Presence Of Clear Zones Research Articles

Identification and Study of Thermophilic Bacteria Found in the Sulfur-Infused Hot Springs of South Lampung

Thermophilic bacteria are microorganisms with the capability to produce a specific enzyme known as amylase, which exhibits resistance to heat and is referred to as thermostable. This study aims to isolate, characterize, and assess the enzyme-producing activity of bacteria from the hot springs of Way Belerang, Lampung, specifically amylase. The research procedures encompassed sample collection, isolation of thermophilic bacteria incubated in NA media for 48 hours, amylum testing with iodine after a 24-hour incubation in NA media containing starch/amylose, biochemical tests including TSIA, SIM, SC, urea test, sugar fermentation test, and catalase test. The results of the study yielded two isolates successfully isolated from the hot spring source, and Gram staining indicated that both isolates were Gram-negative bacteria. The assessment of bacterial activity in producing amylase was demonstrated by the presence of clear zones, with the largest bacterial diameter measuring 1.04 mm as measured using a vernier caliper. Characterization of the bacteria identified them as belonging to the Pseudomonas genus for isolates AB2 and AB3. Thus, the bacteria obtained in this study produce amylase enzymes that can be utilized in various industrial applications, including the food, beverage, textile, detergent, paper, pharmaceutical, and other sectors.

Antibacterial potential marine sponge extract and bacteria symbionts Callyspongia vaginalis from Kendit Waters Against the bacteria Vibrio harveyi

Vibrio harveyi is the reason for Vibriosis disease in shrimp which causes mortality above 80%. The diseases caused by these bacteria can be treated using antibiotics, but the use of antibiotics in fisheries and aquaculture is currently estimated to harm the environment. This research was conducted to determine the screening of sponge symbiont bacteria, the antibacterial potential of sponge extract, and secondary metabolites in sponge extract. This method includes an antibacterial test, antagonist test, and qualitative phytochemical test. The test results showed that the sponge extract of Callyspongia vaginalis had the potential against V. harveyi, this was indicated by the presence of clear zones at concentrations of 10%, 20%, 80%, which indicated that these concentrations had weak inhibition criteria, while at a concentration of 40% and 60% have medium barriers criteria. The screening test results for symbiont bacteria showed that only 4 active isolates showed the presence of antibacterial activity marked by the appearance of an inhibition zone around the paper disc and categorized as having weak inhibitory power. Phytochemical tests on this sponge extract showed it had secondary metabolites: flavonoids, alkaloids, terpenoids, and steroids.

Potential application of bioemulsifier RAG-1 as an anti-staling agent in flat bread quality.

Bread undergoes physicochemical processes known as 'staling', which limits shelf life and quality. Despite the fact that several chemical emulsifiers have been employed to combat this issue, they may offer risks to human health. In this investigation, the effects of bioemulsan, a natural bioemulsifier (BE), on bread quality and staleness were examined. The yield of emulsan generated by Acinetobacter calcoaceticus RAG-1 was 1.49g/L. The presence of clear zones around colonies, high emulsification value of 100%, and remaining surface tension below 40 mN/m after heating (at 250°C for 15-20min) verified emulsan thermal stability. BE-supplemented bread had a greater moisture percentage than the control, resulting in reduced crumb hardening and improved bread quality during storage as measured by moisture content. The first day after adding 0.5% emulsan, the hardness rose from 90.45 N (for the control) to 150.45 N. Texture analysis showed that although the hardness increased during storage, adding emulsan allowed obtaining bread with clearly softer crumb after 2 and 3days of baking, especially at 0.5% level (from 215.6 N for the control to 150.5 N for 0.5% BE-enriched bread after 2days, and from 425.7 to 210.25 N after 3days). Based on the sensory evaluation results, emulsan did not lead to any unpleasant changes on bread organoleptic parameters. Therefore, using bioemulsifier RAG-1 as a green emulsifier and anti-staling agent found to be more promising.

