Phylogenetic Relationship of Bacterial Species Involved in Bioremediation of Hydrocarbon Polluted Soils

The bio-stimulation of hydrocarbon degrading microbial population in soil using agricultural wastes was carried out. Top soil (0-25 cm depth) from three points were bulked to form composite soil samples, 6 kg each of the composite soil sample was weighed and transferred into 150 plastic buckets with drainage holes at the base. The soil in each plastic bucket was spiked with 300 ml crude oil and amended with different concentrations of agro-wastes and allowed for duration of 30, 60 and 90 days. The soil samples were then collected and analysed for both total heterotrophic bacterial counts and crude-oil utilizing bacterial counts. Data collected were subjected to a three-way analysis of variance and significant means were separated using Least Significant Difference Test at 5% probability level. The result showed that the application of the amendments increased the bacterial counts in the soil at different treatment levels. However, a higher proliferation rate was observed with bacteria counts exposed to higher waste concentrations compared to their counterparts exposed to lower waste concentrations. The phylogenetic relationship of the hydrocarbon degrading bacterial species shows that the identified bacteria were in two clusters: cluster 1 consist of Bacillus cereus, Bacillus thuringiensis, Bacillus altitudinis, Pseudomonas aeruginosa, Proteus mirabilis and Proteus penneri, while cluster 2 consist of Serratia marcescens, Providencia rettgeri and Enterobacter asburiae. The bacterial species obtained shows a greater relationship, this imply that the similarity of the bacterial species could be the reason for their high proficiency in degrading the hydrocarbons in the soil.

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Characterization of Indigenous Bacterial Communities in Crude-Oil Impacted Sites at Obagi Town, Onelga, Rivers State, Nigeria

Fine Focus ◽

10.33043/ff.2.1.7-14 ◽

2016 ◽

Vol 2 (1) ◽

pp. 7-14

Author(s):

Chioma Blaise Chikere ◽

Chinyere Augusta Ajuzieogu ◽

Michael Chukwugoziem Miller

Keyword(s):

Crude Oil ◽

Heterotrophic Bacteria ◽

Bacterial Species ◽

Polluted Soil ◽

Soil Samples ◽

Isolation And Characterization ◽

Significant Difference ◽

Rivers State ◽

Hydrocarbon Utilizing Bacteria

Hydrocarbon utilizers are expected to be indigenous in crude-oil polluted environments. The isolation and characterization of hydrocarbon utilizers is often a key strategy in bioremediation of hydrocarbon-polluted environments. In this study, crude-oil polluted soil samples from Obagi town, Onelga, Rivers state were enumerated and characterized for putative hydrocarbon utilizing bacterial populations. Biochemical characterization identified five bacterial species representative of five genera: Bacillus, Pseudomonas, Acinetobacter, Micrococcus and Staphylococcus. Amongst the genera of bacteria isolated, Bacillus had the highest frequency of occurrence (40%). The mean count of total heterotrophic bacteria was 1.7 X 107 cfu/g, while hydrocarbon utilizing bacteria (HUB) count mean density was 1.0 X 107 cfu/g for the three soil samples. Statistical analyses revealed no significant difference at p>0.05 between Total Heterotrophic Bacterial (THB) and Hydrocarbon Utilizing Bacterial (HUB) counts, suggesting that most of the bacteria present in the sampled sites were hydrocarbon utilizers. Findings from this study suggest the presence of indigenous putative hydrocarbon utilizing bacteria in the crude-oil polluted soil of Obagi town. Hence, a promising potential exists for future bioremediation studies on the site.

