research & reviews journal of microbiology and biotechnology impact factor

Research & Reviews: Journal of Microbiology and Biotechnology

E- ISSN: 2320 - 3528 P- ISSN: 2347 - 2286

• +447389646377
• Journal h-index : 14
• Journal cite score : 4.50
• Journal impact factor : 2.8
• Average acceptance to publication time (5-7 days)
• Average article processing time (30 - 45 days) Less than 5 volumes 30 days 8 - 9 volumes 40 days 10 and more volumes 45 days

Welcome to the Journal

Index Copernicus Value: 131.69,  PubMed NLM ID: 101707674

Journal of Microbiology and Biotechnology (JMB) is a peer reviewed, international open access journal that makes significant contributions in the field of Microbiology and Biotechnology such as Antibiotics, Antifungals, Antiviral compounds, Bioactive Compounds, Functional Foods Biodegradation, Bioremediation Cell Culture, Biomedical Engineering, Cellular Nutrition in Health, Disease Fermentation, Food Technology, Foodborne Pathogens, Food Safety, Functional Genomics, Systems Biology, Host-Microbe Interactions, Pathogenesis Infection, Immunity Microbial Ecology, Diversity Microbial Genetics, Physiology, Metabolism Microbial Genomes, Metagenomics, Microbiomes, Plant Microbiology, Protein Engineering, Evolution Synthetic Biology, Metabolic Engineering, Whole Cell Biocatalysis, Bioprocess Engineering, Molecular Biology and Omics, Microbial Cell Biology, Bioactive Compounds, Chemical Biology, Biocatalysis, Fermentation Technology, Food Microbiology and Biotechnology, Soil Microbiology, Animal Biotechnology, Plant Biotechnology, Bioprocess and Metabolic Engineering, Environmental Microbiology and Engineering, Clinical Microbiology, Immunology and Virology, and Molecular Genetics, Medical Microbiology.

Journal of Microbiology and Biotechnology (JMB) is an open access journal which means that all content is freely available without charge to the user or his/her institution. Users are allowed to read, download, copy, distribute, print, search, or link to the full texts of the articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author.

Archived and indexed in popular scientific databases like PubMed, the journal accepts research articles, review articles, commentaries, letters to the editor, case reports and short communications. The journal adopts single blind peer reviewing to ensure quality and uses Editorial tracking system for manuscript submission, review and tracking its status. Review process is performed by the editorial board members of Journal of Microbiology and Biotechnology & outside experts. Two independent reviewers approval followed by editor's approval is required for acceptance of any citable manuscript. manuscripts as an e-mail attachment to the Editorial Office at [email protected] or online at https://www.scholarscentral.org/submissions/research-reviews-microbiology-biotechnology.html

Fast Editorial Execution and Review Process (FEE-Review Process):

Journal of Microbiology and Biotechnology is participating in the Fast Editorial Execution and Review Process (FEE-Review Process) with an additional prepayment of $99 apart from the regular article processing fee. Fast Editorial Execution and Review Process is a special service for the article that enables it to get a faster response in the pre-review stage from the handling editor as well as a review from the reviewer. An author can get a faster response of pre-review maximum in 3 days since submission, and a review process by the reviewer maximum in 5 days, followed by revision/publication in 2 days. If the article gets notified for revision by the handling editor, then it will take another 5 days for external review by the previous reviewer or alternative reviewer.

Acceptance of manuscripts is driven entirely by handling editorial team considerations and independent peer-review, ensuring the highest standards are maintained no matter the route to regular peer-reviewed publication or a fast editorial review process. The handling editor and the article contributor are responsible for adhering to scientific standards. The article FEE-Review process of $99 will not be refunded even if the article is rejected or withdrawn for publication.

The corresponding author or institution/organization is responsible for making the manuscript FEE-Review Process payment. The additional FEE-Review Process payment covers the fast review processing and quick editorial decisions, and regular article publication covers the preparation in various formats for online publication, securing full-text inclusion in a number of permanent archives like HTML, XML, and PDF, and feeding to different indexing agencies.

Respiratory microbiomes

Epigenetics.

