Spread Plate Method Research Articles

Isolation and treatments of Aeromonas hydrophila and Staphylococcus lentus implicated in the seasonal autumn mortalities of farm-raised Cyprinus carpio, Basrah governorate, Iraq

Isolation and treatments of Aeromonas hydrophila and Staphylococcus lentus implicated in the seasonal autumn mortalities of farm-raised Cyprinus carpio, Basrah governorate, Iraq

Ochratoxin A and citrinin in green coffee and dietary supplements with green coffee extract

The aim of this study was to determine the degree of mold contamination and mycotoxin levels in commercially available green coffee products and dietary supplements with green coffee extract. The study included 34 samples from green coffee products: raw beans (n = 16), ground coffee (n = 15) and instant coffee (n = 3), as well as 22 samples from dietary supplements in form of capsules (n = 19), tablets (n = 2) and sachets (n = 1). Total mold count was determined with spread-plate method. Anamorphic mold were identified based on their microscopic morphology and the type of sporulation. Concentrations of mycotoxins, ochratoxin A and citrinin, were quantified by means of HPLC-fluorescence detection. Molds, typically Aspergillus spp. and Penicillium spp., were found in 94% of green coffee beans, 100% of ground and instant coffee samples, and 55% of dietary supplement samples. None of the samples contained detectable levels of citrinin. Ochratoxin A (0.4 ng/g) was detected in only one sample of raw green coffee beans, but in up to 40% and 67% of ground and instant coffee samples, respectively. Mean concentrations of ochratoxin A in ground and instant coffee samples were 3.28 ng/g and 4.09 ng/g, respectively, and maximum concentrations amounted to 6.65 ng/g and 7.44 ng/g, respectively. Ochratoxin A (mean concentration 9.60 ng/g, maximum level 31.4 ng/g) was also detected in up to 58% of the supplement capsules, but in none of tablets and sachets.

In vitro bactericidal, antidiabetic, cytotoxic, anticoagulant, and hemolytic effect of green-synthesized silver nanoparticles using Allium sativum clove extract incubated at various temperatures

AbstractThe current research aimed to evaluatein vitrobiological activities of green-synthesized silver nanoparticles using theAllium sativumclove extract. The stability of green-synthesized silver nanoparticles was evaluated via storage at 4°C, room temperature (37°C), and calcined at 300°C, 500°C, and 700°C. The antibacterial effect was evaluated using agar well, spread plate, biofilm reduction, and cell proliferation inhibition assays. The cytotoxic and antidiabetic effects were determined via brine shrimp lethality, protein kinase inhibition, and α-amylase inhibition assays. DPPH scavenging, iron-chelating, anticoagulant, and hemolytic effects were evaluated. The highest inhibition ofKlebsiella pneumoniaewas observed when freshly prepared, calcined (300°C), and stored nanoparticles (4°C and 37°C) were applied (9.66, 9.55, 7.33, and 6.65 mm) against freshly prepared and calcined at 700°C which showed the highest inhibition ofPseudomonas aeruginosa(8.55 and 7.66 mm). Cell viability assay, biofilm reduction assay, and spread plate method showed a significant antibacterial effect of freshly prepared silver nanoparticles. Freshly prepared and calcined nanoparticles at 300°C and 500°C possessed strong antioxidant and iron-chelating activity. Among all the synthesized silver nanoparticles, freshly prepared and calcined nanoparticles (300°C and 500°C) increases the prothrombin time. Silver nanoparticles possessed significant anticoagulant properties and less toxic at least concentration toward human RBCs. In brine shrimp lethality assay, freshly prepared nanoparticles showed a stronger toxic effect and caused high mortality of larvae. Protein kinase inhibition assay revealed that freshly prepared nanoparticles had the highest zone of inhibition (18.0 mm) at 50 µg/disc. Green-synthesized nanoparticles would be used as potential therapeutic agents to overcome both infectious and noninfectious diseases.

