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India is an agricultural country. Nearly 70% of the populations thrive in rural areas, engaged in agriculture making the backbone of our economy. The Trichoderma genus is widely distributed and provides an alternative to chemical pesticides in crops and plants growth. Trichoderma species are free living and commonly found in soil and root ecosystems. Root colonization by Trichoderma also regularly promotes growth and development of plants. They also perform beneficial and significant role in enhancing crop productivity, fight against pathogens, resistant to abiotic stress and increase of nutrients uptake from soil. The present study was undertaken to investigate quantitative and qualitative estimation of phosphate solubilizer by using nineteen isolates of Trichoderma grown on Pikovskaya’s Agar and NBRIP media. It was found that all isolates were capable of differentially utilizing 5 g/L TCP in NBRIP broth. This was indicated by the soluble phosphate concentrations and increase in acidity of growth medium. Out of 19 isolates, all isolates were capable in phosphate solubilization. However, the P- solubilizing efficiency was found higher in case of the isolate number-59 and 89. While isolate number-102 was the strongest and isolate number-23 was the weakest P-solubilizer. Minimum Phosphate solubilization concentration was observed in 3rd day (0%) that simultaneously increased on 7th day (52.63%) followed by 9th day (100%) and remaining all days 13th, 17th and 21st days also observed 100% efficiency.

BackgroundEnvironmental water samples are increasingly recognised as an important reservoir of antimicrobial resistance (AMR) genes. Polymerase chain reaction (PCR) and next generation sequencing (NGS) offer a potentially inclusive surveillance platform for a wide range of AMR genes. However, molecular methods are dependent upon the extraction of DNA of high yield and quality. Current options for DNA extraction from complex environmental matrices for downstream molecular applications are either expensive or low yielding. We present here a novel magnetic bead-based DNA extraction method, for the detection of antimicrobial resistance genes (ARGs) from river water in Malawi, named MagnaExtract. MethodsMagnaExtract involves initial filtration of 250ml freshwater, followed by an overnight incubation of the filter in 15ml buffered peptone water (BPW), common procedure in microbiology laboratories. 200{micro}l is then taken for a boil (95{degrees}C) and spin step and mixed with magnetic beads to bind DNA. Following washes with ethanol, the DNA is eluted in nuclease-free water. To determine the effectiveness of this method, 98 freshwater samples were collected from two rivers in Southern Malawi, and DNA was isolated using the MagnaExtract method, two commercial Qiagen (Germany) kits; PowerWater and DNeasy Blood and tissue, alongside a boil and spin of BPW, and a boil and spin from bacterial isolate grown on agar media. All samples were screened with a high-resolution melt (HRM) PCR panel previously validated for the detection of third generation cephalosporin and carbapenem ARGs. We compared the DNA yield obtained using all extraction methods, as well as the identification of each ARG. ResultsDNA yield using MagnaExtract was statistically greater than both boil and spin methods and DNeasy Blood & Tissue (Qiagen, Germany). DNA yield was slightly lower than using PowerWater (Qiagen) but the difference was not statistically significant. MagnaExtract was the only method to identify ARGs in all 98 water samples compared with PowerWater (n=82), DNeasy (n=95) boilate of BPW (n=75) and boilate of bacterial isolate (n=87). The most commonly detected ARG was OXA-48 (n=93). In addition, we found overnight incubation in non-selective enrichment broth (BPW) to promote the growth of bacteria harbouring extended spectrum beta lactamase (ESBL) genes and reduction in the detection of carbapenemase genes. ConclusionThe MagnaExtract approach offers a simple, affordable, high yielding DNA extraction method for the detection of ARGs isolated from river water samples.

This work reports for the first time the anticandidal activity of the dichloromethane (DCM) extract from the Algerian invasive red marine algae A. armata Harvey. In a preliminary assessment, the DCM extract was screened for its antifungal activity against two strains of Candida albicans, namely (IP 444) and (ATCC 10231). As a well-established approach, the anticandidal activity was expressed as zone of inhibition by agar well and disk diffusion methods. The minimum inhibitory concentration (MIC) was also determined for the two fungal strains. Within this framework, the DCM extract of the red algae A. armata has a strong inhibition effect against the considered Candida albicans strains. In addition, Candida albicans IP444 (MIC = 0.58 mg/mL) was more sensitive to DCM algal extract when compared to Candida albicans ATCC 10231 (MIC = 2.34 mg/mL). GC–MS analysis suggested that brominated compounds and fatty acids are responsible for anticandidal activity of dichloromethane extract of A. armata. These preliminary results may constitute a future applied of lipophilic extract of the red algae A. armata as a promising source of natural compounds with antifungal properties.

The aim of this study was to evaluate the phytochemicals, antioxidant, antibacterial and anti-quorum sensing (QS) properties of methanol extract of Muntingia calabura L. leaves against biofilm-forming strain Pseudomonas aeruginosa. The antioxidant potential of M. calabura methanol leaf extract was evaluated using a radical scavenging assay. Since the findings were so promising, the study aims to screen the phytochemical profiles using qualitative and quantitative approaches and to expand the investigation to anti-QS activity. The antibacterial activity was assessed by determining the minimum inhibitory concentration against pathogenic Gram-negative and Gram-positive biofilm-forming bacterial strains using an agar well diffusion plate method. Muntingia calabura methanol leaf extract was most potent against P. aeruginosa. The QS controlled virulence factors in P. aeruginosa is significantly inhibited by M. calabura leaf extract. The microscopy images revealed a major reduction in P. aeruginosa biofilm formation. Interestingly, M. calabura leaf extract was not toxic to Vero cell line. These findings make M. calabura a strong candidate for new antimicrobial and anti-QS herbal agents.

