Liquid Culture Medium Research Articles

Phase Contrast Image-Based Rapid Antimicrobial Susceptibility Testing of Bacteria in Liquid Culture Media

Currently, the world is facing the problem of bacterial resistance, which threatens public health, and bacterial antimicrobial susceptibility testing (AST) plays an important role in biomedicine, dietary safety and aquaculture. Traditional AST methods take a long time, usually 16-24 h, and cannot meet the demand for rapid diagnosis in the clinic, so rapid AST methods are needed to shorten the detection time. In this study, by using an in-house built centrifuge to centrifuge bacteria in a liquid medium onto the inner wall of the bottom surface of a counting plate, and using a phase contrast microscope to track bacterial growth under the effect of different antibiotic concentrations, the results of the minimum inhibitory concentration (MIC) of bacteria under the effect of antibiotics can be obtained in as early as 4 h. We used a combination of E. coli and tigecycline and obtained MIC results that were consistent with those obtained using the gold standard broth micro-dilution method, demonstrating the validity of our method; due to the time advantage, the complete set can be used in the future for point of care and clinical applications, helping physicians to quickly obtain the MIC used to inhibit bacterial growth.

Identification of Nocardia and non-tuberculous Mycobacterium species by MALDI-TOF MS using the VITEK MS coupled to IVD and RUO databases.

Identification of Nocardia and Mycobacterium species by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) is still a challenging task that requires both suitable protein extraction procedures and extensive databases. This study aimed to evaluate the VITEK MS Plus system coupled with updated RUO (v4.17) and IVD (v3.2) databases for the identification of Nocardia spp. and Mycobacterium spp. clinical isolates. Sample preparation was carried out using the VITEK MS Mycobacterium/Nocardia kit for protein extraction. From 90 Nocardia spp. isolates analysed, 86 (95.6%) were correctly identified at species or complex level using IVD and 78 (86.7%) using RUO. Only two strains were misidentified as other species pertaining to the same complex. Among the 106 non-tuberculous Mycobacterium clinical isolates tested from a liquid culture medium, VITEK MS identified correctly at species or complex level 96 (90.6%) isolates in the IVD mode and 89 (84.0%) isolates in the RUO mode. No misidentifications were detected. Although the IVD mode was unable to differentiate members of the M. fortuitum complex, the RUO mode correctly discriminated M. peregrinum and M. septicum. The robustness and accuracy showed by this system allow its implementation for routine identification of these microorganisms in clinical laboratories.

Culture of Mycobacterium avium subsp. paratuberculosis: challenges, limitations and future prospects.

Mycobacterium avium subsp. paratuberculosis (MAP) causes paratuberculosis (Johne's disease) in ruminants and is suspected to be involved in the development of Crohn's disease and several autoimmune disorders. As such, sensitive and specific MAP detection methods are required to confirm infection in animals and identify potential sources of animal and human exposure. Despite recent developments in immunological and nucleic acid-based detection methods, culture-based detection of MAP remains the 'gold standard' against which the sensitivity and specificity of other detection methods are measured. However, not all culture-based approaches are equivalent in terms of detection capability, which can lead to errors in the evaluation of other detection methods. This review will provide an overview of the chronological development of culture methods for MAP, and will consider the unique growth requirements of MAP, the merits of solid versus liquid culture media, the relative performance of the commonly used MAP culture media, and sample preparation/decontamination protocols for different sample types. The limitations of current MAP culture methods and prospects for improvements are discussed.

