Clear Zone Research Articles

Purification, and characterization of detergent-compatible serine protease from Bacillussafensis strain PRN1: A sustainable alternative to hazardous chemicals in detergent industry

Owing to vast therapeutic, commercial, and industrial applications of microbial proteases microorganisms from different sources are being explored. In this regard, the gut microbiota of Monopteruscuchia were isolated and examined for the production of protease. All the isolates were primarily and secondarily screened on skim milk and gelatin agar plates. The protease-positive isolates were characterized morphologically, biochemically, and molecularly. Out of the 20 isolated strains,6 belonging to five different genera viz.Bacillus,Priestia,Aeromonas,Staphylococcus, and Serratia demonstrated proteolytic activity. Bacillussafensis strain PRN1 demonstrated the highest protease production and, thus, the largest hydrolytic clear zones in both skim milk agar (15 ± 1 mm) and gelatin (16 ± 1 mm) plates. The optimized parameters (time, pH, temperature, carbon, nitrogen) for highest protease activity and microbial growth of B.safensis strain PRN1 includes 72 h (OD600 = 0.56,1303 U/mL), pH 8 (OD600 = 0.83, 403.29 U/mL), 40 °C (OD600 = 1.75, 1849.11 U/mL), fructose (OD600 = 1.22, 1502 U/mL), and gelatin (OD600 = 1.88, 1015.33 U/mL). The enzyme was purified to homogeneity using salt-precipitation and gel filtration chromatography. The sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that the purified enzyme was a monomer of a molecular weight of ∼33 kDa. The protease demonstrated optimal activity at pH 8 and 60 °C. It was strongly inhibited by phenylmethylsulfonyl fluoride (PMSF), demonstrating that it belongs to the serine-proteases family. The compatibility of the enzyme with surfactants and commercial detergents demonstrates its potential use in the detergent industry. Furthermore, the purified enzyme showed antibacterial and blood-stain removal properties.

ANALYSIS OF ECO ENZYME CHARACTERISTICS WITH VARIATION OF “TAPE” YEAST CONCENTRATIONS

Eco enzyme is a multipurpose liquid which produced from organic waste. The liquid made with the ratio of raw materials in the form of brown sugar: organic waste: water is 1:3:10 which is processed using a fermentation process for 90 days. One method to shorten the fermentation time is adding yeast as catalyst throughout the fermentation process. This research aims to analyze the characteristics of the eco enzyme with variations in “tape” yeast concentrations. Raw material used in this research included organic waste (orange and melon peels with a ratio of 1:1), brown sugar, water, and “tape” yeast (yeast concentrations of 1, 2, 3, 4 and 5%w/v) through fermentation process for ten days. The eco enzyme products were tested to determine the pH value, TDS value, acetic acid content, aroma, color and protease enzyme activity. The analysis results showed that the pH value of eco enzyme products was in the range of 3.9 to 4.1, TDS (Total Dissolved Solids) value of 1339 to 1405 ppm, acetic acid content of 0.81 to 1.08%w/w, had bright yellow and deep yellow color with a fermented aroma. The selected eco enzyme product in this research was on eco enzyme with yeast concentration of 3 %w/v which had pH value of 3.9 TDS value 1403 ppm, acetic acid content of 0.81%w/w, and the color was bright yellow with fermented aroma. However, there was no clear zone around the colony in the test indicating that there was no protease enzyme activity.

Analysis of Amylase Activity in Bacteria Isolated from Hot Spring of Pentadio Resort

The hot spring of Pentadio Resort is one of the geothermal areas likely to be the place of bacteria or microorganisms that can produce amylolytic enzymes. This research aims to analyze bacteria in the Pentadio Resort hot springs that possess the potential to produce amylase enzymes, analyze the characteristics of bacterial isolates that produce amylase enzymes in Pentadio Resort hot springs and determine the types of bacteria that produce amylase enzymes in Pentadio Resort hot springs. The research employs a qualitative and descriptive method. The research involved several methods, including sample collection, media preparation, isolation and selection of thermophilic bacterial isolates, purification of thermophilic bacterial isolates, selection of isolates producing amylase enzymes, identification of selected isolates (motility test, biochemical test, gram staining), and molecular identification. The research yielded seven isolates, and isolate code A6 demonstrated the potential to produce amylase enzymes with a clear zone size of 23,26 mm, bacillus, gram positive, and positive motility. Subsequently, molecular identifidation was carried out in which, based on the results of phylogenetic tree reconstractiopisalate A6 was related to the bacteria Bacillus cereus strain MD 152, which belongs to the Bacillus cereus bacteria

