threat to the well-being of a large population, especially infants and toddlers. The objective of this study was to ascertain the existence of microorganisms in unprocessed raw camel milk by employing a molecular-based technique in combination with a histological examination of bacteria. The identification of microbial species was achieved by employing PCR amplification and sequencing of 16s rRNA gene fragments. Various micorganisms found includes the probiotic Lactobacillus species, Staphylococcus succinic, Macrococcus casealyticus, Bacillus cohnii, and Salinicoccus kunmingensis. To prevent microbial contamination in raw milk, it is necessary to adequately heat or pasteurise the milk and to wash and sterilise the udder before milking the camel. This is because raw milk contains microbes that cause multiple diseases. Moreover, in the current era of the COVID-19 pandemics, ensuring proper sanitary conditions in milk and its derivatives might potentially mitigate the transmission of various diseases among consumers shortly. Keywords: camel, microbiota, 16s rRNA gene, PCR.

Experimental Investigation on Influence of Binder Constituents on Workability, Strength and Microstructure of SCGC

Abstract: The present study investigates the influence of bacteria on varying binder constituents to enhance the workability, strength, and microstructure properties of self-consolidating geopolymer concrete (SCGC). Geopolymer concrete is a sustainable alternative to conventional cement-based concrete, as it utilizes industrial by-products and reduces carbon emissions. The bacterial strains, known for their ability to promote mineral precipitation and improve concrete properties, are introduced in varying proportions to the geopolymer binder constituents. The goal of this experimental investigation is to better understand how Bacillus Cohnii bacteria affect the workability and strength of alkali-activated Self-consolidating Concrete (SCC) mix compositions. Three mix formulations with variable binder contents in the range of 400 kg/m3 and 450 kg/m3 were the subject of experimental research. This research work aims on investigating various binder constituents on Strength, Workability and Microstructure. Three different SCGC mixes were prepared by varying the percentage fly ash and Alcofine in range of 0%, 10%, 30% and 0%, 5%, 5% to the quantity of GGBFS respectively. Alcofine increases final setting time. For all mixes, ambient curing was done. As per EFNARC recommendations, the fresh qualities of SCGC were determined using the Slump test, T50 slump test, L-Box test, V- funnel test, and J-ring test. By conducting compression tests, split tensile tests, and flexure tests after 7, 14 and 28 days, the strength characteristics of SCGC were identified. The SCGC's microstructure and chemical composition were both determined by SEM and EDS analyses, respectively. According to test results, an alkaline solution sample with a 13M concentration and 450 kg/m3 of binder produced the highest compressive, flexural, and split tensile strengths after 28 days producing values of 58.32 MPa, 3.5 MPa, and 4.27 MPa, respectively.

Bio-concrete: Unveiling self-healing properties beyond crack-sealing

Do microbes categorically heal the cracks in concrete or only seal the cracks - this study aims to answer, based on a few experimental findings, highlighting the self-healing concepts in cement-based materials. Alkaliphilic spore-forming Bacillus cohnii microbes (105-1010 cells/ml) are used in the cement-concrete (bio-concrete) in addition to varying nutrient amounts. The healing in the form of regaining its structural efficiency is evaluated by its mechanical (compressive, flexural) strength and durability properties. Bio-mineralized calcium carbonate seals 37% of 20–50 nm dimensioned pores and 87% of 50–200 nm dimensioned pores resulting in 43–48% total voids reduction within 28 days of the healing period. Microbes only degrade the nutrients to precipitate healing agents (calcium carbonate) without affecting the C–S–H structure in concrete. Statistical analysis reveals that calcite deposition mostly influences the strength of self-healed bio-concrete whereas experimental findings show 5.8–6.3 wt% calcium carbonate deposition augments the 40–60% compressive strength of bio-concrete compared to reference concrete. The complete-to-60% sealing of cracks of 0.02–2 mm width is examined using visual inspection of concrete skins. Microbes reduce porosity, sealing cracks intensifies mechanical strength, and the durability of the concrete validates self-healing rather than only sealing the cracks.

