Manual Of Systematic Bacteriology Research Articles

Optimization of cold active amylase production by mesophilic Bacillus cereus RGUJS2023 under submerged fermentation

Aim: To produce the highest amount of cold-active alpha-amylase within a short time using mesophilic bacteria with optimized media to save the energy consumption cost and obtain higher enzyme production. Methodology: Amylase producing twenty-three strains were isolated on starch peptone agar plates. Among them, one strain, A5 was selected on the basis of highest clear (12 mm) zone on starch peptone agar plates. It was characterized and identified following Bergey's Manual of Systematic Bacteriology. Enzyme was characterized as Cold alpha amylase. All physico-chemical parameters (temperature, pH, Inoculum size) including carbon, nitrogen, metal ion and amino acid sources were optimized for maximum production of the enzyme. The optimized media was used for enhancing the cold active amylase production. Results: The strain, A5 was identified as Bacillus cereus RGUJS2023 by 16S rRNA sequencing analysis for further experiments. This strain showed the highest activity (9.922± 0.143 U ml-1) on the basal starch peptone media. Though, crude enzyme showed its activity at 4°C to 48°C temperature, but the temperature was 28°C. The highest cold active enzyme was produced (18.87±0.06 U ml-1) at 16 hr of bacterial growth at 35 °C with a pH 6.5 in the optimized media containing 0.5% starch, 0.1% peptone and 0.03% MgSO4.7H2O as a carbon, nitrogen and metal ion sources, respectively, with addition of 0.03% arginine. Interpretation: The cold active alpha amylase could be used commercially for the benefit of pharmaceutical and starch processing industries. Key words: Bacillus cereus RGUJS2023, Cold-active amylase, Starch hydrolysis

Lactic acid bacteria as an exopolysaccarides (EPS) producing starter from pakoba fruit (Syzygium sp.), endemic species at Minahasa, North Sulawesi

: Pakoba fruit ( sp.) is one of the medicinal plants of the Minahasa people and is an endemic species in North Sulawesi. Pakoba can only be found in the Minahasa area and very popular among Minahasa people. The taste is sour, making this fruit as the main ingredient to be processed into rojak, sweets, dodol, jam or syrup. Pakoba fruit is widely used for treatment in the simplest way because this fruit contains many bioactive compounds. The study aim to determine the presence of lactic acid bacteria (LAB) in Pakoba fruit and their potential in producing exopolysaccarides.: From the total of 35 producing-acid bacteria, 17 isolates were confirmed as Lactic Acid Bacteria (LAB) isolates with the characteristics of bacill cells, gram positive, catalase negative, non-motile, non-spore forming, gas production, mesophilic, aciduric, can ferment carbohydrates. : Based on Bergey's Manual of Systematic Bacteriology, the seventeen isolates were identified as member of genus. The seventeen isolates also showed the ability to produce exopolysaccharides in the range of 102-1570 mg /L.

Lactic Acid Bacteria as an Exopolysaccharides (EPS) Producing Starter from Pakoba Fruit (Syzygium sp.), Endemic Species at Minahasa, North Sulawesi

Pakoba fruit (Syzygium sp.) is one of the medicinal plants of Minahasan folks and it is an endemic species in North Sulawesi. Pakoba can only be found in Minahasa areas and is very popular among people in Minahasa. The taste is sour, making this fruit as the main ingredient to be processed into rojak, sweets, dodol, jam, or syrup. Pakoba fruit is widely used for treatment in the simplest way because this fruit contains many bioactive compounds. Pakoba fruit can act as a source of probiotics because it contains good microorganisms, namely Lactic Acid Bacteria (LAB). Some types of LAB are able to synthesize exopolysaccharides (EPS), which are polysaccharide polymers considered essential for health. Therefore this research aims to determine the presence of lactic acid bacteria (LAB) in Pakoba fruit and their potential to produce exopolysaccharides. The presence of lactic acid bacteria in Pakoba fruit was detected through the pour plate method on MRSA media (de Man, Rogosa and Sharpe Agar) + CaCO3 1%. The culture was incubated at 37°C for 48 hours. Growing colonies are observed morphological features of colonies, cells, biochemical and physiological properties. The isolates believed to be LAB, were then tested for its potential to produce EPS. From the total of 35 producing-acid bacteria, 17 isolates were tested as lactic acid bacteria group that had characteristics of bacilli cells, gram-positive bacteria, catalase-negative, non-motile, non-spore-forming, gas production, mesophilic, aciduric, can ferment carbohydrates. Based on Bergey’s Manual of Systematic Bacteriology, the seventeen isolates were identified as members of the Lactobacillus genus. The seventeen isolates also showed the ability to produce exopolysaccharides in the range of 102-1570 mg/L.

