Colony Pigmentation Research Articles

Diagnostic Validity of Cica Beta Test 1 for the Detection of Extended Spectrum Beta-Lactamase (ESBL) Producing Gram Negative Bacteria by Comparing with Phenotypic Method

Background: Detection of extended spectrum beta lactamase producing bacteria is an important issue in the clinical settings. Objective: The purpose of the present study was to validate the Cica Beta Test 1 for detection of extended spectrum beta-lactamase (ESBL) producing bacteria. Method: This analytical type of cross-sectional study was carried out in the Department of Microbiology and Immunology at Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka from January 2006 to December 2006 for a period of one (01) year. All the patients presented with the clinical features of urinary tract infection and surgical as well as burn wound infection at any age with both sexes were selected as study population. All bacteria were isolated and identified by their colony morphology, staining characters, pigment production, motility and other relevant biochemical tests. Phenotypic confirmation of ESBLs producing isolates were done by inhibitor potentiated disc diffusion test according to CLSI recommendation. The Cica Beta Test 1 was performed according to the manufacturer’s instructions. Result: A total number of 288 Gram negative bacteria were isolated. Among these isolates Cica Beta test 1 was positive in 97 strains and phenotypic confirmatory test was positive in 89 strains. The test sensitivity of Cica Beta Test 1 was 100% (95% CI 95.9% to 100.0%). Specificity of the test was 96.0% (95% CI 92.2% to 98.2%). The positive predictive value (PPV) and negative predictive value (NPV) were 92.7% (95% CI 84.5% to 95.7%) and 100.0% (95% CI 98.0% to 100.0%) respectively. The accuracy of the test was 97.2% (95% CI 95.1% to 99.1%). Area under ROC curve = 0.980 (95% CI 0.964 to 0.996); p value 0.0001. Conclusion: In conclusion, Cica Beta Test 1 is very high sensitivity and specificity for the detection of ESBL from Gram negative bacteria.

Isolation and Identification of Endophytic Fungi from UiTM Reserve Forest, Negeri Sembilan

Endophytic fungi are those living inside the host plant without causing any apparent negative effect on the host plant. Two isolates endophytic fungi from leaves and two isolates from root at Universiti Teknologi MARA (UiTM) Reserve Forest, Negeri Sembilan were successfully isolated and identified by morphology and molecular characteristic. Samples were surface sterilized and sub-cultured to obtain a pure culture. Characteristics of the isolates such as colony appearance, mycelial texture, conidia/spores and pigmentation were studied to explore their morphology. Isolates were also subjected to a PCR-based genotyping test. There were noticeable differences in morphological characteristics among the four isolates. Microscopic analysis showed four isolates consist of septa and conidia/spores. The pigmentation result showed that colony in A1leaf samples demonstrated an orange color on potato dextrose agar (PDA) media, colony in A1root demonstrate a black texture in PDA media while hairy colonies in the others two isolates showed a white color on PDA media. Based on molecular analyses the fungal genera showed 99-100% similarity with the related fungi recorded in the GenBank. Both morphology and molecular sequencing of internal transcribed spacer (ITS) regions of endophytic fungi showed that three isolates (A1root, C2leaf, and C3root) were grouped in Basidiomycota while one isolate (A1leaf) belonged to Ascomycota. The endophyte funguses were identified as Daldinia sp. (A1leaf), Polyporales sp. (A1root,) Lentinus sp. (C2leaf,) and Rigidoporus sp. (C3root). Overall, the new discoveries of isolated endophyte fungal have dyeing potential of fungal pigments which offer a viable alternative to natural vegetable and harmful synthetic dyes.

Identification of a Novel Partitivirus of Trichoderma harzianum NFCF319 and Evidence for the Related Antifungal Activity.

