Journal of Microbiology

Journal Articles

Inhibition of Virulence Associated Traits by β-Sitosterol Isolated from Hibiscus rosa-sinensis Flowers Against Candida albicans: Mechanistic Insight and Molecular Docking Studies: Pallvi Mohana, Atamjit Singh, Farhana Rashid, Sharabjit Singh, Kirandeep Kaur, Rupali Rana, Preet Mohinder Singh Bedi, Neena Bedi, Rajinder Kaur, Saroj Arora; J. Microbiol. 2024;62(12):1165-1175. Published online November 6, 2024; DOI: https://doi.org/10.1007/s12275-024-00174-5

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Abstract: The emerging drug resistance and lack of safer and more potent antifungal agents make Candida infections another hot topic in the healthcare system. At the same time, the potential of plant products in developing novel antifungal drugs is also in the limelight. Considering these facts, we have investigated the different extracts of the flowers of Hibiscus rosa-sinensis of the Malvaceae family for their antifungal efficacy against five different pathogenic Candida strains. Among the various extracts, the chloroform extract showed the maximum zone of inhibition (26.6 ± 0.5 mm) against the Candida albicans strain. Furthermore, the chloroform fraction was isolated, and a sterol compound was identified as β-sitosterol. Mechanistic studies were conducted to understand the mechanism of action, and the results showed that β-sitosterol has significant antifungal activity and is capable of interrupting biofilm formation and acts by inhibiting ergosterol biosynthesis in Candida albicans cells. Microscopic and molecular docking studies confirmed these findings. Overall, the study validates the antifungal efficacy of Candida albicans due to the presence of β-sitosterol which can act as an effective constituent for antifungal drug development individually or in combination.

Fleagrass (Adenosma buchneroides Bonati) Acts as a Fungicide Against Candida albicans by Damaging Its Cell Wall: Youwei Wu, Hongxia Zhang, Hongjie Chen, Zhizhi Du, Qin Li, Ruirui Wang; J. Microbiol. 2024;62(8):661-670. Published online July 3, 2024; DOI: https://doi.org/10.1007/s12275-024-00146-9

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Abstract: Fleagrass, a herb known for its pleasant aroma, is widely used as a mosquito repellent, antibacterial agent, and for treating colds, reducing swelling, and alleviating pain. The antifungal effects of the essential oils of fleagrass and carvacrol against Candida albicans were investigated by evaluating the growth and the mycelial and biofilm development of C. albicans. Transmission electron microscopy was used to evaluate the integrity of the cell membrane and cell wall of C. albicans. Fleagrass exhibited high fungicidal activity against C. albicans at concentrations of 0.5% v/v (via the Ras1/cAMP/PKA pathway). Furthermore, transmission electron microscopy revealed damage to the cell wall and membrane after treatment with the essential oil, which was further confirmed by the increased levels of β-1,3-glucan and chitin in the cell wall. This study showed that fleagrass exerts good fungicidal and hyphal growth inhibition activity against C. albicans by disrupting its cell wall, and thus, fleagrass may be a potential antifungal drug.

Antimicrobial Efficacy of Allium cepa and Zingiber officinale Against the Milk‑Borne Pathogen Listeria monocytogenes: Abirami Arasu , Nagaram Prabha , Durga Devi , Praveen Kumar Issac , Khaloud Mohammed Alarjani , Dunia A. Al Farraj , Reem A. Aljeidi , Dina S. Hussein , Magesh Mohan , Jehad Zuhair Tayyeb , Ajay Guru , Jesu Arockiaraj; J. Microbiol. 2023;61(11):993-1011. Published online December 4, 2023; DOI: https://doi.org/10.1007/s12275-023-00086-w

