Growth In Different Media Research Articles

Multi-metal tolerant Porostereum umbrinoalutaceum isolated from Pleurotus ostreatus fruit body showing root colonization, growth promotion of chilli (Capsicum annuum L.) plant and antagonism against fungal pathogens.

Mushroom associated microbes could be utilized to improve crop productivity providing nutrients, plant growth promoting substances, production of hydrolytic enzymes and protecting plant from biotic and abiotic stress. An endophyte designated as KUFC101 was isolated from fruit body of Pleurotus ostreatus and identified as Porostereum umbrinoalutaceum based on nuclear-rRNA gene sequence analysis. Growth in different culture media, metal tolerance, biochemical characterization and effect on chilli plant growth promotion were studied. The isolate showed best growth in Malt extract medium and least growth in synthetic media. It could tolerate toxic metals (Mg, Ca, Fe, Cu, Mn, Zn and Cd each at 100ppm concentration). It produced amylase, cellulase, chitinase, pectinase, catecholate type of siderophore and indole acetic acid, and inhibited growth of Alternaria solani and Penicillium citrinum. It could colonize in the rhizosphere of chilli plant and influence growth of chilli plant by improving biomass and metabolite content. Porostereum umbrinoalutaceum KUFC101 could be utilized in formulation of biofertiliser under sustainable agricultural system.

Plastic Debris in the Aquatic Environment: An Emerging Substratum for Antimicrobial Resistant (AMR) Biofilms.

Plastic pollution has quadrupled over the past years and has become a global concern due to its direct impact on life forms. The present study analysed whether the plastic debris in aquatic environments could act as the substratum for the antimicrobial-resistant (AMR) bacteria to form biofilm for survival. We have collected various plastic debris (n = 32) from six sites of the Periyar River, the drinking water source for the entire city and one of the most polluted rivers in Kerala (India). The chemical composition of plastics was screened via FTIR analysis and found that they comprised two types, viz., polyethylene and polypropylene. Bacteria isolated from the samples were screened for the AMR characteristics towards eight different classes of antibiotics. All isolates showed 100% resistance towards colistin and obtained the MAR index value of 0.1-0.4 range. Six representative bacterial isolates with high multiple antibiotic resistance (MAR) index were selected and identified by 16sRNA sequencing as Lysinibacillus mangiferihumi, Bacillus pumilus, Bacillus safensis, Bacillus cereus, Bacillus altitudins and Bacillus pumilus. In vitro biofilm formation was experimented on the purchased plastic samples in artificial media and river water using two selected strains, Bacillus pumilus and Bacillus cereus. Significant variations were observed in biofilm growth in different media (P < 0.05) regardless of plastic types (P > 0.05). The extracellular polymeric substances (EPS) and the characteristic holes on the surface morphology were visualized in SEM analysis, thus indicating the conditioning of the plastics by the isolates for biofilm formation.

Secondary metabolites and transcriptomic analysis of novel pulcherrimin producer Metschnikowia persimmonesis KIOM G15050: A potent and safe food biocontrol agent

