Type Culture Research Articles

The sex-specific effects of early life adversity and chronic psychosocial stress during adulthood on bone are mitigated by Mycobacterium vaccae NCTC 11659 in mice.

Introduction Chronic stress is a major burden in our society and increases the risk for various somatic and mental diseases, in part via promoting chronic low-grade inflammation. Interestingly, the vulnerability for chronic stress during adulthood varies widely among individuals, with some being more resilient than others. For instance, women, relative to men, are at higher risk for developing typical stress-related diseases, including depression and post-traumatic stress disorder (PTSD). Moreover, the experience of early life adversity (ELA) may increase an individuals' vulnerability for chronic stress during adulthood (CAS), possibly due to its association with chronic inflammation. Because one severe consequence of stress-induced immune activation is a dysregulated endochondral ossification, delayed long-bone growth and bone regeneration following fracture, the aim of this study was to investigate the sex-specific effects of ELA alone or in combination with CAS on bone. As enhancement of an individuals' immunoregulatory potential by repeated administrations of a heat-inactivated preparation of Mycobacterium vaccae NCTC (National Collection of Type Cultures) 11659 has been shown to promote stress resilience in mice, we further aimed to investigate if M. vaccae NCTC 11659 also protects against the negative effects of ELA/CAS on bone. Methods Male and female C57BL/6N mice were subjected to ELA using a maternal separation (MS) model. CAS was induced by either using the chronic subordinate colony housing (CSC) procedure in males, or by using the social instability paradigm (SIP) in females. The effects on bone were evaluated by µCT, histological and gene expression analysis. M. vaccae NCTC 11659 was administered repeatedly s.c. prior to CAS. Results No additive effects of ELA and CAS on bone could be detected. Female mice seem to be more susceptible to ELA while male mice to CAS. Importantly, repeated M. vaccae NCTC 11659 administrations were able to mitigate the negative consequences of stress on bone in both sexes. Conclusion Our results support the hypotheses that the negative effects of ELA and CAS on bone are highly sex-dependent. Moreover, repeated s.c. administrations with immunoregulatory microorganisms might be a future therapeutic option for stress-related bone disorders.

Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling.

Madin-Darby Canine Kidney (MDCK) cells are a key cell line for influenza vaccine production, due to their high viral yield and low mutation resistance. In our laboratory, we established a tertiary cell bank (called M60) using a standard MDCK cell line imported from American Type Culture Collection (ATCC) in the USA. Due to their controversial tumourigenicity, we domesticated non-tumourigenic MDCK cells (named CL23) for influenza vaccine production via monoclonal screening in the early stage of this study, and the screened CL23 cells were characterised based on their low proliferative capacity, which had certain limitations in terms of expanding their production during cell resuscitation. It was thus our objective to enhance the proliferation efficiency of MDCK cells for influenza vaccine production following cell resuscitation, with a view to improving the production of non-tumourigenic MDCK cells for vaccines and enhancing the production of influenza virus lysate vaccines from MDCK cells through genetic intervention. We concentrated on the protein thrombosponin-1 (THBS1), which was markedly differentiated in the proteomics data of the two MDCK cells. By integrating this finding with related studies, we were able to ascertain that THBS1 exerts a significant influence on the level of cell proliferation and apoptosis. Consequently, our objective was to investigate the impact of THBS1 expression on MDCK cell apoptosis by verifying the differences in THBS1 expression between the two MDCK cells and by interfering with THBS1 expression in the MDCK cells. We found that the knockdown of THBS1 significantly increased the proliferation and apoptosis of CL23 cells without causing significant changes in cell migration and invasion, and its overexpression significantly decreased the proliferation of M60 cells and increased cell migration, invasion, and apoptosis. In addition, the TGF-β/Smad pathway target genes transforming growth factor-β1 (TGF-β1), mothers against decapentaplegic homolog 2 (Smad2), and mothers against decapentaplegic homolog 3 (Smad3), were significantly down-regulated in CL23 cells after THBS1 knockdown and up-regulated in M60 cells after overexpression, with consistent expression identified at both the mRNA and protein levels. The treatment of cells with TGF-β activators and inhibitors further demonstrated that THBS1 regulated MDCK cell proliferation and apoptosis through the TGF-β/Smad signalling pathway. Finally, we found that THBS1 also regulated H1N1 influenza virus replication. These findings enable a comprehensive understanding of the regulatory mechanisms of THBS1 regarding MDCK cell proliferation and apoptosis functions and the effects of influenza virus replication.