Optimization of spore production and activation conditions of concrete crack healing bacteria and research on crack repair effect

Currently, research on the spore production rate and spore activation rate of microorganisms is mostly used in the pharmaceutical and food industries, with little research on their application in repairing concrete cracks. The spore production ability of mineralized microorganisms and the spore activation ability in the concrete environment are key influencing factors in ensuring the repair effect of concrete cracks. The effects of various factors on the spore production rate and spore activation rate of Bacillus alcalophilus (Bacillus cohnii) were studied using a single factor controlled variable method in this paper, and the optimal spore production culture conditions and activation conditions were determined. Furthermore, the mineralization reaction after microbial spore activation and the repair effect of concrete cracks were investigated. The results showed that under optimal spore production culture conditions, the spore production rate reached 93.9 %. The spore activation rate of microorganisms was as high as 94.5 % under optimal spore activation conditions. The rate of microbial spore activation was 78.6 % at room temperature (20 ℃) without heat shock treatment. Calcite calcium carbonate produced by the mineralization reaction after microbial spore activation could be used to repair subsequent concrete cracks. The optimal concentration of calcium acetate required for microbially induced carbonate deposition was 100 g/L. The concrete crack with a width of 1.12 mm could be repaired after 28 days of repair age, and the concrete strength recovery rate was 70.81 % when the microbial self-healing agent prepared by Bacillus alkalophilus (Bacillus cohnii) was incorporated into concrete.

Microbial Induced Calcite Precipitation (MICP) Potential of Ureolytic Bacillus sp. Isolated from the Soil of Eroded Ecosystems for Stabilizing and Improving the Fertility of Eroded Soils

The loss of soil from lands due to erosion has a negative effect on ecosystems and food security. Bacillus due to high catabolic capability is an appropriate candidate for application in biocementation process. The aim of this study is isolate and characterize Bacillus sp. with biocementation capability from various ecosystems. The isolates were separated from 400 samples, and characterized by biochemical and molecular methods include the amplification and sequencing analysis of gyrA and 16S rRNA genes. Growth in presence of urea, in different salinity, pH, and temperature, also scanning electron microscope (SEM), X-ray diffraction (XRD), and wind tunnel analysis were applied to determine biocementation ability. A total number of 195 isolates were recovered from environmental samples, of which 25 isolates (12.82%) were identified as urease-positive Bacillus which belonged to 10 species consisting of Bacillus subtilis five strains (20%), Bacillus vallismortis and Bacillus seohaeanensis four strains (16%) each, Bacillus mobilis, Bacillus pseudofirmus, Bacillus cohnii, Bacillus cereus, Bacillus alkalinitrilicus two strains (8%) each, and Bacillus sphaericus and Bacillus megaterium one strain (4%) each. Moreover, 15 urease-positive isolates (7.7%) belonging to Ralostenia, Actinomycete, and Halomonas genera were identified. Optimum conditions for microbial induced calcite precipitation (MICP) by isolates are 30 °C, pH 9, and 6% salinity. The highest rate of calcium carbonate formation and urease activity recorded in B. subtilis with 24.15 mg/mL of calcium carbonate and 4.40 × 103 unit/L of urease, followed by B. mobilis and B. alkalinitrilicus with 22.85 mg/mL of calcium carbonate and 3.93 × 103 unit/L of urease. After MICP the lowest soil loss ratio at a flow rate of 90 km/h, was observed in B. subtilis 100-fold reduction, followed by B. seohaeanensis, B. cereus, B. vallismortis, with 90, 85, 80-folds reduction, respectively. Results indicate that the diversity of Bacillus sp. offers the potential ability for adaptation to harsh and untapped environments, also showed that the use of MICP on the soil surface can have a very significant role in reducing soil losses due to wind erosion.