Isolation, Identification and Characterization of Thermostable Amylase Producing Bacteria from Budamada Hot Spring, Misrak Badewacho, Hadiya Zone, Southern Ethiopia

Thermophilic microorganisms are microorganisms that are stable over a wide range of harsh environmental conditions. Thermophillic amylase enzyme producing bacteria live at 45°C–80°C. Thermostable microbial amylases are more simplified and economical than other sources. The present research was aimed to isolate, identify and characterize thermostable amylase producing bacterial isolated from Budamada hot spring of Misrak Badawacho woreda. Thirty samples were collected from the source. Totally 18 isolates were isolated and 10 isolates were screened based on their ability to hydrolyze starch. Different biochemical tests were done to identify isolates based on Bergey’s Manual of systematic bacteriology. The results revealed that three enzymes from Bacillus spp. (SED2), Pseudomonas spp. (SED5) and Bacillus spp. (SED7) have maximum activity at 4% starch, 65˚C and pH of 8.5. Lactobacillus spp. (W20-0) and Bacillus cereus (SED12-1) at 8% starch, 75 and 65˚C, 8.5 and 7.5 pH respectively. Bacillus subtilis (W26-2) Bacillus licheniformis (W20-1), Lactobacillus spp. (SED10) at 6%starch; 35, 65, 45˚C; 6.5, 4.5, 4.5 PH., Bacillus spp.(SED8-3)&Bacillus spp. (W9) at 4% & 2%, 55˚C, 8.5 &4.5 pH and have maximum activity at 4% starch, 65˚C and pH of 8.5 and at 8% starch, 75 and 65˚C, 8.5 and 7.5 pH respectively., Lactobacillus spp. (SED10) at 6% starch; 35, 65, 45˚C; 6.5, 4.5, 4.5 PH. Bacillus spp. (SED8-3) and Bacillus spp.(W9) at 4% and 2%, 55˚C, 8.5 &4.5 PH. The amylase produced by bacterial isolates from Budamada hot spring was thermostable and performed from 65oC up to 75˚C. For most of the enzymes the optimum temperature was observed to be 65˚C; whereas optimum substrate concentration and pH for most of them were 4% starch and 8.5pH respectively. They were active at a various starch conditions (2%-10%), pH of 4.5-8.5, and temperature of 35-75°C.Generally, the bacteria isolated were thermophilic and produced thermostable amylase. Therefore, Budamada hot spring could be the source of thermophilic microorganisms and thermozymes as well.

Isolation and characterization of novel carotenoid pigment from marine Planococcus maritimus MBP-2 and their biological applications