Vancomycin-resistant Enterococcus – a study on its prevalence from different clinical samples in a rural medical college hospital

Background: Vancomycin-resistant Enterococcus (VRE) is an important cause of nosocomial infection with Enterococcus faecium causing most of the VRE infections. Widespread use of glycopeptides in health care facilities has led to the development of VRE and enterococcal infections with high-level resistance to aminoglycosides, beta-lactamase production and glycopeptide (including vancomycin) resistance are difficult to treat and often pose a therapeutic challenge to health care facilities. Aims and Objectives: This study aimed to determine the antibiotic susceptibility pattern of Enterococcus species from various clinical specimens and to find out the occurrence rate of vancomycin-resistant enterococci. Materials and Methods: This study was conducted at the Department of Microbiology of Tamralipto Government Medical College and Hospital, East Midnapore, West Bengal. A total of 688 clinical samples were the total sample size taken. Isolation and identification of Enterococcus spp. were done by standard microbiological procedures such as culture, Gram staining, and suitable biochemical tests were conducted. Antibiotic susceptibility testing was done by the Kirby–Bauers disc diffusion method on Mueller–Hinton agar and results were interpreted according to Clinical and Laboratory Standards Institute (CLSI) guidelines 2023. Teicoplanin sensitivity was performed for those isolates showing resistance to vancomycin. For strains showing resistance to vancomycin by the Kirby–Bauers disc diffusion method, vancomycin minimum inhibitory concentrations (MIC) were performed by E-test (Hi media) with MIC breakpoints between 4 and 32 as per CLSI criteria. This was compared with the control strain of American Type Culture Collection Enterococcus faecalis 29212 as per CLSI 2023 guidelines. Results: A total of 48 Enterococcus isolates were obtained from 688 clinical samples; 31 (8.05%) were detected from 385 urine samples, 9 (6.72%) were detected from 134 blood samples, 5 (5.15%) were detected from 97 pus/wound swab, and 3 (4.17%) were detected from 72 bronchoalveolar lavage fluid samples. Among the 48 Enterococcus isolates, 13 (27.08%) were vancomycin-resistant out of which nine were E. faecalis and four were E. faecium. Enterococcus species showed maximum resistance toward ciprofloxacin followed by ampicillin and maximum sensitivity toward teicoplanin and linezolid. Conclusion: Implementation of strict infection control measures, antimicrobial policies, and proper surveillance are required to identify, contain, and treat VRE infections to reduce mortality and morbidity.

Antibacterial Potential of Selected Traditional Medicinal Plants for Wound Healing in Sekela District, Northwestern Ethiopia

ABSTRACT This study evaluated the antibacterial activity of medicinal plants (MPs) used to treat wounds. Methanol and ethyl acetate crude extracts of four medicinal plants were examined for antibacterial efficacy against gram-positive and gram-negative American Type Culture Collection (ATCC) strains. Plant crude extracts were produced using the maceration technique and analyzed using qualitative phytochemical tests. The antibacterial properties of plant products were evaluated using the agar disc diffusion assay. The broth microdilution method was used to determine the minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations. The data were analyzed using Analysis of Variance (ANOVA). The methanol extract of Rumex nervosus had the highest yield (21%). The methanol extracts of all tested MPs tested positive for alkaloids, phenolics, and tannins. The methanol and ethyl acetate extracts exhibited a dose-dependent increase in the growth inhibition zone against all ATCC. The methanol extract of Plantago lanceolata demonstrated the highest antibacterial activity (16.67 ± 1.15) against Staphylococcus epidermidis. S. aureus and S. epidermidis recorded a relatively lower MIC (6.25 mg mL−1) from methanol extracts of P. lanceolata, while P. lanceolata and R. nervosus recorded a lower MBC (25 mg mL−1) against S. aureus.

Evaluation of a new protocol for rapid identification of Streptococcus pneumoniae in blood cultures using the modified bile solubility test: Gram staining is still standing.

This study aimed to evaluate a new protocol of the bile solubility test performed directly on the blood from positive blood culture bottles to identify Streptococcus pneumoniae rapidly. Seventy-five positive blood cultures (PBC) showing Gram-positive cocci in pairs or chains on Gram stain, including 32 S. pneumoniae isolates and three reference American Type Culture Collection (ATCC) isolates were included to evaluate the performance of a modified bile solubility test (MBST). One milliliter of blood from the PBC bottle was mixed with 0.5 mL of 10% desoxycholate or a saline solution. Both suspensions were analyzed after 10 min of incubation through a Gram stain to detect solubilization. This technique was compared with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identification, performed on PBC following extraction or on colonies after short or standard incubation, and the optochin susceptibility test on colonies. The capsular serotypes were determined for all S. pneumoniae, and the Belgian National Reference Center confirmed the identification. All 32 clinical isolates and the ATCC isolate of S. pneumoniae were solubilized on the desoxycholate-treated slides, while the other species tested remained visually unchanged on both, the test and control slides. The MBST test demonstrated a 100% sensitivity and specificity with a mean turnaround time (TAT) of just 39 min, making it 14 h and 56 min faster than the optochin susceptibility test. This rapid variant of the bile solubility test appears to be a reliable method to identify S. pneumoniae directly from positive blood culture bottles, with a TAT of 39 min. It is a cost-effective, easy-to-perform, and time-efficient technique. Negative results should be interpreted cautiously, as they may result from mixed infections with S. pneumoniae and other Gram-positive cocci.