Evaluating the Properties of Self-healing Concrete: A Review

The development of cracks is a regular phenomenon that causes water and various chemicals to seep into the structure reducing the strength and longevity of the building. The intrusion of external materials might affect the reinforcements when exposed to moisture and CO2 exposure, degrading the structure. Regular maintenance and treatments are needed to fix the cracks and maintain the structure. Research is in progress identifying solutions to these issues with active and self-processes in concrete which aids in repairing the cracks by generating calcium carbonate (CaCO3) particles which could seal the porosity of concrete and micro-cracks. Various bacteria like Bacillus pasteurii, Bacillus subtilis, Bacillus cohnii and Bacillus sphaericus are chosen for the crack sealing treatment as those bacteria could survive in an alkaline medium. Distinct species of bacteria have various growth conditions. Bacterial self-healing treatments were situated in an environment containing various minerals and chemicals at a constant temperature and for a specific amount of time to develop healing properties. This paper investigates the different experimental works studied on self-healing processes in large concrete samples with different sizes and materials. Studies revealed that bacteria enhanced the systemic toughness, flow ability, durability, and flexural modulus of normal concrete. This paper also investigates the different mathematical formulations established to examine the stress-strain behaviour of bacterium, which has been utilized to increase the strength of concrete.

Performance of Bacillus Cohnii Added Microbial Concrete

In the era of rapid urbanization, concrete has been playing a very important role in construction industry but inherently, it is very much vulnerable to formation of cracks, due to which its durability gets decreased. If immediate action is not taken, cracks developed in the section may spread further and increase the cost of repair. In concrete structures, these cracks become an easy path of entry for water, oxygen and carbon dioxide resulting into the decrease in durability of concrete. Numerous researches around the world are on to heal this defect with the help of addition of various bacteria in the form of Bacterial Concrete. This study aims to find out the change in salient properties of concrete experimentally after adding the microbes. Concrete cubes were cast with different bacteria concentrations, tested for salient properties after 7 and 28 days. Test results reveal that addition of bacteria results into increase in compressive strength and decrease in water absorption and porosity of concrete. Findings also indicate that the concrete mix with higher bacteria concentration gives higher compressive strength and lower water absorption and porosity in concrete.

Comparative Study of Mechanical Properties of Concrete Using Commercially Available Bacterial Strains

Bacteria impregnated concrete is a recent advancement in concrete technology which is being studied extensively by different researchers all around the world. This study aims to study the effect of bacteria on different parameters of concrete and compare the results from compressive test, split tensile test and flexure test with that of a normal mix concrete. The main objective of this research was to study the mechanical properties of concrete upon adding commercially available Bacillus Subtilis (5 billion cells/gm) and laboratory cultured Bacillus Cohnii (105 cells/ml) to the concrete mix. Both the bacterial concretes are mixed with calcium lactate and are cured in Calcium Chloride solution. This particular study confirms that commercially available Bacillus Subtilis imparts better concrete properties compared to laboratory cultured Bacillus Cohnii.

Studies on Compressive Strength Microstructural Analysis of Self-Compacting Mortar with Bacteria

This research proposed bacteria-treated concrete, which is highly desirable because the mineral precipitation induced from microbial activities is pollution free and natural. This study investigates the effects of Bacillus Licheniformis, Bacillus Cohnii and mixed bacteria on compressive strength, ultrasonic pulse velocity test, rapid chloride penetration test, microstructural analysis and bacteria cell concentration of 10 5cell/ml of normal and self-compacting mortars made with and without fly ash for 7 and 28 days. With self-compacting mortar, the compressive strength of Bacillus Licheniformis improved by 30.3 % and by adding Bacillus Cohnii in 28 days, the ion penetration value decreased. Finally, it was observed that the deposition of calcium carbonate in mortar improved compressive strength using SEM-EDS and XRD with compound polymorphisms of calcite, vaterite, quartz, and Hautirite.