Metschnikowia persimmonesis, a novel endophytic yeast strain isolated from Diospyros kaki calyx, possesses strong antimicrobial activity. We investigated its potential use as an environmentally safe food biocontrol agent through genomics, transcriptomics, and metabolomics. Secondary metabolites were isolated from M. persimmonesis, followed by chemical structure elucidation, PUL gene cluster identification, and RNA sequencing. Pulcherrimin was isolated using 2 M NaOH, its structure was confirmed, and the yield was quantified. Biocontrol efficacy of M. persimmonesis on persimmon fruits and calyx was evaluated by assessing lesion diameter and disease incidence. Following compounds were isolated from M. persimmonesis co-culture with Botrytis cinerea and Fusarium oxysporum: fusaric acid, benzoic acid, benzeneacetic acid, 4-hydroxybenzeneacetic acid, 4-(-2-hydoxyethyl)-benzoic acid, cyclo (Leu-Leu), benzenemethanol, 4-hydroxy-benzaldehide, 2-hydroxy-4-methoxy-benzoic acid, 4-hydroxy-benzoic acid, lumichrome, heptadecanoic acid, and nonadecanoic acid. Exposing M. persimmonesis to different growth media conditions (with or without sugar) resulted in the isolation of five compounds: Tyrosol, Cyclo (Pro-Val), cyclo(L-Pro-L-Tyr), cyclo(Leu-Leu), and cyclo(l-tyrosilylicine). Differentially expressed gene analysis revealed 3264 genes that were significantly expressed (fold change ≥2 and p-value ≤0.05) during M. persimmonesis growth in different media, of which only 270 (8.27%) showed altered expression in all sample combinations with Luria–Bertani Agar as control. Minimal media with ferric ions and tween-80 triggered the most gene expression changes, with the highest levels of PUL gene expression and pulcherrimin yield (262.166 mg/L) among all media treatments. M. persimmonesis also produced a higher amount of pulcherrimin (209.733 mg/L) than Metschnikowia pulcherrima (152.8 mg/L). M. persimmonesis inhibited the growth of Fusarium oxysporum in persimmon fruit and calyx. Toxicity evaluation of M. persimmonesis extracts showed no harmful effects on the liver and mitochondria of zebrafish, and no potential risk of cardiotoxicity in hERG-HEK293 cell lines. Thus, M. persimmonesis can be commercialized as a potent and safe biocontrol agent for preserving food products.

Morphological, Biochemical and Molecular Identification Bacteria Streptomyces spp. Causing Common Scab Disease on Potato in Iraq

This study was carried out at the University of Anbar - College of Agriculture - Department of Plant Protection. The study aimed to isolate and diagnose the bacterium that causes scabies on potatoes, Streptomyces spp., depending on the phenotypic characteristics, biochemical interactions, and molecular methods. The results of isolation from different regions and on PDA medium showed that it was obtained On 45 pure isolates of the pathogenic bacteria Streptomyces spp, and the results of growth in different media showed that the best medium for growth was PDA and Nutrient agar. The results of phenotypic diagnosis and biochemical tests revealed that 8 isolates belong to the type Streptomyces scabies and two isolates belong to the type Streptomyces acidiscabies. DNA was extracted from 10 isolates of pathogenic bacteria, and the results of molecular diagnosis using the 16S rRNA gene showed that light bands of 1250 pairs were obtained. The nitrogenous base sequences of these isolates were determined and deposited in GenBank under accession codes (OQ8191, OQ819106, OQ815918, OQ789975, OQ789966, OQ815949, OQ815950).

Cell wall composition in Cryptococcus neoformans is media dependent and alters host response, inducing protective immunity.

Cryptococcus neoformans is a basidiomycete fungus that can cause meningoencephalitis, especially in immunocompromised patients. Cryptococcus grows in many different media, although little attention has been paid to the role of growth conditions on the cryptococcal cell wall or on virulence. The purpose of this study was to determine how different media influenced the amount of chitin and chitosan in the cell wall, which in turn impacted the cell wall architecture and host response. Yeast extract, peptone, and dextrose (YPD) and yeast nitrogen base (YNB) are two commonly used media for growing Cryptococcus before use in in vitro or in vivo experiments. As a result, C. neoformans was grown in either YPD or YNB, which were either left unbuffered or buffered to pH 7 with MOPS. These cells were then labeled with cell wall-specific fluorescent probes to determine the amounts of various cell wall components. In addition, these cells were employed in animal virulence studies using the murine inhalation model of infection. We observed that the growth of wild-type C. neoformans KN99 significantly changes the pH of unbuffered media during growth. It raises the pH to 8.0 when grown in unbuffered YPD but lowers the pH to 2.0 when grown in unbuffered YNB (YNB-U). Importantly, the composition of the cell wall was substantially impacted by growth in different media. Cells grown in YNB-U exhibited a 90% reduction in chitosan, the deacetylated form of chitin, compared with cells grown in YPD. The decrease in pH and chitosan in the YNB-U-grown cells was associated with a significant increase in some pathogen-associated molecular patterns on the surface of cells compared with cells grown in YPD or YNB, pH 7. This altered cell wall architecture resulted in a significant reduction in virulence when tested using a murine model of infection. Furthermore, when heat-killed cells were used as the inoculum, KN99 cells grown in YNB-U caused an aberrant hyper-inflammatory response in the lungs, resulting in rapid animal death. In contrast, heat-killed KN99 cells grown in YNB, pH 7, caused little to no inflammatory response in the host lung, but, when used as a vaccine, they conferred a robust protective response against a subsequent challenge infection with the virulent KN99 cells. These findings emphasize the importance of culture media and pH during growth in shaping the content and organization of the C. neoformans cell wall, as well as their impact on fungal virulence and the host response.