MPN Drop Agar Method for Determination of Heterotrophic Microorganisms in Soil and Water Samples Using Tissue Plate as a Carrier

The most probable number (MPN) method is a culture-based method commonly used in the field of environmental microbiology to examine microbial populations in liquid substrates. The MPN tests require a wide range of special laboratory equipment, a lot of laboratory space, and skilled staff, which together limit their applicability. This paper presents a modified MPN method, which reduces the experimental requirements by applying tissue plate as a carrier. The modified MPN method introduces a fast-filled tissue plate with 5 × 5 squares as agar carrier, instead of the commonly used set of glass tubes. Further, self-refilling automatic syringe Socorex® was implemented to apply samples to the plate. The response of the modified MPN method was tested on eight selected bacterial strains as well as on soil and water samples. Simultaneously, all the strains and samples were tested by standard spread plate method. High linear correlation between the two methods was found, which makes a new modified MPN method a useful alternative within the field of environmental microbiology.

Studies on comparative antimicrobial activities of Aerva lanata and Momordica charantia leaf extracts

Screening and comparison of antimicrobial action of leaf extract of Aerva lanata and Momordica charantia. Ethyl acetate and methanolic extracts of leaves of plants were screened for antimicrobial activity using the cup plate method and the spread plate method against gram positive and gram negative reference organisms (Bacillus substilis and Escherichia coli).The standard antibacterial agent used for reference is Choramphenicol and the results were calculated as zone of inhibition. Methanolic extract showed comparatively broader and better antimicrobial spectrum than ethyl acetate extract in selected plants. Plant extracts showed dose dependent action, results were similar to the action of the standard Choramphenicol. Extracts of Aerva lanata and Momordica charantia demonstrated antimicrobial activity on tested microorganisms. Methanolic extracts showed higher antimicrobial potential than ethyl acetate extract. Aerva lanata extracts showed better response than Momordica charantia extracts in the cup plate method antibacterial activity with Bacillus substilis and Escherichia coli.

Susceptibility of Mature Staphylococcus Biofilms to Chinese Herbal Decoction Sanhuang Jiedu: An In Vitro Study.

Background External socking and washing with the Chinese herbal Sanhuang Jiedu decoction (SHJD) can effectively control local limb infections with bone and implant exposure. However, the antibiofilm activities of this decoction in vitro have not yet been investigated. Therefore, the aim of this study was to examine the effects and characteristics of SHJD on the mature biofilms of multidrug-resistant staphylococci on a titanium surface. Methods Biofilm-forming methicillin-resistant Staphylococcus epidermidis ATCC 35984 and S. aureus ATCC 43330, and non-biofilm-forming S. epidermidis ATCC 12228 were selected as the experimental strains. The mature biofilms were prepared on titanium surfaces. The five experimental groups were based on dilution concentrations (DC) of SHJD: the control group (biofilm incubated with 0.85% NaCl solution), the SHJD (DC:1/8) group (initial SHJD solution was diluted 1/8), the SHJD (DC:1/4) group, the SHJD (DC:1/2) group, and the SHJD (DC:1/1) group (initial SHJD solution). The effects of SHJD on the mature biofilms were observed with the bacterial spread plate method, crystal violet (CV) staining, scanning electron microscopy, and confocal laser scanning microscopy. Results After culture in tryptic soy broth for 72 h, ATCC 43300 and ATCC 35984 produced mature biofilms and ATCC 12228 did not. The optical density value of ATCC 12228 was 0.11 ± 0.02, significantly lower than that of ATCC 35984 (0.42 ± 0.05) or ATCC 43300 (0.41 ± 0.03) (P < 0.05). The mature biofilms of ATCC 43300 and ATCC 35984 clearly disintegrated when incubated for 12–24 h with SHJD (DC:1/1) or SHJD (DC:1/2), showing only scattered bacterial adhesion. In the SHJD (DC:1/4) group, although many residual bacterial colonies still clustered together, presenting a biofilm structure, it was very looser than that in the SHJD (DC:1/8) group in which the biofilm was similar to that in the control group. For ATCC 12228, only colony adhesion was observed, and the number of colonies decreased as the concentration of SHJD or the culture period increased. The quantitative results for the bacterial spread plate and CV staining showed significant differences between the SHJD groups (P < 0.05). Conclusion SHJD has antibiofilm activity against multidrug-resistant Staphylococcus strains. It weakens or disrupts already-formed mature biofilms on titanium surfaces in a concentration- and incubation time-dependent manner.

