Spread Plate Research Articles

Isolation and characterization of potential multiple extracellular enzyme-producing bacteria from waste dumping area in Addis Ababa

Extremozymes are innovative and robust biocatalysts produced by various microorganisms from harsh environments. As thermophilic organisms can only develop in a few places, studying them in geothermal environments has provided new insights into the origins and evolution of early life and access to significant bio-resources with potential biotechnology applications. The work aimed to isolate and identify likely multiple extracellular enzyme-producing thermophilic bacteria from an Addis Ababa landfill (Qoshe). The streaking approach was used to purify 102 isolates acquired by serial dilution and spread plate method. The isolates were morphologically and biochemically characterized. Thirty-five cellulases, 22 amylase, 17 protease, and nine lipase-producing bacteria were identified using primary screening methods. Further secondary screening using Strain safety evaluation; two bacterial strains (TQ11 and TQ46) were identified. Based on morphological and biochemical tests, they were found to be gram-positive and rod-shaped. Furthermore, molecular identification and phylogenic analysis of selected promising isolates confirmed the identity of the isolates, Paenibacillus dendritiformis (TQ11) and Anoxybacillus flavithermus (TQ46). The results indicated that, multiple extracellular enzyme-producing thermophilic bacteria isolated from a waste dumping area in Addis Ababa offer useful features for environmental sustainability in a wide range of industrial applications due to their biodegradability and specialized stability under extreme conditions, increased raw material utilization, and decreased waste.

Solation, Screening and Optimization for Laccase production by Fusarium spp. under submerged fermentation

Laccases are oxidases with broad substrate specificity and ability to oxidize various phenolic and nonphenolic compounds. This study was carried out to isolate and characterizes laccase producing fungi from the environmental samples. Soil samples were collected from different locations. Suspensions of the samples (1g in 10 ml sterile physiological saline) were serially diluted, inoculated onto Czapek Dox Agar (CZA) containing antibiotic chloramphenicol and incubated for 7 days at 30 o C. The fungal isolates were characterized macroscopically and microscopically. Screening was done using Czapek-Dox agar was supplemented with 0.1% tannic acid, 0.5% Bromophenol blue, 0.05 ml in 25 ml of Methanol Guaiacol the pure culture of the single colonies from the spread plate method were inoculated on the media and incubated at room temperature for 5-6 days. After 5-6 days the plates were checked for clear transparent zone.

Formulation of Effective Microorganism [EM] to Analyse its Impact on Municipal Wastewater Management

Discharging wastewater from households, industries, and other sources into the environment without treatment hurts the ecosystem as well as people’s quality of life by causing negative impacts on all living species, including health concerns to humans such as diarrhoea which kills 3,50,000 Indian children annually and only 33% of India’s urban wastewater is treated. The majority of energy is dedicated solely to the aeration process, which regulates the degradation of organic matter (sludge) found in wastewater, which is one of the most critical issues in wastewater treatment. As a result, the goal of this research is to develop long-term strategies for degrading organic materials using Effective Microorganisms Technology (EM). EM, also known as Effective Microorganism, is a carefully selected and proportioned liquid culture of fermenting microorganisms. In this study, three microorganisms were isolated from various sources, including soil, yogurt, and milk by serial dilution and spread plate technique, each isolate was made into a pure culture after incubation. They were identified by the VITEK method as Pseudomonas aeruginosa, Lactobacillus sporogenes, and Candida albicans. The microbial consortium was then formed by incubating all three organisms in a nutrient broth that had been modified. Each microorganism culture and formulated microbial culture were added to sewage samples taken in Erlenmeyer flasks. All samples were tested for physico-chemical parameters such as pH, BOD, COD, TDS, TSS, SO4 ²– and Cl- after a three-day incubation period at room temperature. The standard deviation method was used for statistical analysis and Microsoft Power BI was used to visualize the results. Lactobacillus sporogenes was the only culture that showed a significant reduction in all parameters except one when compared to other cultures that showed a reduction in most parameters except a few. As a result, Lactobacillus proved to be effective in wastewater treatment when compared with other cultures. Further research is required to determine its true potential.