Effect of Plectranthus scutellarioides (L.) Leaf Extract as Natural Antibacterial Against Staphylococcus aureus and Escherichia coli Isolated From Dairy Cattle with Subclinical Mastitis

Mastitis is one of the causes of low milk production in dairy cows, and can be caused by bacteria such as Escherichia coli and Staphylococcus aureus. Plectranthus scutellarioides (L.) leaf has antibacterial compounds such as flavonoids, eugenol, polyphenols, steroids, tannins, and so on. The type of this research is experimental laboratory research that aims to test the antibacterial activity of the P. scutellarioides (L.) leaf extract with ethanol pro analysis as the solvent against E. coli and S. aureus bacteria isolated directly from the sample of dairy cow’s milk which CMT score is (+) 3, by measuring its inhibition ability indicated by the presence of clear zone using the agar-well diffusion method. The extract concentrations tested are 5%, 10%, 20% and 40%. The result of the antibacterial test of P. scutellarioides (L.) leaf extract with ethanol p.a. as the solvent showed antibacterial activity against E. coli and S. aureus bacteria isolated directly from cow's milk samples with subclinical mastitis at certain concentrations.

Pengaruh Waktu Fermentasi Madu dengan Bawang Putih Tunggal (Allium sativum L.) terhadap Daya Hambat Bakteri Propionibacterium acnes

Honey is a substrate that has widely used by the people of Indonesia for treatment, one of which is used to treat acne. Honey has antimicrobial inhibitory power, antimicrobial ability is also found in other natural ingredients, one of it is Single Garlic (Allium sativum L.). Community trend towards the two natural ingredients, one of which is by combining Honey and Single Garlic (Allium sativum L.) using the method of soaking Single Garlic (Allium sativum L.) in Honey. The purpose of the study is to determine the inhibitory power of honey bacteria from soaking / fermentation against propionibacterium acnes bacteria.Testing use the disc paper diffusion method with a sample of fermented Honey and a single Garlic (Allium sativum L.) against the length of honey fermentation time, positive control (Clindamycin 300 mg), and negative control (aquadest). Sample Honey and Garlic Sample (Allium sativum L.) were fermented length time at weeks 1,2,3 and 4. The results showed that the length of time honey fermentation can increase the inhibitory power of the growth of Propionibacterium acnes bacteria by the presence of clear zones around the disc paper. The diameters of honey fermentation in weeks 1,2,3 and 4 are 0.416 mm, 0.65 mm, 0.775 mm and 1.8 mm. The conclusion that the length of fermentation time of Honey and Single Garlic (Allium sativum L.) will increase in diameter in the clear zone of its inhibitory power known in the fourth week to have a maximum inhibitory power of 1.8 mm. Keywords: Fermented honey, Single Garlic,Inhibitory power of bacteria Propionibacterium acnes

Isolation and selection of amylase-producing microbes isolated from ragi tape and cassava tape available on the markets

Amylase has an important role in biotechnology development and occupies an important position in the world enzyme market, as a biocatalyst in various industrial fields. This study has the goal to find microbial isolates that have the ability to produce amylase enzymes. The study was conducted in two stages, namely: 1) Isolation and selection of microbes that can produce amylase enzymes using starch as substrate, was incubated for 4-7 days at 30°C. Microbial isolates that can produce amylase enzymes are characterized by the presence of clear zones around the colony after the addition of an iodine solution of 1% in the overgrown media of microbes, 2) Test the activity of amylase enzymes using a dinitrosalicylic acid reagent test. The activity of the amylase enzyme is determined by measurement using a spectrophotometer at a wavelength of 540 nm. The sample used comprised of 7 types of ragi tape and 2 samples from cassava tape that has been fermented for 5-7 days. The results obtained in the first stage were 65 microbial isolates, 16 of which had clear zones, consisting of 7 isolates from ragi tape samples and 9 isolates from cassava tape samples. In the enzyme activity test, there are several isolates that have the potential to produce amylase enzymes, these include R5I4 (0.897 ± 0.018 U/mL), R2I5.1 (0.814 ± 0.011 U/mL), R5I3 (0.727 ± 0,042 U/mL) (derived from cassava ragi tape samples) and T2I2.2 (0.812 ± 0.013 U/mL), T2I6.1 (0.817 ± 0.010 U/mL), T2I2.1 (0.735 ± 0.023 U/mL), T1I4 (0.755 ± 0.020 U/mL) (derived from cassava tape samples). The isolate with the highest enzyme activity is the R5I4 which has the value enzyme activity of 0.897 ± 0.018 U/mL and with a fairly high or moderate category, while the lowest enzyme activity is the T1I1.1 isolate of 0.284 ± 0.020 U/mL.