We have reported 15 agarose gel band patterns of double-stranded RNA (dsRNA) from Trichoderma spp. We describe herein that band pattern IX in Trichoderma harzianum NFCF319, which appeared to be a single band but consisted of two dsRNAs of similar size, was identified as a novel mycovirus, designated Trichoderma harzianum partitivirus 1 (ThPV1). The larger segment (dsRNA1) of the ThPV1 genome comprised 2,289 bp and contained a single open reading frame (ORF) encoding an RNA-dependent RNA polymerase (RdRp). The smaller segment (dsRNA2) consisted of 2,245 bp with a single ORF encoding a capsid protein (CP). Evaluation of the deduced amino acid sequence and phylogenetic analysis indicated that ThPV1 is a new member of the genus Betapartitivirus in the family Partitiviridae. Curing of virus infection by single-sporing generated 31 virus-free single-spore clones. No significant differences in growth rate, conidia production, or pigmentation were observed between ThPV1-infected and -cured isogenic strains. In addition, comparison of the newly ThPV1-transmitted isolates with their ThPV1-cured parental strain showed no significant difference in colony morphology or pigmentation. However, inhibition of growth in co-cultured Pleurotus ostreatus and Rhizoctonia solani by T. harzianum was increased in ThPV1-containing strains compared with ThPV1-cured isogenic strains. Moreover, β-1,3-glucanase activity was significantly increased in the ThPV1-containing strains. However, no difference in chitinase activity was observed, suggesting that ThPV1 regulates the activity of a specific fungal enzyme.

Aspirin Suppresses PGE2 and Activates AMP Kinase to Inhibit Melanoma Cell Motility, Pigmentation, and Selective Tumor Growth In Vivo.

There are conflicting epidemiologic data on whether chronic aspirin (ASA) use may reduce melanoma risk in humans. Potential anticancer effects of ASA may be mediated by its ability to suppress prostaglandin E2 (PGE2) production and activate 5'-adenosine monophosphate-activated protein kinase (AMPK). We investigated the inhibitory effects of ASA in a panel of melanoma and transformed melanocyte cell lines, and on tumor growth in a preclinical model. ASA and the COX-2 inhibitor celecoxib did not affect melanoma cell viability, but significantly reduced colony formation, cell motility, and pigmentation (melanin production) in vitro at concentrations of 1 mmol/L and 20 μmol/L, respectively. ASA-mediated inhibition of cell migration and pigmentation was rescued by exogenous PGE2 or Compound C, which inhibits AMPK activation. Levels of tyrosinase, MITF, and p-ERK were unaffected by ASA exposure. Following a single oral dose of 0.4 mg ASA to NOD/SCID mice, salicylate was detected in plasma and skin at 4 hours and PGE2 levels were reduced up to 24 hours. Some human melanoma tumors xenografted into NOD/SCID mice were sensitive to chronic daily ASA administration, exhibiting reduced growth and proliferation. ASA-treated mice bearing sensitive and resistant tumors exhibited both decreased PGE2 in plasma and tumors and increased phosphorylated AMPK in tumors. We conclude that ASA inhibits colony formation, cell motility, and pigmentation through suppression of PGE2 and activation of AMPK and reduces growth of some melanoma tumors in vivo This preclinical model could be used for further tumor and biomarker studies to support future melanoma chemoprevention trials in humans. Cancer Prev Res; 11(10); 629-42. ©2018 AACR.

Prediction of solar irradiance using ray-tracing techniques for coral macro- and micro-habitats