Impact of electro-activated whey on growth, acid and bile resistance of Lacticaseibacillus rhamnosus GG and Lactobacillus acidophilus ATCC 4356

The aim of this work was to study the impact of electro-activated whey containing lactulose on the growth and survival Lacticaseibacillus rhamnosus GG and Lactobacillus acidophilus ATCC 4356 in acidic and bile salts containing media. Electro-activated whey was compared to whey and MRS alone and supplemented with lactulose. The results showed that OD600 was the highest for all these bacteria when grown in the electro-activated medium. At the same time, the obtained results showed that the initial growth phase was the most delayed in this medium. The OD600 results were verified by the bacteria plating method on nutrient agar. The obtained data showed that for each given bacteria, no significant difference was observed according to the CFU/mL results. Thus, it has been suggested that electro-activated whey could have a significant effect of bacterial fitness by enhancing their activity even at an equivalent population in each medium. A study of the stability of the probiotic bacteria for 14 days refrigerated storage at pH 4.6 and in the presence of bile salts revealed that the growth substrate did not significantly affect bacterial survival during this storage period and that all the tested probiotic bacteria remained close to 109 CFU/mL. The 16S rRNA gene sequencing of Lacticaseibacillus rhamnosus GG after 24 h growth in different media showed highly significant difference in upregulated and downregulated genes between the electro-activated whey and the regular sweet whey even when it was supplemented with lactulose. The obtained results support the hypothesis that electro-activated whey has evident prebiotic effect compared to lactulose.

Characterisation of the E. coli and Salmonella qseC and qseE mutants reveals a metabolic rather than adrenergic receptor role.

Catecholamine stress hormones (norepinephrine, epinephrine, and dopamine) are signals that have been shown to be used as environmental cues, which affect the growth and virulence of normal microbiota as well as pathogenic bacteria. It has been reported that Escherichia coli and Salmonella use the two-component system proteins QseC and QseE to recognise catecholamines and so act as bacterial adrenergic receptors. In this study, we mutated the E. coli O157:H7 and Salmonella enterica serovar Typhimurium genes encoding QseC and QseE and found that this did not block stress hormone responsiveness in either species. Motility, biofilm formation, and analysis of virulence of the mutants using two infection models were similar to the wild-type strains. The main differences in phenotypes of the qseC and qseE mutants were responses to changes in temperature and growth in different media particularly with respect to salt, carbon, and nitrogen salt sources. In this physiological respect, it was also found that the phenotypes of the qseC and qseE mutants differed between E. coli and Salmonella. These findings collectively suggest that QseC and QseE are not essential for E. coli and Salmonella to respond to stress hormones and that the proteins may be playing a role in regulating metabolism.