Bivalent copper ions presence triggers removal and homeostatic mechanisms in the metal-resistant microorganism Apiotrichum loubieri M12

Microorganisms, especially those habiting mining environments, are of great importance for the retention of toxic metals in the environment. This work aimed to isolate a copper removing-microorganism from sediments of an Acid Mine Drainage-affected environment and to study the cellular responses trigger by metal presence. Apiotrichum loubieri M12 was able to tolerate and remove Cu(II) from liquid culture media, reaching a 30–35% removal capacity when it was exposed to 40 μg mL−1 Cu(II) after 48 h. Analysis of the biomass exposed to the metal through SEM-EDS showed copper presence on the cell surface and variations in the proportion of other biomass constituent elements. Proteomics revealed that the presence of Cu(II) induces differential expression of intracellular proteins involved in a wide variety of metabolic processes. Interestingly, a specific response to the metal was detected in cell-free supernatants, in which copper binding proteins were identified. A large number of proteins with metal ion binding sites were detected both at intra and extracellular levels. The microorganism responds not only by adjusting intracellular protein expression, but also by adjusting expression of proteins in the extracellular space.

Utilization of Corn Steep Liquor for the Production of Fairy Chemicals by Lepista sordida Mycelia.

There are various potential practical uses of fairy chemicals (FCs) in the fields of agriculture, cosmetics, and medicine; however, the production costs of FCs are very high. To enable the practical use of FCs, more efficient and inexpensive methods of culturing the mycelia of FCs-producing fungi and producing FCs need to be developed. The purpose of the present study was to determine methods of reducing the production costs of FCs and mycelia of the FCs-producing fungus Lepista sordida. We investigated the effects of four food industrial by-products, i.e., corn steep liquor (CSL), rice bran, wheat bran, and Japanese liquor lees, as nutritional additives in the liquid culture medium of the fungus. We found that CSL was more effective than the other tested additives in increasing the production of FCs and mycelia. Medium containing 1% CSL was optimal for increasing the mycelial yield while medium containing 6% CSL was optimal for increasing the production of FCs. The reason for this difference in the optimal CSL concentration was considered to be related to the stress on the mycelia caused by the amount of nutrients in the liquid medium. These results are expected to facilitate the practical use of FCs and the mycelia of FCs-producing fungi.

Development of liquid culture media mimicking the conditions of sinuses and lungs in cystic fibrosis and health.

The respiratory tract is a compartmentalised and heterogenous environment. The nasopharynx and sinuses of the upper airways have distinct properties from the lungs and these differences may shape bacterial adaptation and evolution. Upper airway niches act as early colonisation sites for respiratory bacterial pathogens, including those, such as Pseudomonas aeruginosa, that can go on to establish chronic infection of the lungs in people with cystic fibrosis (CF). Despite the importance of upper airway environments in facilitating early adaptation to host environments, currently available in vitro models for study of respiratory infection in CF focus exclusively on the lungs. Furthermore, animal models, widely used to bridge the gap between in vitro systems and the clinical scenario, do not allow the upper and lower airways to be studied in isolation. We have developed a suite of culture media reproducing key features of the upper and lower airways, for the study of bacterial adaptation and evolution in different respiratory environments. For both upper and lower airway-mimicking media, we have developed formulations that reflect airway conditions in health and those that reflect the altered environment of the CF respiratory tract. Here, we describe the development and validation of these media and their use for study of genetic and phenotypic adaptations in P. aeruginosa during growth under upper or lower airway conditions in health and in CF.

Contribution to optimization and standardization of antibacterial assays with silver nanoparticles: the culture medium and their aggregation

The antimicrobial activity of silver nanoparticles is determined by their size and specific properties, as well as by the chemical composition of the exposure medium in which the nanoparticles are suspended. When the antibacterial tests are carried out in a culture medium, aggregation of the nanoparticles is produced, decreasing their effectiveness. This study proposes the addition of surfactants to the culture medium to prevent the aggregation of silver nanoparticles and optimizes the concentrations of these surfactants. The aggregation of silver nanoparticles was studied by dynamic light scattering (DLS) after dispersion in three liquid culture media (Mueller-Hinton (MH), Luria-Bertani (LB) and Brain Heart Infusion) in which four different surfactants (SDS, Triton X100, Tween 80 and CTAB) were added at concentrations of 0, 0.1, 0.5, 1, 1.5 and 2%. Results showed that, the optimal culture media to prevent aggregation of silver nanoparticles were MH and LB with higher concentrations of Tween 80 and Triton X100 surfactants; being MH + 2% of Tween 80 and MH + 1% Triton X100 the best combinations obtained because the results obtained were closest to the sizes of nanoparticles in ultrapure water. In addition, it has been verified that the optimal medium + surfactant combinations chosen did not affect the viability of Escherichia coli bacteria. Nanoparticle aggregation was not observed by single particle inductively coupled plasma mass spectrometry (SP-ICP-MS) when nanoparticles were incubated for long incubations periods (24 h) in the optimal medium chosen.