Heavy metal resistance (Cu, Zn, Fe, Cr, Pb, Cd) and removal capacities of 19 selected actinobacteria strains were performed in order to investigate their applicability as bioremediators for polluted areas. Streptomyces sp. WR1L1S8 was used in the present investigation as heavy metals removing strain, which was isolated from inner tissu of the marine brown algae Fucus sp., and identified based on polyphasic and molecular characterization. The results showed that WR1L1S8 could successfully remove Fe2+, Cu2+ and Zn2+ by 60.74%, 42% and 39.02%, respectively, using Atomic Absorption Spectroscopy (AAS) analysis. The strain WR1L1S8 had a considerable tolerance to initial Cu2+ with a minimum inhibitory concentration value recorded at 250 mg L−1. Rotatable Central Composite Design (CCD) was used to optimize the copper removal yield using nutrient broth in batch experiment studying the influence of four operating parameters: incubation time, stirring speed, inoculum size and percentage of seawater. The optimal conditions were found at incubation time of 9 days under 114.9 rpm of a stirring speed, 18 agar cylinders as inoculum and 80.75% of seawater reaching a removal capacity of 75.24% which correspond to 29.65% increase compared with the non-optimized conditions. WR1L1S8 exhibited a promising potential for copper removal from aqueous solutions.

Aims Escherichia albertii is an emerging diarrheagenic pathogen causing food- and water-borne infection in humans. However, no selective enrichment broths for E. albertii have ever been reported. In this study, we tested several basal media, selective supplements and culture conditions which enabled selective enrichment of E. albertii. Methods and Results We developed a selective enrichment broth, novobiocin–cefixime–tellurite supplemented modified tryptic soy broth (NCT-mTSB). NCT-mTSB supported the growth of 22 E. albertii strains, while inhibited growth of other Enterobacteriaceae at 37°C, except for Escherichia coli and Shigella spp. Enrichment of E. albertii was improved further by growth at 44°C, a temperature that suppresses growth of several strains of E. coli/Shigella. Combined use of NCT-mTSB with XR-DH-agar, xylose–rhamnose supplemented deoxycholate hydrogen sulphide agar, enabled isolation of E. albertii when at least 1 CFU of the bacterium was present per gram of chicken meat. This level of enrichment was superior to those obtained using buffered peptone water, modified-EC broth, or mTSB (with novobiocin). Conclusions Novobiocin–cefixime–tellurite supplemented modified tryptic soy broth enabled effective enrichment of E. albertii from poultry samples and was helpful for isolation of this bacterium. Significance and Impact of Study To our knowledge, this is the first report of selective enrichment of E. albertii from poultry samples.

BACKGROUND: Periprosthetic joint infection is a major complication of total joint arthroplasty, with treatment requiring a two-stage exchange procedure and 6 weeks of systemic antibiotics. However, depending on the infection site, intravenous delivery of antibiotics like vancomycin (VCM) can have poor tissue transferability, thus reducing their therapeutic effect. OBJECTIVE: This study demonstrates the 24-week in vivo release profile and antibacterial activity of VCM from calcium phosphate cement impregnated with VCM (CPC/VCM) and compares them with those from polymethylmethacrylate impregnated with VCM (PMMA/VCM). METHODS: Rats were implanted with the test specimens between the fascia and quadriceps. After implantation for 24 weeks, the test specimens were removed and residual VCM was extracted to calculate the concentration of VCM released into rat tissues. We also examined the antibacterial activity of releasable VCM from the removed test specimens by placing them directly onto the surface of agar. RESULTS: CPC/VCM released greater concentrations of VCM for a longer period of time within the 24 weeks than PMMA/VCM. Moreover, CPC/VCM released 1.4 to 26.1-fold more VCM than PMMA/VCM. Using Staphylococcus aureus, antibacterial activity was logarithmically correlated with VCM concentration across the entire concentration range tested (12.5–800 μg/mL). While the area within which inhibition was observed—the inhibition zone—for both CPC/VCM and PMMA/VCM formed and gradually shrank with time after implantation, that for CPC/VCM was significantly larger than that for PMMA/VCM in each week after implantation. CONCLUSION: CPC/VCM releases greater amounts of VCM with antibacterial activity for longer periods of time than PMMA/VCM, suggesting that CPC is effective for facilitating the release of antibiotics for local action in patients with established postoperative infection.

Selenium (Se) is an essential trace element for human beings and animals. Traditional plant Se enrichment technology suffers from selenium pollution. Herein, environmentally friendly Se-agarose (Se-Agar) hybrid hydrogels are prepared by simply mixing agar with different Se species including selenocarrageenan (SeCA), selenite and Se yeast under heating and stirring for 0.5 h without any other reagent. Such Se-Agar hybrid hydrogels with excellent biocompatibility were used as natural substrates for the cultivation of Se-enriched mung bean sprouts. Compared with Se yeast, SeCA and selenite show a better Se enrichment effect on mung bean sprouts. Furthermore, the growth indices including plant weight and plant height of mung bean sprouts were investigated with different concentrations and sources of Se. Notably, the Se-Agar hybrid hydrogels could be easily regenerated and reused for multiple cycles. The results indicated that Se-Agar hybrid hydrogels as recyclable natural substrates offer a simple, sustainable and affordable strategy for plant Se enrichment.

