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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.

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.

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.

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 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.

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.

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.

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.

Bacterial infections are a major cause of chronic infections and mortality that continues to threaten public health worldwide. Various medicinal plants used in folk medicine have demonstrated wound healing and antibacterial properties. The aims of the study are investigation phytochemical and antibacterial activity of ethyl acetate fraction from jarak pagar roots ( Jatropha curcas Linn.). The roots were treated with maceration method using ethanol 70% and subsequently fractioned to ethyl acetate fraction. The fractions were used for phytochemical screening and antibacterial assay using Kirby and Bauer agar difussion method. The results showed that ethyl acetate fraction jarak pagar roots contained alkaloid, flavonoids, saponins, steroids, poliphenol dan tannins. The antibacterial activity of jarak pagar roots againts Staphylococcus aureus and Escherichia coli shows that the activity of this fraction with concentration 8 mg/mL has strong inhibitory potency against the growth S. a ureus with inhibitory zone is 14,3 mm and has moderate inhibitory potency against E. c oli with inhibitory zone is 9 mm. The increase concentrations of ethyl asetate fraction jarak pagar roots shows high inhibition diameter of bacterial growth.

Plantago major L. (Plantaginaceae) é uma planta utilizada no tratamento de feridas e tosse devido ao seu potencial anti-inflamatório, cicatrizante e antimicrobiano. Visando o emprego da planta na formulação de produtos para uso oral, o objetivo deste estudo foi avaliar a atividade antimicrobiana desta espécie in vitro. Extratos etanólico e hidroalcoólico, preparados com partes aéreas de P. major, foram submetidos à triagem fitoquímica e análise de cromatografia em camada delgada (CCD). A atividade antimicrobiana dos extratos e de tintura comercial foi testada frente a Enterococcus faecalis e Pseudomonas aeruginosa por difusão em ágar. As análises fitoquímicas demonstraram a presença de substâncias fenólicas, cumarinas e triterpenos, além de baixo teor de fenóis e flavonoides nos extratos. Os sinais observados na CCD demonstraram ausência do marcador químico, ácido 5-O-cafeoilquínico, porém o número de classes de substâncias encontradas e a presença de um provável derivado de ácido 5-O-cafeoilquínico condizem com exemplares de P. major. Na avaliação da atividade antimicrobiana, os extratos alcoólicos e tintura não foram capazes de inibir o crescimento bacteriano, entretanto não deve ser descartada a sua ação anti-inflamatória devido as substâncias encontradas e o seu uso em sinergismo como outros agentes antimicrobianos para o desenvolvimento de produtos de uso oral.

The aim of the study has been to develop a biodegradable film from marine polysaccharides. The optimization of polysaccharides quantity for the composite film was sought by empirical response surface methodology. The Box–Behnken Model Design was applied to optimize the concentration of chitosan (1.0-2.0% (w/v), agar (1.0-2.0% (w/v) and glycerol (0.1-0.5% (w/v) as independent variables to achieve the goal. The overall desirability function fits with the quadratic model (0.862043) at a significant level (p < 0.05) for the optimum concentration of chitosan (1.5% (w/v), agar (2.0% (w/v) and glycerol (0.41% (w/v) to obtain the minimum water vapor permeability (7.25 10-10g m m-2 Pa-1 s-1) and maximum tensile strength (12.21 Ma P), elongation at break (7.32%) and puncture resistance (16.18 N) in the optimized composite film. The absolute residual errors of experimental and predicted responses were between 1.24 and 3.56% acceptable levels. Attenuated total reflection–Fourier transform infrared spectroscopy confirmed the intermolecular non-covalent hydrogen bond between the hydroxyl groups of agar and glycerol with the amino group of chitosan. 3D atomic force microscopy images revealed that the chitosan, agar and glycerol film has layer-by-layer smooth surface properties due to homogenous interaction among the polysaccharides; this provides the film with good mechanical properties and with functional application. Chitosan was found to be responsible for the lower level of water vapor permeability and higher puncture resistance of the film. Tensile strength and elongation at break were influenced by agar and glycerol. The whiteness of the film was negatively affected with the concentration of chitosan.

To assess the cariogenic potential of almond milk, soya milk, coconut milk and bovine milk, was evaluated to check ability to enable Streptococcus mutans association formed, acid manufacturing, as well as their ability to buffer pH transforms. The baseline non stimulated whole salivary sample (2.5-5 ml) will be collected from the students in the morning at least 1 hr after breakfast. The salivary samples (pre-test and post-test) will be collected and tested for the CFUs. Salivary samples are transported in a plastic container to the microbiological laboratory. They were then tested for the number of CFUs for S. mutans using mitis salivarius bacitracin agar. Soya milk promoted much more biofilm development, whereas sugar free almond milk promoted a little. When pH test was performed, sugar free almond milk had the lowest cation exchange capacity, whereas bovine milk had the maximum cation exchange capacity.