Depolymerization of lignin by extracellular activity of Pycnoporus cinnabarinus, to obtain cellulose

Abstract Cellulose can be used to produce biofuels and many other products like pharmaceutical goods, food supplements, cosmetics, bio-plastics, etc. Lignocellulosic materials, like O. ficus indica residuals, are a heterogeneous biopolymer formed mainly by lignin, hemicellulose and cellulose. Lignin provides protection to the plants against chemical and microbial degradation, but it can be degraded by white rot fungi species, like Pycnoporus cinnabarinus. Since cellulose molecules are arranged in regular bundles enveloped by hemicellulose and lignin molecules, it is necessary to brake lignin and hemicellulose molecules to recover cellulose for its use in bioprocess. In this work, a biotechnological process for cellulose recovery from cactus waste through depolymerization of lignin by P. cinnabarinus, is presented. The delignification is carried out by aerobic culture in batch stirred bioreactors, with a liquid culture medium enriched with nutrients and minerals with O. ficus indica residuals as the unique carbon source, during eight-day span under continuous feeding of oxygen. A factorial design of experiments (DOE) for eight sets of factor values was selected for this study. The factors were: particle size, pH level, and process temperature. For each experiment, biomass, total reducing carbohydrates (TRC) and dissolved oxygen (DO) concentrations were measured every 24 h. At the end of each experiment, the percentage of delignification, and cellulose recovery was measured by Infrared (IR) spectroscopy. Up to 67% of delignification and 22% of cellulose recovery were obtained by the process. These results were analyzed by a factorial DOE in order to maximize each response individually and to optimize both responses together. The delignification of Opuntia ficus indica thorns has not been previously reported to our knowledge.

A novel PGPF Penicillium olsonii isolated from the rhizosphere of Aeluropus littoralis promotes plant growth, enhances salt stress tolerance, and reduces chemical fertilizers inputs in hydroponic system.

The hydroponic farming significantly enhances the yield and enables multiple cropping per year. These advantages can be improved by using plant growth-promoting fungi (PGPF) either under normal or stress conditions. In this study, the fungal strain (A3) isolated from the rhizosphere of the halophyte plant Aeluropus littoralis was identified as Penicillium olsonii based on sequence homology of its ITS region. The A3 fungus was shown to be halotolerant (up to 1 M NaCl) and its optimal growth was at 27°C, but inhibited at 40°C. In liquid culture medium, the A3 produced indole acetic acid (IAA) especially in the presence of L-tryptophan. Tobacco plants grown under hydroponic farming system were used to evaluate the promoting activity of the direct effect of A3 mycelium (DE) and the indirect effect (IDE) of its cell-free culture filtrate (A3CFF). The results showed that for the two conditions (DE or IDE) the tobacco seedlings exhibited significant increase in their height, leaf area, dry weight, and total chlorophyll content. Interestingly, the A3CFF (added to the MS liquid medium or to nutrient solution (NS), prepared from commercial fertilizers) induced significantly the growth parameters, the proline concentration, the catalase (CAT) and the superoxide dismutase (SOD) activities of tobacco plants. The A3CFF maintained its activity even after extended storage at 4°C for 1 year. Since the A3 is a halotolerant fungus, we tested its ability to alleviate salt stress effects. Indeed, when added at 1:50 dilution factor to NS in the presence of 250 mM NaCl, the A3CFF enhanced the plant salt tolerance by increasing the levels of total chlorophyll, proline, CAT, and SOD activities. In addition, the treated plants accumulated less Na+ in their roots but more K+ in their leaves. The A3CFF was also found to induce the expression of five salt stress related genes (NtSOS1, NtNHX1, NtHKT1, NtSOD, and NtCAT1). Finally, we proved that the A3CFF can reduce by half the chemical fertilizers inputs. Indeed, the tobacco plants grown in a hydroponic system using 0.5xNS supplemented with A3CFF (1:50) exhibited significantly higher growth than those grown in 0.5xNS or 1xNS. In an attempt to explain this mechanism, the expression profile of some growth related genes (nitrogen metabolism (NR1, NRT1), auxin (TRYP1, YUCCA6-like), and brassinosteroid (DET2, DWF4) biosynthesis) was performed. The results showed that all these genes were up-regulated following plant treatment with A3CFF. In summary the results revealed that the halotolerant fungus P. olsonii can stimulates tobacco plant growth, enhances its salt tolerance, and reduces by half the required chemical fertilizer inputs in a hydroponic farming system.