Characterization and pathogenicity of a Pythium aphanidermatum isolate causing ‘damping off’ in pepper seedlings

&lt;p&gt;The production of the pepper seedling (&lt;em&gt;Capsicum annuum&lt;/em&gt;) is affected by the fungal complex that causes the ‘damping-off’, in which some species of the oomycete &lt;em&gt;Pythium&lt;/em&gt; spp., stand out. The objective of the present study was to identify the causal agent of the death of pepper plants and evaluate its pathogenicity in pepper seeds and seedlings. A fast and aggressive growing oomycete was isolate from pepper plants, morphologically identified as P. aphanidermatum based on its sexual and asexual reproduction structures and, by molecular techniques. This isolate had a high degree of&lt;em&gt; in vitro&lt;/em&gt; pathogenicity in pre-emergence and post-emergence in chile, showing 100% mortality. In addition, it presented a high rate of mycelial growth in different culture media (V8-Agar, Corn Agar, Corn Potato Agar, Potato Dextrose Agar, Czapek &amp;amp; Oat Agar), being in V8-Agar medium the only medium where it developed reproduction structures sexual and asexual. The isolation presented a mycelial growth rate of 58.3 ± 0.3 mm / day at 26 ± 2 °C in PDA medium. Due to its rapid growth and its high degree of pathogenicity &lt;em&gt;in vitro&lt;/em&gt;, it is an unusual and aggressive isolate for pepper seedlings.&lt;/p&gt;

The prodigiosin change on the surface of Serratia marcescens detected by flow cytometry.

The potential of flow cytometry for the study of changes in prodigiosin on the cell surface of Serratia marcescens is of academic and practical interest. This is because S. marcescens can produce prodigiosin, a secondary metabolite, with potential use as a cancer-cell inhibitor. In this study, three groups of bacterial cultures with different carbon sources were compared, and the effect of the addition of cAMP to the sucrose-based culture was studied. Both cellular morphology and DNA content were detected by flow cytometry, rendering a broad description of the bacterial behavior. It is the first use of flow cytometry to investigate the dynamics of prodigiosin on the surface of S. marcescens during growth in different media. The fluorescence intensity is related to the DNA content, the forward-scattered light is related to cell volume, and the side-scattered light is related to the surface morphology, especially the surface prodigiosin. These may contribute to the potential development of a bacterial metabolic monitoring strategy using both DNA content analysis and bacterial morphology based on flow cytometry technique.

Characterization of a new strain of Metarhizium novozealandicum with potential to be developed as a biopesticide

ABSTRACT The fungal species Metarhizium novozealandicum, that occurs only in New Zealand and Australia has been poorly studied. In this work, a new strain of M. novozealandicum isolated from a larva of Wiseana sp. is described based on morphology, genomic multilocus (ITS, EF-1α and β-tubulin) phylogeny, growth in different culture media and insecticidal activity. The isolate AgR-F177 was clustered in the same clade with M. novozealandicum. AgR-F177 colonies developed faster on Sabouraud Dextrose Agar (SDA) than on Potato Dextrose Agar (PDA) when incubated at 25°C, with no growth observed at 30°C on either media. Conidia yield on an oat-based medium in semisolid fermentation was 7.41 x 108conidia/g of substrate and a higher yield of 1.68 x 109conidia/g of substrate was obtained using solid fermentation on cooked rice. AgR-F177 formed microsclerotia (MS) in liquid fermentation after 7 days reaching the maximum yield of 3.3 × 103 MS/mL after 10 days. AgR-F177 caused mortality in Wiseana copularis, Costelytra giveni and Plutella xylostella larvae with efficacies up to 100%, 69.2%, and 45.7%, respectively. The ease of production of AgR-F177 with different fermentation systems and its pathogenicity against different insect pests reveal its potential as a new biopesticide.