L-Asparaginase – A prospective anticancer enzyme on MCF-7 cell lines extracted from soil Microflora

This study aims at investigating the soil bacteria for L asparaginase enzyme production and its anti-cancer effect on MCF 7 cell line. In this study, 30 soil samples were screened for bacterial producing L asparaginase enzyme. The soil samples were collected from various places in Ethiraj College, Chennai. Isolation was done using spread plate technique, biochemical and microscopic characterization for the potential isolates revealed the bacterial genera as Bacillus, Escherichia and Pseudomonas species. Of the thirty samples collected, six isolates produced pronounced L asparaginase enzyme. These samples were further subjected to crude enzyme extraction and estimation. The isolates S2 and Ni5 showed maximum production of enzyme with 1090 units/ml of activity. Dialysis was carried out to concentrate the enzyme and molecular characterization was performed using SDS PAGE to separate the proteins according to the molecular weight which was found to be approximately 100 to 116 kDa. Anti-cancer activity was evaluated on MCF 7 tumour cell line. The cell viability was dose dependent and IC50 of S2 was 62.5 μg and Ni5 31.2 μg respectively.

Detection of microorganisms with antibacterial activities from different industrial wastes and GC-MS analysis of crude microbial extract.

Many soil microorganisms' i.e., bacteria and fungi produce secondary metabolites called antibiotics. These are used for the treatment of some of the bacterial, fungal and protozoal diseases of humans. There is a need for isolation of a broad spectrum of antibiotics from microorganisms due to the emergence of antibiotic resistance. In the present study two antibiotic producing bacteria Klebsiella pneumoniae and Bacillus cereus were isolated from pharmaceutical and poultry feed industry of Hattar, Haripur Pakistan. Total 10 waste samples were collected from different industries (Marble, Ghee, Soap, Mineral, Steel, Poultry Feed, Pharmaceutical, Qarshi, Cosmetic and Glass). Thirty-three bacterial strains were isolated from industrial wastes of these ten different industries. Fourteen out of thirty-three bacterial strains exhibited antimicrobial activities against at least one of the test microbes considered in this study including Escherchia coli, Staphylococcus aureus and Salmonella typhi. The bacteria were isolated by standard serial dilution spread plate technique. Morphological characterization of the isolates was done by Gram staining. Nine bacterial isolates out of fourteen were initially identified as B. cereus and five as K. pneumoniae through biochemical characterization. The antibacterial activities were tested by well diffusion method. Maximum number of antibiotic producing bacteria were isolated from pharmaceutical and poultry feed industry based on the results of primary screening, the most potential isolates S9, S19, S20, S22 and S23 were selected for secondary screening. The maximum activity against E. coli and S. aureus was recorded by bacterial isolate S19 i.e zones of inhibition of 6.5mm and 9mm while S20 showed 7.5mm and 6mm zones respectively. Molecular identification was carried out on the basis of 16S rRNA sequence analysis. Finally, the isolates were identified as B. cereus accession number LC538271and K. pneumoniae accession number MT078679. Analysis of bacterial extract S20 through GC-MS indicated the presence of 8 compounds of diverse nature and structure. Present study suggests that wastes of pharmaceutical and poultry feed industry may have antibiotic producing bacteria. These bacteria could be utilized for the production of antibiotics. B. cereus and K. pneumoniae isolated from wastes of poultry feed and pharmaceutical industries have the potential to produce antibiotics and could be used to control the microbial growth.

Ability Analysis Of Waste Milkfish (Chanos chanos) As Alternative Medium Of Nutrient Agar On Escherichia coli And Staphylococcus aureus Growth