Light distribution on coral reefs is very heterogeneous at the microhabitat level and is an important determinant of coral thermal microenvironments. This study implemented a solar load model that uses a backward ray-tracing method to estimate macroscale and microscale variations of solar irradiance penetrating the ocean surface and impacting the surfaces of coral colonies. We then explored whether morphological characteristics such as tissue darkness (or pigmentation) and thickness may influence the amount of light captured and its spectral distribution by two contrasting coral colony morphologies, branching and massive. Results of global horizontal irradiance above and below the sea-surface and at the surface of coral colonies were validated using spectrometer scans, field measurements, and empirical correlations. The macroscale results of horizontal, irradiated, and shaded irradiance levels and solar altitude angles for PAR, UVA and UVB compared very well with the spectrometer-based observations (typically within < 5%). In general, a comparison between the model results and field and empirical measurements indicated that the contributions of clouds, turbidity, and tides to variations in irradiance at various depth (up to 5 m) were typically within 5–10% of each other. Moreover, the effect of colony darkness or pigmentation on light microenvironment was notably more pronounced for the massive species than branching colony. This study provided insights that species with thinner tissue have the ability to intercept more light with the difference in terms of irradiance levels between 0.1 mm and 0.8 mm tissue thickness for both massive and branching colonies were approximately 2 W m−2, which was quite unlikely to influence the overall coral heat budgets.

Screening for Bioflocculant-Producing Bacteria from the Marine Environment of Sodwana Bay, South Africa

Abstract Flocculants are chemicals that mediate flocculation process, by aggregating colloids from suspension to form floc. Chemical flocculants are hazardous to the environment, which inform the search for safer and eco-friendly alternatives from microorganisms. Bacterial strains were isolated from water and sediment samples collected from Sodwana Bay, South Africa, and physiological properties of the bacterial strains were observed. Flocculation test using kaolin clay suspension was done on all isolates and the ones that showed flocculating activity were identified molecularly using 16 rRNA gene sequence analysis. Forty marine bacteria isolates were gotten from sediments and water samples collected from Sodwana Bay. Most of the isolates exhibited a range of colony pigmentation (pink, creamy, yellow, and white). After purification of individual isolates, they were screened for their potential to produce bioflocculant. The result revealed that isolates marked SOD3, SOD10, SOD12, SOD26, SOD27, SOD28, SOD32, SOD33 and SOD34 produced bioflocculants as shown by the flocculating activities from their crude extract. All these isolates showed good flocculation of kaolin clay suspension above 60% (flocculating activity) except SOD12. These bioflocculant producing isolates were identified as Pseudoalteromonas sp, Alcaligenes faecalis, Bacillus subtilis, Bacillus cereus, Bacillus stratosphericus. The results showed Sodwana Bay, South Africa as a reservoir of bacteria with potential to produce flocculants. However, further studies on the optimisation of culture conditions for bioflocculant production, extraction, characterisation and application of isolates is on the way to underscore the biotechnological importance of these microbes as producers of substitutes to harmful chemical flocculants commonly used in water and wastewater treatment.

Identification of purple blotch pathogen of shallot by PCR using specific primer for Alternaria genus

Purple blotch (Alternaria porri), having symptom similar to stemphylium blight (Stemphylium vesicarium), is one of the important diseases that significantly lowered shallot yield in Indonesia. However, stemphylium blight has never been reported. Purple blotch pathogens from farmers’ crops in Bantul and experimental crop in Centre of Innovative Agricultural Technology Universitas Gadjah Mada (CIAT-UGM) Sleman Regencies were identified to observe whether Stemphylium sp. was found with A. porri. Thirteen isolates obtained showed variability in colony texture, colour and pigmentation. DNA fingerprinting through Polymerase Chain Reaction of BOX and Enterobacterial Repetitive Intergenic Consensus region dispersed those isolates into two main groups. Identification using Alternaria-specific primers, Dir1ITSAlt/Inv1ITSAlt revealed that 10 isolates were Stemphylium sp. and 3 isolates were Alternaria sp. A. porri was only identified from CIAT-UGM. This finding emphasise that the Alternaria-specific primer was able to amplify Stemphylium sp. Besides, stemphylium blight disease may have been occurred in farmers’ crops in Bantul.

Morphological and Transcriptomic Analysis Reveals the Osmoadaptive Response of Endophytic Fungus Aspergillus montevidensis ZYD4 to High Salt Stress.