Microbial Load of Domestic Water Sources Treated with Moringa oleifera and Jatropha curcas Seed Powder

Aim: Microbial load of domestic water sources treated with Moringa oleifera and Jatropha curcas seed powder.&#x0D; Study Design: The container test method was used for the treatments. One gram (1.0 g) each of the plant seed (Moringa oleifera and Jatropha curcas seeds) powder was weighed and added separately into 1000 ml of water sample. The mixture was stirred rapidly for 3 minutes and allowed to stand undisturbed for 1 hour and 3 hours, after which the top water was decanted.&#x0D; Place and Duration of Study: Advanced Research Laboratory, Department of Microbiology, Gregory University Uturu, from May to September 2017.&#x0D; Methodology: Tenfold serial dilutions were used for processing of the domestic water samples, after which 0.5 ml of the water sample was cultured on the media using the spread plate method. This was incubated appropriately and other standard microbiological methods were employed to determine microbial loads and characterize the isolates.&#x0D; Results: The microbial counts were generally high, 0.6×101 – 2.5×102 cfu/ml (borehole), 0.8×101 – 6.3×103 cfu/ml (well), 2.0×101 – 1.4×104 cfu/ml (stream), while the total potential pathogenic bacteria counts (TPPBC) were the least in occurrence. Treatment with Moringa oleifera and Jatropha curcas seed powders showed a significant decrease in the microbial load. After treatment with 1.0g of the seed powder for one hour, an apparent decrease in the microbial load was noticed. When allowed for three hours (3 hrs), the counts further reduced to no growth for potential pathogenic bacteria (TPPBC) especially for water samples that had low counts.&#x0D; Conclusion: Moringa and Jatropha seeds powder showed efficiency as a biocoagulant and thus can be used for water treatment. Moringa seed powder had a greater potential to serve as an alternative coagulant for water treatment. The intervention improved the quality of water and will provide significant benefits to the health of the consumer rural populace.

A luminescent Zn(II) coordination polymer for selective metal ions detection and treatment activity on tuberculosis by inhibiting the ropA gene expression in Mycobacterium

ABSTRACT By utilising the mixed-ligand approach, a novel coordination polymer based on Zn(II) ions as nodes have been synthesised by the Zn(NO3)2 · 6H2O reacting with 1,4-H2NDC in the presence of a second N-donor ligand L, and its chemical formula is {[Zn(L)1.5(1,4-NDC)]·H2O} n (1, 1,4-H2NDC = 1,4-naphthalenedicarboxylic acid and L = N,N′-di(4-pyridyl)suberoamide). The as-synthesised complex 1 reveals outstanding water stability and it could be utilised as a platform of the multi-responsive recyclable luminescent sensor for the Al3+ and Fe3+ in water. The compound was synthesised in this research for the tuberculosis treatment. The biological function of the compound was assessed, and its detailed mechanism was explored. The Mycobacterium bacterial numbers in lung tissue was determined with spread plate method. In addition to this, the real-time reverse transcription-polymerase chain reaction (RT-PCR) method was conducted to determine the inhibition of compound against the ropA expression in Mycobacterium.