BackgroundNumerous types of carotenoids with other structures are produced by organisms like bacteria, fungi, algae, yeast, higher plants, etc. In the present study, carotenoid pigment-producing, orange or red-pigmented organisms were isolated from scraping from the ship hull. Material and MethodsAccording to Bergey's manual of systematic bacteriology, the isolated bright orange pigmented organism was identified based on morphology, biochemical and physiological characters, strain myelin basic protein MBP-2 and identified as Planococcus maritimus. ResultsOrange pigment was extracted by solvent extraction method and characterized by using UV–Vis spectroscopy, scanning electron microscope, Transmission electron microscope, and Fourier-transform infrared spectroscopy. The Maximum absorbance was obtained at 447 nm, justify the presence carotenoid pigment in MBP-2 and FTIR results reveals the same. The antibacterial activities were performed against gram positive bacteria, Staphylococcus aureus (ATCC 25923) was 6 ± 0.12 mm, Bacillus cereus (ATCC 14579) was 12.3 ± 1.02 mm and gram negative Escherichia coli was 5.8 ± 0.91 mm, Pseudomonas aeruginosa (ATCC 39315) was 5.6 ± 1.12 mm and Klebsiella pneumoniae (ATCC 13313) was 11.6 ± 1.01. Furthermore antifungal activities were tested against Aspergillus niger (5 ± 2.15 mm), Aspergillus flavus (25 ± 0.85 mm), Pencillium commune( 14.8 ± 1.15 mm), Pencillium digitatum (12.0 ± 1.8) mmand showed sensitive to some of the bacterial and fungal pathogens. The in-vitro cytotoxic activity of pigment showed 19.46 ± 1.46% of cell death against DLA cells at 20 μl/ml of the drug concentration. ConclusionThe isolated orange carotenoid is a good source to treat antimicrobial, antifungal and anticancer activates and further study to be implemented.

METABOLISM AND FUNCTIONAL HETEROGENEITY OF FERMENTED MILK ORIGIN LACTIC ACID BACTERIA FOR LACTOSE INTOLERANCE

Lactic acid bacteria (LAB) are cosmopolitan in distribution with multiple ecological niches. LAB shows diverse applications in improvement of health by biochemical interference or imuno modulation to overcome several clinical circumstances. Lactose intolerance (LI) is one such situation, where individual show lactose maldigestion after the consumption of dairy products. It is necessary to overcome such a condition by employing indigenous beneficial bacteria or their products. Therefore, in this study we have isolated and characterized LAB from fermented milk samples, from remote villages of districts of south India. Traditionally fermented milk samples (68) were collected, cultivated on MRS medium, identified by biochemical and carbohydrate metabolic activity and correlated with Bergey's manual of systematic bacteriology. When matched with other LAB, Lactiplantibacillus isolates were able to reduced pH of medium significantly and reached pH of 4.6 in 48 hours. On MRS agar, 450 different bacterial isolates were isolated, recognized as presumptive LAB and classified up to the level of genera as Lactiplantibacillus (285), Lactococcus (70), Pediococcus (19), Streptococcus (20), and Enterococcus (16). Later, β-galactosidase screening was carried out using MRS/X-gal agar medium. Out of 450 LAB isolates only Lactiplantibacillus isolates were potential β-galactosidase producers. GRAS organisms such as LAB are multifaceted diverse group of bacteria localized in varieties of fermented foods/in the intestine and recognized as probiotics. Distinct contribution of LAB in health care and disorder management made this organism as a choice for alternate therapy; hence functionality of LAB can be promoted for LI management.

Biocontrol potential of bacteria belonging to the Bacillus subtilis group against pests and diseases of agricultural interest through genome exploration.

The usage of microorganisms as biocontrol agents and biofertilizers has been recommended and recognized as an ecologically correct alternative to maintaining the productivity and safety of crops. Thus, the objectives of this work were to characterize twelve strains belonging to Invertebrate Bacteria Collection of Embrapa Genetic Resources and Biotechnology by molecular, morphological, and biochemical methods and to evaluate the pathogenicity of these strains against pests and diseases of agricultural interest. The morphological characteristic of the strains was performed according to the principles of Bergy's Manual of Systematic Bacteriology. The genomes of the 12 strains were sequenced in Macrogen, Inc. (Seoul, Korea) using the HiSeq2000 and GS-FLX Plus high-performance platforms. In the determination of antibiotic sensibility profiles, disc-diffusion methods (Cefar Diagnótica Ltda) were adopted©. Selective bioassays were carried out with insects of the Lepidoptera (Spodoptera frugiperda, Helicoverpa armigera, and Chrysodeixis includens), Coleoptera (Anthonomus grandis), Diptera (Aedes aegypti) and Hemiptera (Euschistus heros) orders, and with the nematode Caenorhabditis elegans. In addition, the antagonistic action of the phytopathogens Fusarium oxysporum f. sp. vasinfectum and Sclerotinia sclerotiorum against the strains under study, and in vitro assays of phosphate solubilization were also performed. Sequencing of the complete genome of the 12 strains determined that all of them belonged to the Bacillus subtilis sensu lato group. In the strains genome were detected genic clusters responsible for encoding secondary metabolites such as surfactin, iturin, fengycins/plipastatin, bacillomycin, bacillisin, and siderophores. Due to the production of these compounds, there was a survival reduction of the Lepidoptera order insects and a reduction in the phytopathogens mycelial growth. These results show that the species of group B. subtilis s.l. can become promising microbiological alternatives to pest and disease control.