Halophilic fungi have evolved unique osmoadaptive strategies, enabling them to thrive in hypersaline habitats. Here, we conduct morphological and transcriptomic response of endophytic fungus (Aspergillus montevidensis ZYD4) in both the presence and absence of salt stress. Under salt stress, the colony morphology of the A. montevidensis ZYD4 changed drastically and exhibited decreased colony pigmentation. Extensive conidiophores development was observed under salt stress; conidiophores rarely developed in the absence of salt stress. Under salt stress, yellow cleistothecium formation was inhibited, while glycerol and compatible sugars continued to accumulate. Among differentially expressed unigenes (DEGs), 733 of them were up-regulated while 1,619 unigenes were down-regulated. We discovered that genes involved in the accumulation of glycerol, the storage of compatible sugars, organic acids, pigment production, and asexual sporulation were differentially regulated under salt stress. These results provide further understanding of the molecular basis of osmoadaptive mechanisms of halophilic endophytic fungi.

Spiribacter aquaticus sp. nov., a novel member of the genus Spiribacter isolated from a saltern.

A moderately halophilic bacterium, designated strain SP30T, was isolated from a solar saltern located in Santa Pola, Alicante, on the East coast of Spain. It was a Gram-stain-negative, strictly aerobic bacterium, able to grow in 7.5-25 % (w/v) NaCl and optimally in 12.5 % (w/v) NaCl. Phylogenetic analyses, based on 16S rRNA gene sequences, showed that the novel isolate is a member of the genus Spiribacter, with the most closely related species being Spiribacter roseus SSL50T (99.9 % sequence similarity) and Spiribacter curvatus UAH-SP71T (99.4 % sequence similarity). The 16S rRNA gene sequence similarity with the type species Spiribacter salinus M19-40T was 96.6 %. The DNA-DNA relatedness value between strain SP30T and S. roseus SSL50T and S. curvatus UAH-SP71T was 40 and 55 %, respectively; these values are lower than the 70 % threshold accepted for species delineation. The major fatty acids were C16:0, C18 : 1ω7c, C19 : 0 cyclo ω8c and C12 : 0. Similarly to other species of the genus Spiribacter, strain SP30Twas observed as curved rods and spiral cells. Metabolic versatility was reduced to the utilization of a few organic compounds as the sole carbon and energy sources, as with other members of Spiribacter. However, it differed in terms of colony pigmentation (brownish-yellow instead of pink) and in having a higher growth rate. Based on these data and on the phenotypic, genotypic and chemotaxonomic characterization, we propose the classification of strain SP30T as a novel species within the genus Spiribacter, with the name Spiribacter aquaticus sp. nov. The type strain is SP30T (=CECT 9238T=LMG 30005T).

Evidence for Nitric Oxide Synthase Activity in Staphylococcus xylosus Mediating Nitrosoheme Formation

Staphylococcus xylosus is used as a starter culture in fermented meat products and contributes to color formation by the reduction of nitrate to nitrite. Nitrite is a food additive that is chemically turned to nitric oxide (NO) in meat but its safety has been questioned. The objective of this study was to determine the ability of NO synthase (NOS) of S. xylosus C2a to produce NO. For this purpose, a nos deletion mutant (Δnos) in S. xylosus was constructed and NO production was evaluated in a test based on its ability to form nitrosomyoglobin and nitrosoheme. Production of NO was abrogated in the Δnos mutant under aerobic conditions and reduced about 35-40% comparing to the wild type C2a under limited oxygenation. This mutant was sensitive to oxidative stress. The expression of genes encoding catalase was modulated in the mutant with an up-regulation of katA and a down-regulation of katB and katC. The Δnos mutant displayed high colony pigmentation after prolonged growth on agar medium. Finally, the Δnos mutant showed no growth in minimal medium. Growth was not restored in the minimal medium by complementation with nos, but was restored by either addition of phenylalanine or complementation with pdt, a gene that encodes a prephenate dehydratase involved in phenylalanine biosynthesis and co-transcribed with nos. Our findings clearly demonstrate NOS-mediated NO production in S. xylosus, a meat-associated coagulase-negative Staphylococcus.