Aims: This study was undertaken to assess the abundance of multiple-antibiotic-resistant (MAR) Salmonella strains in fecal samples of Rhinoceros unicornis of the Kaziranga National Park (KNP), India. Study Design: Antibiotic-resistance profile of the Salmonella isolates from fecal samples of rhinoceros was carried out by replica plating on Muller Hinton Agar (MHA) plates containing antibiotics. The presence of class 1 integrons in metallo-β-lactamase (MBL) producing Salmonella isolates was determined by multiplex PCR. Place and Duration of Study: The study was carried out on rhinos of KNP situated in the Golaghat district of Assam, India in April 2015. Methodology: Fresh rhino fecal samples (designated as 1R, 2R, 3R, 4R and 5R) were dilution plated onto MacConkey agar. Purified bacterial colonies were then streaked separately on bismuth sulphite (BS) agar plates. All black colonies which are characteric growth of Salmonella were selected and used to make master plates on Luria Agar. To determine the antibiotic-resistance profile of the isolates, master-plates of purified single colonies of Salmonella spp. were replicate-printed on plates containing antibiotics from the β-lactam, aminoglycoside, or quinolone groups. To detect the presence of an integron, a conserved segment polymerase chain reaction was used. Results: 97.6% of the Salmonella isolates tested were resistant to >1 antibiotics (multidrug resistant or MAR). A total of 100 isolates from two fecal samples, 4R and 5R, were found to be imipenem resistant; 52 of them tested positive for the presence of MBLs. Five of the twenty MBL producing strains contained class 1 integrons. Conclusion: Because Salmonella is usually spread by drinking contaminated water, it is likely that water bodies in KNP are contaminated with MAR Salmonella strains. In case of Salmonellosis outbreak among Indian one-horned rhinos, our antibiogram will assist the veterinarians to choose the appropriate regimen of antibiotics for the rhinos in the KNP.

Hyperglycemia is a condition of increasing blood glucose levels exceeding the normal value in patients with Diabetes mellitus (DM). This study is a screening to describe that respon of hyperglycemic mice to the administration of lactobacillus isolated from sap as an alpha glukosidase inhibitor. The final aim of this study was to measure the activity of alpha glukosidae inhibitor from lactobacillus isolated from sap in hyperglycemic wistar rats. The cultivation of de man rogosa sharpe Agar (MRSA) and the isolated lactobacillus was carried out using the pout plate methode.Lactobacillus isolated from sap produced in West Lombok district was carried out given to white rats for treatment. This study is a quasi-experimental study using 25 hyperglycemic white rats divided into 5 (five) treatments. The mean blood sugar levels after alloxan induction were 374.6, 347.5, 373.2, 348.4. The mean blood sugar levels at P1, P2 and P3 respectively from the first day to the third day were P1 247.2, 225.8, 209.6, P2 was 241.2, 141.8, 99.6 and P3 were 138,8, 112, 84.6.

Background and Objectives: Accuracy and promptness in the detection of methicillin resistance are of key importance in ensuring correct antibiotic treatment in infected patients and control of MR staphylococci in the hospital environment. The aim of this study was to detect MRSA phenotypically by oxacillin screen agar and Oxacillin MIC method and to evaluate cefoxitin disc diffusion test as a screening tool for MRSA detection. Materials and Methods: In the present study, a total of 50 isolates of from various clinical samples collected were used for the detection of Methicillin resistant (MRSA). Methicillin resistance was determined by oxacillin disc diffusion, cefoxitin disc diffusion the oxacillin screen agar test and MIC. Result: Out of 50 isolates 21 (42%) isolates were detected as MRSA based on MIC method, which is considered as gold standard method for the detection of MRSA. All the isolates of MRSA were 100% susceptible to vancomycin and linezolid. rnIn the present study, cefoxitin diffusion method has given 100% sensitivity and specificity in concordance with MIC method. However, the oxacillin screen agar method showed 95.24% sensitivity and 96.55% specificity. Conclusion: As per our study and previous reports elsewhere on phenotypic detection of MRSA, cefoxitin is more potent inducer of the regulatory system and an accurate surrogate marker for the detection of MRSA in the routine susceptibility testing. Keywords: MRSA, Cefoxitin, Drug Resistance.