The effect of bacteria Bacillus Cohnii on the synthesised calcium silicate hydrate (C–S–H) with various calcium to silica ratio in nanoscale

Calcium silicate hydrate (C–S–H) is the main cement hydration product, providing the most mechanical strength to concrete structure. As per the previous studies of bacteria-based self-healing concrete using non-ureolytic bacteria, one of the main issues is that the amount of externally added calcium salt was restricted by the properties of carriers, limiting the content of precipitated calcium carbonate. In addition, previous studies barely investigated the potential impact of microbial activities on the individual cement hydration products. Therefore, in this study, for the first time, we investigated the individual impact of bacteria on synthesised C–S–H gel and the potential of biomineralisation of C–S–H gel. In light of results obtained in this study, we evidenced that C–S–H could be biomineralised to produce a considerable amount of calcium carbonate. Decalcification of C–S–H gel and microbial carbon-capture were simultaneously occurred prior to the precipitation of calcium carbonate, resulting in increase of calcium and bicarbonate ions in the solution. The amount of precipitated CaCO3 is closely in response to Ca/Si ratio in C–S–H, higher Ca/Si essentially led to more precipitates, whilst the remaining structure and mass of C–S–H gel that could not be further biomineralised was generally similar.

Evaluation of the crack-healing performance and durability of bacteria integrated alkali-activated fly ash composites

This study aims to introduce Bacillus Cohnii endospores into an alkali-activated fly ash-based composite (AAFC) to improve the crack-healing properties and durability while maintaining an environment-friendly strategy. Few important parameters such as water absorption capacity, sulfate resistivity, rapid chloride ions penetration, and its migration through bacteria impregnated AAFC (Bio-AAFC) are considered for the assessment of the healed structure's activities. The cracks up to 0.59 mm width dimension are completely plugged up and 12% of mechanical strength of healed-AAFC is regained within 28 days, determined in the microscopic study, strength gains calculation, and velocity of ultrasonic pulse waves assessment. Bio-mineral precipitation into Bio-AAFC produces durable concrete, particularly in chloride and sulfate-laden environments. Due to calcium/sodium carbonate precipitation in pores and voids, the microstructure of the biosystem became compact and dense, resulting in excellent performances. It is the microbial activity for the biomineralization process that causes mainly calcium carbonate deposition to reduce pore size (35% average 510 nm pore diameter) in Bio-AAFC samples, thereby verifying the bacterial endospores can persist for a long time inside AAFC. Specifically, Bio-AAFC, having magnificent durability and crack-healing abilities explores the recycling of industrial dry wastes in a promising way.

Impact of ureolytic and nonureolytic bacteria on self healing of artificial cracks in biofortified concrete

ABSTRACT Two species of ureolytic bacteria (Bacillus sphaericus, Bacillus megaterium), and one non-ureolytic (Bacillus cohnii) were examined alone and in combination for their impact on crack repair and compressive strength in biofortified concrete. A combination of Bacillus cohnii and Bacillus megaterium achieved the maximum compressive strength (47.29 MPa) in 1 OD cells/ml after 28 days of curing in conventional 30 MPa strength of concrete. Further fly ash-blended (25%) concrete with Bacillus cohnii and Bacillus megateriume exhibited the compressive strength of 42.42 MPa at after 28 days of curing. The combination mix of Bacillus cohnii and Bacillus megaterium completely healed artificial cracks of 0.3 mm width by the formation of calcium carbonate. Further, XRD and SEM with EDS analysis confirmed that all three bacteria were able to make calcium carbonate precipitate in two phases of calcite and aragonite. The maximum amount of calcium carbonate mineral was deposited in the co-culture mix of bacteria Bacillus cohnii and Bacillus megaterium as confirmed by XRD test.