Antimicrobial and antioxidant effects of phlorotannin coating in spoilage prevention of tomato during storage

The microbial flora present on vegetables and fruit is commercially significant as they cause spoilage and potential health risks in consumers. Synthetic chemicals used to combat food microbes posed serious health and environmental hazards leading to the search for natural alternatives. In this study, phlorotannins from three Indian brown seaweeds, Sargassum myriocystum, Turbinariaornata, and Padina tetrastromatica were evaluated for antimicrobial, antioxidant, and fruit spoilage prevention potentials. The crude phlorotannin of Padina with the highest content (0.114±0.0005g kg-1) and antioxidant activity (92.65±3.43%) was purified by column chromatography. The Padina water fraction I with the highest antioxidant potential was characterized by Fourier Transform Infra-Red spectroscopy and a 5 % solution was applied as a coating on tomatoes segregated into control chilled (CC), treated chilled (TC), control non-chilled (CNC) and treated non chilled (TNC). Spoilage parameters were studied over twenty days, using a swab and spread plate methods to assess the microbial load on tomatoes. Compared to the control sets, the treated sets exhibited a low number of microbial colonies and retained a fresh look and yellow color. The antibacterial and antioxidant properties of phlorotannin coating as a natural alternative for preservatives, effectively reduced bacterial load, inhibiting the early decay of tomatoes

A novel image cytometry-based Lactobacillus bacterial enumeration method for the production of kettle sour beer

The demands for a variety of craft beer flavors have been increasing in the United States. To meet this rising demand, breweries have been experimenting with kettle sour beer that utilizes lactic acid-producing bacteria for fermentation. The current standard bacterial quantification method is insufficient for rigorous quality control, thus there is a need for a better method to monitor lactobacilli concentration in a kettle sour environment. In this work, an automated Lactobacillus counting method was developed using fluorescence-based image cytometry. Three commonly used species were cultured, the concentrations were measured using image cytometry and evaluated against the standard spread-plating method. This procedure was undertaken in vitro at different dilutions and the method was repeated with two species in a kettle sour environment at different time points. Both the in vitro and fermentation experiments were repeated three times. Results demonstrated that the new method was not significantly different when compared to the standard plating method in either controlled settings or within the kettle sour fermentation. The proposed method provides a rapid tool to monitor and control lactobacilli growth in kettle sour beer production, and allows for standardization of the products due to the availability of near instantaneous information for quality control.

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

Aim: To determine the bacterial and physicochemical analysis of topsoil from an electronic waste (e-waste) dumpsite within Port Harcourt metropolis using standard procedures.&#x0D; 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.&#x0D; 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.&#x0D; 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.&#x0D; 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.

Análise microbiológica da água de equipamentos odontológicos

Nos últimos anos, a relevância dos processos de esterilização e antissepsia tem sido bastante discutida entre os profissionais da área de saúde, incluindo cirurgiões-dentistas. No entanto, na prática odontológica, a qualidade da água utilizada nos equipamentos, aparentemente inofensiva, pode ser considerada um fator de risco para a saúde do paciente. Diante disso, o objetivo deste estudo foi verificar a presença de contaminação em equipamentos odontológicos e avaliar a qualidade da água dos reservatórios desses equipamentos, por meio de análise microbiológica. Foram coletadas 12 amostras de água diretamente dos reservatórios das cadeiras odontológicas e 12 amostras da seringa tríplice de seis clínicas multidisciplinares do Departamento de Odontologia da Universidade Federal do Rio Grande do Norte (UFRN). O plaqueamento foi realizado com base no método Spread Plate, em meio ágar BHI (Brain Heart Infusion) com incubação subsequente em estufa bacteriológica programada de 37,5°C a 38°C, por um período de 24 a 48 horas. Os resultados demonstraram que 83% das placas incubadas apresentaram crescimento expressivo de fungos filamentosos e bactérias que, devido ao odor de tutti-frutti e com base nas características macroscópicas, sugere-se que sejam do gênero Pseudomonas. Nesse contexto, os achados deste estudo atestam a urgência da elaboração de uma rotina de cuidados sanitários, viabilizando a descontaminação dos sistemas de água dos equipamentos bem como de seus reservatórios, atendendo aos requisitos de segurança do paciente, que deve ser inerente aos consultórios odontológicos.