High Burden of Multidrug-Resistant Bacteria Detected in Different Water Sources Can Direct the Spread of Antibiotic Resistance Genes in the Environment

Antibiotic-resistant bacterial infections are of global concern nowadays. Environmental sources like water and soil are playing a key role in spreading antibiotic-resistance genes to humans, animals, and other environments. Objective: The purpose of this study was to identify and report the presence of multidrug-resistant bacteria (MDRs) in environmental water sources that can direct the spread of resistant genes to other bacteria/environments. Methodology: Environmental water samples were collected from 2 livestock farms and a fish pond. Bacterial isolation and identification were carried out by following Burgey’s manual of systematic bacteriology. Antibiotic susceptibility testing was done using the disk diffusion method and CLSI guidelines. Multiple antibiotic-resistant indexes were calculated. Whole genome sequences of previously reported bacteria were downloaded from NCBI to detect the resistance genes associated with phenotypic drug resistance and compared using the bioinformatics approach. Results: Microbial load was significantly high in all water sources. Following Genera were the most common: Klebsiella, Escherichia, Proteus, Serratia, Acinetobacter, Enterobacter, Pseudomonas, Bacillus, Lactobacillus, and Staphylococcus. Out of 10 classes of antibiotics, resistance against 8 classes were identified. Multiple Antibiotic Resistance (MAR) index range of isolated strains was between 0.4 and 0.9. Key Findings: Resistance against beta-lactam antibiotics was highest in our isolated strains with a MAR index of greater than 0.4 altogether. Conclusion: High burden of multidrug-resistant bacteria were detected in all water samples which can trigger the silent pandemic of antibacterial resistance.

Microbial degradation of quinoline by immobilized bacillussubtilis

Quinoline containing effluent discharged from pharmaceuticals, dyes and pesticide industries pollute the environment affecting human health due to its toxicity and nauseating odor. Quinoline is carcinogenic, mutagenic and toxic to the environment. Biotreatment is a viable process as quinoline gets easily degraded by microbes. We isolated a bacterium from the soil suspension of paper mill effluent and it was analyzed by Institute of Microbial Technology, Chandigarh from biochemical, morphological and physiological tests. The test results match with Bacillus subtilis as per the Bergey's manual of systematic Bacteriology. Quinoline mineralization by free and immobilized cells was studied. Adsorption technique was used for immobilization. 100 ppm of quinoline was completely degraded in 45 h by the free cells whereas cells immobilized on coconut shell carbon takes only 30 h. Cells on coconut shell carbon are capable of degrading even 300 ppm quinoline completely in 70 h. It was also found that cells immobilized on activated coconut shell carbon were more effective than cells on foam pieces. The optimum pH for efficient quinoline degradation was 9. The stability and recyclability of immobilized cells for 3 months at different concentrations for quinoline degradation was tested by performing an experiment. The excellent storage stability and recyclability of immobilized Bacillus subtilis can be critically used for the mineralization of heterocyclic organics in wastewater.