Introduction Gingivitis is an oral condition characterized by inflammation and bleeding of the gingiva (gums), largely caused by Porphyromonas gingivalis. Oral hygiene options for controlling P. gingivalis include mouthwash containing Commiphora myrrha (myrrh), which has been shown to be effective against the microbe. Silver nanoparticles (SN) have been studied for their antibacterial effect in different oral health applications, including mouthwash. This was an in vitro laboratory study of the anti-microbial actions of myrrh and SN against P. gingivalis. Methods We compared the anti-microbial properties against P. gingivalis of four solutions: a) placebo solution, b) myrrh solution (MS), c) MS mixed with silver nanoparticles (MSN), and d) SN suspension alone. Sixteen agar plates were divided into four groups of four plates, and each group was treated with one of the solutions/suspensions. The solution/suspension was administered on the agar disc diffusion method, and inhibition zones (IZs) were measured after 24 (time 1), 48 (time 2), and 72 h (time 3). To characterize MSN and SN, Fourier-transform infrared spectroscopy (FT-IR) was used. UV–Vis spectroscopy and energy dispersive X-ray (EDX) were used to further characterize MSN. Results After 24 h, the median IZ for the MS plates was 16 mm, and the median IZ for MSN plates was 15 mm. At time 2, the MS median IZ was 15 mm, but the MSN median IZ increased to 18 mm, and the interquartile ranges (IQRs) did not overlap. At time 3, the median IZs was similar again, with MSN and MS having IZs of 16 mm and 15 mm, respectively. SN alone showed no anti-microbial activity. Conclusions Our findings show that MSN displayed superior anti-microbial activity against P. gingivalis compared to MS and SN after 48 h of incubation, but not after 24 h. Also, the increased anti-microbial activity had ceased by 72 h.

Nanotechnology has proven as progressive technology that enables to contribute, develop several effective and sustainable changes in food products. Incorporating nanomaterials like TiO2, SiO2, Halloysite nano clay, Copper sulfide, Bentonite nano clay, in carrageenan to develop innovative packaging materials with augmented mechanical and antimicrobial properties along with moisture and gas barrier properties that can produce safe and healthy foods. Intervention of carrageenan-based bio-nanocomposites as food packaging constituents has shown promising results in increasing the shelf stability and food quality by arresting the microbial growth. Nanomaterials can be incorporated within the carrageenan for developing active packaging systems for continuous protection of food products under different storage environments from farm to the fork to ensure quality and safety of foods. Carrageenan based bio nanocomposite packaging materials can be helpful to reduce the environmental concerns due to their high biodegradability index. This review gives insight about the current trends in the applications of carrageenan-based bio nanocomposites for different food packaging applications.

Antifungal susceptibility testing is an essential tool for guiding antifungal therapy. Reference methods are complex and usually only available in specialised laboratories. We have designed an expanded agar-based screening method for the detection of azole-resistant Aspergillus fumigatus isolates. Normally, identification of resistance mechanisms is obtained only after sequencing the cyp51A gene and promoter. However, our screening method provides azole resistance detection and presumptive resistance mechanisms identification. A previous agar-based method consisting of four wells containing voriconazole, itraconazole, posaconazole and a growth control, detected azole resistance to clinical azoles. Here, we have modified the concentrations of voriconazole and posaconazole to adapt to the updated EUCAST breakpoints against A. fumigatus. We have also expanded the method to include environmental azoles to assess azole resistance and the azole resistance mechanism involved. We used a collection of A. fumigatus including 54 azole-resistant isolates with Cyp51A modifications (G54, M220, G448S, TR53 , TR34 /L98H, TR46 /Y121F/T289A, TR34 /L98H/S297T/F495I), and 50 azole susceptible isolates with wild-type Cyp51A. The screening method detects azole-resistant A. fumigatus isolates when there is growth in any of the azole-containing wells after 48h. The growth pattern in the seven azoles tested helps determine the underlying azole resistance mechanism. This approach is designed for surveillance screening of A. fumigatus azole-resistant isolates and can be useful for the clinical management of patients prior to antifungal susceptibility testing confirmation.

Electrolyzed waters, containing mainly hypochlorous acid, are used in dental practice because of their high microbicidal effect. For wider use, three neutral electrolyzed water-based gels, namely, HOCl-containing aqueous gels were prepared with a thickening/gelling agent in this study. We evaluated their microbicidal effects against four strains and storage stabilities indicated by available chlorine concentration. Immediately after preparation, all gels (70 ppm) could completely remove microbes by a 3-min treatment. The gel prepared with xanthan gum remarkably reduced its available chlorine concentration even under shaded and refrigerated storage conditions, failing to maintain its microbicidal effect following 1-day storage, whereas other gels, prepared with carboxyvinyl polymer or agar, maintained effective concentration (>20 ppm), with high microbicidal effects following 9-day and 21-day storage, respectively. Neutral electrolyzed water-based gels might be useful to remove oral microbes. Based on our results, agar is the most suitable thickening/gelling agent from the viewpoint of storage stability.

This research aimed to isolate protease-producing bacteria from Sidoarjo shrimp paste for extracting collagen from milkfish scales. This study began with isolation, followed by screening and purification of protease-producing bacterial isolates. Further confirmation of the isolates’ proteolytic indices and the crude protease production, the enzymes’ efficacy in extracting collagen from milkfish scales were tested, followed by pathogenicity and identification using 16S rRNA molecular technique. The study has successfully isolated 15 proteolytic bacterial isolates using skimmed milk agar, but only isolates of TR-10, TR-4.1.1, and TR-15.1 exhibited prospective proteolytic activity based on their corresponding proteolytic indices of 2.96 ± 0.06, 3.10 ± 0.10, and 3.71 ± 0.48. Although the proteolytic activity of isolates TR-10 (0.22 ± 0.05 U/mL) and TR-15.1 (1.07 ± 0.14 U/mL) was high in a salt medium using peptone as the nitrogen source, only the former showed satisfactory activity to extract soluble collagen from milkfish scales. Based on the 16SrRNA, the TR-10 isolate was identified as Bacillus megaterium. The non-pathogenicity of the TR-10 bacterium signified its promising role as a protease source for the halal collagen extraction from milkfish scales.