Peach palm plantlet growth in different culture media in a temporary immersion system

ABSTRACT: Peach palm is a domesticated palm commercially important for the production of fruits and hearts of palm. Somatic embryogenesis, an effective technique for mass propagation, was successfully established for this species. Furthermore, a temporary immersion system improved plant regeneration. However, production can be further improved by understanding the peach palm’s growth dynamic and modifications of culture media. The aims of this study were to evaluate the growth of plantlets cultured in different culture media in a temporary immersion system and to correlate the results with nutrient uptake during the growth period. Somatic embryo-derived young plantlets approximately 1 cm in length were cultivated for 12 weeks in a twin flask system containing MS, Y3 or N6 salts, Morel and Wetmore vitamins and 3% sucrose, with a monthly medium refreshment. Growth was measured and mineral analysis of the plantlets was carried out after 12 weeks of culture. The Y3 and MS salts were the most appropriate for the plant growth. Number of roots was 52.52% higher and the root size was 40.42% between the N6 and MS medium and the root number in Y3 medium was 37.74% greater than in MS medium, which is important for post acclimatization survival. K and Na are important elements for peach palm. N is not required at such a high concentration as in Murashige and Skoog formulation. The Chu (N6) medium did not generate high quality plantlets, possibly due to the absence of some micronutrients, like Mo, Cu and Co.

Phenotypic and molecular characterisation of Sporothrix globosa of diverse origin from India.

Sporotrichosis is one of the neglected tropical diseases causing subcutaneous chronic granulomatous lesion by thermally dimorphic fungi belonging to Sporothrix species. Sporothrix brasiliensis, Sporothrix mexicana and Sporothrix globosa are the common pathogenic species. In Asian countries, S. globosa constitutes nearly 99.3% of all Sporothrix species. We studied 63 cases of sporotrichosis of geographically diverse origin from India and Sporothrix isolates were characterised for its growth in different media, temperatures, ability to assimilate sugars and antifungal susceptibility profile. Molecular characterization was performed by sequencing of the calmodulin (CAL), beta tubulin (BT) and translational elongation factor 1-alpha (TEF-1α) and typing by fluorescent amplified fragment length polymorphism (FAFLP). In patients who presented with fixed (49.2%), lymphocutaneous lesions (23.8%), in 26.9% the details were not known, none had systemic dissemination. All the isolates tested were Sporothrix globosa and that could grow up to 35°C and unable to grow at and beyond 37°C. The assimilation of sucrose, ribitol and raffinose helps in identifying S. globosa. Sequences of CAL or BT or TEF-1α can differentiate S. globosa from other species in the complex. FAFLP results exhibited low genetic diversity. No correlation was noted between genotypes and clinical presentation, or geographic distribution. Itraconazole, terbinafine and posaconazole showed good in vitro antifungal activity against S. globosa whereas fluconazole and micafungin had no activity. S. globosa of Indian origin is relatively less pathogenic than other pathogenic Sporothrix species as it does not cause systemic dissemination and in the diagnostic laboratory, incubation of the cultures below 37°C is essential for effective isolation.

Correlation between protein concentration and bacterial cell size can reveal mechanisms of gene expression

Classically, gene expression is modeled as a chemical process with reaction rates dependent on the concentration of the reactants (typically, DNA loci, plasmids, RNA, enzymes, etc). Other variables like cell size are in general ignored. Size dynamics can become an important variable due to the low number of many of these reactants, imperfectly symmetric cell partitioning and molecule segregation. In this work we measure the correlation between size and protein concentration by observing the gene expression of the RpOD gene from a low-copy plasmid in Escherichia coli during balanced growth in different media. A positive correlation was found, and we used it to examine possible models of cell size dynamics and plasmid replication. We implemented a previously developed model describing the full gene expression process including transcription, translation, loci replication, cell division and molecule segregation. By comparing with the observed correlation, we determine that the transcription rate must be proportional to the size times the number of plasmids. We discuss how fluctuations in plasmid segregation, due to the low copy number, can impose limits in this correlation.

Nitrogen Depletion Blocks Growth Stimulation Driven by the Expression of Nitric Oxide Synthase in Tobacco.