Antibacterial Activity Test Of Leaves Bidara (Ziziphus mauritiana Lam) Ethanolic Extracts Against Staphylococcus aureus

Infection is the most common ilnness in public. One of the triggers of infection is bacteria. Bidara leaf (Ziziphus mauritiana L) is a plant that has antibacterial compounds is phenolics, saponins and triterpenoids/steroids. The study aims to test antibacterial potency Staphylococcus. antibacterial activity testing was carried out using paper disc difussion methode, the etanol extract show antibacterial activity against Staphylococcus Aureus which is characterized by the presence of clear zone on the media. The results of ethanol extract of bidara leaves with concentrations of 30%, 50% and 70% against Staphylococcus aureus bacteria, the average diameter of inhibitory zones respectively were 8,39 mm, 10,64 mm and 12 mm with the positive control diameter of inhibition zone was 23,46 mm, the negative control diameter of inhibition zone was 0 mm.

Seaweed-Associated Fungi from Sepanjang Beach, GunungKidul, Yogyakarta as Potential Source of Marine Polysaccharides-Degrading Enzymes

Brown algae (Phaeophyceae) and red algae (Rhodophyta) are a group of seaweed that scattered all over the ocean. In addition, previous studies have reported the biotechnological potential of its associated fungi. However, there are only a few studies related to the extracellular enzyme of seaweed-associated fungi. The purposes of this research were to isolated brown algae and red algae associated fungi from Sepanjang Beach, GunungKidul, Yogyakarta, Indonesia, and screen its enzyme production. Padina sp., Asparagopsis sp., and Chondrophycussp. were collected from Sepanjang Beach, GunungKidul, Yogyakarta. Swab tap method on PDA medium and STD medium was conducted to isolate seaweed-associated fungi, while starch agar medium, agar medium, alginate agar medium, and carrageenan agar medium, was utilized to screen the enzyme activity with addition of povidone-iodine 10% reagent. The presence of clear zone around the colony indicated enzyme.

Bakteri Simbion Akar Mangrove Avicennia Sp. Sebagai Pendegradasi Pewarna Tekstil

Textile dye is a material that has a negative effect on environmental quality, one of the textile dyes is violet crystal. Biodegradation is one of the methods that used in waste treatment and can take advantage of microbial activity to degrade waste. Mangroves are ecosystems that are found along coastal waters in tropical and subtropical areas. Microorganisms such as bacteria are able to live and develop in mangrove roots. This study aims to isolate and identify the bacteria that live on mangrove roots morphologically and are able get degrade textile dyes. This research was conducted using experimental laboratory methods. Samples were isolated from mangrove roots obtained in Wonorejo Surabaya sterilely. The process of isolation and verification of bacterial morphology was carried out at the Microbiology Laboratory of Hang Tuah University, Surabaya. The method of isolation was carried out using the spread method, and the purification of the bacteria was carried out using the streak method. The results obtained 10 bacterial isolates from the results of isolation and morphological examination, while 2 isolates were able to degrade the violet crystal textile dye, called AB.1.2 and AB.1.5 which were indicated by the presence of clear zones.