The role of bacterially induced calcite precipitation in self-healing of cement paste

Self-healing processes in cementitious materials have been intensively studied in recent years. They present a promising way to increase the durability of concrete and extend the service life of concrete structures, thus mitigating the impacts of the construction industry on public budgets and the environment. The role of bacterially induced calcite precipitation (BICP), the basis of one of the processes, is not fully understood yet, even though many researchers have reported improved mechanical properties or water tightness of cracked cementitious composites self-healed in the presence of bacteria. Therefore, the authors of this paper are focused on detailed characterization of BICP with the purpose of determining the direct impact of bacteria on the self-healing in cement composites. Three different bacterial strains, namely Sporosarcina pasteurii (SP), Bacillus cohnii (BC) and Bacillus pseudofirmus (BP) were studied. Their viability and metabolic activity in optimal and sub-optimal conditions were compared. Several analytical methods (XRF, TGA, TCD, FTIR) were used to characterize the attainable BICP depending on the particular strains. BP proved to be the most suitable bacterium, showing excellent viability and sufficient calcite production, even in sub-optimal conditions (10 °C temperature). Microscopic analysis of the self-healing products in artificially cracked cement paste showed that the application of BP with precipitation media lead to complete sealing of the crack by very dense calcite crystals (less than 1 μm), while porous structure of larger crystals (up to 30 μm) was formed in reference samples not treated with the bacteria, where only spontaneously induced calcite precipitation (SICP) occurred. As a result, it was proved that the bacteria directly contribute to the self-healing of cementitious materials and that BICP leads to the formation of a superior crack-sealing material.

Development of Bacterium for Crack Healing and Improving Properties of Concrete under Wet–Dry and Full-Wet Curing

Concrete cracking is inevitable, coupled with increased permeability, exacerbating the adverse impacts of atmospheric conditions and chemical attacks. Calcium carbonate precipitation resulting from certain microorganisms’ metabolism is a novel approach that can self-heal the cracks and improve concrete properties. In this study, the development and effect of bacteria Bacillus cohnii on crack healing, regained compressive strength after pre-cracking, sorptivity, water absorption, and concrete microstructures were investigated. For this purpose, a Bacillus cohnii bacterial concentration of 105 cells/mL was used as a water replacement in the concrete mixtures. Two methods subsequently cured the prepared concrete specimens: wet–dry (W-D) cycle and full-wet (F-W). In the wet–dry cycle, the cast specimens were immersed in water for 24 h and then kept at room temperature for 24 h, which was considered as one cycle; this process was repeated for 28 days. In the full-wet curing, specimens were immersed in water for 28 days. However, the curing water was changed every 24 h to facilitate the essential oxygen supply for bacterial activity to precipitate calcium carbonate. The results revealed that 90% and 88% surface healing was noticed in full-wet and full-dry pre-cracked specimens at 28 days.

Comparative performance of different bacteria immobilized in natural fibers for self-healing in concrete

Carrier compounds besides the type of bacteria for microbial induced calcite precipitation are equally important in self-healing concrete. This study investigates the potential utilization of natural fibers namely, coir, flax, and jute fibers to carry bacterial spores for self-healing in concrete. The comparison is drawn for introducing calcite precipitation bacteria namely Bacillus subtilis KCTC-3135T, Bacillus cohnii NCCP-666, and Bacillus sphaericus NCCP-313 in concrete along with calcium lactate pentahydrate and urea as organic nutrients. The findings from visual, microstructural, and mechanical properties of self-healing concrete suggest that natural fibers were capable of substantial immobilization potential for bacterial spores. Flax fibers provided better protection to bacteria with improved crack-healing and regain in compressive strength, whereas coir fibers resulted in higher compressive strength. It is also observed that B. sphaericus NCCP-313 precipitated more calcium carbonate as a healing product with uniform healing in the entire length of cracks in concrete. Consequently, approximately 75–85% and 60–65% healing rates were attained by fiber immobilized bacteria in 7 days and 28 days pre-cracked specimens, respectively.

Optimization of Sporulation and Germination Conditions of Functional Bacteria for Concrete Crack-Healing and Evaluation of their Repair Capacity.