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Abstract

Listeria monocytogenes is an important food-borne pathogen that causes listeriosis and has a high case fatality rate despite its low incidence. Medicinal plants and their secondary metabolites have been identified as potential antibacterial substances, serving as replacements for synthetic chemical compounds. The present studies emphasize two significant medicinal plants, Allium cepa and Zingiber officinale, and their efficacy against L. monocytogenes. Firstly, a bacterial isolate was obtained from milk and identified through morphology and biochemical reactions. The species of the isolate were further confirmed through 16S rRNA analysis. Furthermore, polar solvents such as methanol and ethanol were used for the extraction of secondary metabolites from A. cepa and Z. officinale. Crude phytochemical components were identified using phytochemical tests, FTIR, and GC–MS. Moreover, the antibacterial activity of the crude extract and its various concentrations were tested against L. monocytogenes. Among all, A. cepa in methanolic extracts showed significant inhibitory activity. Since, the A. cepa for methanolic crude extract was used to perform autography to assess its bactericidal activity. Subsequently, molecular docking was performed to determine the specific compound inhibition. The docking results revealed that four compounds displayed strong binding affinity with the virulence factor Listeriolysin-O of L. monocytogenes. Based on the above results, it can be concluded that the medicinal plant A. cepa has potential antibacterial effects against L. monocytogenes, particularly targeting its virulence.

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Cultural Perspectives on the Sustainable Use and Added Value of Plant-Based Food Dyes—A Case Study from Bulgaria
Mihail Chervenkov, Teodora Ivanova, Yulia Bosseva, Dessislava Dimitrova
Sustainability.2024; 16(20): 9049. CrossRef

Identification and Functional Analysis of Acyl‑Acyl Carrier Protein Δ9 Desaturase from Nannochloropsis oceanica: Ruigang Yang , Hui Wang , Lingyun Zhu , Lvyun Zhu , Tianzhong Liu , Dongyi Zhang; J. Microbiol. 2023;61(1):95-107. Published online January 31, 2023; DOI: https://doi.org/10.1007/s12275-022-00001-9

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Abstract: The oleaginous marine microalga Nannochloropsis oceanica strain IMET1 has attracted increasing attention as a promising photosynthetic cell factory due to its unique excellent capacity to accumulate large amounts of triacylglycerols and eicosapentaenoic acid. To complete the genomic annotation for genes in the fatty acid biosynthesis pathway of N. oceanica, we conducted the present study to identify a novel candidate gene encoding the archetypical chloroplast stromal acyl-acyl carrier protein Δ9 desaturase. The full-length cDNA was generated using rapid-amplification of cDNA ends, and the structure of the coding region interrupted by four introns was determined. The RT-qPCR results demonstrated the upregulated transcriptional abundance of this gene under nitrogen starvation condition. Fluorescence localization studies using EGFP-fused protein revealed that the translated protein was localized in chloroplast stroma. The catalytic activity of the translated protein was characterized by inducible expression in Escherichia coli and a mutant yeast strain BY4389, indicating its potential desaturated capacity for palmitoyl-ACP (C16:0-ACP) and stearoyl-ACP (C18:0-ACP). Further functional complementation assay using BY4839 on plate demonstrated that the expressed enzyme restored the biosynthesis of oleic acid. These results support the desaturated activity of the expressed protein in chloroplast stroma to fulfill the biosynthesis and accumulation of monounsaturated fatty acids in N. oceanica strain IMET1.

Direct current exerts electricidal and bioelectric effects on Porphyromonas gingivalis biofilms partially via promoting oxidative stress and antibiotic transport: Peihui Zou , Peng Li , Jia Liu , Pei Cao , Qingxian Luan; J. Microbiol. 2022;60(1):70-78. Published online November 26, 2021; DOI: https://doi.org/10.1007/s12275-022-1238-5

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Abstract

Low electric current can inhibit certain microbial biofilms and enhance the efficacy of antimicrobials against them. This study investigated the electricidal and bioelectric effects of direct current (DC) against Porphyromonas gingivalis biofilms as well as the underlying mechanisms. Here, we firstly showed that DC significantly suppressed biofilm formation of P. gingivalis in time- and intensity-dependent manners, and markedly inhibited preformed P. gingivalis biofilms. Moreover, DC enhanced the killing efficacy of metronidazole (MTZ) and amoxicillin with clavulanate potassium (AMC) against the biofilms. Notably, DC-treated biofilms displayed upregulated intracellular ROS and expression of ROS related genes (sod, feoB, and oxyR) as well as porin gene. Interestingly, DC-induced killing of biofilms was partially reversed by ROS scavenger N-dimethylthiourea (DMTU), and the synergistic effect of DC with MTZ/AMC was weakened by small interfering RNA of porin gene (si-Porin). In conclusion, DC can exert electricidal and bioelectric effects against P. gingivalis biofilms partially via promotion of oxidative stress and antibiotic transport, which offers a promising approach for effective management of periodontitis.