Bioremediation of toxic metals in mining site of Zamfara metropolis using resident bacteria (Pantoea agglomerans): A optimization approach

BackgroundVarious clean-up techniques for heavy metals have been suggested and practiced for its biosorption from the contaminated or pollutant soil by using chemical and physical methods. But most of the methods are hazardous to the environment and expensive. This study was on how to determine the potential of resident bacteria in the removal of heavy metals from contaminated soils in Abare situated in Anka Local Government of Zamfara State, Nigeria. Thus, this study employed bioremediation technique for removal of heavy metals. ResultsThe preparation of Culture media and Isolation of bacteria of the different contaminated soils were achieved by spread plate method. Whereas, concentrations of the heavy metals (Lead (Pb), Copper (Cu) and Iron (Fe)) were determined by Atomic absorption spectrophotometer (AAS. Pantoea agglomerans was used for biosorption experiment. The concentrations of Pb ranged between 1.328 ± 0.493 to 2.326 ± 2.093 mg/L, Cu 0.234 ± 0.117 to 1.054 ± 1.486 mg/L and Fe 18.498 ± 11.462 to 27.754 ± 57.510 mg/L. The optimum temperature for biosorption condition was found to be 35 °C. More so, the optimum pH of (7) was observed for maximum biosorption of Pb and Cu ions by Pantoea agglomerans which may be attributed to homeostatic phenomenon and the availability of metal binding sites on the biosorbents. Metal uptake biosorption percentage revealed that Pantoea agglomerans absorbed 99.6% of Pb, 60% of Cu and 96% of Fe. ConclusionThis study revealed that Pantoea agglomerans potential for bioremediation of the three metals.

Bacteria Associated with Petroleum Hydrocarbon Contaminated Soil from NNPC Depot

The microorganisms associated with soil polluted with petroleum hydrocarbon were isolated at NNPC depot Ibadan, in Ido Local Government Areas of Oyo State. Soil samples were taken from three different points along the point of discharge. The spread plate method was used to isolate the microorganisms found in these rivers and were later identified. From the research, seven different species of microorganism were isolated, which are Pseudomonas sp, Bacillus sp., Micrococcus sp. Staphylococcus aureaus sp., Enterobacter sp., Klebsiella sp. and Escherichia coli. Pseudomonas species were found to be more prominent in the samples collected in a research work. This suggested that the isolates are resistant to the petroleum hydrocarbon and can be recommended as candidates for the clean-up of petroleum hydrocarbon contaminated soil.

ISOLASI ACTINOMYCETES SIMBION SPONGE PULAU BARRANG CADDI MAKASSAR SEBAGAI PENGHASIL SENYAWA ANTIMIKROBA

Sponges as a residence for microorganisms make it a High Microbial Abundance Sponge (HMAS) where 40-60% of the biomass of the sponge consists of microorganisms. One of the bacteria that originates from the sea and has long been known to be breed is Actinomycetes. Actinomycetes that are symbiotic with sponges are very numerous and rich in the discovery of new chemotypes with interesting therapeutic activities that are not found in terrestrial organisms. The purpose of this study was to determine the antimicrobial activity of Actinomycetes isolates obtained from Barrang Caddi Island sponges. Actinomycetes isolation was carried out by pour and spread plate methods. From seven isolates actinomycetes obtained from sponge collections from Barrang Caddi Island, Makassar City, South Sulawesi are able to inhibit test microbes in initial screening with antagonistic test.