The in vitro effects on mycelial growth of the antagonistic fungi Trichoderma asperellum (ICC 012) and Trichoderma gamsii (ICC 080) were studied based on the active ingredients of 12 commonly used herbicides: amidosulfuron, dicamba, fluazifop-P-butyl, fluorochloridone, foramsulfuron, isoxaflutole, metribuzin, pendimethalin, prosulfocarb, pyridate, S-metolachlor and tembotrione. The fungal growth rates were evaluated at 20 °C and 25 °C using PDA plates that included 100%, 75%, 50%, 25%, 12.5%, 6.25% and 0% of the recommended herbicide application rates. These herbicides were classified into four categories based on inhibition of mycelial growth in the toxicity testing. Growth inhibition of Trichoderma spp. was seen for all of these herbicides at almost all doses and at both temperatures, although some major differences were seen. Lower herbicide concentrations in the agar also reduced the inhibitory effects, but generally not linearly. Temperature had no significant effects on the levels of inhibition of mycelial growth. Amidosulfuron, dicamba and isoxaflutole showed the least inhibition of mycelial growth of both Trichoderma spp., and the lowest doses with amidosulforone (6.25%, 12.5%) even significantly stimulated mycelial growth of both Trichoderma spp. Fluazifop-P-butyl, pyridate and tembotrione showed moderately harmful to harmful effects on mycelial growth, with 70% to 80% inhibition at the recommended herbicide application rates. Fluorochloridone, S-metolachlor, prosulfocarb, metribuzin and pendimethalin inhibited mycelial growth the most, in descending order of inhibition. At 50% to 100%, fluorochloridone, prosulfocarb and S-metolachlor had fungicidal effects. Fluorochloridone and prosulfocarb had significant inhibitory effects (> 75%) even at the lowest concentration of 6.25%. Of these herbicides tested, the least adverse effects were seen for amidosulfuron, dicamba and isoxaflutole, which are thus likely to be compatible with Trichoderma spp. in the field.

The purpose of this research is to obtain hydrocarbon degrading bacteria that work synergistically in a consortium. Consortium microorganisms is mixture of microbial populations in the form of communities that have mutualistic relationships and doesn’t inhibition the growth of other microbes. In this study, isolates were obtained from the rhizosphere of soil contaminated with petroleum. The isolates obtained were tested for synergism to determine the relationship between bacterial isolates. Synergism testing was carried out using the spread plate method on agar media. The results of this study showed that isolate number one showed antagonistic properties to other bacterial isolates by forming a clear zone around the disc paper. A total of eight bacterial isolates showed the greatest percentage of synergism, namely ≥ 80% so that the eight rhizosphere bacterial isolates could be used as materials for mixed culture.

Currently, the majority of fresh apricots destined for the production of dried apricots undergo sulphur oxide fumigation before drying to protect the fruit against fungal spoilage. To eliminate the use of sulphite, packaging assisted with essential oil is a promising strategy to increase shelf-life of dried apricots since it does not impact its flavor characteristics. In this study, three essential oils were selected: clove, lemongrass and thyme. They were screened for antifungal activity against Eurotium spp. with different methods: micro- and macro-dilution and agar-diffusion. Growth/no-growth data were used to develop models for all three methods. Clove exerted the strongest antifungal activity with an inhibitory concentration of 0.075%, 0.035% and 0.05% through respectively micro-dilution, macro-dilution and agar diffusion. For thyme the following values were obtained: 0.775%, 0.070% and 0.100%. This means that the antifungal activity of thyme is 10 times lower in micro-dilution and 2 times lower in macro-dilution and agar diffusion compared to clove. Through micro-dilution, lemongrass was found to have the second highest antifungal activity (0.25%). When used in the volatile atmosphere of dried apricots and in macro-dilution, the antifungal activity of lemongrass was the lowest, with respective values of > 0.200% and 0.105% for G/NG prediction.

Urinary tract infection (UTI) has been reported most often in recent days. In a clinical aspect there are various reasons for this cause. Out of 100 samples tested (Clinical samples from Rashi Diagnostic Center- Bangalore NABL laboratory), under the analytical aspect several comparative analysis was done. Urine was tested within half an hour of the collection. Physical characteristics of Urine sample were studied initially. Laura M is the device used to analyses the Uro-dip. Sugar and the presence of albumin was also found. Sediment of the centrifuged sample was subjected to gram staining and direct microscopic analysis. Pus cells, epithelial cells, yeast cells and rod shaped bacteria. It was found that epithelial cells are visualized in female samples and very less or not visualized in male samples. Bilirubin content, Bile salt content and ketone bodies were analyzed and compared in each of the samples. Sample is then subjected to culturing were the T streak is done on the Mac Conkey agar and Blood agar. This will differentiate between the lactose and Non-Lactose fermenting bacteria. Each of the colony is subjected to Gram staining. Yeast cells are also reported often. Though the yeast cells are the commensals they are considered non- pathogenic. But it was found that the yeast with pseudo-hyphae are pathogenic in nature. Both Lactose and Non-Lactose fermenting bacterial colonies from Blood agar is subjected to various biochemical methods to identify the organism. The most commonly noticed non-lactose fermenting organism was Pseudomonas spp. And Lactose fermenting organisms were Escherichia coli, Enterobacteria spp and Klebsiella spp. Each Colony from the culture plate was picked and streaked on the MHA agar and Kirby Bauer method has been performed (Disc diffusion method) under the McFerland standard. Zone of inhibition of each organism towards the antibiotic in the disc was recorded.