SCREENING OF ACIT SULPHATE SOILS FUNGI FROM SOUTH KALIMANTAN AS SOURCE OF EXTRACELULAR ENZYMES

Kalimantan acid sulphate land has the potential to be developed into productive land, with good land optimization. Utilization of rhizosphere microorganism diversity, especially mold can potentially provide a solution in optimizing agricultural land, namely the ability to produce extracellular enzymes. This study aims to determine the potential of mold originating from acid sulphate fields in producing extracellular enzymes (pectinase, chitinase, glucanase, cellulase, and phosphatase). The study was conducted in June-July 2019 at the Microbiology Laboratory, Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development. Screening of extracellular enzyme-producing fungi was carried out on selection media. The results obtained by some isolates have the ability to produce extracellular enzymes. Indications of the ability of mold to produce extracellular enzymes are the presence of clear zones in the selection medium. In pectinase, chitinase and glucanase testing all isolates showed negative results. Potential isolates in producing extracellular enzymes include Penicillium sp. Paddy 4.1 (cellulolytic index 2.43), Clonostachys sp. KRMT 17.9 and Penicillium singorense KLK 13.7 (proteolytic indices 3.97 and 3,00, respectively). The difference in index values ​​indicates the variation in the level of enzyme activity.

Isolation and Characterization of Thermophilic Bacteria as Cellulolytic Enzyme Producer from the Hot Spring of Ie Seuum Aceh Besar, Indonesia

Cellulase enzymes can be isolated from thermophile bacteria obtained from the hot spring Ie Seuum, Aceh Besar. This research aimed to recover and characterize the isolates morphologically and biochemically followed by determination of the thermophile bacterial isolates potential as cellulolytic enzyme producers,. The sampling method in this research was conducted by a purposive sampling at temperature of 70 oC, 60 oC and 50 oC. Isolation of thermophilic bacteria was carried out on nutrient agar (NA) media. There were four isolates of thermophilic bacteria isolated recovered at 70 oC, five isolates at 60 oC, and seven isolates at 50 oC. Of the 18 isolates obtained, 15 of them were able to produce cellulase enzymes. Cellulase enzyme production can be determined by the presence of clear zones around bacterial colonies on CMC media after addition of 1% congo red drops and wash with 1 M NaCl. The highest five Cellulolytic Index (CI) values ​​were obtained from isolates ISB75; ISB64; ISB52; ISB54; ISB56 that were 1.23; 2.22; 1.39; 1.59; 1.10, respectively. Biochemical tests carried out on 5 isolates with the highest cellulolytic index values showed that the bacterial isolate were suspected to be from the genera of Bacillus sp.

Characteristics of Physicochemistry , Microbiology and Antibacterial Activities from Fermentation of Viscera Fish Sauce

The fermentation process involved the hydrolysis of lactic acid bacteria, single amino acid and peptide were produced and were accounted for their antibacterial activity. This study aims to determine the physical and chemical characteristics, total lactic acid bacteria, total plate count, and antibacterial activity of tuna viscera sauce. The research results showed physicochemical characteristics including color (L* 8.3, a* 1.3 and b* 5.7), viscosity of 10.38 cP, pH 5.00, salt content of 13.21%, total acid 0.74% and TVBN 28.00 mgN/gr) Proximate analysis were also identified, resulting in moisture 62.87%, ash 1.37%, protein 23.18% and carbohydrate 0.42%. The total lactic acid bacteria and total plate count respectively were 2.3 log CFU/gr and 2.3 × 101 CFU/gr. The results of the antibacterial activity tested on three pathogenic bacteria, Vibrio pharahaemolyticus, Salmonella typhimurium, and Escherichia coli, showed inhibition with the presence of clear zones.