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Bacteriological Analysis of Top Soil from an Electronic Waste Dumpsite in Port Harcourt Metropolis

South Asian Journal of Research in Microbiology ◽

10.9734/sajrm/2020/v7i330173 ◽

2020 ◽

pp. 28-34

Author(s):

Janet Olufunmilayo Williams ◽

C. J. Ugboma ◽

Faith Ibiene

Keyword(s):

Waste Disposal ◽

Electronic Waste ◽

Bacterial Count ◽

Soil Samples ◽

Coliform Bacteria ◽

Percentage Composition ◽

Central Processing ◽

Port Harcourt ◽

Top Soil ◽

Significant Difference

Aim: To determine the bacterial and physicochemical analysis of topsoil from an electronic waste (e-waste) dumpsite within Port Harcourt metropolis using standard procedures. Place and Duration of Study: The study area was Kaduna street, beside Fruit and Vegetable Garden Market, Mile l which is located in Port Harcourt, Rivers State, Nigeria. The coordinates are 4o47’57.5” N 7o00’02.7” E. The duration of study was between March and September, 2019. Methodology: The waste disposed were mainly television sets, computer monitors, radio sets, stoves, laptops and central processing units. Soil samples were cleared off top debris, collected within 5cm of the top soil from four (4) different points of the dumpsite and a control was collected from area devoid of waste disposal, 20m away from the dumpsite. The five samples were kept in clean sterile polythene bags. Contamination observed from soil samples was attributed to the waste disposal. The total heterotrophic bacterial count was performed using l gram of soil from e-waste dumpsite in a 9-fold serial dilution using a spread plate method, in duplicates on dried nutrient agar plates and incubated at 30°C for 24hours. Centrimide agar plates were used to obtain Pseudomonas isolates and incubated at 37°C for 24 hours while MacConkey agar plates were used to isolate coliform bacteria, incubated at 0c for 48 hours. The physicochemical parameters were determined using standard methods. Results: Seven (7) bacterial genera were isolated from the topsoil of the e-waste dumpsite and they were Staphylococcus, Pseudomonas, Kluyvera, Bacillus, Micrococcus, Chromobacterium and Pectobacteria species. Staphylococcus spp. had the highest percentage composition of 42.3% and Kluyvera spp, the lowest percentage composition of l.9% of bacterial isolates found in the topsoil of the electronic waste dumpsite. The total heterotrophic bacterial count ranged from l.30 x l06 to l.97 x l06 cfu/g, total coliform count was 3.05 x l03 to 7.98 x l03 cfu/g and total Pseudomonas count ranged from l.00 x l02 to 2.88 x l03 cfu/g with a significant difference at .05 probability level to the control samples. The temperature ranged from 27.67±0.580C to 28.00±l.000C with a control of 29.00±l.000C, pH value ranged from 6.33±0.58 with a control of 7.00±0.00. The pH values were lower than the control indicating that the soil was slightly acidic to neutral. Moisture content had 4% with a control of 2.7%, an organic matter of l7.33±0.58 with a control of 4.47±0.58. Conclusion: The presence of the isolated organisms could cause public health risk or environmental hazard. Proper education and legislations on handling of e-waste in the society should be intensified to forestall waste related problems along the food chain.

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The Effects of Enrichment Nutrients on the Growth of Indigenous Bacteria Species in Spent Lubricating Oil Contaminated Water

Journal of Applied Life Sciences International ◽

10.9734/jalsi/2020/v23i330152 ◽

2020 ◽

pp. 39-48

Author(s):

Tudararo-Aherobo Laurelta

Keyword(s):