Exploring the Antimicrobial and Antitumor Potentials of Streptomyces sp. AGM12-1 Isolated from Egyptian Soil.

The occurrence of extensive antibiotics resistant bacteria increased the demands for mining out new sources of antimicrobial agents. Actinomycetes, especially Streptomyces sp. have grasped considerable attention worldwide due to production of many useful bioactive metabolites. In the present study, a total of 52 actinomycetes were isolated from agricultural soil samples in Beni-Suef, Egypt. All isolates were characterized based on colony morphology, mycelium coloration, and pigment diffusion. They were screened for their capabilities to show antimicrobial activities against different indicator microorganisms, and only 20 isolates have shown significant antimicrobial activities against at least one of the tested indicator microorganisms. The isolate AGM12-1 was active against all tested microorganisms and showed a marked antitumor activity with IC50 3.3 and 1.1 μg/ml against HCT-116 and HepG-2 cell lines respectively. It was genotypically characterized as Streptomyces sp. with the presence of PKS Π biosynthetic gene cluster. Mannitol, ammonium sulfate, pH 7, 2% inoculum size and incubation for 11 days at 30°C were the optimum conditions that used to maximize the production and hence allowed purification of one active antimicrobial compound to homogeneity using high performance liquid chromatography with a molecular mass of m/z 488.05. Nuclear magnetic resonance structural elucidation showed that this compound was a diketopiperazine derivative.

Carotenoid production and phenotypic variation in Azospirillum brasilense

We assessed the occurrence of phenotypic variation in Azospirillum brasilense strains Sp7, Cd, Sp245, Az39 and phv2 during growth in rich media, screening for variants altered in colony pigmentation or extracellular polysaccharide (EPS) production. Previous studies showed that EPS-overproducing variants of Sp7 appear frequently following starvation or growth in minimal medium. In contrast, no such variants were detected during growth in rich media in the tested strains except for few variants of phv2. Regarding alteration in colony pigmentation (from pink to white in strain Cd and from white to pink in the others), strain Sp7 showed a relatively high frequency of variation (0.009–0.026%). Strain Cd showed a lower frequency of alteration in pigmentation (0–0.008%), and this type of variation was not detected in the other strains. In A. brasilense, carotenoid synthesis is controlled by two RpoE sigma factors and their cognate ChrR anti-sigma factors, the latter acting as negative regulators of carotenoid synthesis. Here, all tested (n = 28) pink variants of Sp7 carried mutations in one of the anti-sigma factor genes, chrR1. Our findings indicate that, in A. brasilense, phenotypic variation is strain- and environment-dependent and support the central role of ChrR1 in regulation of carotenoid production.

Chryseobacterium sp. PLI2 and Aeromonas hydrophila co-infection in pacu, Piaractus brachypomus (Cuvier, 1817) fries cultured in West Bengal, India

The present study reports the first isolation of Chryseobacterium sp. and co-infection with Aeromonas hydrophila in pacu, Piaractus brachypomus fries cultured in West Bengal, India. The infected pacu fries showed white patches on gill, tail rot, skin discoloration, scale loss, skin peeling, emaciation, and kidney inflammation. The yellow pigmented colony from the kidney of pacu on selective cytophaga agar was identified as Chryseobacterium sp. based on colony pigmentation, cell morphology, phenotypic characterization, and 16S rDNA gene sequencing. Phylogenetically, the members of the genus Chryseobacterium clustered together as a separate clade and the strain PLI2 (NCBI accession number KP898212) branched with a non-type strain C. indologenes VIT-CMJ1 (accession number KJ437473) with high node value (100). Yet, the strain PLI2 was distinctly different from closely related type strain C. indologenes LMG 8337 (accession number LN681561) with 97% DNA homology and low (80) node value or other type strains of NCBI GenBank database. The challenge experiments indicated that Chryseobacterium sp. PLI2 can cause substantial mortalities (25%) in abraded and bath (2.2×107/mL) treated pacu fries. The kidney of challenged pacu became pale and inflamed. Most likely, this report is a new record on the isolation of Chryseobacterium sp. from the kidney of P. brachypomus. Statement of relevance•Co-infection of Chryseobacterium sp. and A. hydrophila in Piaractus brachypomus caused nephropathy and large scale mortalities.•The record of a new bacterial disease Chryseobacterium sp. infection in Indian aquaculture is a serious cause for concern.