Water is essential to the life of every living organism. The water used by the consumer must be safe for the preservation of his health. The objective of this study was to assess the quality of well water used in households in the city of Korhogo. To do this, water samples were taken from the wells of different households. Parameters such as pH, temperature, electrical conductivity and total dissolved solids were measured at the sampling sites. Nitrates and ammonium were determined by spectrophotometric methods using sodium salicylate and indophenol blue, respectively. The membrane filtration technique has been used for the enumeration of total coliforms, Escherichia coli, fecal enterococci, sulfite-reducing anaerobic bacteria, yeasts and moulds. The agar incorporation method was used for the enumeration of mesophilic aerobic germs. The non-conformity relative to pH was 73%. All wells were contaminated with nitrates and ammonium. Non-compliance rates of 80% and 53% were obtained for nitrates and ammonium respectively. Non-compliance with mesophilic aerobic germs, total coliforms, E. coli and fecal enterococci was 100% for each indicator. No spore of sulphite-reducing clostridia has been identified. Contamination rates of 80% (mould) and 67% (yeast) were recorded. The moulds isolated from the various water samples belong to the genus Aspergillus. The quality of well water in households is not suitable for human consumption. These waters require treatment or household residents must switch to other safe sources of drinking water.

Heat treatment is one of the most widely used processing technologies in the dairy industry. Its primary purpose is to destroy microorganisms, both pathogenic and spoilage, to ensure the product is safe and has a reasonable shelf life. In this study microwave volumetric heating (MVH) was compared with a conventional tubular heat exchanger (THE), in terms of the effects of each at a range of temperatures (75°C, 85°C, 95°C, 105°C, 115°C, and 125°C) on indigenous microflora viability and the germination of inoculated Bacillus licheniformis endospores in reconstituted skim milk. To assess the heat treatment-related effects on microbial viability, classical agar-based tests were applied to obtain the counts of 4 various microbiological groups including total bacterial, thermophilic bacterial, mesophilic aerobic bacterial endospore, and thermophilic aerobic bacterial endospore counts, and additional novel insights into cell permeability and spore germination profiles post-heat treatment were obtained using real-time flow cytometry (FC) methods. No significant differences in the plate counts of the indigenous microorganisms tested, the plate counts of the inoculated B. licheniformis, or the relative percentage of germinating endospores were observed between MVH- and THE-treated samples, at equal temperatures in the range specified above, indicating that both methods inactivated inoculated endospores to a similar degree (up to 70% as measured by FC and 5 log reduction as measured by plate counting for some treatments of inoculated endospores). Furthermore, increased cell permeability of indigenous microflora was observed by FC after MVH compared with THE treatment of uninoculated skim milk, which was reflected in lower total bacterial count at a treatment temperature of 105°C. This work demonstrates the utility of FC as a rapid method for assessing cell viability and spore inactivation for postthermal processing in dairy products and overall provides evidence that MVH is at least as effective at eliminating native microflora and inoculated B. licheniformis endospores as THE.

Streptomyces are known for their ability to produce various secondary metabolites used in biotechnology, human medicine and agriculture. Understanding of surface properties is very interesting in the control of interfacial phenomena. The objective of this study was to investigate the effect of consistency and composition of growth medium on the physicochemical properties of the surface of Streptomyces strains. To achieve this objective, Six Streptomyces strains belonging to bioprocess and bio-interfaces laboratory are cultivated in two media Bennett (rich) and GBA (minimum). Both media are tested in solid (agar) and liquid (broth) mode. The wettability θw, electron donor character ˠ (-), electron acceptor character ˠ (+) and Surface free energy ΔGiwi are determined using contact angle measurements. On the two solid media Bennett and GBA, Streptomyces strains develop a hydrophobic surface (96.9°

Summary Neoagarobiose (NA2) derived from agar marine biomass is a rare reagent that acts as an anti‐melanogenesis reagent and moisturizer. Here, for the economical manufacturing of NA2, we developed the co‐secretory production system of endo‐type β‐agarases (DagA) and exo‐type β‐agarases (EXB3) in Corynebacterium glutamicum. For this purpose, we first developed a secretory system of DagA via Tat pathway. To improve the secretion efficiency, we coexpressed two Tat pathway components (TatA and TatC), and to improve the purity of secreted DagA in the culture supernatant, two endogenous protein genes (Cg2052 and Cg1514) were removed. Using the engineered strain (C. glutamicum SP002), we confirmed that DagA as high as 1.53 g l‐1 was successfully produced in the culture media with high purity (72.7% in the supernatant protein fraction). Next, we constructed the expression system (pHCP‐CgR‐DagA‐EXB3) for the simultaneous secretion of EXB3 via Sec‐pathway together with DagA, and it was clearly confirmed that DagA and EXB3 were successfully secreted as high as 54% and 24.5%, respectively. Finally, using culture medium containing DagA and EXB3, we successfully demonstrated the conversion of high‐concentration agar (40 g l‐1) into NA2 via a two‐stage hydrolysis process.