S1.5d Spectrum of etiological agents of mycotic keratitis: A n 11- year review

S1.5 Mycotic Keratitis, September 21, 2022, 11:00 AM - 12:30 PMBackgroundTo assess the spectrum of etiological agents of mycotic keratitis over an 11-year-period at a tertiary eye care center in southern India.MethodsA retrospective review was made of microbiological data relating to corneal scrapings performed over a period of 11 years (January 2011-December 2021) on 1200 individuals who presented with suspected microbial keratitis. Each individual underwent corneal scraping and the scraped materials were subjected to meticulous microbiological analysis that included direct microscopy (Gram-stain and lactophenol cotton blue wet mount) and culture on multiple solid and liquid culture media.ResultsA total of 404 fungal isolates were recovered from the corneal scrapings of 1200 patients with suspected microbial keratitis. Of the 404 fungal isolates, Fusarium spp (133) were the predominant isolates, followed by Aspergillus flavus (104), Curvularia spp (24), Aspergillus fumigatus (17), Bipolaris sp (7), Alternaria sp (2), Colletrotrichum sp (2) Cylindrocarpon lichenicola (2) Exserohilum sp (1) and Drechslera sp (1); there were also 111 filamentous fungus isolates that defied identification in spite of various efforts made to induce sporulation. Of the 404 culture-proven cases of mycotic keratitis, 381 patients confirmed ocular trauma while engaged in agricultural activity.Conclusion Fusarium spp., followed by Aspergillus spp., were the most common organisms found in mycotic keratitis patients in this specific geographical area. Additional efforts are required to spread awareness among villagers about the dangers of not promptly treating mycotic and other forms of microbial keratitis so that blindness and visual disability caused by corneal scarring in rural areas can be reduced.

Effective single and contest carcinogenic dyes adsorption onto A-zeolite/bacterial cellulose composite membrane: Adsorption isotherms, kinetics, and thermodynamics