Caribbean massive corals not recovering from repeated thermal stress events during 2005-2013.

Massive coral bleaching events associated with high sea surface temperatures are forecast to become more frequent and severe in the future due to climate change. Monitoring colony recovery from bleaching disturbances over multiyear time frames is important for improving predictions of future coral community changes. However, there are currently few multiyear studies describing long‐term outcomes for coral colonies following acute bleaching events. We recorded colony pigmentation and size for bleached and unbleached groups of co‐located conspecifics of three major reef‐building scleractinian corals (Orbicella franksi, Siderastrea siderea, and Stephanocoenia michelini; n = 198 total) in Bocas del Toro, Panama, during the major 2005 bleaching event and then monitored pigmentation status and changes live tissue colony size for 8 years (2005–2013). Corals that were bleached in 2005 demonstrated markedly different response trajectories compared to unbleached colony groups, with extensive live tissue loss for bleached corals of all species following bleaching, with mean live tissue losses per colony 9 months postbleaching of 26.2% (±5.4 SE) for O. franksi, 35.7% (±4.7 SE) for S. michelini, and 11.2% (±3.9 SE) for S. siderea. Two species, O. franksi and S. michelini, later recovered to net positive growth, which continued until a second thermal stress event in 2010. Following this event, all species again lost tissue, with previously unbleached colony species groups experiencing greater declines than conspecific sample groups, which were previously bleached, indicating a possible positive acclimative response. However, despite this beneficial effect for previously bleached corals, all groups experienced substantial net tissue loss between 2005 and 2013, indicating that many important Caribbean reef‐building corals will likely suffer continued tissue loss and may be unable to maintain current benthic coverage when faced with future thermal stress forecast for the region, even with potential benefits from bleaching‐related acclimation.

MoVrp1, a putative verprolin protein, is required for asexual development and infection in the rice blast fungus Magnaporthe oryzae

Endocytosis is a crucial cellular process in eukaryotic cells which involves clathrin and/or adaptor proteins, lipid kinases, phosphatases and the actin cytoskeleton. Verprolin proteins, such as Vrp1 in Saccharomyces cerevisiae, are conserved family proteins that regulate actin binding and endocytosis. Here, we identified and characterized MoVrp1 as the yeast Vrp1 homolog in Magnaporthe oryzae. Deletion of the MoVRP1 gene resulted in defects in vegetative growth, asexual development, and infection of the host plant. The ∆Movrp1 mutants also exhibited decreased extracellular peroxidase and laccase activities and showed defects in colony pigmentation, hyphal surface hydrophobicity, cell wall integrity, autophagy, endocytosis, and secretion of avirulent effector. Our studies provided new evidences that MoVrp1 involved in actin cytoskeleton is important for growth, morphogenesis, cellular trafficking, and fungal pathogenesis.

Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain Reaction.