Background Agar has been commonly used as one of the materials to fabricate magnetic resonance imaging phantoms in the past few decades. In this study, eleven agar gel phantoms with different iron (III) oxide (Fe2O3) masses were prepared. This study was aimed to evaluate the signal-to-noise ratio (SNR) uniformity and stability of agar gel phantoms with and without the addition of Fe2O3 at two different time points (TPs). Fe2O3 powder was used as a relaxation modifier to manipulate and produce various SNR, T1 and T2 values. These phantoms were scanned using turbo spin echo pulse sequence to produce T1- and T2-measurement images. The SNR was then computed by plotting 1, 3 and 25 regions of interest on the images using ImageJ software. The T1 and T2 relaxation equations were then fitted to the experimental results of SNR versus TR and SNR versus TE curves for the determination of saturation (SNRo), T1 and T2 values. Results The results demonstrated that the agar gel phantoms were able to maintain SNR uniformity but not SNR stability after 4 weeks of phantom preparation. The change in the water content and microstructure of the phantoms have no significant effect on T2 relaxation but on T1 relaxation. The T1 and T2 of the agar gel phantoms were minimally affected although there was a systemic increase in the content of the Fe2O3 powder. Conclusions It can be concluded that the agar gel phantoms exhibited the characteristics of SNR uniformity, but they showed instability of SNR at TP2. The Fe2O3 in powder form is not an effective relaxation modifier to reduce the T1 and T2 when it is introduced into the agar gel phantoms. Dissolved nanosized particles should be the focus of future studies.

In-home or food service antimicrobial treatment options for fresh produce are limited. Hot water treatments for whole (unpeeled) produce have been proposed, but data to support this practice for onions are not available. Separate cocktails of rifampin-resistant Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella were cultured on agar and suspended in sterile water. The outer papery skin at the equator or root or stem ends of the whole yellow onions was spot inoculated at 6 log CFU per onion. After drying for 30 min and, in some cases, storage at 4°C for 6 days, onions were immersed in water at ca. 100°C for 5 s or 85°C for 10 to 180 s. No significant difference (P > 0.05) in the mean decline of Salmonella was found on onions that were exposed to hot water after drying the inoculum for 30 min or after storage at 4°C for 6 days. Exposure of whole onions at 100°C for 5 s reduced E. coli O157:H7 and L. monocytogenes populations by >5 log CFU per onion at all inoculum sites and Salmonella populations by >5 log CFU per onion at the stem end and equator but not consistently at the root end. Mean root-end reductions of ≥5 log CFU per onion of E. coli O157:H7, L. monocytogenes, and Salmonella were achieved consistently when the root end was fully immersed in 85°C hot water for 45 or 60 s except in a small number of cases (4 of 57; 7%) when the root end was oriented upward and above the water line during treatment. When onions were held at 85°C for 180 s with the root end above the water line in an uncovered water bath, no significant declines in Salmonella populations were observed; significant mean declines in Salmonella were achieved (mean, 5 log CFU per onion; range, 3.49 to 6.25 log CFU per onion) when the water bath was covered. Short exposure to hot water can significantly reduce pathogens on the surface of whole onions. Reductions are more consistent when the root end is submerged and when the water bath is covered. HIGHLIGHTS

There is evidence that Staphylococcus aureus colonisation is linked to severity of atopic dermatitis. As no gold standard for S. aureus sampling on atopic dermatitis skin lesions exists, this study compared three commonly used methods. In addition, effectiveness of standard skin disinfection to remove S. aureus colonisation from these inflamed skin lesions was investigated. In 30 atopic dermatitis patients, three different S. aureus sampling methods, i.e. detergent scrubbing, moist swabbing and tape stripping, were performed on naïve and disinfected skin lesions. Two different S. aureus selective media, mannitol salt agar and chromID agar, were used for bacterial growing. Quantifying the S. aureus load varied significantly between the different sampling methods on naïve skin lesions ranging from mean 51 to 1.5 × 104 CFU/cm2 (p p S. aureus load (p S. aureus selection agar plates to implement further clinical studies for the effectiveness of topical anti-staphylococcal antibiotics. Other disinfection regimes should be considered in atopic dermatitis patients when complete de-colonisation of certain skin areas is required, e.g. for surgical procedures.

The availability of non-synthetic media from natural ingredients is needed to answer the needs in laboratories where the price of nutrient media is quite expensive and there are limited supplies of material ware houses. Cowpea (Vigna unguiculata L.) and pigeonpea (Cajanus cajan L.) are the local foods of people of NTT (East Nusa Tenggara) which have a high enough nutritional content which has the potential to be developed into cheap, easy and simple non-synthetic media in making. The purpose of this study was to determine whether the agar media contained nutrient from cowpea and pigeonpea water can be used as a alternative for nutrient agar for the growth of Escherchia coli and Staphylococcus aureus bacteria. This research is a true experiment with posttest-only control design. The growth rate of S. aureus bacteria on pigeonpea medium, cowpea medium, nutrient agar medium, were 164 CFU/mL (SD=3,13), 161 CFU/mL (SD=3,02) and 164 CFU/mL (SD=3,21), respectively. The average growth of E. coli on cowpea medium, pigeonpea medium, and nutrient agar control medium were 163 CFU/mL (SD=2,79), 167 CFU/mL (SD-2,63) and 164 CFU/mL (SD=2,75) respectively. Test results ANOVA between pigeonpea medium, cowpea medium and nutrient medium in order to obtain p value = 0.145 (p> 0.05) for the growth of E. coli bacteria and p value = 0.393 (p> 0.05) for growth S. aureus. It was concluded that there was no difference between the number of bacterial colonies of E. coli and S. aureus on three medium. The pigeonpea medium and cowpea can be used to grow and alternative nutrient agar in order to grow bacteria E. coli and S. aureus.