Epigenetics is the study of how your behaviors and environment can cause changes that affect the way your genes work. Unlike genetic changes, epigenetic changes are reversible and do not change your DNA sequence, but they can change how your body reads a DNA sequence.

Genomics is an interdisciplinary field of biology focusing on the structure, function, evolution, mapping, and editing of genomes. A genome is an organism's complete set of DNA, including all of its genes as well as its hierarchical, three-dimensional structural configuration. In contrast to genetics, which refers to the study of individual genes and their roles in inheritance, genomics aims at the collective characterization and quantification of all of an organism's genes, their interrelations and influence on the organism. Genes may direct the production of proteins with the assistance of enzymes and messenger molecules. In turn, proteins make up body structures such as organs and tissues as well as control chemical reactions and carry signals between cells. Genomics also involves the sequencing and analysis of genomes through uses of high throughput DNA sequencing and bioinformatics to assemble and analyze the function and structure of entire genomes. Advances in genomics have triggered a revolution in discovery-based research and systems biology to facilitate understanding of even the most complex biological systems such as the brain.

Bacteriology

Bacteriology is the branch and specialty of biology that studies the morphology, ecology, genetics and biochemistry of bacteria as well as many other aspects related to them. This subdivision of microbiology involves the identification, classification, and characterization of bacterial species.

Virology will feature articles on human, animal, plant, insect, bacterial, and fungal viruses. The journal will also publish articles on molecular aspects of the control and prevention of viral infections with vaccines and antiviral agents and on the use of viruses as gene therapy vectors, as well as research on other agents such as prions. The approaches and techniques used are expected to encompass many disciplines, including molecular genetics, molecular biology, biochemistry, biophysics, structural biology, cell biology, immunology, morphology, genetics and pathogenesis.

Parasitology

Articles covering host-parasite relationships and parasitic diseases will be considered, as well as studies on disease vectors. Articles highlighting social and economic issues around the impact of parasites are also encouraged. As an international, Open Access publication, Journal of Parasitology Research aims to foster learning and collaboration between countries and communities.

Infectious Diseases & Therapy

Several topics include.

Antibiotics, Antifungals, and Antiviral compounds Bioactive Compounds and Functional Foods Biodegradation and Bioremediation Cell Culture and Biomedical Engineering Cellular Nutrition in Health and Disease Fermentation and Food Technology Foodborne Pathogens and Food Safety Functional Genomics and Systems Biology Host-Microbe Interactions and Pathogenesis Infection and Immunity Microbial Ecology and Diversity Microbial Genetics, Physiology, and Metabolism Microbial Genomes and Metagenomics Microbiome Plant Microbiology Protein Engineering and Evolution Synthetic Biology and Metabolic Engineering Whole Cell Biocatalysis and Bioprocess Engineering

Bioengineering

Bionics and biological cybernetics: implantology; bio–abio interfaces Bioelectronics: wearable electronics; implantable electronics; “more than Moore” electronics; bioelectronics devices Bioprocess and biosystems engineering and applications: bioprocess design; biocatalysis; bioseparation and bioreactors; bioinformatics; bioenergy; etc. Biomolecular, cellular and tissue engineering and applications: tissue engineering; chromosome engineering; embryo engineering; cellular, molecular and synthetic biology; metabolic engineering; bio-nanotechnology; micro/nano technologies; genetic engineering; transgenic technology Biomedical engineering and applications: biomechatronics; biomedical electronics; biomechanics; biomaterials; biomimetics; biomedical diagnostics; biomedical therapy; biomedical devices; sensors and circuits; biomedical imaging and medical information systems; implants and regenerative medicine; neurotechnology; clinical engineering; rehabilitation engineering Biochemical engineering and applications: metabolic pathway engineering; modeling and simulation Translational bioengineering

The scope of the journal encompasses studies of microbiomes colonizing humans, animals, plants or the environment, both built and natural or manipulated, as in agriculture. Studies on the development and application of meta-omics approaches or novel bioinformatics tools, on community/host interaction with emphasis on structure-function relationship that would lead to substantial advances in the field will be considered for publication. Microbiome is especially interested in studies that go beyond descriptive omics surveys and include experimental or theoretical approaches that mechanistically support proposed microbiome functions, and establish, if possible, cause and effect relationships. Studies of individual microbial isolates/species in vivo or in laboratory cultures without exploring the mechanisms by which they affect the host or the complex microbiome structures and functions will not be considered. Through this collection of literature Microbiome hopes to integrate researchers with common scientific objectives across a broad cross-section of sub-disciplines within microbial ecology.