Screening of microscopic fungi isolated from charcoal and wooden samples for Protease production

Proteases, otherwise called proteinases or proteolytic chemicals, have a place with a gathering of hydrolases. Microorganisms are the most well-known wellsprings of business chemicals because of their physiological and biochemical properties. Fungus from charcoal and wooden sample were secluded by sequential weakening agar plate method. The fungal disengages were to begin with evaluation of their capacity to create protease based on zone of clearance on skimmed milk agar plate. The segregates that gave excessive positive consequences in screening were identified as Aspergillus fumigatus, Fusarium species, Mucor recemossus, Penicillin citrium with the aid of inspecting the morphology and Lacto - phenol cotton blue (LPCB) staining procedure. Among those disconnects, Aspergillus fumigatus which constrained from charcoal floor, exhibited the maximum noteworthy protease movement confirming the use of the clean sector across the colony that is brooded at room temperature at a pH of round 7 for a hundred and twenty hours at 28 ˚C. Three fungal disengages chosen by beginning tests were exposed to extracellular protease creation in protease creation medium. The outcomes showed that three fungi could be utilized in the creation of protease compounds and in the upscale for modern creation.

Isolation and production of proteases from bacteria isolate from Bekasam and Rusip (Traditional Food from South Sumatera, Indonesia)

The research aims to assay enzyme activity of bacteria from traditional fermentation product (bekasam and rusip). The bacteria were inoculated in skim milk agar (SMA), and will produce clear zone. The best isolates from 9 isolates of bekasam and 9 isolates from rusip will be choosen, and the best result from bekasam and rusip were A3K2 and B3K2, respectively. Index proteolitic of A3K2 and B3K2 isolates were 10.0 and 7.0, respectively. Optimum time production of protease from A3K2 and B3K2 were 16 h and 32 h, respectively.

Calliandra haematocephala’s phytochemical analysis and antifungal effectiveness against fish pathogen in the lake of Bhopal

Aquaculture has been a major part of our Economy from ancient times and antimycotic resistant fish illnesses have emerged as a major threat in aquaculture from a very long time. As routinely used anti-fungal medications have become less effective and traditional medicinal herbs have been identified as a possible medication source that can counteract antifungal resistance. The current study will look at the antifungal efficacy of leaf and flower extracts of the nutritional and traditional medicinal plant called Calliandra haematocephala against 3 common fish fungal pathogens (Achlya, Saprolegnia and Aphanomyces) that were isolated from Lower Lake of Bhopal (M.P.) and causes illness in fishes present in Lower Lake, Bhopal (M.P.). The leaves and flowers of Calliandra haematocephala were extracted sequentially with ethanol as a solvent, and the presence of possible phytoconstituents was qualitatively evaluated. Flavonoids, phenol, alkaloids, carbohydrates, tannins and saponins were present in both flowers and leaves and steroids were found exclusively in the ethanolic extract of the leaf only not in flower extract. The antifungal activity of ethanolic extract was determined using the agar well diffusion technique. The ethanolic fraction of the flower demonstrated a wide range of activity against fish pathogens, with a maximal inhibitory or clearance activity against Saprolegnia in a amount of 20 μL (4 mm) and in leaves it showed maximum clearance zone in Achlya and Saprolegnia in 20μL (4 mm). In conclusion, Calliandra haematocephala’s extracts consist of potential secondary metabolites that might be employed as antifungal treatments for 3 prevalent fish Pathogens, and the leaf ethanolic extracts have more inhibitory efficacy than the flower ethanolic extract.