Finding of antibiotic compounds pyrrolo [1,2-a]pyrazine-1,4-dione, hexahydro and Cyclo(prolyl-tyrosyl) isolated from the degradation of epoxy

ABSTRACT The investigation focused on the endophytic gram-positive microbial isolate KARE_P3, capable of degrading epoxy resin materials. This isolate produced bioactive secondary metabolic molecules with potent inhibitory effects against Staphylococcus aureus, Microbial Type Culture Collection and GenBank (MTCC 96). The study examined microbial growth and degradation mechanisms at various time points, with samples collected on the 35th and 70th days of fermentation. Three different solvents were used for extraction, and the crude metabolite was analysed using Thin Layer Chromatography (TLC), Gas Chromatography – Mass Spectrophotometry (GC-MS), Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). These analyses revealed numerous secondary metabolites that help microbes adapt to changing environments. Further characterisation using bioautography and GC-MS identified 71 compounds, with 3–5 showing positive results. Important findings include compounds such as pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro 98% and Cyclo(prolyl-tyrosyl), which have diverse pharmaceutical applications. Antibacterial efficiency was studied using an in silico model, showing higher binding energy and inhibition rates of pyrrolo[1,2-a]pyrazine-1,4-dione and Cyclo(prolyl-tyrosyl) against Candida albicans compared to Staphylococcus epidermis. Challenges remain in the efficient sorting, recycling, and recovery of epoxy materials, crucial for future biotechnological progress.

Design and Development of Bio-based <i>Anisomeles indica</i> Kuntze Leaf Essential Oil Nanoemulsion: Antibacterial Activity, Antibiofilm Activity and Antiquorum Sensing Activity Against Skin Pathogens

Background: Plant aromatic oils have the phytochemicals to inhibit bacterial growth. However, an appropriate encapsulation method is required to enhance their activity and stability. Anisomeles indica is a weed and does not affect human health. Hence, A. indica aromatic oil nanoformulation could be used as a medication to treat skin infections. Aim: In this study, aromatic oil nanoformulation of A. indica was prepared and characterised. Further, antibacterial, antibiofilm and quorum quenching properties of nanoformulation were evaluated. Methods: Hydro-distillation process was used to extract aromatic oil. Further nanoformulation was prepared and characterised by the following methods: Ultrasonication method, Dynamic light scattering method, Transmission Electron Microscopy (TEM) analysis, Viscosity determination, pH measurement, Turbidity, % Transmittance and % Creaming index measurements. Agar disc diffusion, well diffusion methods and crystal violet staining assay were used to evaluate the anti-bacterial, antibiofilm, and quorum quenching activity respectively. Results: Nanoformulation 20AIU4 (Oil-in-water) was prepared using A. indica aromatic oil, Tween-20 and water using the ultrasonication method. Nanoformulation contains particles with an average size of 52nm, Polydispersity Index (PDI) value of 1.000, 99.9 % transmittance, and thermodynamically stable for three months. 20AIU4 nanoformulation inhibits the bacterial growth of the Staphylococcus aureus Microbial Type Culture Collection (MTCC) 902 and Pseudomonas aeruginosa MTCC 424 with the inhibition zone of 14.76 ± 0.4nm and 13.50 ± 0.5nm respectively. A. indica aromatic oil nanoformulation was inhibited the (85.71% and 85%) of bacterial attachment and biofilm development (80.55% and 85.71%) against S. aureus MTCC 902 and P. aeruginosa MTCC 424 respectively. Nanoformulation shows anti-quorum sensing activity with inhibit Chromobacterium violaceum MTCC 2656 growth with an inhibition zone of 23.9 ± 0.36nm. Conclusion: The present study reveals that the A. indica aromatic oil nanoformulation 20AIU4 is a potent antibacterial agent that can be used against bacterial skin infections.

Wild-Type p53 Regulates Apoptosis of Human Breast Cancer Cells.