Antagonistic activity of phylloplane yeasts from Moringa oleifera Lam. leaves against Aspergillus flavus UNJCC F-30 from chicken feed

Aspergillus flavus is widely known as an aflatoxin-producing fungus that frequently contaminates feed and affects livestock, which leads to severe health problem for animal and human. Biological agents have been proven to prevent this contamination since they can produce metabolites which have antagonistic activity. In this study, phylloplane yeasts isolated from Moringa oleifera leaf have shown an ability to inhibit the growth of Aspergillus flavus UNJCC F-30 collected from chicken feed. This research was conducted in three stages: (1) yeast isolation (leaf washing and direct method), followed by (2) antagonistic test using dual culture method, and (3) molecular identification using D1/D2 region of 26S rDNA. In the first stage, 38 yeast isolates have been succesfully obtained. These isolates were of different colors: peach pigment (60.5%), the non-pigmented yeast (26.5%), cream (10%), and orange (3%). Antagonistic activity against A. flavus UNJCC F-30 was tested based on growth, sporulation, and the presence of clear zones. Screening result showed that 12 yeast isolates are capable of inhibiting A. flavus UNJCC F-30. Among them, 4 isolates with the code K4, K10, K15, and K26 showed the highest antagonist ability. Molecular identification resulted that the 4 isolates show a similar identity with Aureobasidium pullulans UWFP 993 (100%), Aureobasidium melanogenum QCC:M017/17 (99%), Aureobasidium melanogenum QCC:M017/17 (100%) and Rhodotorula taiwanensis CBS:11729 (99%), respectively. Isolate K10 exhibited the highest percentage of inhibition activity among all isolates which is potential for application as biocontrol agent against A. flavus. As A. pullulans is a common yeast found on leaf surfaces of many Indonesian flora, therefore it can be considered as safe and alternative to reduce fungal contamination from A. flavus in feed chicken.

Phosphate Solvent Fungi Exploration and Identification in West Bali National Park Forest Area

Research on the exploration and identification of phosphate solvent fungi has never been carried out in West Bali National Park (TNBB), therefore researchers aims to explore and identify microscopic fungi to be used as phosphate solvent fungi which later will be taken from each plant soil samples (Lantana camara) that the presence is very common in TNBB. The research was implemented in two stages. The first stage is exploration of soil fungi in the field (TNBB) and identification of fungal species and the second stage is the phosphate solvent fungus test on Pikovskaya media. The results of the identification of the fungi obtained as follow: Aspergillus niger, Aspergillus bertholletius, Aspergillus flavus, Aspergillus isolate 4, Aspergillus isolate 5, Penicillium citrinum, and Trichoderma amazonicum. From the entire types of fungi obtained, there are onlybfour fungi that have the potential as phosphate solvents, namely Aspergillus niger, Aspergillus flavus, Aspergillus bertholletius and Penicillium citrinum with the presence of clear zones on Pikovskaya media. Fungi that has the best potential in the process of phosphate dissolution is Aspergillus niger.
 
 Key words: Rhizosfer, Lantana camara, clear zone, phosphate solvent fungus

In Vitro Assay of Ethanolic Heat Reflux Extract of Nicotiana tabacum L. var Virginia Against Nosocomial Bacteria Pathogen

Tobacco plays an important role in international trade as one of the export commodities. Indonesia is one of the good quality export contributors of tobacco leaves in the world. Nevertheless, tobacco is used only as a raw material for the cigarette industries, and the rise on anti-cigarette regulations prompted the exploration of alternative product from tobacco plants. The content of alkaloids, flavonoids, terpenoids and steroids in tobacco leaves were reported in literatures as antibacterial. Therefore, this study proposed in vitro assay of the ethanolic heat reflux extract (EHRE) of Nicotiana tabacum var. Virginia against nosocomial bacteria pathogen ((Pseudomonas aeruginosa (ATCC 27853), Eschericia coli (ATCC 25922), Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 29212)). Kirby-bauer diffusion method was used for this assay. The concentration of the EHRE for Kirby-bauer assay were 20; 40; 60; 80; and 100%. The presence of clear zones on Kirby-bauer test, against the growth of each nosocomial bacteria pathogen show that tobacco extract has antibacterial effect. Statistical analysis result showed that each extract concentration had significant difference value (p <0,05). This study indicated that the content (alkaloids, flavonoids, terpenoids and steroids) of tobacco leaf extracts (N. tabacum) has potential as antibacterial against nosocomial bacteria pathogen. Nevertheless, optimization of tobacco leaf extract to obtain maximum active ingredient still needs to be done. This study is important for further development of the tobacco leaf extract as antibacterial