ANTIBACTERIAL ACTIVITY OF SILVER NANOPARTICLES SYNTHESIZED USING <i>ELEUSINE INDICA</i> (L.) GAERTN. LEAF EXTRACT ON THE BACTERIA ISOLATED FROM THE VASE SOLUTION

Green synthesis of silver nanoparticles (SNPs) using Eleusine indica (L.) Gaertn. leaf extract as a reducing agent is reported. SNPs were characterized with UV-Vis spectroscopy and transmission electron microscopy (TEM). The SNPs were rod like and spherical in shape with sizes from 3 to 33 nm and an average size of 16.73 nm. Seven bacterial strains were isolated from the vase water, including Bacillus cereus CA1, Alcaligenes faecalis CA2, Micrococcus luteus CA3, Pantoe agglomerans CA4, Pseudomonas aeruginosa CA5, Pseudomonas aeruginosa CA6, and Pantoe agglomerans CA7. Identifications were made according to Bergey’s Manual of Systematic Bacteriology and Bergey’s Manual of Determinative Bacteriology. The SNPs inhibited the growth of bacteria and exhibited significant antimicrobial activity against different isolated bacteria strains. SEM images showed that the SNPs damaged the cell membranes of bacteria, released plasmic contents, and altered the morphology of the cells. The impact of SNPs on gram-negative bacteria was more severe than on gram-positive bacteria. This study revealed that biosynthesized SNPs from Eleusine indica (L.) Gaertn. leaf extract are potential agents in combating bacterial contamination.

Antibiotic resistant bacteria in diverse ecological water samples are a threat to Human Food security

Antibiotic resistance is increasing with every passing day globally and it has become common not only in clinical settings but also in different environments. Aquatic systems are major reservoirs of antibiotic resistance genes in the environment. In this study, we have isolated the antibiotic resistance bacteria from aquatic systems including river Ravi, fish farm and underground drinking water source. Physical, chemical and microbiological parameters were studied, bacteria were isolated, gram staining and genus identification was done by following the Burgey’s manual of systematic bacteriology. While, antibiotic susceptibility testing was done by following the disk diffusion method and CLSI guidelines. Multiple antibiotic resistant indexes were calculated. There was no significant discrepancy in physical and chemical parameters of water samples whereas in microbiological parameters, a variable number of bacteria were found in all 3 water samples indicating the presence of microbial contamination. A total of 33 bacterial strains were identified including Enterobacteriaceae, Pseudomonas, Bacillus, Lactobacillus and Staphylococcus species. Out of 10 classes of antibiotics, resistant against 8 classes were identified. Organisms isolated from river Ravi were the most resistant isolates. Most of the organisms were resistant to Penicillins, Carbapenems and Cephalosporins. MAR index of isolated strains ranges from 0.43 to 0.88. AMR is common in all environmental water samples indicating a threat to human food safety

Isolation and characterisation of endophytic bacteria present in the leaves of Glycosmis pentaphylla (Retz.) Correa

Endophytes are an endosymbiotic group of microorganisms that dwell in plant tissues and are reservoirs of bioactive compounds. However, the researches on this area are limited. The present investigation was undertaken to isolate and characterise endophytic bacteria from the leaves of Glycosmis pentaphylla (Retz.) Correa. An effective surface sterilisation procedure was developed from the experiment to isolate endophytic bacteria from the leaves of the candidate species. There was no bacterial growth observed in the sterility test. A total of 3 endophytic bacteria were isolated from leaves of G. pentaphylla. Isolates showed distinct morphological and biochemical characteristics. Biochemical characterisation of the isolates was performed by following Bergey’s manual of systematic bacteriology. Two isolates were found gram-positive, and one was gram-negative. All three isolates were found positive for the catalase test and negative for the indole test; two isolates (GP-1, GP-3) were positive for the oxidase test; 2 isolates (GP-1, GP-2) were found positive for both citrate and methyl red test. Some plant growth-promoting activities of the isolates were also performed. All the 3 isolates (GP-1, GP-2, and GP-3) were found positive for the ammonia production test; isolate GP-2 was found positive for phosphate solubilisation test; isolate GP-3 was found positive for both IAA production and lipase activity test. The isolated endophytic bacteria survived in different salt concentrations.