Waste milkfish got a lot from the residual processed milkfish production of Sidoarjo. The intestines, liver, and kidneys of waste milkfish contain protein, carbohydrates, and fat. The abundant availabilities were usable as a bacterial growth medium, including Nutrient Agar. Nutrient Agar has a high price and is produced by many foreign companies that can trigger a hard availability in laboratories of domestic healthcare, especially in frontier, outermost, and underdeveloped areas. This research aims to analyze the ability of waste milkfish as an alternative medium for Nutrient Agar on the growth of Escherichia coli and Staphylococcus aureus. The ability has been observed by the characteristics and number of bacterial colonies. This research was held in the bacteriology laboratory of the Medical Laboratory Technology Department, Health Polytechnic of Surabaya health ministry. The research type was True-experimental with a post-test-only control group design. Based on the preliminary test results, the waste milkfish mass variations were found in 7 grams, 7.5 grams, 8 grams, 8.5 grams, and 9 grams. Bacteria have been inoculated by the spread plate method, bacterial colony growth has been calculated by the Total Plate Count method. The data were analyzed by the Kruskal-Wallis test with a Sig. value <0.05 that there were differences in the number of Escherichia coli and Staphylococcus aureus colonies in milkfish waste media with various mass. The results showed that the best ability of waste milkfish as an alternative medium for Nutrient Agar on bacterial growth was found in 9 grams mass variation. the average number of Escherichia coli colonies were 186 × 1013 CFU/mL and Staphylococcus aureus colonies were 188 × 1013 CFU/mL.

Analysis of microbiological quality of Tapioca (abaachaa) sold at different bus stops in Ihiala along Owerri-Onitsha road

This research work was done to ascertain the bacterial and fungal quality of mixed tapioca popularly called Abaachaa sold at five major bus stops in Ihiala Local government area of Anambra State. The samples were bought and collected from the vendors; mostly women using sterile conical flask. Spread plate technique was done after serial dilution of the samples. The bacterial count on nutrient agar showed 3.8×1012, 7.7×1011, 3.8×1012, 6.7×1011, 5.7×1011 cfu/g for Amorka, Uli Center, Total;Post (Okija) bus Stops respectively. Also bacterial count on MacConkey are 3.5×1011, 7.5×1010, 3.5×1011, 7.5×1010, 6.5×1010 were recorded. For the fungal count; 2.7×1010, 7.6×109, 3.8×1010, 5.6×109, 7.6×109 were recorded. Some bacterial species like Bacillus species; Staphylococcus species; Bacillus subtilis; Enterococcus faecalis; Serracia species; Escherichia coli were found. Fugal species like Rhizopus, Penicillium, and Aspergillus species were also isolated. The significance of this study is that it has revealed that the Abaachaa sold at these five bus stops are contaminated and should be eaten with care. Because there was no record of heat treatment in its final preparation both in gravy making and mixing, contamination of the food was noticed. The study recommend that Tapioca (Abaachaa) should be warmed before eaten; though enjoyment of the food is better when taken while it is cold.

Production of Bioplastic by Local Strains of Bacillus subtilis using Watermelon Peels as Substrate

The amount of environmental contamination brought on by the careless disposal of plastic garbage has increased to 400 million tons every year on a global scale. These synthetically generated traditional polymers are not easily biodegradable. This work was therefore undertaken to isolate Bacillus subtilis from soil with potential to produce bioplastic: Poly-ß-hydroxybutyrate (PHB). Samples of soil were collected from various locations (BG = Botanical Garden, FARD = Fine Art Refuse Dumpsite, SHRD = Suleiman Hall Refuse Dumpsite, FVM = Faculty of Veterinary Medicine animal paddock) within Ahmadu Bello University, Zaria, Nigeria. Spread plate technique was used to isolate B. subtilis on nutrient agar, and the isolates' cultural, morphological, and biochemical properties were identified. The isolates of B. subtilis were screened for PHB production using two different methods: the plate assay method and slide technique using Sudan black B dye. The PHB was then produced using submerged fermentation with watermelon peel as sole source of carbon in the production medium. The PHB was extracted using Sodium-hypochlorite method and the quality of the PHB was determined using FT-IR analysis. Four isolates of Bacillus subtilis were obtained from the soil samples (50 %), one each out of the two samples (50 %) per unit location (BG2, FARD3, SHRD2, and FVM4). The screening revealed that all the isolates were PHB producers. The B. subtilis isolate SHRD2 from the students’ dormitory was found to produce the highest PHB yield of 0.98 g/L from the watermelon substrate, whereas isolate from the animal paddock (FVM4) yielded the lowest quantity of the PHB (0.12 g /L). The biopolymer's (bioplastic) identity was confirmed to be PHB based on the peaks in the FT-IR spectra, which displayed wave numbers for a variety of functional groups, including -O-H, C-H, C-O, and C=O. It was concluded that the local isolates of B. subtilis have potentials for PHB production using watermelon peels as source of carbon and energy.