Characterization of Probiotics Isolated from Intestine of Mackerel Fish (Rastrelliger sp.) from Lembata Regency of East Nusa Tenggara

The research aimed to isolate, characterize, and analyze the ability of lactic acid bacteria (LAB) potential as probiotics to produce hydrolase enzyme. The LAB was isolated using MRS agar by the spread plate method. The LAB characterization includes antimicrobial activity, tolerance to low pH, bile salt, salinity, autoaggregation properties, and ability to produce hydrolytic enzymes. The isolate which has the highest ability to inhibit Aeromonas hydrophila is KBP 3.3, while the isolate which inhibits the highest Streptococcus agalactiae is KBP 1.1.1. The KBP 3.3 and KBP 1.1.1 were able to survive at pH 1 for 24 hours with a survival rate of 93.6% and 98.3%. The KBP 3.3 and KBP 1.1.1 are tolerant to 7.5% bile salt concentrations for 24 hours of 99.46% and 99.11%. The KBP 3.3 is tolerant to 0.5 % salinity for 24 hours with the highest survival rate of 113.38%, while KBP 1.1.1 is 94%. The KBP 3.3 and 1.1.1 have autoaggregation properties of 92.18% and 87.84%. The KBP 3.3 produced the highest lipase enzyme, while KBP 1.1.1 produced the protease enzyme.Keywords: hydrolytic enzyme, lactic acid bacteria, mackerel, probiotic

Evaluation of antibacterial activity and molecular characterization of bacteria from Holothuria atra intestine collected from anthropogenic and non-anthropogenic region in Karimunjawa, Indonesia

Abstract. Sulardiono B, Widyorini N, Suprapto D, Ayuningrum D, Rahman A. 2020. Evaluation of antibacterial activity and molecular characterization of bacteria from Holothuria atra intestine collected from anthropogenic and non-anthropogenic region in Karimunjawa, Indonesia. Biodiversitas 21: 3149-3155. Evaluation of antibacterial activity and molecular characterization of bacteria from the intestine of Holothuria atra is needed in the anthropogenic region of Menjangan Besar, Karimunjawa. The research aims to evaluate antibacterial activity and molecular characterization of bacteria from intestine of Holothuria atra collected from anthropogenic and non-anthropogenic regions in Karimunjawa, Indonesia. Sea cucumber samples were collected at Menjangan Besar waters as anthropogenic region (code of HM) and Alang-alang waters as non-anthropogenics region (code of HA), Karimunjawa National Park, Indonesia. The H. atra sample collection was using purposive sampling method. Examination of the bacteria from isolation until molecular characterisation was done at the Tropical Marine Biotechnology Laboratory, Uiversitas Diponegoro. The isolation process was conducted using spread plate method, followed by bacterial isolates purification using streak plate method. Screening of antibacterial activity using the agar plug method, as well as molecular identification was conducted by 16S rDNA PCR amplification. The results of this study indicated that a total of 26 bacterial strains were successfully isolated from sea cucumber intestine, consisted of eleven bacterial isolates from H. atra in Menjangan Besar waters and the rest fifteen isolates from H. atra in Alang-alang waters. The preliminary assay of antibacterial test showed H. atra in both waters have potential bacterial isolates. Those bacterial isolates with antibacterial activity were HM1.2 and HA1.1, which based on molecular identification, showed isolates HM1.2 and HA1.1 had the closest similarity with Bacillus paramcoides and Vibrio alginolyticus consecutively, with BLAST homology 98% and 99%. The accession number for both isolates were LC550090 and LC550089, respectively.