While microbial spore production and germination of bacteria have been widely studied for their applications in animal husbandry, aquatic products, medicine, and food, few studies have investigated their use for the crack-healing of concrete. To effectively heal the cracks in concrete, studies suggest that the rate of sporulation and the germination of bacteria should be sufficiently high. This study investigates the effects of different carbon sources, nitrogen sources, Mn2+ concentrations, and external culture conditions on the sporulation rate and analyzes the effects of the pH value, heat activation, germinants, various cations, and nutrients on the germination of spores. Bacillus cohnii (B. cohnii) is chosen as the bacterium to be mixed in concrete because of its alkalophilic nature. The mineralization activity of spores after germination and the crack-healing capacity of concrete are studied. The optimal culture medium and the optimum external conditions for spore production are obtained. The total cell count and sporulation rate of bacteria obtained on this medium are 3.14 × 109 CFU/mL and 92.6%, respectively, under the optimum external conditions. The optimal pH value for the spore germination of B. cohnii is 9.7. While the cation Na+ strongly stimulates the germination of B. cohnii spores, other cations (such as K+, NH4+, and Ca2+) do not stimulate spore germination. The optimal concentration of Na+ is 200 mM. The germination rate of spores in the control group concrete specimen (room temperature 24°C) was more than 50%, thus suggesting that B. cohnii bacteria can be used in the self-healing of concrete cracks. The mineralization activity test proves that the spores of B. cohnii have a mineralizing function after germination, and the crystals produced by microbial-induced carbonate precipitation (MICP) are of pure calcite. When the crack width of the concrete specimen with spores of B. cohnii is less than 1.2 mm, it can be completely repaired after 28 days of healing.

The effect of temperature on bacterial self-healing processes in building materials

This paper is focused on the bacterial induced calcitation for the crack healing. The bacteria applied for this purpose are from group which is adapted for growth in the high pH environment as a concrete in hydration phase and their metabolic activity leads to create of calcite. In this study, three different bacteria strains (Sporosarcina pasteurii, Bacillus cohnii, Bacillus pseudofirmus) were applied and the influence of various temperatures on their microbial properties was investigated. Our previous experiment indicated that one of the applied bacterial strain in spores form (Bacillus pseudofirmus) are able to survive the temperatures in the range from -20 °C to 140 °C. The experiment described in this paper extends the previous study and determines the effect of different temperatures on the change in growth activity. In this experiment, bacterial activity was determined based on the change of absorbance in 640 nm by spectrophotometric measurements. The experiment was performed at optimal temperature (30 °C) and lower temperature (10 °C) and it used suitable broth for calcitation. The results showed that the beginning of metabolic activity was shifted by 40 to 50 hours. Only Bacillus cohnii showed different results because its metabolic activity was nearly zero at 10 °C.

Biodegradation of aliphatic waste components of oil sludge used micro symbiont of Sponge Niphates sp.

Investigation has been carried out using a sponge Niphates sp. as biodegradation agent of the aliphatic hydrocarbon component of crude oil sludge. The experiment was carried out by colonizing and making suspension of Bacillus pumilus strains GLB197 and Bacillus cohnii strains of DSM 6307, isolated from sponge Niphates sp. Both types of micro symbiont suspension were mixed with sludge waste for 30 days. There are five biodegradation parameters observed, carried out every five days of incubation. Parameters that observed during incubation were pH, gas bubbles and fermentation odor. Meanwhile, the optical density of the media was measured using spectrometer-D20+, Aliphatic components degradation was measured using GC-MS. Observation of biodegradation parameters showed there was a change in pH from 7 to 6, formed gas bubbles, resulting in fermentation odors and showed an increased optical density by an average of 35%. These four parameters occur during the interaction of 10-20 days of interaction in the media. There were 20 types of aliphatic components identified in the crude oil sludge waste. Analysis of four aliphatic components, namely Tridecane, Octadecane, Tricosane, and Nonacosane showed a decrease in the average concentration of 48.11 %.