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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.

Cryptococcus neoformans is a capsule containing yeast pathogen, commonly found in urban environments that causes Cryptococcosis which is a worldwide fungal disease. The current study. This study was aimed to compare the ability of some culture media to detect melanin production by C. neoformans isolates. A total of 202 clinical samples obtained from patients suffering from meningitis, during the period from September 2020 through April 2021. The isolation and identification results revealed that 179 (88.61%) samples were positive for yeast growth. 29 (14.35%). Twenty-nine from 202 of samples were C. neoformans according to the morphological characteristics and the biochemical tests, and 150 (74.25%) samples for Candida spp. All C. neoformans isolates produced blue dry colonies on cryptococcus differential agar. Several differential media were used for the purpose of testing the ability of C. neoformans to produce melanin pigment, these includes: Niger seed agar medium (Staib’s Agar), and the colonies of C. neoformans appeared on this medium in a brown color after 2–5 days of incubation, Tobacco agar where those colonies gave a dark brown color after 48–72 h, and Methyldopa agar which was the best medium for the detection of melanin production because the results were faster from the other media, interestingly the colonies can produce the pigment and can growth very fast within 24hr.

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.

Background Since the last two decades, substantial increase in fungal infections has been observed in immunocompromised hosts. Virulence factors of Candida albicans play a role in adherence, haemolytic activity, phenotypic switching and production of hydrolytic enzymes. The secreted aspartyl proteinases (SAP family) contribute to adhesion, tissue damage and invasion, while phospholipase (PLB) supports the hydrolysis of phospholipids. Very few studies showed the correlation of phenotypic activity and detection of genes contributing to the similar enzyme activity. Therefore, our study aimed at demonstrating correlation between in vitro phenotypic production of phospholipase and proteinase enzymes with genetic level detection of SAP and PLB genes. Methods The present study was carried out on a total of 799 samples over a period of one year. Culturing was carried out on Sabouraud's dextrose agar. Confirmation and speciation of Candida spp. was carried out using cornmeal agar and CHROMagar and by the germ tube test, urease test and automated identification system Vitek-2, and antifungal susceptibility testing was performed on Candida isolates. Phenotypic phospholipase and proteinase activity was analysed, and molecular detection of SAP4 and PLB genes was carried out. Results In our study, we have screened 799 samples for mycological protocol; of which, 269 (33.6%) were Candida species, 44% (119) of which were C. albicans. Proteinase activity was exhibited by 77 (64.7%) and phospholipase activity was exhibited by 73 (61.34%) isolates, while 46.2% exhibited both activities among the C. albicans species. The PLB gene was detected in 97.3% isolates, while SAP4 was detected in 94.7% of C. albicans isolates. Conclusion The study of in vitro expression of virulence factors and gene detection of C. albicans will help to improve the prognosis despite the nature of the sample.

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.

Certain cheeses can be legally produced in the United States using raw milk, but they must be aged for at least 60 d to reduce pathogen risks. However, some varieties, even when aged for 60 d, have been shown to support growth of Listeria monocytogenes or survival of Shiga toxin–producing Escherichia coli (STEC). Thermization, as a subpasteurization heat treatment, has been proposed as a control to reduce the risk of pathogens in raw cheese milk while retaining some quality attributes in the cheese. However, the temperature and time combinations needed to enhance safety have not been well characterized. The objective of this research was to determine and validate decimal reduction values (D-values) for L. monocytogenes and STEC at thermization temperatures 65.6, 62.8, and 60.0°C; a D-value at 57.2°C was also determined for L. monocytogenes only. Nonhomogenized, pasteurized whole-milk samples (1 mL) were inoculated with 8-log cfu/mL L. monocytogenes or STEC (5- or 7-strain mixtures, respectively), vacuum-sealed in moisture-impermeable pouches, and heated via water bath submersion. Duplicate samples were removed at appropriate intervals and immediately cooled in an ice bath. Surviving bacteria were enumerated on modified Oxford or sorbitol MacConkey overlaid with tryptic soy agar to aid in the recovery of heat-injured cells. Duplicate trials were conducted, and survival data were used to calculate thermal inactivation rates. D65.6°C-, D62.8°C-, and D60.0°C-values of 17.1 and 7.2, 33.8 and 16.9, and 146.6 and 60.0 s were found for L. monocytogenes and STEC, respectively, and a D57.2°C-value of 909.1 s was determined for L. monocytogenes. Triplicate validation trials were conducted for each test temperature using 100 mL of milk inoculated with 3 to 4 log cfu/mL of each pathogen cocktail, A 3-log reduction of each pathogen was achieved faster in larger volumes than what was predicted by D-values (D-values were fail-safe). Data were additionally compared with published results from 21 scientific studies investigating L. monocytogenes and STEC in whole milk heated to thermization temperatures (55.0–71.7°C). These data can be used to give producers of artisanal raw-milk cheese flexibility in designing thermal processes to reduce L. monocytogenes and STEC populations to levels that are not infectious to consumers.