The tumor suppressor wild-type p53 is known for its role in inducing apoptosis in tumor cells. This study investigated the relationship between wild-type p53 and protein phosphatase 1 (PP1) and caspase in promoting apoptosis of breast cancer cells. Human breast cancer cell lines MCF-7 and MDA-MB-231 obtained from the American Type Culture Collection were used in this study. Small interference RNAs (Si-RNA) and plasmids were used to regulate wild-type p53 expression in these two tumor cell lines through liposome-mediated transfection. GSK-2830371 (PP1 inhibitor) and zVAD (Caspase inhibitor) were employed to further verify the PP1 activating function of wild-type p53 in Caspase-dependent MCF-7 and MDA-MB-231 apoptosis. PP1 activity was quantitatively detected by phosphorus colorimetric assay. Co-immunoprecipitation (Co-IP), flow cytometry assay, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, Western blot, the real-time reverse transcriptase-polymerase chain reaction (RT-qPCR), and immunofluorescence staining were used to analyze cell apoptosis degree and marker protein expression. The expression level of PP1 in the breast cancer cells was successfully regulated by cell transfection. The phosphatase activity was increased, and obvious apoptotic cytological characteristics were observed in p53-overexpressed breast cancer cells. p53 knockdown/overexpression increased/decreased the level of B cell lymphoma 2 (Bcl-2), and decreased/increased levels of Caspase-3, cleaved Caspase-3, cleaved Caspase-8, Cytochrome C (Cyt-C), Truncated BID (tBid), Bcl-2-associated X (Bax), and cell apoptosis (p < 0.01). The promotion of proteins and apoptosis induced by p53 overexpression was reversed by GSK-2830371 or zVAD. Wild-type p53 might promote Caspase-dependent apoptosis of human breast cancer cells through PP1 activation.

Risk Factors,Diagnosis and Outcome of Proven and Probable Invasive Trichosporonosis in a Tertiary Care Hospital: A Cross-sectional Analytical Study

Introduction: Trichosporon species are basidiomycetous yeast like fungus that is ubiquitous in the environment. They form part of the normal flora in humans. In recent years, they have been causing invasive infections, especially in the immunocompromised hosts. Disseminated cases including trichosporonemia can rapidly progress, leading to increased morbidity and mortality. Aim: To diagnose and identify risk factors and outcomes of proven and probable invasive Trichosporon species infections. Materials and Methods: This cross-sectional analytical study was done at Sri Ramachandra Institute of Higher Education and Research, Chennai, Tamil Nadu, India, over a period of one year (March 2023 till February 2024). All the clinical samples sent for routine diagnostics to the laboratory during the study period and archived samples were included (Total isolates, n=60). European Organisation for Research and Treatment of Cancer/Invasive Fungal Infection Cooperative Group (EORTC/ IFICG) and the National Institute of Allergy and Infectious Disease/Mycoses Study Group (NIAID/MSG) definitions were used to categorise samples as proven invasive and probable invasive trichosporonosis. Phenotypic characterisation was done by colony morphology on Sabouraud Dextrose Agar (SDA), Gram stain, Dalmau technique and urea hydrolysis. Genotypic characterisation was done by Trichosporon genus specific and Trichosporon asahii specific Polymerase Chain Reaction (PCR). The reference strains used as quality control were Trichosporon asahii Microbial Type Culture Collection (MTCC) 6179, Trichosporon jirovecii MTCC 9036 and Candida albicans American Type Culture Collection (ATCC) 90028. All statistical analysis was performed using Microsoft Excel (2016) and descriptive statistics were presented as numbers and percentages. Results: All the 60 (100%) isolates were characterised as Trichosporon asahii. Amongst them 4 (6.7%) were categorised as proven invasive trichosporonosis and 56 (93.3%) were probable invasive trichosporonosis. Some important risk factors for invasive disease were Intensive Care Unit (ICU) admission, antibiotic usage, diabetes and hypertension. All 4 (6.3%) proven cases were isolated from blood and 2 (50%) amongst them succumbed to the disease. Amongst the probable cases, 16 (28.6%) succumbed to the disease. Conclusion: Invasive infections caused by Trichosporon species have high mortality especially amongst immunocompromised hosts. Hence, identification at an early stage by appropriate diagnostic methods and initiating appropriate antifungal agents can result in better outcome of the patient.

Green synthesis of zinc oxide nanoparticles using Enterobacter cloacae microorganism and their application in enhanced oil recovery