The condition of eating places such as street vendors will determine the level of food hygiene, including the water used to wash food utensils. This study aimed to determine the number of microbes and identify Escherichia coli and Salmonella sp. on the water that was used to wash tableware by street vendors around UIN Sunan Ampel Surabaya. Water samples before and after being used to wash tableware were obtained from 4 street vendors. The number of microbes in the sample was tested by the Total Plate Count (TPC) and Most Probable Number (MPN) methods, Escherichia coli was identified using Eosin Methylene Blue Agar (EMB) media, and Salmonella sp. was identified using Salmonella-Shigella agar (SSA) and Triple Sugar Iron agar (TSIA) media. Based on the results of TPC test, the water sample before being used to wash tableware had the lowest and highest values that were 3.0 x 102 Cfu/ml and 9.8 x 105 Cfu/ml, respectively, while the water sample after being used to wash tableware had the lowest and highest values that were 4.0 x 102 Cfu/ml and 3.5 x 105 Cfu/ml, respectively. In the MPN test, the water sample before being used to wash tableware had the lowest and highest values that were 15/100ml and 26 100/ml, respectively, while the water sample after being used to wash tableware had the lowest and highest values that were 72/100 ml and 271/ 100 ml, respectively. Escherichia coli was detected in all samples 1&3 while in sample 2 Escherichia coli was detected in the water before it was detected the genus Enterobacter and in samples 4 the genus Enterobacter was detected and all samples water used to wash tableware of street vendors was positive for Salmonella.

Background Silver nanoparticles (AgNPs) are known to possess antimicrobial properties. Although the antibiofilm activity of AgNPs has been demonstrated in humans, this activity has not yet been elucidated in veterinary medicine. Objectives The purpose of this study was to evaluate the antibiofilm activity of silver nanoparticles against Staphylococcus pseudintermedius. Methods Ten isolates of S. pseudintermedius obtained from dogs with otitis externa were treated with AgNPs, and the antibiofilm activity was measured using a modified microtiter plate and Congo red agar (CRA) method and scanning electron microscopy. Results AgNPs displayed a significant dose‐dependent antibiofilm activity and reduced biofilm formation at concentrations of 20 and 10 µg/ml (p, Silver nanoparticles are known to have antimicrobial effects and antibiofilm acitivity against several pathogens in humans but little is known in dogs. In this study, we demonstrated that silver nanoparticles have anti‐biofilm effects for S. pseudintermedius, the main causative agents of canine otitis externa using modified microtitier palte, Congo red agar methods and scanning electron microscopy.

Aim. To synthesize, study the antimicrobial activity and suggest antimicrobial activity mechanism for the novel derivatives of 6-(1H-benzimidazol-2-yl)-5-methylthieno[2,3-d]pyrimidin-4(3H)-one. Results and discussion. As the result of the targeted modification of 6-(1H-benzimidazol-2-yl)-5-methylthieno[2,3-d]-pyrimidin-4(3H)-one in position 3 with acetamide and 1,2,4-oxadiazol-5-ylmethyl substituents, the compounds, which demonstrated better antimicrobial activity in the agar well diffusion assay than the reference drug Streptomycin, were obtained. To elucidate the mechanism of action of the novel compounds, the docking studies were con-ducted to the active site of the 16S subunit of ribosomal RNA, the proven target for aminoglycoside antibiotics, as well as tRNA (Guanine37-N1)-methyltransferase (TrmD), which inhibitors were considered as a new potential class of antibiotics. Experimental part. By the interaction of 6-(1H-benzimidazol-2-yl)-5-methylthieno[2,3-d]pyrimidin-4(3H)-one with a series of N-arylchloroacetamides and 3-aryl-5-(chloromethyl)-1,2,4-oxadiazoles in DMF in the presence of K2CO3 the target compounds were obtained. The antimicrobial activity was assessed by the agar well diffusion method. The concentration of microbial cells was determined by the McFarland standard; the value was 107 cells in 1 mL of the media. The 18 – 24 hour culture of microorganisms was used for tests. For the bacteria cultivation, Müller-Hinton agar was used, Sabouraud agar was applied for C. albicans cultivation. The compounds were tested as the DMSO solution with the concentration of 100 µg/mL; the volume of the solution was 0.3 mL, the same volume was used for Streptomycin (the concentration 30 µg/mL). The docking studies were performed using Autodock Vina. Crystallographic data for the complexes of Streptomycin with the 16S subunit of ribosomal RNA (1NTB) and its active site, as well as for tRNA (Guanine37-N1)-methyltransferase (EC 2.1.1.228; TrmD) (5ZHN) and its active site were obtained from the Protein Data Bank.Conclusions. It has been determined that 2-[6-(1H-benzimidazol-2-yl)-5-methyl-4-oxothieno[2,3-d]pyrimidin-3(4H)-yl]-N-[4-(ethoxy)phenyl]acetamide, which is the most active as an antimicrobial agent among the compounds tested, also shows the best binding activity towards the active site of tRNA (guanine37-N1)-methyltransferase.