Pantoea agglomerans, Klebsiella pneumoniae, and Shigella flexneri isolated from the Cisadane River as multiresistant bacteria to copper and dyes

Copper pollution in Cisadane is a serious environmental issue that needs to be resolved immediately due to its negative impacts on river ecosystems. Bioremediation utilising indigenous bacteria offers excellent potential to restore copper‐contaminated river water. This study aimed to obtain indigenous copper‐resistant bacteria isolated from the Cisadane River as copper bioremediation agents. Bacteria from Cisadane River water samples were isolated by the spread plate method on Luria Bertani medium containing 3 mM CuSO4. Resistance was determined based on the minimum inhibitory concentration value, while copper concentration was measured using an atomic absorption spec‐ trophotometer. The results presented a total of 13 bacterial isolates with a minimum inhibitory concentration of up to 8 mM CuSO4. Sequence alignment analysis was performed on three selected copper‐resistant bacteria, i.e. isolate IrCis1, IrCis4 and IrCis13, which were identified as Pantoea agglomerans, Klebsiella pneumoniae and Shigella flexneri based on 16S rRNA, respectively. Each isolate accumulated copper at 1.19 mg, 1.34 mg and 0.92 mg/g DW of cells, with copper biosorption potentials of 73.74%, 70.17% and 67.73%, respectively. In conclusion, P. agglomerans strain IrCis1, K. pneu‐ moniae strain IrCis4 and S. flexneri strain IrCis5 isolated from the Cisadane River can be used as copper bioremediation agents.

The Impact of Effluents Derived from Palm Oil Extraction Plants on Soil Microbial Community

In order to know the consequences of the effluent on the microflora of the soil and its respective fertility, the size of the experimental sample was defined, which was represented by five locations, from which ten portions of soil were collected for microbiological analysis. Unimpacted soil samples served as controls. Total counts of heterotrophic bacteria, coliforms, and fungi were assessed using the pour plate and spread plate method. The result showed that the counts of total heterotrophic bacteria, total coliforms and total fungi (3.8x103CFU/mL, 3.5x103CFU/mL and 3.4x102CFU/mL respectively) of the impacted soils were lower compared to the values (3.6x106 CFU/mL, 3.6x106CFU/mL and 2.6x104UFC/mL respectively) of the non-impacted soils that served as control. Bacterial genera involved in margarine production processes include Bacillus spp, Micrococcus spp, Klebsiella spp, Pseudomonas spp, Staphylococcus spp, Salmonella spp. The genera of fungi isolated from soils with effluents from palm oil extracting plants include Aspergillus spp, Rhizopus spp and Mucor spp. The results obtained indicate that effluents from palm oil extracting plants have an immediate negative impact on the microbiota of the soil, since its application could cause changes that affect the microbial communities. Therefore, it is recommended that adequate guidelines be established for the possible pre-treatment and safe discharge of effluents from palm oil mills, in addition to avoiding their effect on the microbiota and soil fertility.

Bacteriological analysis of OKPA (Bambara nut Moi Moi) sold in NKWO market of Okija town near Legacy University Okija.

This research was conducted to ascertain the bacterial content of Okpa(Bambara nut moi moi) sold in Nkwo market in Okija town. This market is strategic as it is a popular market serving Legacy and Madonna Universities and the entire Okija town. Five samples were collected and analyzed using spread plate technique after tenfold serial dilution. Nutrient agar was used to enumerate total colony count while MacCokey agar was used for coliform count. After incubation for 24 hours at 37 ℃, 1.2 x105, 2.1 x 104, 4.5 x 104 and 2.0 x 104 cfu/g were observed for nutrient agar. For the coliform count, 1.1 x 102, 2.1 x 10, 1.2 x 10 and 1.0 102 cf/g were observed on MacConkey agar. The results showed that, though the Okpa were contaminated, the coliform count was not at the level of causing disease. Also, the research revealed that the sources of the pathogenic bacteria are through unhygienic actions of the people that prepares the food and those that hawk them. The research also recommended that adequate sanitary practices and procedures should be adopted during Okpa preparation.