Employing safe and inexpensive methods for the synthesis of biocompatible nanoparticles (NPs) can be very challenging. Green synthesis refers to the process of synthesizing nanoparticles without using toxic and dangerous chemicals. One of the applications of nanoparticles is increasing production from oil reservoirs, known as enhanced oil recovery (EOR). The main aim of the current study is the biosynthesis of zinc oxide (ZnO) nanoparticles (NPs) using Enterobacter cloacae (Persian Type Culture Collection (PTCC): 1798) microorganism, extracted from the formation water of one of the southwestern Iranian reservoirs, as a novel approach in EOR applications. Several analytical methods, including Fourier transform infrared (FTIR), field emission scanning electron microscope (FSEM), X-ray diffraction (XRD), dynamic light scattering (DLS), energy dispersive X-ray spectroscopy (EDS), and zeta potential were used to analyze the produced NPs. The FESEM analysis confirmed the amorphous form of the nanoparticles and estimated their size in the range of 32 to 58 nm. In investigating the effect of synthesized nanoparticles on interfacial tension (IFT) and stability tests, three levels of base fluids (distilled water, seawater, and diluted sea water) and five levels of nanoparticle concentrations (0, 100, 500, 1000, and 2000 ppm) were considered. The IFT analysis showed that an increase in nanoparticle concentration causes a decrease in the IFT. Also, ZnO nanoparticles were chosen at concentrations of 500 and 1000 ppm for wettability alteration and the EOR test through injection into porous media. The results for the EOR test demonstrated a maximum oil recovery factor of 56% for nanofluid injection with a concentration of 1000 ppm with diluted seawater as the base fluid. Furthermore, oil recovery factors of 43% and 49% were achieved by injection of distilled water and seawater with a concentration of 1000 ppm, respectively.

Application of Cyberlindnera fabianii isolate as a potential starter culture in type IV sourdough to improve techno-functional capabilities of barley bread

Indigenous sourdough (SD) yeasts have promising potential applications as starter culture in processing of the baked goods. In the present study, techno-functional capabilities of the predominant yeasts isolated from barley SD were characterized to select a proper starter culture for type IV SD. Accordingly, techno-functional characteristics of the produced barley bread including textural features using a texture analyzer, sensory properties through evaluation of the panelist's scores, mold-free shelf-life using a challenge test, and their antioxidant capacity based on the DPPH free radical scavenging activity were compared to each other. According to the results, crumb hardness of the barley bread produced with BSY 3 isolate was significantly (P < 0.05) lower than those of the other samples. This bread also showed the highest crumb porosity (14.20 %). In addition, methanolic and water-salt soluble extracts of the aforementioned sample had the highest ability to scavenge DPPH free radical (equal to 87.75 % and 32.65 %, respectively). Although the surface growth of Aspergillus niger on the produced samples was significantly higher than that of the control sample containing baker's yeast, while SD breads showed higher mold-free shelf-life. Furthermore, there was no significant difference among the produced samples in terms of the overall acceptability. Based on the sequencing results of the PCR products, the selected yeast isolate was also identified as Cyberlindnera fabianii. Accordingly, the yeast isolate can be used as starter culture in type IV SD with the aim of improving the quality characteristics of the produced barley bread.

XII International Mycological Congress: report of Congress action on nomenclature proposals relating to fungi

Procedures, appointments and outcomes of the Fungal Nomenclature Session (FNS) of the XII International Mycological Congress (IMC12) are summarized, including the composition of the Fungal Nomenclature Bureau and the Nominating Committee of the IMC. Between 124 and 322 mycologists attended the three sessions of the FNS, at which formal proposals to amend Chapter F of the International Code of Nomenclature for algae, fungi, and plants were debated. The 15 proposals considered included eight "from the floor", five of which were withdrawn prior to the FNS. One of the withdrawn proposals was directed to the Editorial Committee for Fungi, in relation to adding examples of best practice when citing living cultures as types. Among the seven proposals published in Taxon, one proposal with a high “no” vote in the Guiding Vote was not re-introduced. Discussion on one proposal led to the authorization of a Special-purpose Committees on “Genomes as Types for Fungi”. For the eight proposals that were put to a vote, two proposals were rejected and six proposals were accepted. The accepted proposals: (1) clarified that a proposal to conserve a name with a conserved type does not require citation of a typification identifier; (2) clarified procedures to protect and reject names of fungi; (3) removed the need to list synonyms of protected names in the Code appendices; (4) clarified that an earlier homonym of a sanctioned name remains unavailable if the sanctioned name is rejected outright; (5) recommended that culture collections or biological resource centres where cultures are lodged should be “public”; and (6) recommended that when original type cultures are lost, neotypification should utilize the progeny of ex-type cultures. Appointments made by the FNS included the Secretary of the Fungal Nomenclature Bureau for IMC13, the officers and members of the Editorial Committee for Fungi, and the officers and members of the Permanent Nomenclature Committee for Fungi. Decisions and appointments of the FNS were ratified in a resolution accepted by the plenary session of the Congress.