Isolation and Characterization of Microbes of Different Kinds of Street Food and Determination of Antibiotic Susceptibility of the Isolates

Aim: Improper personal hygiene can facilitate the transmission of the pathogenic microorganisms found in environment and on people’s hands via food to humans. The present study was undertaken to investigate the microbiological quality of different street food and determining the antibiotic susceptibility of the isolated microorganisms.
 Study Design: Collection of food samples for isolation of Pathogenic Microorganisms, to identify them by using Biochemical test, molecular test (16sr RNA typing), Antibiotic susceptibility was done by using different antibiotics against the isolates.
 Place and Duration of Study: Food samples were collected from street vendors of Sainkpuri area, work was done from December 2020 to April 2021 at Microbiology Department, St. Francis College for Women, Hyderabad.
 Methodology: Five samples of street food were collected in sterilized bottles, tested for the presence of microorganisms by following standard microbiological method used for isolation of microorganisms. The organisms were identified by carrying out various biochemical test according to Bergey’s Manual of Systematic Bacteriology. The Molecular characterization was done based on 16sr RNA typing. Determining the sensitivity of the isolates against different Antibiotics by employing Kirby Bauer technique.
 Results: The organisms isolated from Manchuria and Ragada samples: Lactobacillus delbrueckii; Lactobacillus casei, Klebsiella pneumonia, Enterococcus faecalis and Vibrio spp respectively; Samosa and bonda: Staphylococcus aureus, Lactobacillus delbrueckii and Pantoea dispersa; Pani puri water: Staphylococcus aureus and Providencia vermicola. Antibiotic susceptibility tests showed that most of the isolated microorganisms were sensitive to Ciproflaxin.
 Conclusion: Hence the quality of street food is found to be low due to following factor: lack hygiene conditions in the food preparation, ingredients may also affect quality of the food, and presence of air borne microorganisms in the surrounding areas of service points.

Molecular identification of Streptomyces sp. isolated from peat land of Palangka Raya, Kalimantan Tengah using 16S rRNA gene sequences analysis

Bacterial identification with 16S rRNA accompanied with cloning techniques to obtain a purer sequence are now highly developed. This research aims to identify Streptomyces bacteria from peatlands in Central Kalimantan as a source of new antibiotics compound using 16srRNA and cloning techniques. Streptomyces sp. isolate was studied in detail and characterized. The morphological and biochemical characterization and tentative identification methods was described following the Bergey’s Manual of Systematic Bacteriology. Further identification and phylogenetic study were done using molecular tools. 16S rRNA helped to determine the species using PCR. Results of tabulation data from BLAST® indicate that isolates 2.1 belong to the genus Streptomyces sp. with the percentage of identity reaching 99.73% with Streptomyces sp. VEL 17 gene for 16S rRNA, partial sequences.

Exploration of Plastic-Degrading Bacteria From Marina Beach, Semarang, Central Java

Plastic waste has threatens the environment and affect to the economic and tourism sectors, marine life, coastal ecosystems and human health. World Wide Fund for Nature (WWF) states that 85% of waste in the oceans is plastic. The Ministry of Environment and Forestry also noted that Indonesia experienced an increase in plastic waste from 14% in 2013 to 16% in 2016. By 2020 the volume of plastic waste in Indonesia predicted to reach 67.8 million tons. Plastic waste takes 100-500 years to completely decompose. An alternative solution is to involve microorganisms to decompose plastic polymers. However, plastic waste reducing bacteria isolated from coastal ecosystem has not been much explored. In this study, an exploration of natural bacteria that degrades plastic waste from coastal ecosystems is carried out. Plastic samples were collected from the Marina Beach Semarang, Central Java. Plastic samples were taken from a depth of 0-10 cm in three coastal ecosystems: coastal sand sediments, rocks and mangroves. Samples then isolated and screened to obtain bacteria that have the potential to degrade polyethylene. Selected bacteria were identified by biochemical physiology according to the method of Cappuccino and Sherman and classified to genus level according to Bergey's Manual of Determinative Bacteriology and Bergey's Manual of Systematic Bacteriology. The results showed that three genera of bacteria had high polyethylene degradation potential with the speed of degradation: Enterobacteriaceae 0.0091%; Moraxella spp. 0.0066%; and Pseudomonas spp. 0.0076% per week.