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Antifungal Activity, Synergism with Fluconazole or Amphotericin B and Potential Mechanism of Direct Current against Candida albicans Biofilms and Persisters
Peihui Zou, Jia Liu, Peng Li, Qingxian Luan
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Carolina Montoya, Lina Roldan, Michelle Yu, Sara Valliani, Christina Ta, Maobin Yang, Santiago Orrego
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Molecular Study of Porphyromonas Gingivalis Strains With fimA Genotypes in Periodontitis Patients
Zhraa F. Faruq, Sami Khalaf Jabar
Iranian Journal of Medical Microbiology.2023; 17(6): 663. CrossRef
Drug delivery approaches for enhanced antibiofilm therapy
Tao Wang, Erik Jan Cornel, Chang Li, Jianzhong Du
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Weak direct current exerts synergistic effect with antibiotics and reduces the antibiotic resistance: An in vitro subgingival plaque biofilm model
Peihui Zou, Yanfeng Wang, Pei Cao, Peng Li, Jia Liu, Qingxian Luan
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Bioelectric device for effective biofilm inflammation management of dental implants
Jihyun Lee, Young Wook Kim
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A Highly Efficacious Electrical Biofilm Treatment System for Combating Chronic Wound Bacterial Infections
Fan Zhao, Yajuan Su, Junying Wang, Svetlana Romanova, Dominick J. DiMaio, Jingwei Xie, Siwei Zhao
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Biofilm characterization of Fusarium solani keratitis isolate: increased resistance to antifungals and UV light: Itzel Margarita Córdova-Alcántara , Diana Laura Venegas-Cortés , María Ángeles Martínez-Rivera , Néstor Octavio Pérez , Aida Verónica Rodriguez-Tovar; J. Microbiol. 2019;57(6):485-497. Published online May 27, 2019; DOI: https://doi.org/10.1007/s12275-019-8637-2

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Abstract

Fusarium solani has drawn phytopathogenic, biotechnological, and medical interest. In humans, it is associated with localized infections, such as onychomycosis and keratomycosis, as well as invasive infections in immunocompromised patients. One pathogenicity factor of filamentous fungi is biofilm formation. There is still only scarce information about the in vitro mechanism of the formation and composition of F. solani biofilm. In this work, we describe the biofilm formed by a clinical keratomycosis isolate in terms of its development, composition and susceptibility to different antifungals and ultraviolet light (UV) at different biofilm formation stages. We found five biofilm formation stages using scanning electron microscopy: adherence, germination, hyphal development, maturation, and cell detachment. Using epifluorescence microscopy with specific fluorochromes, it was elucidated that the extracellular matrix consists of carbohydrates, proteins, and extracellular DNA. Specific inhibitors for these molecules showed significant biofilm reductions. The antifungal susceptibility against natamycin, voriconazole, caspofungin, and amphotericin B was evaluated by metabolic activity and crystal violet assay, with the F. solani biofilm preformation to 24 h increased in resistance to natamycin, voriconazole, and caspofungin, while the biofilm preformation to 48 h increased in resistance to amphotericin B. The preformed biofilm at 24 h protected and reduced UV light mortality. F. solani isolate could produce a highly structured extra biofilm; its cellular matrix consists of carbohydrate polymers, proteins, and eDNA. Biofilm confers antifungal resistance and decreases its susceptibility to UV light. The fungal biofilm functions as a survival strategy against antifungals and environmental factors.