Pseudomonas oleovoransNCIMB 6576 and Ralstonia eutropha NCIMB 10442 were used for the production of Polyhydroxyalkanoates (PHA) from industrial waste cooking oils, the bacteria were cultured on tryptone soya broth (TSB) and Tryptone soya agar (TSA).P. oleovoransNCIMB6576 gave a better percentage PHB yield (8.2%) with PS oil as carbon source as compared to 6.45% with TS oil. However, a very low yield (0.64%) was recorded whenP. oleovoransNCIMB6576 was grown on TSB without the oils as carbon source. Ralstonia eutropha NCIMB 10442 gave an appreciable yield of 13.63% and 14.80% with PS and TS oil samples respectively as carbon source with negligible variation in the yields. The results obtained across all experiments were compared with one another. The SEM images from the PHB samples generated from the experiments shows that there is a slight difference in the surface morphologies of the PHB with respect to the oil samples as well as the different bacteria used in the experiment.

The aim of the work is to evaluate ability of gentian violet and malachite green to inhibit 2 Brucella test strains and 8 strains of unwanted organisms and the influence of autoclaving on their activity. The study also aims to evaluate variability of inhibition in different batches of colorants and necessity in adjustment of their concentration.Materials and methods. The study included 3 imported lots of gentian violet and malachite green. Inhibitors were put into nutrient medium containing pancreatic hydrolysates of gelatin and fsh meal, yeast extract, sodium chloride, glucose, sodium pyrosulfate, and agar. The same medium without colorants was used as a control medium. Effect of the colorants was studied with the help of test strains of Brucella abortus 19 ВА, B. melitensis Rev I, Enterococcus faecium VKM B-602, Escherichia coli ATCC 25922, Proteus vulgaris HX 19 222, Pseudomonas aeruginosa 22/99, Serracia plymuthica 1, Shigella flexneri 1a 8516, Staphylococcus aureus ATCC 25923, Yersinia enterocolitica C-187.Results and discussion. We have determined biological parameters of nutrient media containing gentian violet (5,0; 2,5 1,25 mg/L) and malachite green (2.0; 1.0; 0.5 mg/L) when cultivating brucella. Gentian violet is able to inhibit Brucella greatly at concentration of 5 mg/L. S. aureus ATCC 25923 did not show any growth on the media containing malachite green or gential violet (5.0 and 2.5 mg/L). Malachite green inhibited the growth of enterococcus more actively than gentian violet. However, it inhibited swarming growth of Proteus less successfully than gentian violet. Other unwanted organisms were not sensitive to such concentrations of colorants at all. We have demonstrated the necessity to combine colorants with other selective agents, as well as advisability to put gentian violet into sterile media, while malachite green – into the media before its autoclaving. Adjustments to the concentration of gentian violet should be made when producing nutrient medium, as variations in inhibiting activity in different batches of this colorant were revealed.

Background and Objectives. It has been shown that Myrtus communis and Marrubium vulgare have antibacterial activity against bacterial and fungal strains of different diseases. This study aimed to evaluate in vitro the antibacterial activity of aqueous and methanolic extracts of Myrtus communis and Marrubium vulgare leaves against selected periodontal pathogens. Materials and Methods. Clinical strains of Aggregatibacter actinomycetemcomitans and Eikenella corrodens and two reference strains of A. actinomycetemcomitans were tested. The antibacterial activity of each studied plant extract was evaluated using agar diffusion and broth microdilution assays. Results. Both aqueous and methanolic extracts of M. communis exhibited high antibacterial activity against periodontal pathogens as compared to M. vulgare extracts. At concentrations of 2.5-0.32 mg/disc, inhibition zones of the methanolic extract of M. communis ranged from 19.66 ± 0.57 to 12.33 ± 0.57 mm. The methanolic extract of M. vulgare showed at concentrations of 5-0.63 mg/disc inhibition zones ranging from 15.66 ± 0.57 to 12 ± 0.00 mm. Its aqueous extract at concentration of 0.63 mg/disc showed no antimicrobial activity against the clinical and reference strain of A. actinomycetemcomitans. Conclusion. This study showed that methanolic and aqueous extracts of M. communis and M. vulgare have in vitro an antibacterial activity against periodontal pathogens. They could be use as ingredients of an oral antimicrobial agent for prevention or treatment of periodontal diseases. Further research on isolating the compounds from these plant extracts and their toxicity effect could be conducted.

Background: Calcium hydroxide is the gold standard medicament for root canal treatment. Enterococcus faecalis, the primary cause of intraradicular persistent endodontic infection, is often identified even after endodontic treatment. Thymoquinone, an active ingredient of Nigella sativa, has an antimicrobial effect on both gram-negative and positive bacteria, including E. faecalis. Purpose: This study aimed to evaluate the inhibitory effect of calcium hydroxide combined with Nigella sativa extract and determine the best ratio for the combined material. Methods: This is an experimental study comprised of six groups (n = 4 per group) based on the material and its ratio, namely; (1) calcium hydroxide; (2) Nigella sativa extract; and groups of the combination of calcium hydroxide and Nigella sativa extract with a ratio (3) 70:30, (4) 50:50, (5) 30:70, (6) 10:90. The inhibitory effect against E. faecalis was evaluated by the agar well diffusion method in Muller–Hinton agar. Observation of the inhibitory zone was performed on the first, third, and seventh days. The collected data were analysed by a one-way ANOVA and LSD post hoc test. Results: Calcium hydroxide has the highest inhibitory effect, and the combination of Nigella sativa extract with calcium hydroxide ratio 50:50 was second. The inhibitory zone of these two groups was significantly higher than in any other group (p