Despite its importance, pyrazinamide (PZA) is a blind spot in drug susceptibility testing in tuberculosis laboratories. The aim of this study was to set up a reliable agar-based proportion method f...

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.

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.

Aim and Objective: The purpose of this study is to isolate, identify and specification of various the Candida species from various clinical samples in a tertiary care hospital, and to characterize various the isolated Candida species. Materials and Methods: A study was conducted on people of different age groups from January 2019 to December 2019. Candida species isolated from different patients by using Potassium Hydroxide mount and processed by BacTalert 3D (Biomerieux) automated blood culture system. Further culture identification of Candida species were done on Sabouraud dextrose agar (SDA). Speciation of Candida was done using Germ tube test, CHROM agar Candida Medium, Cornmeal agar, Sugar Fermentation test and Sugar Assimilation test. Results: In our study was the most common species isolated, among non albicans Candida i.e. 21 (38.9%); 19(35.2%) of was the most common followed by 9(16.7%) ofand 5(9.3%) of . Maximum number of Candida isolates were obtained from NICU i.e. 27(50.0%) followed by 11 from Med (20.3%), 7 from E/W (13.0%), 2 from BICU (3.7%), 2 from Skin (3.7%), 1 from PICU (1.9%), and 1 from R/R (1.9%). Conclusion: Our study showed that is the most common isolates species. Among , was found to be the most common isolate followed by . Children less than 1 year are most affected with maximum number of Candida species were obtained from NICU department. HiChrom Candida is proven to be more useful as differential agar. Keywords: Candida Albican, C.Tropicalis, C.Glabrata, C.Krusei, Hi-Chrom Agar, Sabouraud Dextrose Agar

Bacterial respiratory infections are most commonly causes of illness for all age group patients in ICU. Most of the patients suffer from urosepsis, postoperative disease and lower respiratory infection when admitted in ICU’s. The main purpose of this study is control in ICU’s infection & determine their antibiotic sensitivity pattern for better treatment of patients. Bacteriological profile and their antibiogram from endotracheal aspirate and bronchoalveolar lavage fluid samples from intensive care unit patients.We performed manual culture of Endotracheal aspirate & BAL fluid specimens collected from ICU’s patients. Samples were cultured on Blood agar and Mac-Conkey agar then incubated at 37C for 24 hours. Any growth were further confirmed by Gram stain and appropriate biochemical tests, and then Antibiotic sensitivity test was done. Among the 123 mechanically ventilated patients and 68 of them developed positive samples. In this study 55.3% (68) pathogenic isolates, 4.1% (5) were non pathogenic isolates and 40.6 (50) were no growth isolates in BAL and endotracheal aspirate samples. In which 67.6% (46) male and 32.4% (22) female patients. Older population falling in the 40-50 years of age group contributes the majority (27.64%) of the infection undergoing treatment for LRT infection in ICU’s patients in TMMC & RC, Moradabad, and in 70-80 years of age group (4.06%) was less contributes of this infection. Among the causative agents, were most common among gram negatives and was common among the gram positive isolates. In our study more resistance gram positive antibiotics were CIP, E, LE, and most sensitive antibiotics were VA, LZ, GEN and TEI. And gram negative bacteria most resistant antibiotics were OF, CPM, CAZ, CIP, CTR, MRP, LE, IPM, PIT and most sensitive antibiotics were AK, GEN, CL, TGC. In this study we conclude that Gram-negative bacilli were predominant in BAL fluid and endotracheal aspirates of ICU patients. Out of 68 multidrug resistance isolates, 1.47% were MRSA, 41.17% were ESBL and 57.36% were MBL isolates higher rate of ESBL production seen in spp., and higher rate of MBL production in spp.High incidence of VAP and the potential MDR pathogens are a real threat in our ICU’s. Combined approach of judicious antibiotic usage and training programs to health care personnel might be of help in combatting high incidence of antibiotic resistance in our institute.