This study reports loading of A-zeolite particles on the bacterial cellulose membrane for single and binary carcinogenic dye uptake from the aqueous solution. The bio based membrane was obtained using Gluconobacter Oxydans NRRL 759 in liquid culture medium. Meanwhile, the incorporation of zeolite particles within the bacterial cellulose matrix and on its surface was implemented via in-situ synthesis. The crystallite structure and the morphology of obtained composite membrane were characterized with X-ray diffraction (XRD) and scanning electron microscopy (SEM) hyphenated with energy dispersive X-ray (EDX). The surface characteristics were intensively investigated by N2 adsorption/desorption isotherm, SBET, pore size distribution, and zeta potential. The results indicate that the treatment of bacterial cellulose with zeolite particles reflected two folds in surface area and three folds in the point of zero charge (pHzpc 6.7) compared with un-modified one. The produced composite membrane was also examined against removal of sole cationic methylene blue "MB", and anionic Congo-Red "CR" or in binary solutions included "MB" with Malachite green "MG" or "CR" with different percentages through batch adsorption technique. Different parameters affecting the adsorption process, such as dye concentration, adsorbent type, contact time, temperature, and pH, were optimized. The highest removal was recorded at optimum conditions for sole "MB" dye (99.2%), "CR" dye (81%), binary mixed "2MB/1CR" (96% after 15 min, 91% after 12 h, respectively), and "2MB/1MG" (98.6% after 30 min). Moreover, linear and nonlinear adsorption isotherm models confirmed that MB adsorption and CR obeys Freundlich isotherm model. Also, the kinetic studies revealed that the dyes' adsorption on the composite membrane could be clarified using linear and nonlinear forms of pseudo-first order, pseudo-second order, and intra-particle diffusion models. Thermodynamic findings emphasized the endothermic and spontaneous adsorption process as the adsorbent/ adsorbate physical interaction decreased the entropy. Eventually, the recyclability test revealed that the composite matrix exhibits excellent removal efficiency up to 5 recycles. The primary adsorption mechanism was proposed and confirmed with Fourier transform infrared (FTIR). Finally, the unprecedented composite membrane can be considered as an economically distinguished adsorbent for the removal of not only sol dyes but also mixtures of anionic and cationic dyes.

Gut and Gill-Associated Microbiota of the Flatfish European Plaice (Pleuronectes platessa): Diversity, Metabolome and Bioactivity against Human and Aquaculture Pathogens.

Similar to other marine holobionts, fish are colonized by complex microbial communities that promote their health and growth. Fish-associated microbiota is emerging as a promising source of bioactive metabolites. Pleuronectes platessa (European plaice, plaice), a flatfish with commercial importance, is common in the Baltic Sea. Here we used a culture-dependent survey followed by molecular identification to identify microbiota associated with the gills and the gastrointestinal tract (GIT) of P. platessa, then profiled their antimicrobial activity and metabolome. Altogether, 66 strains (59 bacteria and 7 fungi) were isolated, with Proteobacteria being the most abundant phylum. Gill-associated microbiota accounted for higher number of isolates and was dominated by the Proteobacteria (family Moraxellaceae) and Actinobacteria (family Nocardiaceae), whereas Gram-negative bacterial families Vibrionaceae and Shewanellaceae represented the largest group associated with the GIT. The EtOAc extracts of the solid and liquid media cultures of 21 bacteria and 2 fungi representing the diversity of cultivable plaice-associated microbiota was profiled for their antimicrobial activity against three fish pathogens, human bacterial pathogen panel (ESKAPE) and two human fungal pathogens. More than half of all tested microorganisms, particularly those originating from the GIT epithelium, exhibited antagonistic effect against fish pathogens (Lactococcus garvieae, Vibrio ichthyoenteri) and/or human pathogens (Enterococcus faecium, methicillin-resistant Staphylococcus aureus). Proteobacteria represented the most active isolates. Notably, the solid media extracts displayed higher activity against fish pathogens, while liquid culture extracts were more active against human pathogens. Untargeted metabolomics approach using feature-based molecular networking showed the high chemical diversity of the liquid extracts that contained undescribed clusters. This study highlights plaice-associated microbiota as a potential source of antimicrobials for the control of human and the aquaculture-associated infections. This is the first study reporting diversity, bioactivity and chemical profile of culture-dependent microbiota of plaice.

Development of liquid culture media mimicking the conditions of sinuses and lungs in cystic fibrosis and health.