Background:Nontuberculous mycobacteria (NTM) are a group of opportunistic pathogens and these are widely dispersed in water and soil resources. Identification of mycobacteria isolates by conventional methods including biochemical tests, growth rates, colony pigmentation, and presence of acid-fast bacilli is widely used, but these methods are time-consuming, labor-intensive, and may sometimes remain inconclusive.Materials and Methods:The DNA was extracted from NTM cultures using CTAB, Chelex, Chelex + Nonidet P-40, FTA® Elute card, and boiling The quantity and quality of the DNA extracted via these methods were determined using UV-photometer at 260 and 280 nm, and polymerase chain reaction (PCR) amplification of the heat-shock protein 65 gene with serially diluted DNA samples.Results:The CTAB method showed more positive results at 1:10–1:100,000 at which the DNA amount was substantial. With the Chelex method of DNA extraction, PCR amplification was detected at 1:10 and 1:1000 dilutions.Conclusions:According to the electrophoresis results, the CTAB and Chelex DNA extraction methods were more successful in comparison with the others as regard producing suitable concentrations of DNA with the minimum use of PCR inhibitor.

Study on the Prevalence and Distribution of Staphylococcus aureus in Raw Cow Milk Originated from Alage Atvet College Dairy Farm, Ethiopia

This investigation was carried out from October 2011-June 2012 to determine the prevalence of Staphylococcus in milk and rate of contamination at farm and milk distribution tank in Alage Agricultural Technical Vocational Educational Training College. Milk samples (row) were cultured on sheep blood agar and incubated. The plates were examined for gross colony morphology, pigmentation and haemolytic characteristics at 24-48 h then presumptive colonies of Staphylococcus aureus was selected and sub cultured on nutrient agar and incubated. Then, bacteria were identified according to its Gram reaction, morphology and the catalase test, tube coagulase test (4 h), haemolysis, pigment production (golden yellow), mannitol and maltose fermentation were used. A total of 170 milk sample of cow were examined for bacteriological status of S. aureus, an overall 28.2% prevalence of Staphylococcus was found. From this, 21.2% was directly from the farm and 35.3% were from milk distributing site to consumers. The total prevalence of Staphylococcus varied among the sample taking site. The prevalence of S. aureus from distributing raw milk site (35.3%) was significantly higher than the prevalence of S. aureus from samples taken directly from the farm (21.2%). From this the study it is considered that Staphylococcosis a potential hazard for the public and contamination rate is high in distribution site which needs improvement of the hygienic status of the personnel’s at distribution site.

Prevalence and antibiotic susceptibility of Pseudomonas aeruginosa isolated from chronic suppurative otitis media

&lt;p class="abstract"&gt;&lt;strong&gt;Background:&lt;/strong&gt; Chronic suppurative otitis media (CSOM) is the most common middle ear infection characterized by recurrent ear discharges or otorrhoea through a tympanic perforation from the middle ear. The organisms isolated from CSOM include both aerobic and anaerobic bacteria. Among the bacteria, &lt;em&gt;Pseudomonas aeruginosa&lt;/em&gt; (&lt;em&gt;P. aeruginosa&lt;/em&gt;) has been particularly blamed for deep seated and progressive destruction of middle ear and mastoid structures through its toxins and enzymes. The present study was done to determine the prevalence and antibiotic susceptibility of &lt;em&gt;P. aeruginosa&lt;/em&gt; isolated from CSOM.&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Methods:&lt;/strong&gt; This was a retrospective study conducted in a tertiary care hospital in South India in March 2016. Data was obtained from the microbiology records from January 2014 to January 2016. Patients who had active ear discharge for at least three months were included in the study. &lt;em&gt;P. aeruginosa&lt;/em&gt; was identified by colony morphology, pigment production, characteristic musty or earthy odor. The identification was confirmed by motility and standard biochemical tests. &lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Results:&lt;/strong&gt; A total of 293 cases of CSOM were included in the study. Microbial growth was seen in 248 (84.6%) cases and remaining 45 cases did not show any growth. Among 248 bacterial growth, 87 (35%) were positive for P. aeruginosa. Most common organism was &lt;em&gt;P. aeruginosa&lt;/em&gt; followed by &lt;em&gt;Staphylococcus aureus&lt;/em&gt;. Highest resistance was seen with Gentamicin and Ofloxacin and least resistance with Imipenem. The increased resistance to Cephalosporin group is warning sign, as these are potent anti-Psuedomals.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusions:&lt;/strong&gt; &lt;em&gt;P. aeruginosa&lt;/em&gt; and Staphylococcus aureus were found to be the common cause of CSOM in the present study. &lt;em&gt;P. aeruginosa &lt;/em&gt;was resistant to commonly used antimicrobials. There was no resistance seen with Imipenem and only two strains were resistant to Piperacillin+tazobactum. Regular evaluation of the microbiological pattern and antibiogram of isolates will help in decreasing the complications of CSOM. &lt;/p&gt;