In vitro antifungal activities of medicinal plants used for treatment of candidiasis in Pader district, Northern Uganda

BackgroundThe emergence of multidrug resistant Candida species to available drugs has led to renewed interest in the use of herbal medicines globally. This study scientifically verified antifungal effectiveness of five commonly used plant species in Pader district, against selected pathogenic candida strains.MethodsPowdered roots of Momordica foetida, Sansevieria dawei and Distimake dissectus; and stem barks of Khaya anthotheca and Mitragyna rubrostipulata were extracted sequentially using petroleum ether and methanol, respectively; and total water extraction at 24.4 °C (maceration), 60 °C (decoction) and boiling water at 87 °C (hot water infusion). Extracts and their combinations, positive controls (amphotericin B, and fluconazole) and negative control (80% dimethyl sulfoxide, verified to be tolerable concentration to the tested Candida species) were screened and verified for their antifungal activity against Candida albicans (ATCC: American Type Culture Collection reference strain 10231, ATCC 90028, 0770a and 0796), C. glabrata (VVc 004, ATCC 2950) and C. tropicalis (ATCC 750 and 0210) using agar well diffusion and broth micro-dilution, respectively.ResultsAqueous extract (24.4 °C) of M. rubrostipulata (ZOI: 18.00 ± 1.00 to 38.33 ± 0.17; MIC: 3.13 ± 0.00 to 20.83 ± 4.17; MFC: 12.50 ± 0.00 to 200.00 ± 0.00), methanol extract of K. anthotheca (10.11 ± 0.31 to 15.11 ± 0.65; 1.04 ± 0.26 to 12.50 ± 0.00; 12.50 ± 0.00 to 100.00 ± 0.00), and combination of aqueous extract (60 °C) of D. dissectus + methanol extract of K. anthotheca (7.89 ± 0.26 to 19.67 ± 0.37; 0.78 ± 0.00 to 50.00 ± 0.00; 12.50 ± 0.00 to 200.00 ± 0.00) exhibited broad spectrum antifungal activities and were fungistatic against all tested Candida species, which comprised 8 clinical/control and susceptible/resistant strains. None of the conventional drugs used demonstrated broad spectrum antifungal activity across all tested Candida species/strains.ConclusionMethanol extract of K. anthotheca, aqueous extract (24.4 °C) of M. rubrostipulata, and combination of aqueous extract (60 °C) of D. dissectus + methanol extract of K. anthotheca could be effective in the treatment of candidiasis. They demonstrated potential broad spectrum antifungal activity against different species and strains of tested Candida than the fluconazole and amphotericin B drugs. Their fungistatic nature showed their ability to inhibit fungal growth. Hence, these extracts/extract combination can offer better treatment option for candidiasis if they are standardized and also their active curative compounds isolated and made into antifungal drugs.

Bioactive Properties of Campomanesia lineatifolia: Correlation Between Anti-Helicobacter pylori Activity, Antioxidant Potential and Chemical Composition.

Helicobacter pylori is found in the stomach of patients with chronic gastritis and peptic ulcers, infecting approximately half of the world's population. Current treatment for H. pylori infection involves a multi-drug therapeutic regime with various adverse effects, which leads to treatment abandonment and contributes to the emergence of resistant strains of H. pylori. Previously, we demonstrated that the essential oil of Campomanesia lineatifolia leaves exhibited an anti-H. pylori activity. In this study, we aimed to evaluate the phenolic content of the phenolic-rich ethanol extract (PEE) from C. lineatifolia and its anti-H. pylori and antioxidant properties. Additionally, the anti-H. pylori activity was assessed in polar and non-polar fractions from PEE, isolated myricitrin (MYR) and a mixture of myricitrin and quercitrin (MYR/QUER) from polar fractions, and aqueous extract (tea) to correlate the responsible fractions or compounds with the observed activity. Broth microdilution assays were performed to assess the anti-H. pylori activity using type cultures (ATCC 49503, NCTC 11638, both clarithromycin-sensitive) and clinical isolate strains (SSR359, clarithromycin-sensitive, and SSR366, clarithromycin-resistant). The antioxidant activity was evaluated using the DPPH assay. The total tannin and flavonoid contents were determined using the hide-powder method, the Folin-Ciocalteu reagent, and the aluminium chloride colourimetric assay, respectively. The tea (MIC 1:100), PEE, polar and non-polar fractions, MYR, and MYR/QUER inhibited the growth of H. pylori strains tested (MIC values ranging from 0.49 to 250 μg/mL). The antioxidant assays revealed that PEE exhibited a higher antioxidant activity (EC50 = 18.47 μg/mL), which correlated to the high phenolic content (tannin and flavonoid, 22.31 and 0.15% w/w, respectively). These findings support the traditional use of C. lineatifolia as a multitarget medicinal plant for treating gastric ulcers and reinforce the potential use of the species as a coadjuvant in therapeutic regimes involving patients with resistant H. pylori infection.