Antimicrobial Activities against Skin Flora from Tart-cherry Extracts (Prnus cerasus)

Purpose: In this study, antibacterial and anti-oxidant activities of Tart-cherry hydrothermal extracts (TCW) and ethanol extracts (TCE) against dermatological bacteria were measured, and the cell viability and anti-inflammatory effects were investigated in RAW264.7 macrophages to determine their characteristics for use as functional cosmetic materials. Methods: Anti-oxidant activity was determined using the DPPH method. The antibacterial activity against dermatophilosis was confirmed by the size of the clear zone formed by DISK diffusion and MIC measurement. The activity against an antibiotic-resistant MRSA strain was also measured. Anti-inflammatory response was identified by measuring cytotoxicity and NO activity in RAW264.7 macrophages, in which inflammatory reactions were induced with LPS. Results: Anti-oxidant activity of the TCE was 85.6%, and of TCW was 88.4% at 500 μg/mL. The size of the clear zone was confirmed using Cutibacterium acnes (18 mm), <i>Staphylococcus epidermis</i> (<i>S. epidermis</i> ) (4 mm), and <i>Staphylococcus aureus</i> (<i>S. aureus</i> ) (16 mm). No antifungal activity against <i>Candida albicans</i> was observed. RAW264.7 macrophages showed a survival rate of >90% at <250 μg/mL of extract. Additionally, a concentration-dependent inhibition of NO production was observed. Conclusion: Tart-cherry extracts possessed high anti-oxidant activity and anti-inflammatory efficacy. In addition to the antibacterial activity against the skin-residing bacteria, <i>S. aureus</i> and <i>S. epidermis</i>, high activity against the acne-causing strain <i>C. acnes</i> suggests a sufficient potential of Tart-cherry extracts as a raw material for the preparation of cosmetics and bioproducts.

Biodeterioration of polyethylene by Bacillus cereus and Rhodococcus equi isolated from soil.

Polyethylene (PE), a non-biodegradable plastic, is widely used in agriculture as a mulch material, which causes serious plastic pollution when it is discarded. Recent studies have described the biodeterioration of PE by bacteria, but it is difficult for a single bacterial species to effectively degrade PE plastic. We isolated two strains with PE-degrading ability, Bacillus cereus (E1) and Rhodococcus equi (E3), from the soil attached to plastic waste on the south side of Mount Tai, China, using a medium with PE plastic as the only carbon source. By clear zone area analysis, we found that E1 mixed with E3 could improve the degradation of PE plastics. The mixture of E1 and E3 was incubated for 110days in a medium containing PE and mulch film as the only carbon source, respectively. After 110days, a decrease in pH and mass was observed. Obvious slits and depressions were observed on the surface of the PE film and the mulch films using scanning electron microscopy. The surface hydrophobicity of both films decreased, and FTIR revealed the formation of new oxidation groups on their surfaces during the degradation process and the destruction of the original CH2 long chains of PE. Besides, we found that surface of the mulch films contained more viable bacteria than the liquid medium. In conclusion, we identified two PE-degrading strains whose mixture can effectively degrade mulch film than pure PE film. Our results provide a reference for understanding PE plastic degradation pathways and their associated degradation processes.

Catharanthus roseus-assisted bio-fabricated zinc oxide nanoparticles for promising antibacterial potential against Klebsiella pneumoniae.

This study emphasized on the synthesis of zinc oxide nanoparticles (ZnO NPs) in an environmentally friendly manner from the extract of Catharanthus roseus leaves and its antibacterial assessment against the pneumonia-causing pathogen Klebsiella pneumoniae. This simple and convenient phytosynthesis approach is found to be beneficial over conventional methods, wherein plants serve as excellent reducing, capping, and stabilizing agents that enables the formation of ZnO NPs without the use of harmful chemicals. The formation of ZnO NPs was confirmed through several characterization techniques such as UV-visible spectroscopy, XRD, FT-IR, SEM, HR-TEM, and EDX. XRD analysis revealed high polycrystallinity with crystallite size of approximately 13nm. SEM and HR-TEM revealed the hexagonal structure of ZnO NPs with the particle size range of 20-50nm. The EDX shows the elemental purity without any impurity. Furthermore, the antibacterial efficacy by the technique of disc diffusion exhibited clear inhibition zones in ZnO NPs-treated discs. In addition, 125µg/mL of ZnO NP concentration showed minimum inhibition by the microbroth dilution method. The potent inhibitory activity was further validated with trypan blue dye exclusion and fluorescence microscopy. Finally, SEM examination confirmed the efficient antibacterial potential of ZnO NPs through disruption of the intact morphology of Klebsiella pneumoniae.