Inaccuracies of the ISO 11731 Method for Environmental Validation of Legionella in Building Water Systems: Opportunities to Improve Sensitivity and Detect Viable but Non-Culturable Legionella

Current environmental diagnostics for the detection of Legionella fail to detect viable but non-culturable Legionella, have sensitivity limitations and are time-consuming (10-14 days to results). The objective of this study was to compare Legionella detection results between the standard ISO 11731 and an innovative Legionella detection method that utilizes a hybrid methodology of traditional microbiology and molecular detection. In this study, four hundred and seventy-six (476) potable building water samples were analyzed with ISO 11731 and the novel method in parallel. Of the 476 total samples that were tested, a discrepancy of 21% was observed when comparing the ISO 11731 method to the novel method. Separating the samples based on hazard control methods yielded a 15.4% discrepancy for chlorinated systems (n = 284) and a 29% discrepancy for monochloraminated systems (n = 192). The data presented here conclusively show inaccuracies in environmental validation for building water systems based on results returned by the standard ISO 11731 method. This is especially evident in systems primarily disinfected with monochloramines. Overall, these data highlight the need for new and innovative methods to overcome the inaccuracies of the traditional ISO 11731 spread plates to prevent disease and injury caused by Legionella.

Determination of Microbiological Quality, Proximate Composition and Physicochemical Parameters of Margarine Produced from Oil Blends of Palm Kernel, Coconut and Melon

The potential of margarine production from blends of melon, coconut and palm kernel oil, and the microbiological qualities and physicochemical parameters of the product were studied. The microbiological evaluation was carried out at the Microbiology Laboratory, Cross River University of Technology, Calabar, Nigeria, using pour plate and spread plate methods to ascertain the viable cell counts. The physicochemical composition, proximate properties were determined using standard methods. The enumeration of total heterotrophic bacterial count revealed low microbial count of 2.6 x 103 CFU/g, total coliform count was 2.5 x 102 CFU/g and total fungal count. 2.4 x 102 CFU/g. The value was within permissible limit of 104 CFU/g. The biochemical characterization and identification revealed Staphylococcus spp, Bacillus spp, Salmonella spp, Escherichia coli, and Micrococcus spp are the suspected bacterial isolates. While the fungal isolates are Aspergillus spp, Fusarium spp, Penicillium spp and Rhizopus spp. The percentage occurrence revealed that Staphylococcus spp had the highest prevalence of 33.3%, followed by Escherichia coli (26.7%), Salmonella spp (20%), Bacillus spp (13.3%), Micrococcus spp (6.7%). While the fungal isolates had the following prevalence; Aspergillus spp (50%), Penicillium spp (16.7%), Rhizopus spp (16.7%) and Fusarium spp (16.7%). Proximate analysis revealed moisture (15.0±0.5), ash (35.0±0.5), crude (17.50±0.25), lipid (22.50±0.03), protein (2.218±0.01), and carbohydrate (7.78±0.01). Results for physicochemical analyses showed that Iodine value was 0.372, saponification value was 22.103 and acid value, 1.159. This result indicates that palm kernel oil, melon oil and coconut oil had the best potential for the production of margarine which would be acceptable due to the low microbial count. However, the presence of Staphylococcus spp and Salmonella spp renders the product a potential health hazard in line with FDA guidelines. Therefore, maximum hygienic conditions during production is advocated and further analysis recommended on these samples to discover their maximum importance and industrial utilization.

In Vitro Degradation and Photoactivated Antibacterial Activity of a Hemin-CaP Microsphere-Loaded Coating on Pure Magnesium.