Specific plate assay for bacterial heparinase

A procedure was developed for detecting heparinase activity on heparin agar plates. The method is based on the differential precipitation of heparin and heparinase-generated heparin fragments by protamine sulfate. Heparinase activity is detected by the presence of clear zones against a white background. This method can be used to screen for the expression of recombinant heparinase and to identify Flavobacterium heparinum mutants expressing heparinase constitutively.

In vitro bone resorption activity produced by a hypercalcemic adenocarcinoma tumor line (CAC-8) in nude mice.

Tumor extracts and conditioned tissue culture media from a canine adenocarcinoma tumor line (CAC-8) propagated in nude mice significantly increased in vitro bone resorption in neonatal mouse calvaria as measured by release of previously incorporated 45Ca. In vitro bone resorption activity was induced in a dose-dependent manner, was not suppressible by indomethacin, and was heat- and acid-stable. Gel exclusion chromatography demonstrated peak bone resorbing activity at a relative molecular mass of approximately 28,000. The parathyroid hormone (PTH) antagonist (8,18norleucine, 34tyrosine) bovine PTH (3-34) amide did not inhibit CAC 8-stimulated or (1-34) bPTH-induced bone resorption. There was an increased number of tartrate-resistant, acid phosphatase-positive cells in calvariae exposed to CAC-8 extract. Ultrastructural evaluation of calvaria revealed hypertrophy and maturation of osteoclasts in calvaria exposed to CAC-8 extract. The maturation effects included close contact to bone surfaces and the presence of clear zones and ruffled borders in osteoclasts. Similar structures were observed infrequently in osteoclasts of control calvaria. These data demonstrate that the tumor line (CAC-8) contained activity capable of stimulating in vitro bone resorption by increasing osteoclast numbers and the activity of existing osteoclasts.

Anticapsin, a new biologically active metabolite: screening and assay procedures.

In addition to its implication in the virulence of Streptococcus pyogenes, the hyaluronic acid capsule produced by this bacterium renders it resistant to infection by bacteriophage. A method employing S. pyogenes and a bacteriophage incorporated into an agar plate was devised as a screen to detect compounds that inhibit the formation of the hyaluronic acid capsule. Filter-paper discs saturated with experimental compounds were applied to the surface of test plates containing host plus phage and control plates of host only. After incubation, inhibition of capsule synthesis was indicated by the presence of clear zones where phage infection and lysis had occurred. Zones of growth inhibition on control plates represented classical antibacterial activity. During the testing of over 6,000 fermentation samples, anticapsin, a unique metabolite, was discovered. Modification of incubation temperature, thickness of agar layers, and host-phage input ratios resulted in a quantitative assay method having a dose-response range of 4 to 160 mug of anticapsin.

Anticapsin, a New Biologically Active Metabolite: Screening and Assay Procedures

In addition to its implication in the virulence of Streptococcus pyogenes, the hyaluronic acid capsule produced by this bacterium renders it resistant to infection by bacteriophage. A method employing S. pyogenes and a bacteriophage incorporated into an agar plate was devised as a screen to detect compounds that inhibit the formation of the hyaluronic acid capsule. Filter-paper discs saturated with experimental compounds were applied to the surface of test plates containing host plus phage and control plates of host only. After incubation, inhibition of capsule synthesis was indicated by the presence of clear zones where phage infection and lysis had occurred. Zones of growth inhibition on control plates represented classical antibacterial activity. During the testing of over 6,000 fermentation samples, anticapsin, a unique metabolite, was discovered. Modification of incubation temperature, thickness of agar layers, and host-phage input ratios resulted in a quantitative assay method having a dose-response range of 4 to 160 mug of anticapsin.