Investigation the biological activities and the metabolite profiles of endophytic fungi isolated from Gundelia tournefortii L.

Endophytic fungi are microorganisms that are considered as a potential source of natural compounds, and can be applied in various industries. The aims of this research were molecular identification of endophytic fungi isolated from the Gundelia tournefortii stems, and investigation their biological activities as well as phenolic and fatty acid profile. Surface sterilized stems of G. tournefortii were placed on potato dextrose agar (PDA) to isolate the fungal endophytes. Genomic DNA was extracted by CTAB method, and PCR amplification was performed by ITS 1 and ITS 4 as primers. The enzyme production of endophytic fungi was determined based on the formation of a clear zone that appeared around the colonies of fungus. The anti-oxidant activity was evaluated by measuring the amount of free radicals DPPH. Also, the total phenol and flavonoid contents were measured obtained by Folin-Ciocalteu and aluminum chloride colorimetric methods, respectively. Moreover, the separation and identification of phenolic acids and fatty acids were done by HPLC and GC, respectively. Phylogenetic analysis was done based on the Internal Transcribed Spacer (ITS) region, and five isolates were identified as following: Aspergillus niger, Penicillium glabrum, Alternaria alternata, A. tenuissima, and Mucor circinelloides. Evaluation of the enzymatic properties showed that P. gabrum (31 ± 1.9 mm), and A. niger (23 ± 1.7) had more ability for producing pectinase and cellulase. The anti-oxidant activity of isolates showed that A. alternata extract (IC50 = 471 ± 29 µg/mL) had the highest anti-oxidant properties, followed by A. tenuissima extract (IC50 = 512 ± 19 µg/mL). Also, the extract of A. alternata had the greatest amount of total phenols and flavonoids contents (8.2 ± 0.4 mg GAL/g and 2.3 ± 0.3 mg QE/g, respectively). The quantification analysis of phenolic acid showed that rosmarinic acid, para-coumaric acid, and meta-coumaric acid (42.02 ± 1.31, 7.53 ± 0.19, 5.41 ± 0.21 mg/g, respectively) were the main phenolic acids in the studied fungi. The analysis of fatty acids confirmed that, in all fungi, the main fatty acids were stearic acid (27.9–35.2%), oleic acid (11.3–17.3%), palmitic acid (16.9–23.2%), linoleic acid (5.8–11.6%), and caprylic acid (6.3–10.9%). Our finding showed that endophytic fungi are a source of bioactive compounds, which could be used in various industries. This is the first report of endophytic fungi associated with G. tournefortii, which provides knowledge on their future use on biotechnological processes.

Aplikasi Edible Coating dari Limbah Kulit Udang dengan Aditif Asap Cair untuk Kemasan Sosis Sapi Antibakteri Ramah Lingkungan

Edible coating is a food preservation technique by coating using antimicrobial substances which can inhibit the growth of bacteria. This research aims to develop an edible coating from shrimp shell waste with liquid smoke additives as antibacterial packaging for processing beef sausages. Chitosan is made in three stages, namely demineralization, deproteination and deacetylation. Beef sausages coated with edible coating were tested using organoleptic and antibacterial tests. The results of this research showed that the highest degree of deacetylation of chitosan was found in the variable concentration of sodium hydroxide (NaOH) 55% with a degree of deacetylation value of 93.59%. The water and protein content in beef sausages coated with edible coating with liquid smoke showed values of 52.45% and 12.65% on the 7th day. This is in accordance with the Indonesian National Standards (SNI). Coating beef sausages using edible coating from 2% chitosan with 8% liquid smoke solvent has antimicrobial properties because in the antibacterial test a clear zone appeared on the beef sausages coated with edible coating.