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.

In this study we reported cultivation of bacteria associated with mangrove soil from the Ngurah Rai Mangrove Forest, Bali. Mangrove soil samples were serially diluted using sterile artificial seawater, spread onto Starch Casein M agar and incubated at 28oC for 28 days. Cultivation of mangrove soil samples yielded 165 bacterial colonies with 68 isolates were selected and purified based on different morphology. Of these 68 isolates, 22 isolates displayed antibacterial activities ranging from weak to strong inhibition against at least one of four bacterial indicators namely Staphyloccocus aureus, Streptococus mutans, Escherichia coli and Klebsiella pneumoniae using perpendicular streak method. Overall, 19 out of 22 bacteria isolates displayed weak antibacterial potential and two isolates exhibited moderate antibacterial activity. The isolate SA4 was the only bacterium with strong antibacterial potential with measured clear distance ≥ 10 mm against the four bacterial isolates. Sequence analysis based on 16S rRNA gene fragment assigned the isolate SA4 as Bacillus subtilis strain BIL/BS-168. Overall, this study confirmed the untapped potential of antibacterial activities from bacteria associated with mangrove soil.

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.

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

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.

In this study, a new method for shoot regeneration via protocorm-like bodies (PLBs) is described. Shoot proliferation using stem elongation under light-emitting diodes (LEDs), and enhanced growth of Anthurium andraeanum ‘Tropical’ plantlets on medium containing silver nanoparticles (AgNPs) was investigated. For shoot regeneration, the micropropagation efficiency via PLB formation was fourfold more efficient than directly from callus after 12 wk of culture. This was possible because each initial callus cluster could be cut into 20 explant slices (1 × 5 × 5 mm in size), each with an average of 42.67 PLBs which are capable of growing into plants. For shoot proliferation, single shoots with 2 cm in size derived from PLB culture were placed under different blue to red LED ratios. This study showed that higher shoot height (4.77 cm), fresh weight (327.33 mg), number of leaves per shoot (6.33), and the number of stem nodes per shoot (5.67 stem nodes) were obtained under 70R:30B as compared to those under other lighting conditions and fluorescent lamp after 8 wk of culture. Especially, the total shoot proliferation (10 shoots) via stem node culture was higher than those in the control. For enhanced plantlet growth, single shoot with a pair of leaves derived from stem elongation under 70R:30B cultured on 1/4 MS medium supplemented with 0.5 mg L−1 NAA and 1 g L−1 activated charcoal, 7 mg L−1 AgNPs, 30 g L−1 sucrose, and 8 g L−1 agar gave the highest growth than other treatments. Plantlets derived from 7 mg L−1 AgNP treatment transferred into greenhouse showed better growth as well as 4-wk earlier flowering than the control treatment.

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.