The respiratory tract is a compartmentalised and heterogenous environment. The nasopharynx and sinuses of the upper airways have distinct properties from the lungs and these differences may shape bacterial adaptation and evolution. Upper airway niches act as early colonisation sites for respiratory bacterial pathogens, including those, such as Pseudomonas aeruginosa, that can go on to establish chronic infection of the lungs in people with cystic fibrosis (CF). Despite the importance of upper airway environments in facilitating early adaptation to host environments, currently available in vitro models for study of respiratory infection in CF focus exclusively on the lungs. Furthermore, animal models, widely used to bridge the gap between in vitro systems and the clinical scenario, do not allow the upper and lower airways to be studied in isolation. We have developed a suite of culture media reproducing key features of the upper and lower airways, for the study of bacterial adaptation and evolution in different respiratory environments. For both upper and lower airway-mimicking media, we have developed formulations that reflect airway conditions in health and those that reflect the altered environment of the CF respiratory tract. Here, we describe the development and validation of these media and their use for study of genetic and phenotypic adaptations in P. aeruginosa during growth under upper or lower airway conditions in health and in CF.

An extended catalogue of ncRNAs in Streptomyces coelicolor reporting abundant tmRNA, RNase-P RNA and RNA fragments derived from pre-ribosomal RNA leader sequences

Streptomyces coelicolor is a model organism for studying streptomycetes. This genus possesses relevant medical and economical roles, because it produces many biologically active metabolites of pharmaceutical interest, including the majority of commercialized antibiotics. In this bioinformatic study, the transcriptome of S. coelicolor has been analyzed to identify novel RNA species and quantify the expression of both annotated and novel transcripts in solid and liquid growth medium cultures at different times. The major characteristics disclosed in this study are: (i) the diffuse antisense transcription; (ii) the great abundance of transfer-messenger RNAs (tmRNA); (iii) the abundance of rnpB transcripts, paramount for the RNase-P complex; and (iv) the presence of abundant fragments derived from pre-ribosomal RNA leader sequences of unknown biological function. Overall, this study extends the catalogue of ncRNAs in S. coelicolor and suggests an important role of non-coding transcription in the regulation of biologically active molecule production.

Test with recombinant tuberculosis allergen (Diaskintest®) in the diagnosis of pulmonary tuberculosis in pulmonological practice

Differential diagnosis of pulmonary tuberculosis and community-acquired pneumonia remains an urgent problem in primary health care despite the improvement of radiological and laboratory diagnostic methods.The aimof study was to evaluate the possibility of using a skin test with recombinant tuberculosis allergen (Diaskintest®, Generium, Russia) for the differential diagnosis of pulmonary tuberculosis and pneumonia.Methods.119 cases of pulmonary tuberculosis first detected in the pulmonology department during differential diagnosis were analyzed. The diagnosis was verified by a liquid media culture in the BACTEC MGIT 960 system (BD, USA). All patients with suspected tuberculosis had a skin test with recombinant tuberculosis allergen upon admission. Statistical analysis was carried out using the statistical software package IBM SPSS Statistics 22.0. The differences were considered statistically significant atp< 0.05.Results.Most patients were of working age and had aggravating social risk factors 49 (41.2%), HIV-infection 75 (63.0%), drug addiction 44 (36.9%), or viral hepatitis C 33 (24.2%). As for the clinical forms, infiltrative and disseminated tuberculosis prevailed. Evaluation of the results of the «fast» methods of the diagnostic minimum showed that a positive result of the test with Diaskintest®was almost three times more likely than the positive result of bacterioscopy of a sputum smear with Ziehl–Neelsen staining.Mycobacterium tuberculosisDNA was detected in sputum using the GeneXpert MTB/RIF cartridge technology in 92.6% patients with a positive result of Diaskintest®skin test and a negative sputum bacterioscopy analysis.Conclusion.Is advisable to use Diaskintest®skin test in patients of a pulmonological hospital in the early stages and confirm the positive result with GeneXpert MTB/RIF to reduce the time for diagnosing tuberculosis and the patient’s bed-days in a non-specialized medical institution.