Diversity of actinomycetes on plant rhizosphere of karst ecosystem of Gorontalo, Indonesia

Abstract. Retnowati Y, Kandowangko NY, Katili AS, Pembengo W. 2024. Diversity of actinomycetes on plant rhizosphere of karst ecosystem of Gorontalo, Indonesia. Biodiversitas 25: 907-915. The objective of this study was to isolate and identify the actinomycetes associated with the rhizosphere of plants in the karst ecosystem. This study was descriptive-quantitative. Soil sampling was conducted at three locations of the karst ecosystem of Gorontalo, i.e., Bangga Hills at Bangga Bubaa Coastal area of Gorontalo District, Hills around Lake Limboto of Gorontalo District, and Oluhuta Beach Hills at Bone Bolango District. Rhizosphere-soil sample was collected from 15-30 cm of soil depth. Isolation of actinomycetes was performed using the plate method. The screening of phosphate-solubilizing bacteria was based on the ability to solubilize phosphates indicated by the clear zone on the Pikovskaya medium. Identification of actinomycetes bacteria based on morphology and molecular characters. The morphology character included the color of aerial and substrate mycelium. Molecular characteristics based on the sequence of 16S rRNA gene compared to sequence data on the gene bank of NCBI. The phylogenetic tree reconstruction was based on the Neighbor-joining algorithm. The results successfully found six isolates, including three actinomycetes isolates, two non-actinomycetes bacteria, and one yeast. They were isolated from the rhizosphere of Malastoma malabathrum, Chromolaena odorata, Cycas rumphii, Leucaena leucocephala, and Jatropha curcas. The actinomycetes isolate showed plant-growth-promoting potential indicated by solubilizing phosphate and Indole 3-Acetic Acid production activities. Based on the molecular analysis, actinomycetes were closely related to Streptomyces aegyptia (MT505707.1:42-1426), Streptomyces sp. GGCR-6 (MH718844.1:5-1391), and Streptomyces carpaticus strain PES-A23 (MH712039).

The impact of different management on pine-based agroforestry system and litter accumulation on the population and activity of cellulolytic bacteria

Abstract. Arfarita N, Nuraini Y, Rinady MVP, Noerhayati E, Prayogo C. 2024. The impact of different management on pine-based agroforestry system and litter accumulation on the population and activity of cellulolytic bacteria. Biodiversitas 25: 924-936. Agroforestry leads to the accumulation of organic matter, and the environmental conditions, litter quality, and decomposing organisms influence the rate at which litter decomposes. Litter decomposition in agroforestry systems is slow due to the high lignin and phenolic substances content, low light intensity, and high humidity under the canopy's shading condition. Therefore, a study was carried out to address this issue to isolate cellulolytic bacteria capable of breaking down plant litter using a qualitative cellulase activity testing method. The complete randomized block design and Tukey's test were used to determine the treatment's significance. Six agroforestry systems were examined: pine-coffee, pine-banana, pine-cardamom, pine-cardamom, mixed garden, and citrus. The PK (Pine-Coffee) plot had the highest canopy cover, litter density, and cellulolytic bacteria. The study identified three cellulolytic bacteria isolates (PK1, PK13, and PK10) from the 50 isolated bacteria of PK plot, producing the largest clear zones on CMC media. These identified isolates belonging to the Bacillus genus were Gram-positive bacteria with rod-shaped cells. The different types of litter in agroforestry systems affect the content of lignin, polyphenols, cellulose, and C/N ratios, which can influence the abundance of cellulolytic bacteria and their potential cellulase activity.

Vancomycin heteroresistance caused by unstable tandem amplifications of the vanM gene cluster on linear conjugative plasmids in a clinical Enterococcus faecium.