Background This study aimed to investigate the occurrence of Legionella species in the respiratory samples of patients with pneumonia symptoms from Ahvaz, Iran by culture and the real-time PCR of 23S-5S rRNA gene spacer region. Methods and results A total of 123 clinical respiratory samples including 63 pleural aspirates, 57 bronchoalveolar lavage (BAL), and 3 sputum were collected from 65 males and 58 females with pneumonia symptoms. All samples were cultured on the Modified Wadowsky-Yee (MWY) agar. The Legionella species was identified by routine bacteriological tests. The presence of the 16S-23S rRNA spacer region gene was investigated by real-time PCR. The Legionella species were differentiated by sequencing of 16S-23S rRNA gene. A total of 2 (1.6%) BAL specimens were positive for Legionella species by culture method. No Legionella spp. were identified in pleural aspirates and sputum samples by the culture method. Using real-time PCR, 9 (7.3%) samples including 6 BAL, 1 sputum, and 2 pleural aspirates were positive for legionella species. These species were detected in 3 (5.2%) females and 6 males (9.2%). The results of sequencing showed that eight species were L. pneumophila while one was L. cherrii. Also, the 2 isolates that were identified by culture method, were confirmed as L. pneumophila by sequencing. Conclusions The results showed that using the real-time PCR has a more efficacy for detecting of Legionella species in respiratory samples. Also, L. pneumophila was the most prevalent species circulating in the southwest region of Iran. So, periodic monitoring programs is recommended to prevent epidemics due to this bacterium.

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.

The seeds of six cultivars of maize were screened by both blotter and agar methods for detection of fungal pathogens, and for testing the transmission of seed-borne fungi using test tube agar. Resu...

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.

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.

Aims: To estimate changes in the number of oxytetracycline -resistant strains, a dynamic population of Bacillus in the gut of Eisenia fetida cultured in processed-cow-dung (PrCD) supplemented with subinhibitory amounts of oxytetracycline, was studied. Study Design: Sterile water containing oxytetracycline was sprayed over PrCD at concentration of 0.012 µg per mg dehydrated PrCD per spraying. The number of oxytetracycline-resistant Bacillus spp. microbiota of E. fetida’s gut was compared to the number of oxytetracycline-resistant Bacillus spp. in its feed (PrCD). For this purpose, Luria Agar (LA) and Bacillus Agar (BA) plates amended with or without oxytetracycline were used. Place and Duration of Study: The study was carried out in the Omics Laboratory of Department of Biotechnology, University of North Bengal situated in Darjeeling district of West Bengal, India during 2017-18. Methodology: E. fetida gut content and PrCD samples were collected on different days of the experiment for bacteriological analysis. Dilution plating on LA yielded the total number of cultivable heterotrophic bacteria. LA plates amended with oxytetracycline (15 µg/ml) were used to acquire the fraction of heterotrophic bacteria resistant to oxytetracycline. BA plates were also utilized to obtain the fraction of resistant Bacillus spp. population. The frequency of mutation was determined using a conventional formula. Results: In the gut of E. fetida reared in PrCD supplemented with sub-inhibitory concentration of oxytetracycline, a rise in the oxytetracycline-resistant Bacillus population was observed. On day 1, the frequency of oxytetracycline-resistant Bacillus spp. (5×10-8) matched the spontaneous mutation frequency, however higher frequencies on days 2 and 7 (1.6×10-4 and 3.5 x 10-6 respectively) suggested significant dissemination of oxytetracycline resistance in the gut environment. Conclusion: Subinhibitory oxytetracycline concentrations in earthworm diet had a significant effect on mutation rates, showing that evolutionary forces on the gut microbiota may be determining their responses to antibiotic stress.

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.

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.