BACKGROUND: Dadiah, traditional yogurt from Indonesia, which is known as a source of probiotics, also contains micronutrients. AIM: This descriptive study aimed to determine whether additional ingredients and processes during the manufacture of Dadiah pudding maintain the iron, zinc, calcium, and total lactic acid bacteria (LAB) contents. METHODS: Dadiah pudding was made using Dadiah originated from Bukittinggi, West Sumatra. Micronutrient levels were analyzed according to Indonesian National Standard 01-2896-1998. The total LAB were counted by inoculating samples on Man, Rogosa, and Sharpe agar with serial dilution, and morphological identification was carried out using gram stain. RESULTS: In 100 g of original Dadiah, mango and chocolate Dadiah pudding contained 347.98, 276.61, and 279.29 mg of calcium; 4.87, 3.75, and 6.31 mg of zinc; 6.53, 6.60, and 9.39 mg of iron; and 6.4 × 10⁹, 6.1 × 10⁹, and 2.4 × 10⁹ CFU/ml LAB, respectively. CONCLUSION: This study found that modifying the original Dadiah into Dadiah pudding has been proved to affect the concentration of calcium, zinc, and iron and to maintain total LAB. We suggest that consuming Dadiah pudding may be a good choice as a food supplementation for pregnant women to optimize the golden period outcomes.

Three species of red algae Gracilaria corticata (J. Agardh), Gracilaria dentata (J. Agardh) and Gracilariopsis longissima (S.G. Gmelin), Steentoft, L. M; Irvine and Farnham (formerly Gracilaria verrucosa (Hudson) were collected from four different sites (Buleji, Hawks Bay, Manora and Paradise Point) of Karachi coast. The G. corticata was the dominant species and the highest yield of agar was compared to other studied species. The physical properties such as gel temperature, melting temperature, density, viscosity and gel strength showed large variations. Interestingly, the gel temperature, melting temperature, density, and gel strength had the highest value in G. corticata samples collected, while gel viscosity recorded the highest value in G. dentata samples. Intensive spectroscopic FTIR analysis was determined in all three species of G. corticata, G. dentata and G. longissima. The bands at 414.7/cm to 3917.2/cm represents stretching and bending vibrations of alcohol O-H, amine N-H, alkane C-H, alkyne C=C, nitriles C=N, carboxyl C=O, nitro aromatic N=O, alkane C-C, nitro methane C-N, aliphatic amines C-N, sulfoxides S=O, alkene C-H alkyl halide C-Cl, C-I groups. The ash content of all studied species (G. corticata, G. dentata and G. longissima) was in the range of 20-30%, while the carbohydrate content was in the range of 22-24%. The results of this study suggested the utilization of our natural resources present in Karachi coast. This could be achieved by determining the quantity and quality of agar in the edible species of Gracilaria/Gracilariopsis.

Tomato paste is fast becoming first choice for many homes in cooking while meeting up with their busy schedules in a cosmopolitan city like Lagos State, Nigeria. The study was aimed at evaluating the antioxidant contents, microbial load and heavy metal contents of both unprocessed and processed tomato products. β-carotene, lycopene and vitamin C contents were analysed accordingly on selected sachet tomato paste. Microbial colony counts were done using both nutrient agar and potatoes dextrose agar. Heavy metals from sampled tomato products were done using Atomic Absorption Spectrometry. The study observed β-carotene ranged between 0.02 – 0.52 mg/g, lycopene ranged between 0.12 – 0.56 mg/g and vitamin C contents of ranged between 12.50 – 75.0 mg/mL. These were more than WHO/FAO and FEPA recommendations. All tomatoes samples showed microbial load higher than limits allowable. Bacillus sp., Salmonella sp., Aspergillus sp. were predominantly isolated from sampled processed tomato products. Cadmium contents were above the permissive limit in some of the tomato paste products. Copper, zinc, lead, nickel, chromium and iron were not detected in all samples evaluated. The study showed few processed tomato products were safe for consumption. Hence, monitoring by regulatory agencies for consumer safety regulations is highly recommended on processed tomato producers.

Polysaccharide biopolymer Agar-Agar extracted from red algae is a natural and biodegradable polymer. It is a combination of agarose (a neutral and linear polymer, with repeated units of agarobiose) and a heterogeneous mixture of agaropectin (a charged sulfated polymer). In this study, a comparative study of structural vibrational and electrochemical properties of agar-agar biopolymer with two different methods HF (Hartree-Fock) and DFT (Density Functional Theory) using a basis set 631+G (d, p) is performed. The comparative structural study of agar-agar biopolymer by HF and DFT method has been carried out to calculate the stability of the molecule. The thermionic properties and Mulliken charge distribution are analysed to deliver a quantitative study of partial atomic charge distribution. The overall vibrational analysis of primal modes of the biopolymer has been studied using FTIR analysis. Based on highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) composition and energies, various chemical parameters of the biopolymer have been evaluated. The Physico-chemical properties of this polysaccharide show a strong correlation with its optimized structure. Agar-agar has its application in the electrochemical, biotechnological, and pharmaceutical fields, as a stabilizer and gelling material.

Background Soleris®E. coli is an automated, growth-based method for detection and semi-quantitative determination of Escherichia coli in foods. The method can be used in dilute-to-specification (threshold) or presence/absence modes. Objective The objective of the study was to validate four modifications to the method: (1) a change in the vial detection window plug composition from agar to agarose to improve plug consistency and robustness, (2) a change in pre-enrichment incubation time for presence/absence testing from 6 h to 18–24 h, (3) a change in vial incubation temperature from 44.5 to 43.5°C, and (4) incorporation of a simple direct-from-vial confirmation test as an alternative to traditional procedures. Methods Elements of the study included inclusivity/exclusivity testing, matrix testing in comparison to the ISO 7251:2005 reference method, reagent stability/lot-to-lot consistency testing, and an independent laboratory study. Results In inclusivity testing, all 55 Escherichia coli strains tested produced positive results. In exclusivity testing, 30 of 31 strains of other bacterial species produced negative results, the sole exception being a strain of Enterobacter cloacae. In internal and independent laboratory matrix testing of mozzarella cheese, condensed milk, pasteurized liquid egg, and frozen green beans, results showed no significant differences in performance of the Soleris and reference methods with two exceptions, one in which the Soleris method produced more positive results, and one in which the reference method produced more positive results. Conclusion Performance characteristics of the modified Soleris E. coli method are consistent with those of the original validated method. Highlights The Soleris E. coli method offers improvements in ease of use and reagent robustness.