Vancomycin heteroresistance is prone to missed detection and poses a risk of clinical treatment failure. We encountered one clinical Enterococcus faecium strain, SRR12, that carried a complete vanM gene cluster but was determined as susceptible to vancomycin using the broth microdilution method. However, distinct subcolonies appeared within the clear zone of inhibition in the E-test assay, one of which, named SRR12-v1, showed high-level resistance to vancomycin. SRR12 was confirmed as heteroresistant to vancomycin using population analysis profiling and displayed "revive" growth curves with a lengthy lag phase of over 13 hours when exposed to 2-32 mg/L vancomycin. The resistant subcolony SRR12-v1 was found to carry an identical vanM gene cluster to that of SRR12 but a significantly increased vanM copy number in the genome. Long-read whole genome sequencing revealed that a one-copy vanM gene cluster was located on a pELF1-like linear plasmid in SRR12. In comparison, tandem amplification of the vanM gene cluster jointed with IS1216E was seated on a linear plasmid in the genome of SRR12-v1. These amplifications of the vanM gene cluster were demonstrated as unstable and would decrease accompanied by fitness reversion after serial passaging for 50 generations under increasing vancomycin pressure or without antibiotic pressure but were relatively stable under constant vancomycin pressure. Further, vanM resistance in resistant variants was verified to be carried by conjugative plasmids with variable sizes using conjugation assays and S1-pulsed field gel electrophoresis blotting, suggesting the instability/flexibility of vanM cluster amplification in the genome and an increased risk of vanM resistance dissemination.

Edible microbial cellulose-based antimicrobial coatings and films containing clove extract

Recently, microbial cellulose-based coatings and films have attracted substantial attention because of their promising uses in numerous fields, such as food packaging. The present work was designed to synthesize active microbial cellulose-based coatings and films with a comprehensive investigation of their antimicrobial and structural properties. Microbial cellulose was synthesized by using a gram-negative bacterium called Acetobacter aceti. The produced microbial cellulose was mixed with sodium alginate, chitosan and starch to obtain two different composite solutions, i.e., microbial cellulose + starch + chitosan (MSC) and microbial cellulose + starch + sodium alginate (MSS). The antimicrobial properties were achieved by incorporating four different concentrations of clove extract into the composite solutions. The resulting composite solutions were tested against S. aureus, Shigella, Salmonella, and E. coli through the agar diffusion assay method. The clove extract was found to be effective in inhibiting the growth of these pathogens, as a clear zone of inhibition was observed at all clove extract concentrations, with a maximum zone of inhibition of 4.0 ± 0.05 cm on E. coli for the MSC solution incorporated with 4% clove extract. The best antimicrobial solutions found were then casted into films by pouring the solutions into petri dishes and drying at 50 °C in a tray drier. The antimicrobial activity was again evaluated for the films. The results indicated that MSC 3% showed a greater zone of inhibition against all pathogens (1.7 ± 0.18 cm). Furthermore, structural and thermal analyses of the formed films were conducted. The results indicated distinctive antimicrobial and structural characteristics of the films.Graphical

Three-dimensional volume rendering ultrasound for assessing placenta accreta spectrum severity and discriminating it from simple scar dehiscence

Prenatal ultrasound discrimination between placenta accreta spectrum and scar dehiscence with underlying nonadherent placenta is challenging both prenatally and intraoperatively, which often leads to overtreatment. In addition, accurate prenatal prediction of surgical difficulty and morbidity in placenta accreta spectrum is difficult, which precludes appropriate multidisciplinary planning. The advent of advanced 3-dimensional volume rendering and contrast enhancement techniques in modern ultrasound systems provides a comprehensive prenatal assessment, revealing details that are not discernible in traditional 2-dimensional imaging. This study aimed to evaluate the use of 3-dimensional volume rendering ultrasound techniques in determining the severity of placenta accreta spectrum and distinguishing between placenta accreta spectrum and scar dehiscence with underlying nonadherent placenta. A prospective, cohort study was conducted between July 2022 and July 2023 in the fetal medicine unit of Dr Soetomo Academic General Hospital, Surabaya, Indonesia. All pregnant individuals with anterior low-lying placenta or placenta previa with a previous caesarean section who were referred with suspicion of placenta accreta spectrum were consented and screened using the standardised 2-dimensional and Doppler ultrasound imaging. Additional 3-dimensional volumes were obtained from the sagittal section of the uterus with a filled urinary bladder. These were analyzed by rotating the region of interest to be perpendicular to the uterovesical interface. The primary outcomes were the clinical and histologic severity in the cases of placenta accreta spectrum and correct diagnosis of dehiscence with nonadherent placenta underneath. The strength of association between ultrasound and clinical outcomes was determined. Multivariate logistic regression analyses and diagnostic testing of accuracy were used to analyze the data. A total of 70 patients (56 with placenta accreta spectrum and 14 with scar dehiscence) were included in the analysis. Multivariate logistic regression of all 2-dimensional and 3-dimensional signs revealed the 3-dimensional loss of clear zone (P<.001) and the presence of bridging vessels on 2-dimensional Doppler ultrasound (P=.027) as excellent predictors in differentiating scar dehiscence and placenta accreta spectrum. The 3-dimensional loss of clear zone demonstrated a high diagnostic accuracy with an area under the curve of 0.911 (95% confidence interval, 0.819-1.002), with a sensitivity of 89.3% (95% confidence interval, 78.1-95.97%) and specificity of 92.9% (95% confidence interval, 66.1-99.8%). The presence of bridging vessels on 2-dimensional Doppler demonstrated an area under the curve of 0.848 (95% confidence interval, 0.714-0.982) with a sensitivity of 91.1% (95% confidence interval, 80.4-97.0%) and specificity of 78.6% (95% confidence interval, 49.2-95.3%). A subgroup analysis among the placenta accreta spectrum group revealed that the presence of a 3-dimensional disrupted bladder serosa with obliteration of the vesicouterine space was associated with vesicouterine adherence (P<.001). Three-dimensional volume rendering ultrasound is a promising tool for effective discrimination between scar dehiscence with underlying nonadherent placenta and placenta accreta spectrum. It also shows potential in predicting the clinical severity with urinary bladder involvement in cases of placenta accreta spectrum.