In vitro prebiotic activity of millets with selected probiotic microorganisms

Abstract BACKGROUND: Millets are an inexpensive grain that contains many major and minor nutrients, such as carbohydrates, protein, fats, and dietary fibers. As millets are rich in carbohydrates, and carbohydrates tend to be a great prebiotic source for the proliferation of gut microbes (probiotics), this study was designed to evaluate the prebiotic potential of millets. Millets used in this study include pearl millet (Pennisetum glaucum L.), finger millet (Eleusine coracana L.), sorghum (Sorghum bicolor L.), and foxtail millet (Setaria italica L.). METHODS: The prebiotic activity was performed by using them as a sole carbon source in De Man–Rogosa–Sharpe agar media with probiotic strains Lactobacillus plantarum [Microbial Type Culture Collection (MTCC) 1407], Lactobacillus paracasei (Microbial Culture Collection 4490), Lactobacillus rhamnosus (MTCC 1408), and Saccharomyces boulardii. Millets were ground into a fine powder with the grinder, and the powder was used as a carbon source in media for cultivating microorganisms under anaerobic conditions maintained in an anaerobic jar at 37°C. RESULTS: The results revealed that the growth of all microorganisms was confluent on their respective Petri plates under anaerobic conditions as observed on positive control plates. When these selected millets powder were mixed in the same ratio and used as a carbon source to check prebiotic activity, the results showed similar trends as observed with individual millets. CONCLUSION: The study concludes that whole grain millets showed prebiotic activity and thus can be recognized as an essential prebiotic nutritious food. These millets may support the growth of the population of healthy gut microbes, thus playing an important role in promoting digestion. Further, in-vivo and clinical validation is necessary to establish the prebiotic potential of these millets leading to the development of millets based ayurvedic prebiotic formulations.

Влияние основной кислотообразующей микрофлоры бактериальных заквасок на формирование органолептического профиля сыров с низкой температурой второго нагревания

Sensory properties of cheese define consumer choice. Their taste and smell depend on many production and maturation factors. These processes result in a dairy hydrolysis complex. If the technology of production and maturation are violated, the final component composition acquires undesired taste, consistency, and appearance. The taste of cheese depends on a number of factors, one of which is the type of starter microflora. This study assessed the effect of monospecific cultures of acid-forming types (Lactococcus lactis subsp. lactis, Lactococcus cremoris, and Streptococcus thermophilus) on the sensory profile of cheese with low-temperature second thermal treatment. The experiment featured semi-hard Gollandsky cheese with a fat mass fraction of 45 % solids. The experimental cheese underwent sensory evaluation for appearance, taste, smell, consistency, and pattern. The monospecific cultures failed to render the cheese its specific pungency, slight sourness, elasticity, slight brittleness, and a pattern of round or oval eyes. However, each acid-forming starter had a different effect on the sensory properties and defect risks.

Ultrastructural characterization of hippocampal inhibitory synapses under resting and stimulated conditions.

The present study uses electron microscopy to document ultrastructural characteristics of hippocampal GABAergic inhibitory synapses under resting and stimulated conditions in three experimental systems. Synaptic profiles were sampled from stratum pyramidale and radiatum of the CA1 region from (1) perfusion fixed mouse brains, (2) immersion fixed rat organotypic slice cultures, and from (3) rat dissociated hippocampal cultures of mixed cell types. Synapses were stimulated in the brain by a 5min delay in perfusion fixation to trigger an ischemia-like excitatory condition, and by treating the two culture systems with 90 mM high K+ for 2-3min to depolarize the neurons. Upon such stimulation conditions, the presynaptic terminals of the inhibitory synapses exhibited similar structural changes to those seen in glutamatergic excitatory synapses, with depletion of synaptic vesicles, increase of clathrin-coated vesicles and appearance of synaptic spinules. However, in contrast to excitatory synapses, no structural differences were detected in the postsynaptic compartment of the inhibitory synapses upon stimulation. There were no changes in the appearance of material associated with the postsynaptic membrane or the length and curvature of the membrane. Also no change was detected in the labeling density of gephyrin, a GABAergic synaptic marker, lining the postsynaptic membrane. Furthermore, virtually all inhibitory synaptic clefts remained rigidly apposed, unlike in the case of excitatory synapses where ~ 20-30% of cleft edges were open upon stimulation, presumably to facilitate the clearance of neurotransmitters from the cleft. The fact that no open clefts were induced in inhibitory synapses upon stimulation suggests that inhibitory input may not need to be toned down under these conditions. On the other hand, similar to excitatory synapse, EGTA (a calcium chelator) induced open clefts in ~ 18% of inhibitory synaptic cleft edges, presumably disrupting similar calcium-dependent trans-synaptic bridges in both types of synapses.