Molecular identification of non-conventional yeasts and the screening for α-terpineol assimilation

The present research aimed to analyze the potential consumption of a monoterpene compound, α-terpineol, by aroma-producing Yeasts. For this purpose, 23 non-conventional yeasts were selected and identified by molecular tools. Subsequently, a screening test was performed to evaluate their tolerance to α-terpineol, by cultivating them in YM broth, at pH 5.0, for 48 h. A mixture of α-terpineol and ethyl alcohol was added to the broth in concentrations ranging from 2.5 to 10.0 μL/mL (intervals of 2.5). The yeasts that survived any of these concentrations were cultured again, but this time in mineral DP liquid culture medium to assess the consumption of α-terpineol. TLC analysis was conducted to analyze the consumption of α-terpineol. The yeasts studied were identified as Clavispora lusitaniae (n=12), Rhodotorula mucilaginosa (n=8), and Lodderomyces elongisporus (n=3). Furthermore, according to the phylogenetic tree, two of these strains (C. lusitaniae and L. elongisporus) have greater genetic proximity than the yeast R. mucilaginosa. From the 23 yeasts, 6 of them were shown to be tolerant to the concentration of 2.5 μL/mL of α-terpineol. The tolerant strains were identified as C. lusitaniae (n = 1), L. elongisporus (n = 2), and R. mucilaginosa (n = 3). However, the yeast that resisted the highest concentration, R. mucilaginosa, was only 12 hours in all concentrations. It is more likely that the defense mechanism of the yeast was not able to prevent further damage to the membrane as the time in contact with α-terpineol increased. The TLC results showed that the extracts from R. mucilaginosa (CMRP3205) and L. elongisporus (CMRP3192) could be interpreted as potentially promising results of new compounds production by the yeasts.

Tolerance and Cadmium (Cd) Immobilization by Native Bacteria Isolated in Cocoa Soils with Increased Metal Content

Twelve cadmium native bacteria previously isolated in soils of cocoa farms located in the western Colombian Andes (Santander), and tolerant to 2500 µM CdCl2 (120 mg Cd/L), were chosen in order to test their tolerance and Cd immobilization using liquid culture medium (Nutritive broth) at different concentrations of heavy metals. Furthermore, in the greenhouse experiments, the strains Exiguobacterium sp. (11-4A), Klebsiella variicola sp. (18-4B), and Enterobacter sp. (29-4B) were applied in combined treatments using CCN51 cacao genotype seeds grown in soil with different concentrations of Cd. All bacterial strains’ cell morphologies were deformed in TEM pictures, which also identified six strain interactions with biosorption and four strain capacities for bioaccumulation; FT-IR suggested that the amide, carbonyl, hydroxyl, ethyl, and phosphate groups on the bacteria biomass were the main Cd binding sites. In the pot experiments, the concentration of Cd was distributed throughout the cacao plant, but certain degrees of immobilization of Cd can occur in soil to prevent an increase in this level in roots with the presence of Klebsiella sp.

Isolation of Inorganic-Phosphate-Solubilizing Bacteria and Analysis of their Effect on Phosphorus Release from Lagoon Sediments

Internal phosphorus (P) loading from sediments is an important cause of eutrophication, and inorganic-phosphate-solubilizing bacteria (IPB) are known to play a vital role in the process of sediment P release. In this study, 10 IPB strains were isolated from the sediments of a lagoon suffering from macroalgal blooms dominated by Chaetomorpha linum – Rongcheng Swan Lagoon, China – and their taxonomy was determined by 16S rRNA sequence analysis. The phosphate-solubilizing characteristics of the obtained IPB strains were evaluated using a liquid medium, and the ability of IPB strains to dissolve P from sediments and the transformation of inorganic phosphorus (IP) fractions were investigated by inoculating IPB strains into a sediment–water experimental system. The 10 isolated IPB strains belonged to one family, two genera, and four species. All IPB strains in the liquid medium culture dissolved tricalcium phosphate, but the phosphate-solubilizing ability of IPB varied greatly between strains and appeared to be promoted when the strains were grown in mixed-strain cultures. Bacillus megaterium showed the strongest phosphate-solubilizing ability. The IPB inoculation experiment showed a large decrease in the Ca-bound P (HCl-P) concentration of sediments in various IPB inoculation treatments, especially in sediments from the central lake. Our results indicate that, under the action of IPB, there is great potential for the release of HCl-P, which is the primary IP fraction of lagoon sediments, and more attention should be given to the endogenous P pollution in Swan Lagoon Wetland.