Nitric oxide (NO) is a messenger molecule widespread studied in plant physiology. Latter evidence supports the lack of a NO-producing system involving a NO synthase (NOS) activity in higher plants. However, a NOS gene from the unicellular marine alga Ostreococcus tauri (OtNOS) was characterized in recent years. OtNOS is a genuine NOS, with similar spectroscopic fingerprints to mammalian NOSs and high NO producing capacity. We are interested in investigating whether OtNOS activity alters nitrogen metabolism and nitrogen availability, thus improving growth promotion conditions in tobacco. Tobacco plants were transformed with OtNOS under the constitutive CaMV 35S promoter. Transgenic tobacco plants expressing OtNOS accumulated higher NO levels compared to siblings transformed with the empty vector, and displayed accelerated growth in different media containing sufficient nitrogen availability. Under conditions of nitrogen scarcity, the growth promoting effect of the OtNOS expression is diluted in terms of total leaf area, protein content and seed production. It is proposed that OtNOS might possess a plant growth promoting effect through facilitating N remobilization and nitrate assimilation with potential to improve crop plants performance.

Growth and yield performance of oyster mushroom (Pleurotus ostreatus) on water hyacinth as a substrate

Cultivation of oyster mushroom ( Pleurotus ostreatus ) on the aquatic weed water hyacinth is an eco-friendly way of controlling and management of such a problematic weed. In the present work, water hyacinth has been used as a low-cost substrate in combinations with rice-straw of ratio 3:1 for the cultivation of P. ostreatus . The objective of this study was to cultivate oyster mushroom in water hyacinth compost degraded with lingo-cellulolytic fungi. The experiment was performed in control, before fungal treated compost and fungal treated compost. The data was analyzed on various aspects like completion of mycelium growth in different media like Potato dextrose agar media and water hyacinth media, the duration for spawn run, the appearance of pinheads, fruiting bodies and number of fruit bodies produced. The nutrient analysis was done in all types of fruit bodies. Aspergillus flavus treated compost took short duration (24 ± 1 days) and control sample compost took a long duration (27.3 ± 1.53 days) for fruiting. The highest production was recorded in Trichoderma sp. treated compost (68.8%) and lowest in control compost (30.7%). This study has successfully demonstrated the possibility of water hyacinth as a substrate in mushroom production and management of water hyacinth.

Two Rab GTPases play different roles in conidiation, trap formation, stress resistance, and virulence in the nematode-trapping fungus Arthrobotrys oligospora.

Rab GTPases are the largest group of the small GTPases family, which play a pivotal role in the secretion of proteins. Arthrobotrys oligospora is a representative nematode-trapping fungus that can produce adhesive networks to capture nematodes. In this study, the roles of two Rab GTPases AoRab-7A and AoRab-2 were characterized by gene knockout in the fungus A. oligospora. The disruption of AoRab-7A hindered the mycelial growth in different media, the conidiation of ΔAoRab-7A transformants was almost abolished, and the transcription of four sporulation-related genes (AbaA, FluG, Hyp1, and VosA) was downregulated compared to the wild-type strain (WT). Furthermore, the tolerance of the ΔAoRab-7A mutants to sodium dodecyl sulfate (SDS) and H2O2 was also significantly reduced compared to the WT, and the transcription of several genes related to environmental resistance, such as genes for catalase and trehalose synthase, was downregulated. Similarly, the extracellular proteolytic activity was decreased. Importantly, the ΔAoRab-7A mutants were unable to produce traps and capture nematodes. However, the disruption of gene AoRab-2 only affected the conidiation slightly but non-significantly, while other phenotypic traits were unaffected. Moreover, the gene AoRab-7A was also involved in the autophagy induced by nitrogen deprivation in A. oligospora. Our results revealed for the first time that the Rab GTPases are involved in the regulation of mycelial growth, conidiation, trap formation, stress resistance, and pathogenicity in the nematode-trapping fungus A. oligospora.

The Sporothrix schenckii Gene Encoding for the Ribosomal Protein L6 Has Constitutive and Stable Expression and Works as an Endogenous Control in Gene Expression Analysis.