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Antifungal activity of 3-acetylbenzamide produced by actinomycete WA23-4-4 from the intestinal tract of Periplaneta americana: Xia Fang , Juan Shen , Jie Wang , Zhi-li Chen , Pei-bin lin , Zhi-yu Chen , Lin-yan Liu , Huan-xiong Zeng , Xiao-bao Jin; J. Microbiol. 2018;56(7):516-523. Published online June 28, 2018; DOI: https://doi.org/10.1007/s12275-018-7510-z

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Abstract

Actinomycetes are well-known for producing numerous bioactive secondary metabolites. In this study, primary screening by antifungal activity assay found one actinomycete strain WA23-4-4 isolated from the intestinal tract of Periplaneta americana that exhibited broad spectrum antifungal activity. 16S rDNA gene analysis of strain WA23-4-4 revealed close similarity to Streptomyces nogalater (AB045886) with 86.6% sequence similarity. Strain WA23-4-4 was considered as a novel Streptomyces and the 16s rDNA sequence has been submitted to GenBank (accession no. KX291006). The maximum antifungal activity of WA23-4-4 was achieved when culture conditions were optimized to pH 8.0, with 12% inoculum concentration and 210 ml ISP2 medium, which remained stable between the 5th and the 9th day. 3-Acetyl benzoyl amide was isolated by ethyl acetate extraction of WA23- 4-4 fermentation broth, and its molecular formula was determined as C9H9NO2 based on MS, IR, 1H, and 13C NMR analyses. The compound showed significant antifungal activity against Candida albicans ATCC 10231 (MIC: 31.25 μg/ml) and Aspergillus niger ATCC 16404 (MIC: 31.25 μg/ml). However, the compound had higher MIC values against Trichophyton rubrum ATCC 60836 (MIC: 500 μg/ml) and Aspergillus fumigatus ATCC 96918 (MIC: 1,000 μg/ml). SEM analysis showed damage to the cell membrane of Candida albicans ATCC 10231 and to the mycelium of Aspergillus niger ATCC 16404 after being treatment with 3-acetyl benzoyl amide. In conclusion, this is the first time that 3-acetyl benzoyl amide has been identified from an actinomycete and this compound exhibited antifungal activity against Candida albicans ATCC 10231 and Aspergillus niger ATCC 16404.

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An in vitro study of the antifungal activity of Trichoderma virens 7b and a profile of its non-polar antifungal components released against Ganoderma boninense: Lee Pei Lee Angel , Mohd Termizi Yusof , Intan Safinar Ismail , Bonnie Tay Yen Ping , Intan Nur Ainni Mohamed Azni , Norman Hj Kamarudin , Shamala Sundram; J. Microbiol. 2016;54(11):732-744. Published online October 29, 2016; DOI: https://doi.org/10.1007/s12275-016-6304-4

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Abstract

Ganoderma boninense is the causal agent of a devastating disease affecting oil palm in Southeast Asian countries. Basal stem rot (BSR) disease slowly rots the base of palms, which radically reduces productive lifespan of this lucrative crop. Previous reports have indicated the successful use of Trichoderma as biological control agent (BCA) against G. boninense and isolate T. virens 7b was selected based on its initial screening. This study attempts to decipher the mechanisms responsible for the inhibition of G. boninense by identifying and characterizing the chemical compounds as well as the physical mechanisms by T. virens 7b. Hexane extract of the isolate gave 62.60% ± 6.41 inhibition against G. boninense and observation under scanning electron microscope (SEM) detected severe mycelial deformation of the pathogen at the region of inhibition. Similar mycelia deformation of G. boninense was observed with a fungicide treatment, Benlate® indicating comparable fungicidal effect by T. virens 7b. Fraction 4 and 5 of hexane active fractions through preparative thin layer chromatography (P-TLC) was identified giving the best inhibition of the pathogen. These fractions comprised of ketones, alcohols, aldehydes, lactones, sesquiterpenes, monoterpenes, sulphides, and free fatty acids profiled through gas chromatography mass spectrometry detector (GC/MSD). A novel antifungal compound discovery of phenylethyl alcohol (PEA) by T. virens 7b is reported through this study. T. virens 7b also proved to be an active siderophore producer through chrome azurol S (CAS) agar assay. The study demonstrated the possible mechanisms involved and responsible in the successful inhibition of G. boninense.