Photoactivated sterilization has received more attention in dealing with implant-associated infections due to its advantages of rapid and effective bacteriostasis and broad-spectrum antibacterial activity. Herein, a micro-arc oxidation (MAO)/polymethyltrimethoxysilane (PMTMS)@hemin-induced calcium-bearing phosphate microsphere (Hemin-CaP) coating was prepared on pure magnesium (Mg) via MAO processing and dipping treatments. The morphology and composition of the coating were characterized via scanning electron microscopy, Fourier transform infrared spectrometer, X-ray diffractometer and X-ray photoelectron spectrometer. Corrosion behavior was evaluated through electrochemical and hydrogen evolution tests. The release of Fe3+ ions at different immersion times was measured with an atomic absorption spectrophotometer. Antibacterial performance and cytotoxicity were assessed using the spread plate method, MTT assay and live/dead staining experiment. The results showed that the corrosion current density of the MAO/PMTMS@(Hemin-CaP) coating (4.41 × 10-8 A·cm-2) was decreased by two orders of magnitude compared to that of pure Mg (3.12 × 10-6 A·cm-2). Photoactivated antibacterial efficiencies of the Hemin-CaP microspheres and MAO/PMTMS@(Hemin-CaP) coating reached about 99% and 92%, respectively, which we attributed to the photothermal and photodynamic properties of hemin with a porphyrin ring. Moreover, based on the release of Fe3+ ions, the MC3T3-E1 pre-osteoblasts' viability reached up to 125% after a 72 h culture, indicating a positive effect of the coating in promoting cell growth. Thus, this novel composite coating holds a promising application as bone implants.

Diazotrophic Bacterial Response to Herbicide Toxicity: In vitro Analysis

Diazotrophic bacteria play critical role in biological nitrogen fixation (BNF), and the application of herbiced significantly affect growth and activities of these bacteria. To investigate this, 1.0 ml of broth stock culture containing 2.9 x 105 cfu/ml and 2.6 x 106 cfu/ml of Azotobacter and Bradyrhizobium, respectively were inoculated into 10.0 ml of their respective growth medium [ TDC broth, and Yeast extract mannitol broth (YEMB)] containing 0, 0.5, 1.0, 1.5 and 2.0 % v/v of the respective herbicides, atrazine, glyphosate, paraquat and 2,4-D. Thereafter, plate counts of the diazotrophs for each concentration was made at 24, 48, 72 and 120 h intervals using spread plate method on TDC agar and YEMA after incubation at room temperature (30 ± 2 ℃) for 72 h. The LC50 of the respective herbicides for Azotobacter and Bradyrhizobium was determined at 120 h using Probit analysis. Results showed that all tested concentrations except control, retarded diazotrophic bacterial population growth. Growth reduction increased progressively with increased concentrations of herbicides (P < 0.05). In general, herbicides were found to suppress the growth of diazotrophs by 29.7 – 100 %. The LC50 indicated symbiotic Bradyrhizobium displayed greater sensitivity to tested herbicides than free-living Azotobacter (P < 0.05). Conclusively, herbicides suppressed diazotrophic bacterial growth.

Antibacterial Effectiveness Test of Lemongrass Leaf (Cymbopogon Citratus) Boiled Water In Decreasing The Number of Escherichia Coli Bacterial Colonies

The presence of disease can be caused by a microorganism (virus, bacteria, fungus and parasite), and the environment that can infect a person either directly or through intermediaries. Environmental components such as air, water, food, and others are the natural transfer paths of disease. So that humans must maintain personal hygiene, the environment and equipment that is often used such as cutlery to avoid the threat of pathogenic microorganisms. Contamination in the washing process of cutlery by water containing Escherichia coli bacteria will damage the quality of food and cause diarrhea. One of the efforts to reduce the number of germs is to disinfect using chemicals such as chlorine. However, the use of chemicals actually has a negative impact on health and the environment, we need alternatives that do not cause pollution and are safe for humans such as the use of natural materials. One of them is lemon grass which can be used as an antibacterial.
 