Bacterial virulence

Bacterial virulence is the ability of the bacteria to produce disease. The virulence of the microorganisms is measured as severity of disease. The method followed bacteria to cause disease is Adhesion, Colonization, Invasion, Toxins. This results when the balance disturbs between bacterial virulence and host resistance.

Related Journals of Bacterial Virulence

International Journal of Microbiology, Journal of Bacteriology, Bacteriological Reviews, Bacteriophage

Biomolecule

Biomolecule is a molecule that is present in living organisms. There are four classes of biomolecule, each have their own characteristics monomers like fatty acids, monosaccharide, amino acids and nucleotides and corresponding polymers like polysaccharide, nucleic acid and polypeptide.

Related Journals of Biomolecule

Biomolecule and Therapuetics, Biomolecules

Animal biology

Animal biology is related to zoology which deals with animal kingdom, their interaction with ecosystem and it also deals with the study of embryology, classification, structure, physiological, Evolutionary, Classification, Ethology, Biogeography, Invertebrate zoology, Vertebrate zoology and Zoography.

Related Journals of Animal Biology

Journals of Microbiology, Journal of Animal Science and Technology, Laboratory Animal Research, The Journal of Venomous Animals and Toxins Including Tropical Diseases, Experimental Animals, Veterinary Microbiology Journals, Research in Microbiology.

Plant biology

Plant biology is related to botany which deals with plant science. It deals with the study about plant structure, classification in plant kingdom, structure, growth, reproduction, metabolism, chemical products, diseases, evolutionary relationships and plant taxonomy. Few branches of Plant biology are Horticulture, Mycology, Phycology, Plant morphology, and Plant systematics.

Related Journals of Plant Biology

Applications in Plant Sciences, BMC Plant Biology, Frontiers in Plant Science, International Journal of Plant Genomics, Molecular Plant, The Plant Cell, Plant and Cell Physiology

Microbial Cell Biology

Cellular world is divided into two types, eukaryotes and prokaryotes. All microorganisms are prokaryotes. There are different shapes of cells like rod shape, spherical and curved. The size of cell is very small which ranges from 0.2 µm and 700 µm in diameter.

Related Journals of Microbial Cell Biology

Microbial Drug Resistance, Microbial Biotechnology, Microbial Informatics and Experimentation, Microbiology Journal Articles, Microbial Cell Factories, Microbiological Research Journals, Microbial Ecology in Health and Disease, Journal of Microbiology, Microbiology Journals, Medical Microbiology Journals, International Journal of Microbiology

Bioactive Compounds

A Bioactive compound can cause an effect in living organism. Bioactive compounds are not essential to human since the body can function properly without them. These are found in both plants and animals. These are used to prevent cancer and heart diseases. Bioactive compounds are experiencing wide range of applications like geo-medicine, food industry, Nano-biosciences and many more.

Related Journals of Bioactive Compounds

Journal of Bioactive and Compatible Polymers, Current Bioactive Compounds, Open Bioactive Compounds Journal, Bioactive Carbohydrates and Dietary Fibre

Microbial Ecology

Microbial Ecology is also called as environmental microbiology study about ecology of microorganism relating with one another. They play primary role in regulating biogeochemical systems in ecology. Besides these includes nitrogen fixation, sulfur metabolism, methane metabolism and carbon fixation.

Related Journals of Microbial Ecology

FEMS Microbiology Ecology, Infection Ecology & Epidemiology, Microbial Ecology in Health and Disease, BMC Ecology, Journal of Medical Microbiology, Infectious Diseases in Obstetrics and Gynecology, Microbiological Journals, Journals of Microbiology, International Journal of Microbiology Research.