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Review

Minireview] The molecular mechanism of azole resistance in Aspergillus fumigatus: from bedside to bench and back: Xiaolei Wei , Yuanwei Zhang Zhang , Ling Lu; J. Microbiol. 2015;53(2):91-99. Published online January 28, 2015; DOI: https://doi.org/10.1007/s12275-015-5014-7

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Abstract

The growing use of immunosuppressive therapies has resulted in a dramatic increased incidence of invasive fungal infections (IFIs) caused by Aspergillus fumigatus, a common pathogen, and is also associated with a high mortality rate. Azoles are the primary guideline-recommended therapy agents for first-line treatment and prevention of IFIs. However, increased azole usage in medicinal and agricultural settings has caused azole-resistant isolates to repeatedly emerge in the environment, resulting in a significant threat to human health. In this review, we present and summarize current research on the resistance mechanisms of azoles in A. fumigatus as well as efficient susceptibility testing methods. Moreover, we analyze and discuss the putative clinical (bedside) indication of these findings from bench work.

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Journal Articles

Optimization of Antifungal Lipopeptide Production from Bacillus sp. BH072 by Response Surface Methodology: Xin Zhao , Ye Han , Xi-qian Tan , Jin Wang , Zhi-jiang Zhou; J. Microbiol. 2014;52(4):324-332. Published online February 17, 2014; DOI: https://doi.org/10.1007/s12275-014-3354-3

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Abstract

Antifungal lipopeptide produced by Bacillus sp. BH072 was extracted from fermentation liquor and determined as iturin A by liquid chromatography-mass spectrometry (LC-MS). For industrial-scale production, the yield of iturin A was improved by optimizing medium components and fermentation conditions. A one-factor test was conducted; fermentation conditions were then optimized by response surface methodology (RSM) to obtain the following: temperature, 29.5°C; pH 6.45; inoculation quantity, 6.7%; loading volume, 100 ml (in 500 ml flasks); and rotary speed, 150 rpm. Under these conditions, the mass concentration of iturin A was increased from 45.30 mg/ml to 47.87 mg/ml. The following components of the medium were determined: carbon sources (glucose, fructose, sucrose, xylose, rhamnose, and soluble starch); nitrogen sources (peptone, soybean meal, NH4Cl, urea, and ammonium citrate); and metal ions (Zn2+, Fe3+, Mg2+, Mn2+, Ca2+, and K+). The effects of these components on iturin A production were observed in LB medium. We selected sucrose, soybean meal, and Mg2+ for RSM to optimize the conditions because of several advantages, including maximum iturin A production, high antifungal activity, and low cost. The optimum concentrations of these components were 0.98% sucrose, 0.94% soybean meal, and 0.93% Mg2+. After iturin A production was optimized by RSM, the mass concentration reached 52.21 mg/ml. The antifungal specific activity was enhanced from 350.11 AU/mg to 513.92 AU/mg, which was 46.8% higher than the previous result. The present study provides an important experimental basis for the industrial-scale production of iturin A and the agricultural applications of Bacillus sp. BH072.

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Note] Antifungal Chitinase against Human Pathogenic Yeasts from Coprinellus congregatus: Yeeun Yoo Hyoung T. Choi; J. Microbiol. 2014;52(5):441-443. Published online February 17, 2014; DOI: https://doi.org/10.1007/s12275-014-3257-3

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Abstract

The inky cap, Coprinellus congregatus, produces mushrooms which become autolyzed rapidly to generate black liquid droplets, in which no cell wall is detected by microscopy. A chitinase (Chi2) which is synthesized during the autolytic phase of C. congregatus inhibits the growths of Candida al-bicans and Cryptococcus neoformans up to 10% at the con-centration of 10 μg/ml, about 50% at concentration of 20 μg/ml, and up to 95% at the concentration of 70 μg/ml. Upon treatment these yeast cells are observed to be severely de-formed, with the formation of large holes in the cell wall. The two yeast species show no growth inhibition at the concen-tration of 5 μg/ml, which means the minimum inhibitory concentrations for both yeast species are 10 μg/ml under these experimental conditions.