 This experimental study aimed to determine the antibacterial effectiveness of lemongrass leaves with concentrations of 10%, 15%, 20% and 25% in reducing the number of Escherichia coli bacterial colonies after 5 minutes, 10 minutes and 15 minutes of contact, using the spread plate method with EMBA media. The results showed a decrease in the number of Escherichia coli bacterial colonies after treatment using boiled water from lemongrass leaves from a concentration of 10% after 5 minutes of contact. Based on the ANOVA analysis, it was found that p-value 0.05 there was a significant difference in each variation of concentration and contact time. The boiled water of lemongrass leaves with the highest level of effectiveness is in concentration of 25% with a contact time of 15 minutes.

Isolation and Characterization of Resistant Bacterial Species Isolated From Shallow Well Water Situated Close to Graves as a Public Health Menace in Osogbo, Osun State

This study was designed to evaluate the physicochemical and bacteriological qualities of well water situated close to graves. Total heterotrophic bacterial count (THBC), total coliform count (TCC) and fecal coliform count (FC) were done on nutrient agar, MacConkey and Eosine methylene blue agar, respectively, using spread plate method at incubation conditions of 37℃ for 24 hours for THBC and TCC whereas 44.5℃ for 24-48 hours for fecal coliforms. Membrane filtration was used for Vibrio spp. count on TCBS agar while conventional biochemical method was performed for bacterial identification. Isolated bacteria were subjected for antimicrobial resistant testing using 8 families of antibiotics. Bacteria with MAR index value of ≥75 were sent for genomic identification and sequencing. The average value of the ten water samples analyzed over the period of study had high total suspended solids (mg/l), phosphate (mg/l); magnesium ion (mg/l) contents and a very low dissolved oxygen mg/L content. The THB count was (5.0×107-1.35×108 CFU/ml), TCC count (3.55×105- 1.04×106 CFU/ml), FC count (2.10×105-6.90×105 CFU/ml) while Vibrio spp. count was 45-144 CFU/100 ml and the MPN estimated reading for coliform was also high. The percentage occurrence and MAR index value of the bacteria isolated from the well water, respectively were Vibrio spp. 23%; 75, Klebsiella spp. 20%; 100, Bacillus spp. 14%; 100, Staphylococcus spp. 14%; 62.5, E. coli 12%; 62.5, Pseudomonas spp. 12%; 75, Glycomyces spp. 3%; 100 and Proteus spp. 3%; 50. The closeness of the well to the grave makes the decomposing leachates with high organic, inorganic, biological and poisonous metals sink into the underground aquifers. Stamford Journal of Microbiology, Vol.12 (1) 2022: 47-53

Isolation and Characterization of Antibiotic Producing Fungi from Soil

This study was aimed to isolate and characterize antibiotic producing fungi from the soil environment within Ahmadu Bello University main campus Samaru, Zaria. Soil samples were collected from five different locations within Ahmadu Bello University main campus for the Isolation of fungi. Spread plate method involving serial dilution technique was used for the Isolation using Saboraud Dextrose Agar (SDA). Six species of fungi were isolated from the soil samples and then characterized microscopically and macroscopically. The fungi were Aspergillus niger, Aspergillus fumigatus, Aspergillus sp, Penicillium sp, Fusarium sp (P14) and Fusarium sp (P15). Sensitivity test using Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae as test pathogens was employed to determine the ability of the fungal isolates to produce antimicrobials. All the fungal isolates were found to inhibit the growth of at least one of the test pathogens except Fusarium sp (P14). Aspergillus niger produces zones of inhibition of 9mm, 5mm and 6mm against Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae, Aspergillus fumigatus produces zones of inhibition of 5mm against Escherichia coli and Klebsiella pneumoniae. Aspergillus sp produced zones of 6mm against Staphylococcus aureus and Escherichia coli, Penicillium sp which produces a zone of 10mm, 7mm and 6mm against Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae. The findings of this study show that antibiotic-producing fungus are prevalent in the soil of Ahmadu Bello University's main campus in Samaru, Zaria, and that these strains could be used by pharmaceutical companies to produce antibiotics from local sources.