Unraveling the impact of fog on driver behavior in highway tunnel entrances: A field experiment

Objectives This research aims to (1) study the visual and psychological characteristics of drivers in foggy tunnel entrances, (2) compare driver behavior inside and outside foggy tunnels, and (3) propose improvement ideas based on the study results. Methods A realistic vehicle trial was conducted. Eight participants completed trials in both foggy and clear zones on the same roadway. Drivers’ physiological and psychological properties were analyzed using gaze, saccade, pupil, and heart rate as primary indicators. Pupil area change rate and heart rate variability (HRV) were used as secondary indicators. Results Visibility and tunnel area had a significant impact on drivers’ visual and psychological traits. For instance, drivers in the foggy zone exhibited longer fixation duration, larger pupil area, faster heart rate, and greater HRV. Moreover, the foggy zone inside the tunnel heightened drivers’ visual discomfort and psychological tension. Conclusion The study suggests a high risk associated with driving in the entrance area of highway tunnels during foggy conditions, particularly after entering the tunnel. This situation significantly increases drivers’ visual and psychological load. Furthermore, the effectiveness of retro-reflective facilities diminishes. Therefore, employing more self-illuminated or actively illuminated visual guiding facilities is recommended to aid drivers in acquiring traffic information.

Production and Optimization of a Lipase-Producing Bacteria Enterobacter hormachei Isolated from an Oil-Contaminated Soil

Lipase enzymes possess a wide range of industrial applications. Thus, the capacity of lipase-producing bacteria to proliferate on tributyrin agar medium was used to screen for them among various sources. The lipase producing bacteria with highest zone of clearance on the screening media were preserved on the agar slants. The preserved slants were characterized by16SrRNA gene sequencing. The nucleotide sequence so obtained by the 16SrRNA gene sequence was then put through phylogenetic analysis and homology search using the NCBI’s BLAST program. The sequence of lipase producing bacteria showed maximum resemblance with Enterobacter hormaechei bacterial strain. These bacterial strains were produced by inoculating the culture in the inoculum media and allowing it to enrich over night. Subsequently, 3% of the inoculum from the inoculum media was added to the production media, which was then incubated for 48 hours in a rotary shaker. After production the media was centrifuged and supernatant was extracted and used further for optimization, Optimization of the physiochemical parameters of the bacterial strain like inoculum pH, incubation period, inoculum size was found using one factor at a time (OFAT) approach and medium parameters like different carbon source, nitrogen source, substrate, minerals, salts (11 factors) were screened using Plackett-Burman (PB) design which is a full factorial design. The Lipase activity was found by using a titrimetric method using olive oil and Arabic gum mixture as substrate mixture. The maximum lipase activity was found for inoculum pH of 5, 48 hours of incubation time, and 5% inoculum size. The results of the PB design showed the significant parameters to be glucose (carbon source), peptone (nitrogen source), KH2PO4 (salt), and NaCl (mineral). The organism of the soil sample containing bacterial strain showed maximum lipase activity of 70 U/ml and protein concentration of 4.3 μg/mL at the optimized conditions. After centrifuging the culture media that had been improved, the supernatant was collected and partially purified using dialysis and the ammonium sulfate precipitation procedure. Following precipitation, the supernatant’s activity was measured to be 74 U/mL. After being gathered, the pallet was dialyzed in a dialysis bag and added to a buffer. Both the protein content and the lipase activity were estimated. The protein concentration determined by Lowry’s technique was found to be 6.2 μg/mL, and the lipase activity was reported to be 85.22 U/mL.