Sporothrix schenckii is one of the causative agents of sporotrichosis, a worldwide-distributed mycosis that affects humans and other mammals. The interest in basic and clinical features of this organism has significantly increased in the last years, yet little progress in molecular aspects has been reported. Gene expression analysis is a set of powerful tools that helps to assess the cell response to changes in the extracellular environment, the genetic networks controlling metabolic pathways, and the adaptation to different growth conditions. Most of the quantitative methodologies used nowadays require data normalization, and this is achieved measuring the expression of endogenous control genes. Reference genes, whose expression is assumed to suffer minimal changes regardless the cell morphology, the stage of the cell cycle or the presence of harsh extracellular conditions are commonly used as controls in Northern blotting assays, microarrays, and semi-quantitative or quantitative RT-PCR. Since the biology of the organisms is usually species specific, it is difficult to find a reliable group of universal genes that can be used as controls for data normalization in experiments addressing the gene expression, regardless the taxonomic classification of the organism under study. Here, we compared the transcriptional stability of the genes encoding for elongation factor 1A, Tfc1, a protein involved in transcription initiation on Pol III promoters, ribosomal protein L6, histone H2A, β-actin, β-tubulin, glyceraldehyde 3-phosphate dehydrogenase, UAF30, the upstream activating factor 30, and the transcription initiation factor TFIID subunit 10, during the fungal growth in different culture media and cell morphologies. Our results indicated that only the gene encoding for the ribosomal protein L6 showed a stable and constant expression. Furthermore, it displayed not transcriptional changes when S. schenckii infected larvae of Galleria mellonella or interacted with immune cells. Therefore, this gene could be used as control for data normalization in expression assays. As a proof of concept, this gene was used to assess the expression of genes encoding for glycosidases involved in the protein N-linked glycosylation pathway, a histidine kinase whose expression is regulated during the fungal dimorphism, and a glycosidase that participates in sucrose assimilation.

Influence of the Culture Media and the Organic Matter in the Growth of Paxillus ammoniavirescens (Contu & Dessi)

The genus Paxillus is characterized by the difficulty of species identification, which results in reproducibility problems, as well as the need for large quantities of fungal inoculum. In particular, studies of Paxillus ammoniavirescens have reported divergent results in the in vitro growth while little is known of its capacity to degrade organic matter. For all the above, and assuming that this variability could be due to an inappropriate culture media, the aim of this study was to analyse growth in different culture media (MMN, MS, and 1/2 MS) and in the case of MMN in presence/absence of two types of organic matter (fresh litter and senescence litter) to probe the saprophytic ability of P. ammoniavirescens. We also evaluated the effects of pH changes in the culture media. Growth kinetics was assessed by weekly quantification of the area of growth in solid culture media over 5 wk, calculating the growth curves and inflection points of each culture media. In addition, final biomass after 5 wk in the different culture media was calculated. Results showed that best culture media are MS and 1/2 MS. Moreover, an improvement in growth in culture media containing decomposing fall litter was observed, leading to confirm differences in the culture media of this species with others of the same genus. Further, we established that all growth media suffered a significant acidification after fungal growth.