Fermentation of soymilk by Lactobacillus acidipiscis isolated from Chinese stinky tofu capable of efficiently biotransforming isoflavone glucosides to dihydrodaidzein and dihydrogenistein.

The soy isoflavone microbial metabolites dihydrodaidzein (DHD), dihydrogenistein (DHG), equol and 5-hydroxy-equol are generally more biologically active than their precursors daidzein and genistein. Bacteria responsible for isoflavone metabolism have been isolated and identified. Fermented soymilk is a potential functional food; however, there are few lactic acid bacteria capable of metabolizing soy isoflavones. A newly isolated Gram-positive facultative anaerobic bacterium, which was named Lactobacillus acidipiscis HAU-FR7, was isolated from the traditional Chinese fermented soy product 'stinky tofu'. Bacterium strain HAU-FR7 can grow under aerobic conditions and can also convert most of the daidzin and genistin in soymilk into DHD and DHG, respectively. The concentrations of DHD and DHG produced were 183 and 134 μmol L-1 , respectively, after fermentation for 24 h. Strain HAU-FR7 does not produce the biogenic amines cadaverine, putrescine, histamine or tyramine, and an antibiotic susceptibility test showed that HAU-FR7 is sensitive to nine of the ten tested antibiotics, except for vancomycin. Moreover, the 1,1-diphenyl-2- picrylhydrazyl free radical scavenging capacity of soymilk fermented with HAU-FR7 was significantly higher than that of unfermented soymilk. A facultative anaerobic lactic acid bacterium, designated Lactobacillus acidipiscis HAU-FR7, is capable of reducing the soy isoflavone glucosides daidzin and genistin in soymilk to DHD and DHG efficiently, even in the presence of atmospheric oxygen. The biotransformation activity of HAU-FR7 grown in soymilk is higher than that in de Man-Rogosa-Sharpe liquid culture medium. © 2022 Society of Chemical Industry.

New tool for Gram-negative and Gram-positive bacterial strains differentiation through GCMS direct identification of exogenous VOC metabolites in headspace vials

In this paper, a chromatographic method for the determination of volatile organic compounds (VOCs) as a tool for Gram-positive and Gram-negative bacterial strains identification was described. Direct Headspace gas chromatography coupled with mass spectrometry (Headspace-GC-MS) has been used for the qualitative determination of VOCs emitted by certain bacterial strains (Salmonella Typhimurium, Escherichia coli, Serratia rubidae, Staphyloccocus aureus, Enterococcus casseliflavus, and Enterococcus faecalis). In the first stage of the study, pathogenic bacteria were cultured in broth-specific liquid culture medium directly (lauryl sulfate broth) into headspace vials. Subsequently, the VOC emissions were analyzed by airborne screening in the headspace vials. Thus, the objective was to analyze all the resulting volatile organic compounds in order to select only those compounds, which can be exclusively associated with specific pathogenic bacteria. Qualitative analysis by Headspace-GC-MS has proven to be a non-invasive, accurate, and fast method for identifying certain bacterial strains, based on VOCs emission. A considerable number of volatile organic compounds have been determined in the headspace air, a significant difference being observed between the VOCs emitted by bacterial cultures compared to the culture medium but also between the types of bacterial cultures themselves. The study presents preliminary results, which prove that the identification of the studied pathogenic bacteria is possible, based on the determination of certain types of VOCs in the headspace air of these cultures. This method can be used successfully for the rapid identification of bacterial culture types compared to classical methods.