In silico encrypted peptide from green alga Tetraspora sp. CU2551 showed high antimicrobial activities

The green alga Tetraspora sp. CU2551 has been previously identified as a biohydrogen producer for an alternative option for sustainable energy production development. Its biomass still contains a source of valuable biomolecules, especially proteins. Crude algal protein showed significant antimicrobial activity against Gram-positive bacteria (Bacillus subtilis TISTR 1248 and Staphylococcus aureus TISTR 746) and Gram-negative bacteria (Escherichia coli TISTR 074 and Pseudomonas aeruginosa TISTR 2370), representing a clear zone of 19.0 ± 0.1, 18.3 ± 0.5, 17.7 ± 0.5, and 19.0 ± 0.5, respectively. Further investigation revealed that encrypted peptides from pepsin-hydrolyzed crude proteins increased the inhibition activity against these bacteria by 3.4–10.5 %. The hydrolyzed peptides were fractionated using a Sephadex G-25 column, and peptides in fraction 17 showed the highest antimicrobial activity. Through peptide identification using LC-MS and prediction of obtained peptide primary sequences with four bioinformatic tools (DBAASP, AMPDiscover, iAMPpred, and AMPscanner), five peptides were selected and synthesized for activity evaluation. The PT11 peptide (RYGAAMAVGIACA) demonstrated the highest antimicrobial activity among the five peptides tested and compared favorably to previously reported algal encrypted peptides. The findings of this research demonstrate that the encrypted PT11 peptide derived from Tetraspora sp. CU2551 has promising potential as an antimicrobial agent. Additionally, we suggest that PT11 might be used for other things besides this, like animal feed.

Development and evaluation of agro-waste biodegradation formulation

In the study, twenty strains of cellulose degrading microbes (Pseudomonas spp., Bacillus spp., Aspergillus spp. etc.) were isolated from the gut of termites and earthworms collected from different locations. The cellulolytic activity of the isolates of Aspergillus spp. were determined based on the formation of clear zone on CMC (Carboxymethyl cellulose) media. The strain C18 of Aspergillus flavus isolated from the gut of termite produced the largest zone with maximum cellulolytic index of 7.64. All the purified strains were subjected to in vitro cellulase activity through DNS (3,5-dinitrosalicylic acid) method which showed that strain C18 of Aspergillus flavus produced the maximum amount of glucose (0.507 mg ML ) and the -1 cellulase enzyme ( IU ML ) was recorded as 0.187 IU ML . Out of twenty microbial strains, ten best -1 -1 microbial strains were selected and subjected to in vitro FPA (filter paper assay). Aspergillus flavus strain C18 was found most effective with maximum glucose (1.059 mg ml ) and cellulase enzyme (0.196 -1 IUML ) production after 9 days of inoculation and therefore the consortium was made from two most -1 effective Aspergillus flavus strains (C12 and C18) which were found best effective in degradation of wheat straw substrate i.e., 61 per cent degradation of initial substrate weight. After evaluation of each strain, effective strains (C12 and C18) were used to prepare consortium formulations for commercial use in the field. These cellulolytic microbes have significant role in agro-waste management by degradation without harming the land and environment.. KEYWORDS :Agro-waste, Aspergillus flavus, biodegradation, cellulolytic microbes, formulation