Cultivation of Aurantiochytrium sp. Using Pickle Seasoning Liquid Waste and Application of the Raw Biomass for Fish Aquaculture

AbstractAurantiochytrium sp. strain L3W is a heterotrophic microorganism that produces docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), which are essential for the growth of marine fish. In this study, pickle seasoning liquid waste was used for culturing strain L3W and the raw biomass of strain L3W was then fed to red sea bream fingerlings (Pagus major) to confirm its usability as a source of DHA and EPA. The strain L3W was cultured on the three pickle seasoning liquid waste samples, Akimurasaki, Hiroshimana, Shisohijiki, diluted with sand-filtered seawater at initial pH values of 4 and 7 under unsterile conditions. The growth of strain L3W was highest at 3.71 g/L on Hiroshimana at an initial pH of 7 with DHA and EPA production at 71.4 and 6.8 mg/g-biomass, respectively. Preparing the raw biomass of strain L3W by the 200 L-scale cultivation using the American Type Culture Collection 790By+ medium because of the DHA and EPA contents close to that produced in the Hiroshimana medium with an initial pH of 7, the raw biomass was spiked to the diet at 3%, 5%, and 10%. The final DHA and EPA contents in the whole body were increased by 4.26 and 3.03 times, respectively, by adding the strain L3W biomass at 10%. These results confirmed the feasibility of a carbon cycling scenario in which the strain L3W is cultivated using liquid food waste with the resultant biomass is utilized as a source of DHA and EPA for fish aquaculture. Graphical Abstract

Importance of Ultraviolet-C (UV-C) Emitter Configuration for the Attenuation of Staphylococcus aureus and Candida auris Pathogens.

The relative importance of different ultraviolet-C (UV-C) emitter configurations on the attenuation of vegetative bacterial and fungal pathogens has not been assessed. We hypothesized that emitter configuration would impact the efficacy of UV-C attenuation of Staphylococcus aureus ( S. aureus) and Candida auris (C. auris) pathogens. American Type Culture Collection (ATCC) S. aureus (ATCC 6538) and C. auris (ATCC MYA-5001) carriers (ReadyNowTM Test Carriers, Stratix Labs Corporation, Saint Paul, MN) were mounted on an aluminum stand along with three calibrated radiometers (International Light Technologies model ILT1270, Peabody, MA). Five UV-C emitter configurations were assessed, including three emitters with a triangular configuration about the stand and each rotating 360° (1), one emitter facing the stand and rotating 360° (2), three emitters facing the stand in a linear configuration and each rotating 5° (3), one emitter facing the stand and rotating 5° (4), and one emitter facing the stand and rotating 90° (5). Three serial experiments were conducted. The first experiment involved the establishment of the minimally effective irradiation dose (mean and standard deviation mJ/cm2) required to achieve no growth (6-log reduction (LR)) with direct exposure to pathogen carriers positioned at the center of the lamp. We then assessed the relative efficacy of delivery of the minimally effective dose via the five emitter configurations in attenuating polycarbonate and textured pathogen carriers. Polycarbonate carriers were positioned at 25.5 and 69.5 inches from the floor and oriented vertically to the emitters. Textured plastic pathogen carriers were positioned at 47.5 or 58.5 inches from the floor and with a 45° or horizontal orientation to the emitters. Standard carriers (1"x0.9") were used for both pathogens and large carriers (1"x3") for C. auris,the latter to address the potential for cell clustering. With standard carriers, the minimally effective dose was 27.01± 0.15 mJ/cm2 for S. aureus but was not achieved for C. auris. The minimally effective dose for large C. auris carriers was 596.62 ± 27.98 mJ/cm2. With standard carriers, all configurations achieved a >6 log reduction for S. aureus, and none achieved a >6 log reduction for C. auris.All configurations achieved a > 6 log reduction when 596.62 ± 27.98 mJ/cm2 was delivered to large C. auris carriers. Changing to textured plastic carriers (standard for S. aureus and large for C. auris) and varying height (47.5-69.5 inches) from the floor and orientation to the emitters (45° and horizontal), the mean ± standard deviation for S. aureus and C. auris log reductions with delivery of the minimally effective dose was 4.44 ± 2.02, 2.58 ± 2.37, 3.55 ± 2.67, 2.33 ± 2.47, and 3.00 ± 2.64 for configurations one through five, respectively. Configuration one achieved a significantly greater LR than configurations two (adjusted P = 0.0018) and four (adjusted P = 0.023). There were 22% (6/27) of sites ≥ 100 colony-forming units (CFU) following cleaning but before UV-C vs. 0% (0/27) of sites ≥ 100 CFU after surface disinfection cleaning and nine minutes of configuration three UV-C treatment (P = 0.023). The choice of UV-C emitter configuration can impact S. aureus and C. auris attenuation when there is indirect exposure to the pathogen. Emitter configuration should be considered as an important parameter for future UV-C technological assessments.

Complete genome sequence of Sphingobacterium thalpophilum BAA-1094.

We report the complete genome of Sphingobacterium thalpophilum BAA-1094 obtained from the American Type Culture Collection. The genome has one circular chromosome (5.7 Mbp). The rather low average nucleotide identity of 76.5% to the type strain Sphingobacterium thalpophilum DSM 11723 suggests the need for reclassification.