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Research Support, Non-U.S. Gov'ts

NOTE] Construction and Characterisation of an Antifungal Recombinant Bacillus thuringiensis with an Expanded Host Spectrum: Qin Liu , Jong Yul Roh , Yong Wang , Jae Young Choi , Xue Ying Tao , Jae Su Kim , Yeon Ho Je; J. Microbiol. 2012;50(5):874-877. Published online November 4, 2012; DOI: https://doi.org/10.1007/s12275-012-2201-7

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Abstract: A novel antifungal Bacillus thuringiensis strain 19–22, ssp. kurstaki (H3a3b3c), was characterised. This strain included cry1Aa, cry1Ab, cry1Ac, and cry1D, which have high insecticidal activities against lepidopteran larvae other than Spodoptera exigua. To expand the host spectrum, a cry1E gene whose product is active against S. exigua was introduced into the isolate. The transformant successfully expressed the Cry1E protein without any loss of its original antifungal activities. These results indicate that this recombinant strain exhibits dual activities and may be used as an integrated control agent to control plant diseases and insect pests.

Antifungal Activity of Leuconostoc citreum and Weissella confusa in Rice Cakes: Eunjong Baek , Hyojin Kim , Hyejung Choi , Sun Yoon , Jeongho Kim; J. Microbiol. 2012;50(5):842-848. Published online November 4, 2012; DOI: https://doi.org/10.1007/s12275-012-2153-y

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Abstract: The antifungal activity of organic acids greatly improves the shelf life of bread and bakery products. However, little is known about the effect of lactic acid fermentation on fungal contamination in rice cakes. Here, we show that lactic acid fermentation in rice dough can greatly retard the growth of three fungal species when present in rice cakes, namely Cladosporium sp. YS1, Neurospora sp. YS3, and Penicillium crustosum YS2. The antifungal activity of the lactic acid bacteria against these fungi was much better than that of 0.3% calcium propionate. We found that organic acids including lactic and acetic acid, which are byproducts of lactic fermentation or can be artificially added, were the main antifungal substances. We also found that some Leuconostoc citreum and Weissella confusa strains could be good starter species for rice dough fermentation. These results imply that these lactic acid bacteria can be applicable to improve the preservation of rice cakes.

NOTE] Antifungal Activity of Extracellular Hydrolases Produced by Autolysing Aspergillus nidulans Cultures: Melinda Szilágyi , Fruzsina Anton , Katalin Forgács , Jae-Hyuk Yu , István Pócsi , Tamás Emri; J. Microbiol. 2012;50(5):849-854. Published online November 4, 2012; DOI: https://doi.org/10.1007/s12275-012-2001-0

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Abstract: Carbon-starving Aspergillus nidulans cultures produce high activities of versatile hydrolytic enzymes and, among these, ChiB endochitinase and EngA β-1,3-endoglucanase showed significant antifungal activity against various fungal species. Double deletion of engA and chiB diminished the antifungal activity of the fermentation broths and increased conidiogenesis and long-term viability of A. nidulans, but decreased the growth rate on culture media containing weak carbon sources. Production of ChiB and EngA can influence fungal communities either directly due to their antifungal properties or indirectly through their effects on vegetative growth. Our data suggest saprophytic fungi as promising future candidates to develop novel biocontrol technologies.

Antifungal Activity and Mechanism of Fengycin in the Presence and Absence of Commercial Surfactin Against Rhizopus stolonifer: Yang Tao , Xiao-mei Bie , Feng-xia Lv , Hai-zhen Zhao , Zhao-xin Lu; J. Microbiol. 2011;49(1):146-150. Published online March 3, 2011; DOI: https://doi.org/10.1007/s12275-011-0171-9

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Abstract: The antifungal activity and mechanism of fengycin in the presence and absence of commercial surfactin against Rhizopus stolonifer were investigated. The MIC (minimal inhibitory concentration) of fengycin without commercial surfactin added was 0.4 mg/ml while the MIC of fengycin with commercial surfactin added was 2.0 mg/ml. Fengycin acted on cell membrane and cellular organs and inhibited DNA synthesis. The antifungal effect of fengycin was reduced after commercial surfactin was added. All these results suggest that the fungal cell membrane may be the primary target of fengycin action and commercial surfactin may reduce the antifungal activity of fengycin.

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