Statement of problem: The use of medical plants has expanded as a result of increased resistance of microbes to chemical medications, as well as their negative effects and excessive costs. Ocimum basilicum (basil) and Origanum Syriacum (Thyme), were studied for their antifungal activities against Candida albicans (C. albicans) species. Purpose: The goal of this study is to see if two plant extracts, Ocimum basilicum (basil) and Origanum Syriacum (Thyme), have antifungal effects on different denture base materials (long cycle heat-polymerized methyl methacrylate PMMA, short cycle heat-polymerized PMMA, and thermoplastic monomer free microcrystalline polymer karadent). Material and methods: Plant extracts (basil and thyme) were tested for antifungal activity using the pores method. The effect of plant extracts on various denture base surfaces was determined using a disc diffusion test. Fifty-four discs of 6mm diameter and 2mm thickness were prepared from long cycle heat-polymerized methyl methacrylate PMMA, short cycle heat-polymerized PMMA, and thermoplastic monomer free microcrystalline polymer karadent and distributed into three groups:-ve control, basil, and thyme. The diameter of growth inhibition zones was determined after these discs were placed on a Sabouraud Dextrose agar surface and incubated for 24 hours at 37 C. Results: Both Origanum syriacum, and Ocimum basilicum were efficient against Candida albicans. Origanum Syriacum extracts showed a superior antifungal effect compared to the Ocimum basilicum extract with long and short curing cycle acrylic resin, but in Karadent the values of Ocimum basilicum were higher than those of Origanum syriacum extract.

Manuka (Leptospermum scoparium) has been studied as a valuable shrub as its leaves are rich in bioactive components such as β-caryophyllene. This study investigated the extractability of hydrophobic deep eutectic solvent (HDES) namely menthol:lactic acid HDES for the total phenolic content (TPC) and antioxidant activity from β-caryophyllene-rich Manuka leaves. Results were also compared with ethanol and n-hexane extracts. The ethanolic extract possessed the highest amount of TPC (86.08 ± 6.22 mg GAE/g DW) among tested extracts. The extraction of TPC using HDES was at a sufficient level (79.82 ± 0.75 mg GAE/g DW), while n-hexane presented poor yield (2.74 ± 0.70 mg GAE/g DW). Higher 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and ferric reducing antioxidant power were found to be in ethanolic extract. The antibacterial effects of Manuka extract obtained by HDES was also assessed and the results were compared with the antibacterial effect of HDES, lactic acid, commercial Manuka essential oil, steam distilled Manuka oil, and β-caryophyllene pure oil. E. coli, Pseudomonas aeruginosa and Staphylococcus epidermidis were tested using agar disc diffusion method. Results revealed that lactic acid, HDES, and Manuka-HDES extract showed toxicity towards the tested bacteria indicating that the bactericidal effect of the Manuka extract was due to the lactic acid as a forming compound. This study verified the possibility of extracting and characterising different bioactive compounds from Manuka leaves using menthol:lactic acid HDES in a single step and thus it could be explored for broader industrial applications.

New porous activated carbons with a high surface area as an anode material for lithium-ion batteries (LIBs) were synthesized by a one-step, sustainable, and environmentally friendly method. Four chemical activators—H2SO4, H3PO4, KOH, and ZnCl2—have been investigated as facilitators of the formation of the porous structure of activated carbon (AC) from an agar precursor. The study of the materials by Brunauer–Emmett–Teller (BET) and scanning electron microscopy (SEM) methods revealed its highly porous meso- and macro-structure. Among the used chemical activators, the AC prepared with the addition of KOH demonstrated the best electrochemical performance upon its reaction with lithium metal. The initial discharge capacity reached 931 mAh g−1 and a reversible capacity of 320 mAh g−1 was maintained over 100 cycles at 0.1 C. High rate cycling tests up to 10 C demonstrated stable cycling performance of the AC from agar.

Viewing the considerable potential of marine agar as a source for the sustainable production of energy as well as nature-derived pharmaceutics, this work investigated the catalytic activity of three novel GH50 agarases from the mesophilic marine bacterium Microbulbifer elongatus PORT2 isolated from Indonesian coastal seawaters. The GH50 agarases AgaA50, AgaB50, and AgaC50 were identified through genome analysis; the corresponding genes were cloned and expressed in Escherichia coli BL21 (DE3). All recombinant agarases hydrolyzed β-p-nitrophenyl galactopyranoside, indicating β-glycosidase characteristics. AgaA50 and AgaB50 were able to cleave diverse natural agar species derived from Indonesian agarophytes, indicating a promising tolerance of these enzymes for substrate modifications. All three GH50 agarases degraded agarose, albeit with remarkable diversity in their catalytic activity and mode of action. AgaA50 and AgaC50 exerted exolytic activity releasing differently sized neoagarobioses, while AgaB50 showed additional endolytic activity in dependence on the substrate size. Surprisingly, AgaA50 and AgaB50 revealed considerable thermostability, retaining over 75% activity after 1-h incubation at 50 °C. Considering the thermal properties of agar, this makes these enzymes promising candidates for industrial processing.