Effect of Various Compounds Blocking the Colony Pigmentation on the Aflatoxin B1 Production by Aspergillus flavus.

Aflatoxins and melanins are the products of a polyketide biosynthesis. In this study, the search of potential inhibitors of the aflatoxin B1 (AFB1) biosynthesis was performed among compounds blocking the pigmentation in fungi. Four compounds—three natural (thymol, 3-hydroxybenzaldehyde, compactin) and one synthetic (fluconazole)—were examined for their ability to block the pigmentation and AFB1 production in Aspergillus flavus. All compounds inhibited the mycelium pigmentation of a fungus growing on solid medium. At the same time, thymol, fluconazole, and 3-hydroxybenzaldehyde stimulated AFB1 accumulation in culture broth of A. flavus under submerged fermentation, whereas the addition of 2.5 μg/mL of compactin resulted in a 50× reduction in AFB1 production. Moreover, compactin also suppressed the sporulation of A. flavus on solid medium. In vivo treatment of corn and wheat grain with compactin (50 μg/g of grain) reduced the level of AFB1 accumulation 14 and 15 times, respectively. Further prospects of the compactin study as potential AFB1 inhibitor are discussed.

PG1058 Is a Novel Multidomain Protein Component of the Bacterial Type IX Secretion System.

Porphyromonas gingivalis utilises the Bacteroidetes-specific type IX secretion system (T9SS) to export proteins across the outer membrane (OM), including virulence factors such as the gingipains. The secreted proteins have a conserved carboxy-terminal domain essential for type IX secretion that is cleaved upon export. In P. gingivalis the T9SS substrates undergo glycosylation with anionic lipopolysaccharide (A-LPS) and are attached to the OM. In this study, comparative analyses of 24 Bacteroidetes genomes identified ten putative novel components of the T9SS in P. gingivalis, one of which was PG1058. Computer modelling of the PG1058 structure predicted a novel N- to C-terminal architecture comprising a tetratricopeptide repeat (TPR) domain, a β-propeller domain, a carboxypeptidase regulatory domain-like fold (CRD) and an OmpA_C-like putative peptidoglycan binding domain. Inactivation of pg1058 in P. gingivalis resulted in loss of both colonial pigmentation and surface-associated proteolytic activity; a phenotype common to T9SS mutants. Immunoblot and LC-MS/MS analyses of subcellular fractions revealed T9SS substrates accumulated within the pg1058 mutant periplasm whilst whole-cell ELISA showed the Kgp gingipain was absent from the cell surface, confirming perturbed T9SS function. Immunoblot, TEM and whole-cell ELISA analyses indicated A-LPS was produced and present on the pg1058 mutant cell surface although it was not linked to T9SS substrate proteins. This indicated that PG1058 is crucial for export of T9SS substrates but not for the translocation of A-LPS. PG1058 is a predicted lipoprotein and was localised to the periplasmic side of the OM using whole-cell ELISA, immunoblot and LC-MS/MS analyses of subcellular fractions. The structural prediction and localisation of PG1058 suggests that it may have a role as an essential scaffold linking the periplasmic and OM components of the T9SS.