Bacterial Species ◽

Organic Wastes ◽

Lubricating Oil ◽

Contaminated Water ◽

Cow Dung ◽

Pseudomonas Sp ◽

Bacterial Isolates ◽

Mineral Salts ◽

Bacterial Counts ◽

Significant Difference

Aim: This study was conducted to compare the effects of enrichment nutrients, NPK (Nitrogen, Posphorus, Potassium) and organic wastes on the growth of indigenous bacterial species in spent lubricating oil contaminated water. Six bacterial species which were isolated from spent lubricating oil impacted soils (Pseudomonas sp., Bacillus sp., Actinomyces sp., Acinetobacter sp., Enterobacter sp., and Micrococcus sp.,) and showed profuse utilization of spent lubricating oil on screening, were used for this study. Place and Duration of Study: The study was conducted in the laboratory of the Department of Environmental Management and Toxicology, Federal University of Petroleum Resources, Effurun, between 2018 and 2019. Methodology: The study and was conducted using Mineral Salts Medium broth, spent lubricating oil substrate and NPK (20:10:10), Chicken droppings and Cow dung as nutrient sources (biostimulants). The effect of the biostimulants on the growth of the bacterial isolates was assessed weekly for 14 days by measuring the turbidity, bacterial counts and pH. Results: Pseudomonas sp. recorded the highest count of 1.16E+19 CFU/ml, 2.53E+17 CFU/ml and 1.74E+14 CFU/ml for biostimulation with NPK, Chicken droppings and Cow dungs respectively. The treatment with NPK enhanced the bacterial isolates most, of the three treatments used at the end of the test period. The pH values obtained for the test cultures at the end of the study, ranged from 6.52±0.02 for Enterobacter sp. in Cow dung treated cultures to7.85±0.03 for Pseudomonas sp.in NPK treated cultures. The values were within the optimum biodegradation range of 6.50 -8.50. There was significant difference between the bacterial counts obtained with the cultures treated with NPK and Chicken droppings (P=0.006), between NPK and cow dungs (P = 0.031) and between NPK and the control (P = 0.033). The study affirms the benefits of using organic wastes in the bioremediation process of hydrocarbon contaminated sites; it enhances the nutrients required by the bacteria for the remediation process and it’s a waste management strategy for disposing these organic wastes at very minimal costs and in an ecofriendly manner.

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An investigation of sudden death in farmed infantSiamesecrocodiles during winter and spring in Hainan, China

Indian Journal of Animal Research ◽

10.18805/ijar.v0iof.6999 ◽

2017 ◽

Author(s):

Jiping Zheng ◽

Jifeng Zeng ◽

Guiying Guo ◽

Jiahao Jiang ◽

Nuo Yang ◽

...

Keyword(s):

Micrococcus Luteus ◽

Bacterial Species ◽

Abdominal Cavity ◽

Edwardsiella Tarda ◽

Internal Organs ◽

Antimicrobial Drugs ◽

Highly Sensitive ◽

One Year ◽

Enterobacter Asburiae ◽

Proteus Penneri

Sudden deaths without any specific clinical sign occurred among one year old young ones on Siamese crocodile (Crocodylus siamensis) farms in Hainan province, China, during winter and spring in 2012 - 2013. The autopsy showed that the internal organs seemed normal except full of clear ascites in abdominal cavity. There were total six bacterial species isolated, Edwardsiella tarda, Proteus penneri, Proteus vulgaris, Enterobacter asburiae, Citrobacter freundii and Micrococcus luteus from either the heart bloods or the spleens and livers. The virulence of isolated strains was tested by intraperitoneal inoculation in female Chinese edible frogs (Rana rugulosa or Hoplobatrachus rugulosus). The six-day mortality revealed that E. tarda was the most pathogenic, whereas P. penneri and E. asburiae exhibited no infection at all, and the rest demonstrated similar inconspicuous toxicity. All these findings suggest that E. tarda would be the principal suspect causing death of Siamese crocodile. The sensitivities of the E. tarda isolates were tested against 20 kinds of antimicrobial drugs. The results showed that ceftazidime, norfloxacin and amikacin were highly sensitive to the isolates of E.tarda.

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Physicochemical Parameter of Palm Oil and Soil from Ihube Community, Okigwe, Imo State Nigeria

International Letters of Natural Sciences ◽

10.18052/www.scipress.com/ilns.62.35 ◽

2017 ◽

Vol 62 ◽

pp. 35-43 ◽

Cited By ~ 10

Author(s):

Christian Ebere Enyoh ◽

Ezechiel Amarachukwu Ihionu ◽

Andrew Wirnkor Verla ◽

Patricia Ngozi Ebosie

Keyword(s):