Antibacterial Activity Of Essential Oil Of Syzygium cumini Leaf Against Pathogenic And Spoilage Bacteria Isolated From Cheese

local cheese has been reported to be a high risk food in Nigeria due to the possible transmission of numerous spoilage and pathogenic bacteria. This study therefore investigated not only the spoilage and pathogenic bacteria associated with local cheese sold within Ilorin metropolis but also microbiological food safety of the food through the use of Syzygium cumini oil as a preservative. Samples of wara were collected from selected seller across the metropolis and analysed for the presence of different bacterial species using spread plate method on appropriate selective growth media. All the bacterial isolates were identified using biochemical test. Invitro therapeutic efficacy of S.cumini oil aginst foodborne isolates was carried out using Kirby-Bauer method. The most six prevalent bacterial isolates were Bacillus subtilis, Proteus vulgaris, Escherichia coli, Listeria monocytogene, Salmonella typhimurium and Staphylococcus aureus were isolated from the wara. Hydrodistilled leaves of Syzygium cumini yielded 0.32% (v/w) of essential oil. Examination of the oil by GC and GC/MS uncovered that the greater part of the oil was comprised by sesquiterpenes. Other outstanding constituents were Cis-Beta-Farnesene (1.40%), Beta-Ocimene (2.11%), Beta-bisabolol (1.48%), Beta-Ocimene (2.11%), and D-Limonene (2.89%). Antibacterial action was measured base on the zone of inhibition and minimum inhibitory concentration. The oil repressed the development of Proteus vulgaris at 25% concentration, Escherichia coli at 12.5% concentration, Listeria monocytogene at 6.25% concentration, Salmonella typhimurium at 12.5% concentration and Staphylococcus aureusat 6.25% concentration while no action was recorded against B. subtilis. Highest activity, represented by diameter of zone of clearance around the loaded wells was recorded for Staphylococcus aureus. Accordingly, the oil can be an elective method for averting spoilage in stored wara in this manner expanding its time span of edibility.

Identification Of Cu Resistant Bacteria From Tin Mining-Affected Sea Sediment

Tin mining activity at sea has a various negative influence on the environment. One of them is heavy metal contamination that can affect the life of fisheries biota. This research conducted to isolate and test the Cu heavy metal resistance of marine sediment bacteria that are affected by tin mining activity. Sediment sampling was carried out in the area of tin mining to the mudflat neared to the mangrove area. Bacterial isolation was carried out by spread plate method and bacterial characterization included cell shape, Gram staining and biochemical tests. Furthermore, bacterial isolates tested for resistance to metals with concentrations of 10 ppm, 20 ppm, 40 ppm, 80 ppm and 100 ppm. The results showed that bacterial isolates originating from marine sediments affected by mining activity were resistant to Cu heavy metal at various concentrations. Isolates B6, B8 and A10 showed the highest resistance up to a concentration of 100 ppm

Quarantine Time Validation for UC-MSC Isolation

Background & Aim Umbilical cord tissue is a potential source of Mesenchymal Stem Cells (MSCs). These cells have been widely used especially in pre-clinical and clinical applications resulting its various effectiveness. Banking process of umbilical cord involves transportation from the hospital where the cord is received to the banking facility to be processed further. Prior to processing, pre-sterility test must be conducted and while waiting for the result, the umbilical cord is quarantined. This quarantine process has to be carried out carefully to maintain cell's recovery and viability. Suitable range of time should be chosen to deliver high quality cells. In this study, we aim to determine the maximum time for quarantined umbilical cord to be processed. Methods, Results & Conclusion One umbilical cord (UC2018030001) was used in this study. Medium transportation is used to carry the umbilical cord to banking facility. Prior to processing, pre sterility was done using spread plate method by spreading the medium transport onto TSA agar. After the results were out and passed, the tissues were put into quarantine medium and were processed at day 0, 7, 14, and 21 for MSCs isolation. The tissue was processed using explant method and sub-cultured until 3rd passage. Graphic shown below are the cell doubling level from different passages at various days of quarantine. The doubling level remain constant with maximum deviation at 0.512. Results showed that tissues which was quarantined until day 21 still produce cells normally. The cell number are slowly increasing because the higher the passage, the more pure the cells are so they can propagate easily. This shows normal growth of normal cells. Their doubling level are also on track with no significant difference. In brief, umbilical cord tissue can be quarantined until 21 days and still produce cells normally.