The aim of this study was to assess the effect of antimicrobial photodynamic therapy (aPDT) with curcumin and riboflavin on three-week Enterococcus faecalis biofilm. At first the 15-mm root canals of 65 single rooted extracted human teeth (including maxillary incisors, mandibular and maxillary canines and mandibular premolars) were separated from the crown and were prepared with ProTaper instruments. After autoclave sterilisation, samples were inoculated with E. faecalis suspension, and incubated for three weeks. After ensuring biofilm formation by scanning electron microscopy (SEM) in two teeth, the remaining 63 teeth were randomly divided into seven groups (n = 9): aPDT + curcumin, aPDT + riboflavin, LED, curcumin, riboflavin, 5.25% NaOCl (positive control) and no intervention (negative control). For light source a LED unit with 390-480 nm wavelength (peak of 460 nm), power density of 1000 ± 100 mW cm-2 and mean energy density of 60 J cm-2 was used. The roots were horizontally sectioned into coronal, middle and apical thirds each with 5 mm thicknesses. Dentin chips with equal weight (1 ± 0.005 g) were collected from the root canal walls with Gates Glidden drills and were transferred into microtubes containing 1 mL of sterile saline and vortexed for 30 s. Next, 10 µL of the contents of each tube was serially diluted and eventually, 10 µL of each solution was cultured on BHI agar. The number of colony-forming units was determined. Data were analysed using the Kruskal-Wallis and Friedman tests. The colony reduction was not significantly different between NaOCl and either riboflavin + LED or Curcumin + LED. The 5.25% NaOCl group showed maximum reduction in colony count, compared with the negative control (P = 0.00). Groups with aPDT with Curcumin + LED (P = 0.005), and with riboflavin + LED (P = 0.011) showed significant reduction in colony count in all three canal thirds (P

A novel actinobacterium NJES-13T was isolated from the gut of Antarctic emperor penguin Aptenodytes forsteri. The new isolate produces bioactive gephyromycin metabolites and exopolysaccharides (EPS). Cells were Gram-negative, motile with the peritrichous flagella, and with a faint layer of extracellular slime. Colonies were yellow when grown on marine agar, ISP1, 2, 4 and TSA media. The strain developed clusters of coccoid, and divided by binary fission in the early phase of growth. The cell clusters were gradually disrupted during the stationary phase and formed short rod-shape cells which were interconnected by viscous EPS showing a three-dimensional net-like morphology, and contained polyhydroxyalkanoates (PHA) granules inside the cells. Growth of strain NJES-13T was observed at 15–45 °C, at pH 6.0–9.0 with 0.5–9.0% (w/v) NaCl. The complete genomic size of strain NJES-13T was 3.45 Mb with a DNA G + C content of 67.0 mol%. The combined polyphasic taxonomic characterizations presented in this study unequivocally separated strain NJES-13T from all known genera in the family Dermatophilaceae. Thus, strain NJES-13T represents a novel species of a new genus, for which the name Gephyromycinifex aptenodytis gen. nov., and sp. nov. is proposed. The type strain is NJES-13T (= CCTCC 2019007T = KCTC 49281T). Genetic prediction of secondary metabolite biosynthesis revealed a 44.5 kb-long biosynthetic gene cluster (BGC) of type III polyketide synthase (PKS) as well as four other BGCs, indicating its great potential to produce novel bioactive metabolites derived from the gut microbiota of animals living in the extreme habitats in the Antarctica.

Introduction: Dermatophytes are fungi that can cause infections of the skin, hair & nails due to their ability to invade keratin. Dermatophytosis is the most common superficial fungal infection worldwide; it is common in tropics and subtropical regions. It may present in epidemic proportions in areas of high humidity. Objective: The present study aimed to identify various species causing dermatophytosis & to determine the invitro susceptibility pattern against commonly used systemic antifungal agents in our tertiary care center. Material and Methods: A total of 149 samples were collected of infected skin, hair and nails in a period of 1 year from January 2020 to December 2020. Samples were collected under aseptic condition by skin scrapping, nail and hair clipping by using scalpel or forceps. Identification of the causative pathogen was done by performing slide culture, lacto-phenol cotton blue mount, hair perforation tests and urease tests. We adopted a newly developed agar based disk diffusion assay to test susceptibility of clinically isolated dermatophytes for antifungal susceptibility testing. Results: Microbiological investigations revealed the presence of dermatophytic fungi in 71.8% of the samples. Trichophyton rubrum was the predominant pathogen isolated. The study showed Itraconazole to be most effective antifungal drugs against dermatophytes followed by terbinafine and fluconazole. Conclusions: Further intensive epidemiological and invitro antifungal susceptibility studies of dermatophytes are required which will have more public health significance. Keywords: Dermatophytes, Antifungals, Susceptibility

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.