Palm Oil ◽

Analytical Methods ◽

Physicochemical Analysis ◽

Soil Samples ◽

Palm Tree ◽

Imo State ◽

Top Soil ◽

Significant Difference ◽

C 50

This study assessed the quality of palm oil and soil quality from Ihube community, Okigwe L.G.A of Imo state in October 2015. The soil samples were collected from the top soil and the palm oil were collected immediately after extraction. The soil and palm oil were analyzed using standard analytical methods and the results obtained were compared to standards. The physicochemical analysis of the soil samples showed that ranges of EC (323.33±0.57-480.33±1.53 us/cm); Temperature (28.0±0.26 29.1±0.20 °C); %MC (20.9±0.80-40.8±0.10); %SOM (1.92±0.61-4.85±0.01) which were within the acceptable limits while CEC (5.471-8.1063 mg/kg); iron (0.311-0.731 mg/kg) and lead (0.004-0.054 mg/kg) were is not appreciable. The palm oil result showed that MC (0.32±0.09 %) which is above the acceptable limit, pH (4.71±0.01-4.74±0.01) and viscosity (23.41±0.44-106.23±0.55 mPas) obtained at temperature range 20 °C-50 °C. The FFA result for 10days showed ranges from (2.15±0.01-6.07±0.05 mgNaOH/g) with Fe, (2.00±0.01-5.43±0.51 mgNaOH/g) with Pb and (1.27±0.03-5.04±0.03 mgNaOH/g) without contamination. This was tested using anova at p = 0.05 respectively and showed significant difference between the results. However, the result obtained from these areas shows good quality soil for palm tree cultivation.

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Impacts of Artisanal Crude Oil Refining Activities on Soil Microorganisms

Journal of Advances in Biology & Biotechnology ◽

10.9734/jabb/2018/v20i330077 ◽

2019 ◽

pp. 1-9

Author(s):

Douglas, Salome Ibietela ◽

Cornelius, Blessing Tamunonegiyeofori

Keyword(s):

Crude Oil ◽

Soil Microorganisms ◽

Microbial Population ◽

Soil Samples ◽

Oil Refining ◽

Bacterial Counts ◽

Total Hydrocarbon ◽

Hydrocarbon Content ◽

Microbiological Methods ◽

Total Hydrocarbon Content

Aim: To evaluate the effect of illegal crude oil refining activities on soil microorganisms using standard microbiological methods. Study Design: This study employs laboratory experimental design, statistical analysis of the data and interpretation. Place and Duration of Study: Soil samples were taken once a month for three months (May- July, 2018) from Ke in Degema Local Government Area of Rivers State, Nigeria, where illegal crude oil refining activities are ongoing. Methodology: Using standard microbiological methods, total culturable heterotrophic bacterial counts, total fungal counts, Hydrocarbon utilizing bacterial and fungal counts were analysed to evaluate the effect of the activities. Total hydrocarbon content of the soil samples was also analysed. Results: The populations of the total heterotrophic bacterial, fungal and hydrocarbon utilizing bacterial (HUB) and fungal (HUF) counts of the contaminated soil were enumerated. The mean total heterotrophic bacterial counts in Station 1 around the pot ranged from 2.5 x 105 to 1.8 x 106cfu/g, fungal counts ranged from 2.1 x 103 to 4.4 x 104cfu/g, HUB (HUB) counts ranged from 4.2 x 104 to 6.4 x 105cfu/g and hydrocarbon utilizing fungal (HUF) counts ranged from 1.5 x 103 to 4.0 x 103cfu/g. The results of soil samples taken 20m away from the Pot location ranged from 7.0 x 105 to 8.2 x 106cfu/g for total heterotrophic bacterial counts, fungal counts ranged from 2.3 x 103 to 1.5 x 104cfu/g, HUB ranged from 4.7 x 104 to 5.7 x 105cfu/g and HUF ranged from 2.0 x 103 to 3.5 x 103cfu/g. Also, the results of total heterotrophic bacterial counts for Station 2 ranged from; 4.3 x 105 to 3.3 x 106cfu/g, fungi 2.0 x 103 to 3.3 x 104cfu/g, HUB ranged from 3.8 X 104 to 5.4 x 104cfu/g and HUF 1.6 x 103 to 3.5 x 103cfu/g, while 20m away from the Pot total heterotrophic bacteria ranged from 1.3 x 107 to 6.5 x 107cfu/g, fungi 5.8 x 103 to 1.4 x 105cfu/g, HUB 5.4 x 104 to 1.1x 105cfu/g and HUF 3.1 x 103 to 4.7 x 104cfu/g. While the control samples taken from inside the community where no such activity is on, ranged from 2.6 x 107 to 7.9 x 107cfu/g for total heterotrophic bacterial counts, total heterotrophic fungal counts ranged from 2.8 x 104 to 5.3 x 104cfu/g, HUB 2.0 x 102 to 3.1 x 102cfu/g and HUF 2.0 x 101 to 2.3 x 101cfu/g. twelve bacterial genera were identified and eight fungal genera: Bacillus, Alcaligenes, Flavobacterium, Acinetobacter, Pseudomonas, Micrococcus, Proteus, Serratia, Enterobacter, Streptococcus, Escherichia, Staphylococcus, Penicillum, Aspergillus, Fusarium, Mucor, Rhizopus, Geotrichum, Candida, and Cladosporium. Total hydrocarbon content ranged from 106 to 281mg/kg across the locations. When compared with the control, it was observed that the microbial population and diversity were adversely affected. These variations observed in the microbial population are indicative of the effect of the illegal refinery on the soil microorganisms. Conclusion: The results of this study indicates that the continuous contamination of the soil environment by the activities of illegal crude oil refining, lead to a decrease in microbial population and diversity. This may result in devastating ecological damage, adversely affecting the ecological balance which may affect food chain and in turn animals and humans.