Bacteriological assessments of indoor air in the staff quarters of a tertiary institution in Benin City, Nigeria

Indoor air contains large number of airborne microorganisms such as bacteria and fungi and their estimation is important for use as index of cleanliness for any particular environment and to determine the relation they bear on human health. This study was aimed at determining the bacteriological air quality of the living rooms in some selected Staff Quarters in University of Benin and University of Benin Teaching Hospital. The airborne bacterial loads of indoor air in the living rooms of Twelve (12) Staff Quarters in University of Benin and University of Benin Teaching Hospital both in Benin City were determined using the Settle Plate methods. The houses were categorized as Apartment 1 which represents University of Benin Junior Staff Quarters (JSQ), Apartment 2 which represents University of Benin Teaching Hospital Quarters, and Apartment 3 which represents University of Benin Senior Staff Quarters (SSQ). The living rooms were sampled twice a month between April, 2017 and September, 2017. The airborne samples were studied using standard microbiological methods and Polymerase Chain Reaction and 16S rRNA techniques were used for the gene sequencing. The antibiotic susceptibility pattern and plasmid profile of the characterized airborne bacterial isolates were evaluated using spread plate and agarose gel electrophoresis methods. The temperature and relative humidity of the indoor air environment in the sampled areas in the living rooms were determined using the thermometer and hygrometer respectively. The mean indoor temperature and relative humidity for Apartments 1, 2 and 3 ranged from 26.10 ± 1.190C to 31.20 ± 0.870C and 81 ± 2.70% to 87 ± 1.89% respectively. The mean indoor airborne bacterial counts in Apartment 1 and Apartment 3 ranged between 1.00 x 103 ± 0cfu/m3 to 4.42 x 103 ± 1.42cfu/m3 and 1.09 x 103 ± 0.09cfu/m3 to 5.17 x 103 ± 3.17cfu/m3 respectively. In Apartment 2, the counts ranged from 1.00 x 103 ± 0.00cfu/m3 to 6.99 x 103 ± 4.69cfu/m3. The difference in the airborne bacterial counts obtained in the morning and afternoon period of study in Apartment 1 was statistically significant (P<0.05) while Apartments 2 and 3 showed no significant differences respectively (P>0.05). Ten airborne bacterial isolates were characterized, further characterization by molecular techniques, confirmed them to be identified as Staphylococcus aureus strain S33 R, Bacillus subtilis subsp. strain 168, Oceanobacillus manasiensis strain YD3-56, Streptomyces vietnamensis strain GIM4.0001, Actinosynnema pretiosum strain C-15003, Micrococcus caseolyticus strain 235, Ornithinibacillus composti strain GSS05 and Bacillus ectoiniformans strain NE-14. General and personal hygienic practices are important for healthy living, and living rooms used for close contact interactions and good ventilation.

Efektifitas Ekstrak Lidah Mertua (Sansevieria trifasciata var laurentii) Sebagai Antifungi Candida albicans

Candidiasis is a disease caused by Candida albicans, this disease has a mortality rate of > 25%, mainly affecting adults, with a frequency of women 2 - 3 times more than men (Raningsih et al., 2018). A commonly used fungi drug is ketoconazole. but it has many side effects so it is necessary to find for alternatives to natural ingredients, namely the leaf of lidah mertua (Sansevieria trifasciata var laurentii) because it contains compounds saponins, flavonoids, triterpenoids, and steroids as antifungals The leaf of lidah mertua were extracted by maceration technique. The purpose of this research was to determine the effect of antifungi of ethanol extract the leaf of Lidah Mertua (Sansiviera trifasciata var Laurentii) against the fungus Candida albicans. This research was experimental using the disc diffusion method with a spread plate technique in the inhibition zone formed at concentrations of 25%, 20%, 15%, 10%, 5%, positive control and negative control. The results of the One way Anova test is known that differ significantly. From the results of this research, it can be concluded that the ethanol extract of the leaves of the leaf of lidah mertua (Sansevieria trifasciata var laurentii) has an effect on the antifungal activity of Candida albicans Keywords : Candida albicans, Leaf of Lidah Mertua, Antifungal, Ketokonazole