Kefir Growth in Different Culture Media

Kefir is a food supplement of great therapeutic value and also a probiotic food. The purpose of this project was to analyze kefir growth in different culture media. First, kefir grains were placed to grow in sterilized test tubes. The colony of kefir was divided into 16 samples containing from 9.579g to 10.400g of kefir grains. Growth was controlled by weight before and after filtration, and the pH of each solution was measured. After each measurement, the solution was disposed of and exchanged for the same amount and ingredients. These procedures were repeated for 3 consecutive days. After four days, the percentage of Kefir growth obtained in the solutions were at: whole milk (WM): 77%; whole milk with papaya juice (WMP): 64%; whole milk with orange juice (WMO): 34%; whole milk with lemon juice (WML): 72%; reduced fat milk (RFM): 120%; reduced fat milk with papaya juice (RFMP): 111%; reduced fat milk with orange juice (RFMO): 86%; reduced fat milk with lemon juice (RFML): 49%; skimmed milk (SM): 75%; skimmed milk with papaya juice (SMP): 75%; skimmed milk with orange juice(SMO): 93%; skimmed milk with lemon juice (SML): 86%; powdered milk (PM): 71%; powdered milk with papaya juice (PMP) 34%; powdered milk with orange juice (PMO): 39%; powdered milk with lemon juice (PML): 44%. The changed pHs obtained after each filtration of the solutions were: WM from 6.5 to 4; WMP: from 6.5 to 3.5; WMO: from 6 to 3; WML: from 4 to 3.5; RFM: from 6 to 4; RFMP: from 6 to 4; RFMO: from 5 to 3.5; RFML: from 4 to 3.5; SM: from 6.5 to 4; SMP: from 6 to 3.5; SMO: from 5 to 3.5; SML: from 4 to 3; PM: from 7 to 4; PMP: from 6 to 3.5; PMO: from 6 to 3.5; PML: from 3 to 3. The Kefir colony showed more growth in the reduced fat milk, and reduced fat milk with papaya juice, and substantially growth at skimmed milk with orange juice. The fermentation induced the growth of Kefir grains, the production of lactic acid and the decrease of the pH in the solutions. However if pH decreases more than 2 units, it decreases the Kefir growth. It is necessary more studies to evaluate the adequate initial pH to optimizing Kefir growth.Support or Funding InformationThis research was supported by the CECS ‐ Engineering, Modeling and Applied Social Science Department of the Federal University of ABC. Solutions and amount of Kefir grains. SOLUTION'S COMPOSITION AMOUNT INITIAL OF KEFIR (g) T1 WHOLE MILK 10.002 T2 WHOLE MILK + PAPAYA JUICE 10.232 T3 WHOLE MILK + ORANGE JUICE 10.156 T4 WHOLE MILK + LEMON JUICE 9.69 T5 REDUCED FAT MILK 10.4 T6 REDUCED FAT MILK + PAPAYA JUICE 10.4 T7 REDUCED FAT MILK + ORANGE JUICE 9.971 T8 REDUCED FAT MILK + LEMON JUICE 10.077 T9 SKIMMED MILK 9.912 T10 SKIMMED MILK + PAPAYA JUICE 9.992 T11 SKIMMED MILK + ORANGE JUICE 10.329 T12 SKIMMED MILK + LEMON JUICE 10.367 T13 POWDERED MILK 9.875 T14 POWDERED MILK + PAPAYA JUICE 10.153 T15 POWDERED MILK + ORANGE JUICE 9.579 T16 POWDERED MILK + LEMON JUICE 9.782

Potential of Rhodobacter capsulatus Grown in Anaerobic-Light or Aerobic-Dark Conditions as Bioremediation Agent for Biological Wastewater Treatments

The use of microorganisms to clean up wastewater provides a cheaper alternative to the conventional treatment plant. The efficiency of this method can be improved by the choice of microorganism with the potential of removing contaminants. One such group is photosynthetic bacteria. Rhodobacter capsulatus is a purple non-sulfur bacterium (PNSB) found to be capable of different metabolic activities depending on the environmental conditions. Cell growth in different media and conditions was tested, obtaining a concentration of about 108 CFU/mL under aerobic-dark and 109 CFU/mL under anaerobic-light conditions. The biomass was then used as a bioremediation agent for denitrification and nitrification of municipal wastewater to evaluate the potential to be employed as an additive in biological wastewater treatment. Inoculating a sample of mixed liquor withdrawn from the municipal wastewater treatment plant with R. capsulatus grown in aerobic-dark and anaerobic-light conditions caused a significant decrease of N-NO3 (&gt;95%), N-NH3 (70%) and SCOD (soluble chemical oxygen demand) (&gt;69%), independent of the growth conditions. A preliminary evaluation of costs indicated that R. capsulatus grown in aerobic-dark conditions could be more convenient for industrial application.