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Biodiversity and Phylogenetic Relationship of Total Hydrocarbon Degrading Genes in Selected Bacteria Species

Asian Journal of Biology ◽

10.9734/ajob/2021/v12i330164 ◽

2021 ◽

pp. 19-29

Author(s):

R. B. Agbor ◽

S. P. Antai ◽

S. E. Ubi

Keyword(s):

Phylogenetic Relationship ◽

Niger Delta ◽

Bacterial Species ◽

Tertiary Structures ◽

Niger Delta Region ◽

Uncultured Bacteria ◽

Complete Knowledge ◽

Great Genetic Diversity ◽

Relationship Of ◽

Microbial Attack

Hydrocarbons which forms the bulk of soil and water pollutants in the Niger Delta region of Nigeria differs in their susceptibility to microbial attack and degradation. Considering the importance of hydrocarbons in the economy of any nation, a complete knowledge of the hydrocarbons degrading capabilities of associated microbial species becomes increasingly important and indispensable. The research was aimed at assessing the biodiversity and phylogenetic relationship of hydrocarbon degrading genes in selected bacteria species. Sequences of nucleotides and amino acids of hydrocarbons degrading genes in 12 species of bacteria such as Pseudomonas aeruginosa I & II, P. stutgeni, Thalassosqir spp. I & II, Alcavorox spp., Arthrobacter spp., Martellela spp., P. taenensis, Aneuribacillus species, Rhodococcus spp. and Uncultured bacteria, were retrieved from the National Center for Biotechnology Information (NCBI) and analysed for their variability in physicochemical properties, percentage identity and similarity, G-C content, secondary and tertiary structures, their biodiversity and their phylogenetic relationship using MEGA 6 soft-wares, GOR IV, Phyre, Genscan and SIB Protparam. The analysis showed great genetic diversity and polymorphism in the hydrocarbon degrading genes present in the bacteria species. The phylogenetic tree showed that the genes in most species had undergone mutation. Martella spp had the highest probability or chance of mutating with E-value of 7e-129. Only two of the bacteria species had low instability index indicating that majority of the bacteria species contains or possessed the unstable nature of the hydrocarbon degrading genes. The biodiversity of bacterial species and their genetic distance was ascertained.

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