Studies of some genes of Bacillus subtilis MN99 local strains against plant diseases

The objective of this study was the isolation and characterization of Bacillus subtilis local strains from the soil in Mongolia. These local strains of B. subtilis are showed to have high antagonistic activities against some plant pathogenic fungi and bacteria. Six strains of B. subtilis were isolated and characterized morphologically, physiologically and biochemically according the Bergey’s Manual of Systematic Bacteriology.
 In order to identify species of the isolated strains, we amplified and sequenced 16S rRNA gene, essential funtinal genes bmyB, spoVG and srfAA, which are related to antagonistic activity of these strains. The sequences were aligned using CLASTALW multiple sequence alignment tool. Phylogenetic tree was drawn according to Maximum likelihood”method and “Tamura-Nei” model using “MEGA-X version 10.2.6 program.
 Among all isolates of B. subtilis MN99 and 7/24 strains had higher antagonist activity against plant diseases. According to partial sequence of srfAA (620bp) gene of MN99, the local strain belongs to B. subtilis and partial sequence of bmyB (370bp), spoVG (22bp) gene of MN7/24 strain showed that the it belongs to B. atrophaeus species. All local strains of B. subtilis had bacillomycin synthesis gene, and B. subtilis MN99 strain had only surfactine synthesis gene, while did not have spore formation and hemolysis gene SpoVG.

KARAKTERISASI ISOLAT BAKTERI PENGHASIL SELULOSA DARI BUAH NAGA MERAH (Hylocereus polyrhizus)

Cellulose is a natural polymer widely used for general industrial purposes. The development of science utilizes bacteria from certain fruits to produce biosellulose. Red dragon fruit ( Hylocereus polyrhizus ) contains a lot of sugar which can be bioconverted to cellulose by cellulose- producing lines. The research aimed to obtain bacteria from red dragon fruit with the potential to produce cellulose, then conduct morphological and biochemical characterization. The research was conducted by the isolation of bacteria from the fruit, purification, and screening tests using Hestrin-Schramm medium. The isolates obtained were characterized morphologically through macroscopic and microscopic observation and the biochemically through catalase test, carbohydrate fermentation, starch and gelatin hydrolysis, motility, and IMViC (Indole, Methyl red, Voges- Proskauer, Citrate). The results showed that 4 bacterial isolates had the potential to produce cellulose. Based on Bergey's Manual of Systematic Bacteriology, 3 bacterial genera were obtained, namely Acetobacter, Gluconacetobacter, and Azotobacter. The results of the characteristics of the isolates obtained at IBBN 2-2 were positive in the catalase test, sucrose and lactose fermentation, and methyl red. IBBN 3-1 was positive in the catalase test, sugar fermentation, starch hydrolysis, methyl red, and citrate. IBBN 3-2 was positive in the sucrose and glucose fermentation test, gelatin hydrolysis and 4-1 IBBN was positive in the sucrose fermentation test and gelatin hydrolysis.

Biological characteristics of enteropathogenic Aeromonas species isolated from different Egyptian foods

A total of 185 food samples in Sharkia Governorate, Egypt were analyzed for the presence of Aeromonas spp. The recovered isolates were characterized by biochemical tests based on Bergey’s Manual of Systematic Bacteriology and confirmed by API-20E kit. Of the samples analyzed, 56 (30.27 %) were positive for Aeromonas spp., with the highest incidence in raw milk (55 %) followed by Nile tilapia (48.6 %). A. hydrophila and A. salmonicida were the most predominantly isolated from raw milk and A. caviae was the most prevalent in raw lean beef. A variety of extracellular virulence factors were determined and 83.3 % exhibited β-hemolytic activity; 80 % exhibited lecithinase activity; 75 % exhibited protease activity and 90 % exhibited lipolytic activity. Furthermore, the cytotoxic (act) and the cytotonic (alt) enterotoxins were screened in three selected isolates using PCR assay and the data showed the detection of the two genes with predicted sizes of 232 bp and 442 bp fragments, respectively. These findings indicated a major health risk of infection with enterotoxigenic Aeromonas bacteria for people consuming raw and improper cooked food in Egypt.