The study was conducted to investigate the socio-economic characteristics of the agar farmers, the extent of constraints in agar wood cultivation; and the factors allied with constraints in agar tree farming. Despite the enormous potential of agar (Aquilaria malaccensis Roxb.) production in Bangladesh, it has yet to gain traction due to socioeconomic, technical, and marketing challenges. The cultivation of agar trees is largely centered in the Sylhet region, where a large number of agar farmers engage. This research was carried out in Maulvibazar district's Barlekha upazila (sub-district) to better understand and assess the restrictions and factors that affect agar tree cultivation. A survey was conducted with 120 households during September 2018 to March 2019. Descriptive statistics, rank order, coefficient of correlation, and multiple linear regression were used to examine the data. Out of 12 constraints, the most significant barrier was a lack of funds to engage in agar farming (2.81), which was followed by a lack of technical guidance (2.71). The lack of labor was the least important constraint faced by the farmers (1.43). It was revealed that education, household size, farm size, number of trees per farmer, annual income, experience in agar farming, agar wood cultivation under trees and knowledge of agar cultivation of the farmers were shown to be realized to agar plantation constraints. However, education, farm size, annual revenue, and agar growing experience were identified as major drivers of the degree of agar farming constraints. The study identified various policymakers' potential to address the influencing variables (education, farm size, annual revenue, and agar farming experience) in establishing agar farming. Furthermore, finance facilities, adequate agar farming instruction, and forest department introspection could all help to reduce agar farming obstacles.

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°

Background and Aim: There is a worldwide controversy about the choice of microbial flora for use as process hygiene indicators. This study aimed to evaluate the pertinence of using either coliforms or Enterobacteriaceae (EB) as process hygiene indicators in the pasteurized milk production line. Two flora families and total flora were used as bacterial indicators in some stages of pasteurized milk production line to identify the origin of post-pasteurization contamination and compare the results obtained for each flora. In addition, the bacteriological profile of isolated coliforms and EB was developed. Materials and Methods: One thousand and two hundred samples of pasteurized cow milk and surfaces (pipes and tank) at various processing stages were taken from two dairies in the northern region of Algeria. The total microbial flora (TF), total coliforms (TC), thermotolerant coliforms, and EB were enumerated, following the recommendations of ISO 4833:2006, ISO 4832:2006, and ISO 21528-2:2017 methods, respectively. The bacteriological profile was determined using the API 20E and 10S tests (bioMérieux, France). Furthermore, the cleaning efficiency and disinfection protocol of surfaces were evaluated using contact agar slides 1 (Liofilchem™, Italy). Results: Enumeration of the different indicators shows that the highest contamination rate is recorded by the total flora in the two units, 3.28 and 3.78 log CFU/mL, respectively. EB (–0.60 log CFU/mL) at post-pasteurization stage in Unit 1 and coliforms (0.44 log CFU/mL) at the pasteurized packaged milk stage in Unit 2 are the least significant germ families. The lowest compliance rates of bacterial contamination were reported for total flora (82-85%) at the three sampled sites in Unit 2. In comparison, the highest was reported in Unit 1 (99.8%) and 2 (98%) by the EB indicator. Assessing the surface cleaning and disinfection protocol compliance shows that the tank records the highest non-compliance rates for EB and TF (4% and 3%) in Unit 2. EB are represented in both units by various species. Acinetobacter baumannii in Unit 1 and Enterobacter cloacae in Unit 2 are the common species of the three indicator families. Acinetobacter and Enterobacter in Unit 1, Escherichia, Citrobacter, Enterobacter, Klebsiella, and Hafnia in Unit 2 are the most time persistent bacterial genera along the production line. Stenotrophomonas, Serratia, Salmonella, Enterobacter, and Escherichia are common genera in both units. Conclusion: The results obtained show no difference in the use of EB or TC as hygiene indicators. However, if the objective is to identify the species of bacterial populations, using EBs are the most appropriate.

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.

Aim: In this study, it was aimed to evaluate AYC.2.2 agar for susceptibility testing of Mycobacterium tuberculosis clinical isolates against first-line drugs. Materials & methods: In the present study, 208 M. tuberculosis clinical isolates were tested on AYC.2.2 agar, which was previously validated for the first-line drugs isoniazid, rifampicin, streptomycin and ethambutol. Results: Specificity, sensitivity, positive predictive value, negative predictive value and agreement for isoniazid–rifampicin–ethambutol–streptomycin were 100–100–97.2–99.3%, 94.8–94.8–79.3–94.3%, 100–100–82.1–98.03%, 97.03–98.03–96.7–98.08%, 98.07–98.5–94.7-98.07%, respectively. Conclusion: Results had shown that the newly developed AYC.2.2 agar promises as an alternative medium that can be used to perform susceptibility testing of M. tuberculosis isolates. However, further multicenter studies are needed to be used in routine mycobacteriology laboratories.

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.

Given the economic importance of the dairy industry, product quality should be a constant concern. Some sensorial and technological properties of milk and dairy products are related to the spoilage contaminant microbiota, which is mainly represented by psychrotrophic proteolytic bacteria producing thermoresistant enzymes. Fresh artisanal cheeses are generally produced with raw milk, so it is of great importance to know the microbiota of these products. Therefore, the objective of this work was to count and isolate the psychrotrophic proteolytic bacteria and to evaluate the spoilage potential of these microorganisms isolated from fresh cheeses handcrafted and commercialized in Salinas, MG, Brazil. After the isolation of psychrotrophic proteolytic bacteria present in fresh cheese, the isolates were characterized in terms of Gram staining, shape, and oxidase activity. The proteolytic activity of the isolates was confirmed by a spot halo test using specific agar. The results highlighted the predominance of Gram-positive psychrotrophic proteolytic bacteria when the spoilage microbiota of the fresh cheese samples was evaluated. Considering the proteolytic potential, 34% of the isolates were considered highly proteolytic. The results showed that the initial contamination of fresh cheeses may interfere with the microbiological quality of these products. Devido a importância econômica da indústria de laticínios, a qualidade dos derivados lácteos deve ser uma preocupação constante. Algumas propriedades sensoriais e tecnológicas do leite e derivados estão relacionadas à microbiota deterioradora contaminante, representada, principalmente, por bactérias psicrotróficas proteolíticas que produzem enzimas termoresistentes. Os queijos frescos produzidos artesanalmente são geralmente fabricados com leite cru, por isso, é de grande importância conhecer a microbiota desses produtos. Portanto, o objetivo deste trabalho foi enumerar e isolar as bactérias psicrotróficas proteolíticas além de avaliar o potencial deteriorador desses microrganismos isolados de queijos frescos artesanais comercializados em Salinas, MG, Brasil. Após o isolamento das bactérias psicrotróficas proteolíticas presentes no queijo fresco, os isolados foram caracterizados em termos de coloração de Gram, forma e atividade de oxidase. A atividade proteolítica dos isolados foi confirmada pela formação de halo de hidrólise em meio de cultura específico. Os resultados destacaram a predominância de bactérias proteolíticas psicrotróficas Gram-positivas quando foi avaliada a microbiota deterioradora das amostras de queijo fresco. Considerando o potencial proteolítico, 34% dos isolados foram classificados como altamente proteolíticos. Os resultados mostraram que a contaminação inicial de queijos frescos pode interferir na qualidade microbiológica destes produtos.