The investigation of antimicrobial and antifungal activity of some Artemisia L. species

Background: Throughout history pathogenic microorganisms cause infectious diseases. Medicinal herbs play an important role in human life because they are used for the therapy and production of herbal remedies. Therefore, it is relevant to study the antibacterial activity of medicinal herbs includingArtemisiaL. species.Aim: The purpose of this study was to investigate the antimicrobial and antifungal activity ofArtemisiaL. species extracts and to identify their synergistic antimicrobial effects with erythromycin against skin isolates ofStaphylococcus aureuswith different mechanisms of MLS-resistance.Materials and methods:Artemisia absinthiumL. herb extracts (AAs-4, AAs-7, AAs-9),Artemisia vulgarisL. herb extracts (AV-4, AV-7, AV-9) andArtemisia abrotanumL. herb extracts (AAr-4, AAr-7, AAr-9) were used in this study (solvents – 40%, 70% and 90% ethyl alcohol respectively).The determination of antimicrobial activity of extracts was performed using clinical isolates of antibiotic susceptible and antibiotic resistant microorganisms. Bacterial cultures were identified on the basis of the biochemical microtests “STAPHYtest 16”, “ENTEROtest 24”, “NEFERMENTtest 24” taking into account the complex of their morphological and cultural properties in accordance with the recommendations of the 9thedition of “Bergey’s Manual of Systematic Bacteriology”. Yeast-like fungi cultures were identified on the basis of 40 biochemical tests with the help of the VITEK 2 system using the VITEK 2 YST ID card.The synergism of the antimicrobial activity of extracts with erythromycin was investigated using the clinical isolate ofStaphylococcus aureuswith the non-inductive MLS-resistance mechanism (resistant to erythromycin at minimum bacteriostatic concentration of 125 μg/ml and minimum bactericidal concentration of 250 μg/ml without induction of resistance to clindamycin).Research results: The evaluation ofArtemisiaL. herb extracts activity was carried out using standard strains of microorganisms which are recommended by the WHO.The results of the conducted studies indicate thatArtemisiaL. herb extracts are able to reduce the growth of microorganisms. The investigated extracts showed the potent bacteriostatic action against the cocci or rod-shaped microflora. The antimicrobial activity ofArtemisiaL. herb extracts directly depends on the concentration of ethanol as the solvent.Artemisia vulgarisL. herb extracts (solvents – 70% and 90% ethanol) andArtemisia abrotanumL. herb extract (solvent – 90% ethanol) show synergism of antimicrobial activity with erythromycin in relation toStaphylococcus aureuswith efflux mechanism of MLS-resistance. The obtainedArtemisiaL. herb extracts can be used to create antifungal drugs, as well as antimicrobial drugs (against gram-positive and gram-negative bacteria).

Antimicrobial Activity of Herbal Toothpaste against Organisms Causing Dental Caries

Aim: Dental caries occurs as a result of acid produced by bacteria which destroys the enamel and underlying dentin. A study was carried out to isolate microorganisms that cause dental caries in patients belonging to different age groups and find out the antimicrobial activity of various toothpastes against them.
 Study Design: Collection of samples for isolation of caries causing organisms, to identify them using biochemical tests and test the activity of herbal toothpastes against them.
 Place and duration of study: Samples were collected from Krish Multispeciality Clinic and Prashanth Dental Clinic, Nagole between June’2016 – December’2017 and worked on, at Microbiology Department, St. Francis College for women, Hyderabad.
 Methodology: Ten samples of each age group from 10-20 yrs., 20-40 yrs. and 40-60 yrs. were isolated. The organisms were identified by carrying out various biochemical tests according to Bergey’s Manual of Systematic Bacteriology and molecular characterization based on 16s rRNA typing. Identified organisms were used to test the antimicrobial activity of herbal toothpastes by employing agar well diffusion method. ANOVA is used for statistical analysis.
 Results and Conclusion: Patanjali herbal products like Dantkanti are gaining lot of importance and have become a part of life of common man because of their effectivity. Our studies revealed that out of the various toothpastes tested, Patanjali toothpaste had good antimicrobial activity against all the isolates. On comparative analysis of f-ratio and P significant value (< .05.) of all pairs, it is observed that Patanjali has good antimicrobial activity against all isolates tested. The efficacy of Meswak is almost similar to that of Patanjali.