Biotechnology Research

Biotechnology research is the field use of living organism and living system by applying advance engineering principles to develop the product or modify it. The research of biotechnology includes in Animal Biology, Computational Biology, Environmental Biotechnology, Human Biology, Industrial Biotechnology, Medical Biology and Plant Biology.

Related Journals of BiotechnologyResearch

BMC Biotechnology, Biotechnology Research International, Industrial Biotechnology, ISRN biotechnology, Industrial Biotechnology, Molecular Biotechnology, Agricultural Biotechnology, Whole Journal of Biotechnology, Asian Journal of Biotechnology, Research Journal of Biotechnology, Latest Research Journal of Biotechnology, Current Research Journal of Biotechnology, Biotechnology Research, Biotechnology Articles, Journal of Microbiology and Biotechnology, Biotechnology Journal Articles, Asian Journal of Microbiology and Biotechnology, World Journal of Microbiology and Biotechnology, Applied Journal of Microbiology and Biotechnology, Journal of Microbiology, Biotechnology Food Sciences, British Biotechnology.

Biological Control Agents

Biological Control Agents are used for invasion of plant's natural enemies. This action occurs naturally. Biological control of weeds includes insects and pathogens. Bio control agents include a wide variety of life forms, including vertebrates, invertebrates, fungi, and microorganisms. Biological control agents of plant diseases are most often referred to as antagonists.

Related Journals of Biological Control Agents

Journal of BIOLOGICAL REGULATORS & Homeostatic Agents, Biological agents peer reviewed journals, Journal of Biological Agents & Warfare, Biological warfare agents

*2023 Journal impact factor was established by dividing the number of articles published in 2021 and 2022 with the number of times they are cited in 2023 based on Google Scholar Citation Index database. If 'X' is the total number of articles published in 2021 and 2022, and 'Y' is the number of times these articles were cited in indexed journals during 2023 then, impact factor = Y/X

Articles published in Research & Reviews: Journal of Microbiology and Biotechnology have been cited by esteemed scholars and scientists all around the world. Research & Reviews: Journal of Microbiology and Biotechnology has got h-index 14 , which means every article in Research & Reviews: Journal of Microbiology and Biotechnology has got 14 average citations.

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Journal highlights, google scholar citation report, citations : 705.

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Identifiers

Linking ISSN (ISSN-L): 2320-3528

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Resource information

Title proper: Research and reviews: journal of microbiology and biotechnology.

Country: India

Medium: Online

Record information

Last modification date: 09/05/2023

Type of record: Confirmed

ISSN Center responsible of the record: ISSN National Centre for India

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A Narrative Review on the Advance of Probiotics to Metabiotics

Gromwell ( Lithospermum erythrorhizon ) Attenuates High-Fat-Induced Skeletal Muscle Wasting by Increasing Protein Synthesis and Mitochondrial Biogenesis

Establishment and Characterization of Immortalized Human Dermal Papilla Cells Expressing Human Papillomavirus 16 E6/E7

Molecular and Phenotypic Investigation on Antibacterial Activities of Limonene Isomers and Its Oxidation Derivative against Xanthomonas oryzae pv. oryzae

Nodulation Experiment by Cross-Inoculation of Nitrogen-Fixing Bacteria Isolated from Root Nodules of Several Leguminous Plants

Inhibitory Effects of Latilactobacillus curvatus BYB3 Cell-Free Extract on Human Melanoma B16F10 Cells and Tumorigenic Mice

Anthocyanins Inhibits Oxidative Injury in Human Retinal Pigment Epithelial ARPE-19 Cells via Activating Heme Oxygenase-1

Nasal Immunization Using Chitosan Nanoparticles with Glycoprotein B of Murine Cytomegalovirus

Coffee Husk By-Product as Novel Ingredients for Cascara Kombucha Production

Aims & Scope

Molecular and Cellular Microbiology (MCM)

Environmental Microbiology and Biotechnology (EMB)

Food Microbiology and Biotechnology (FMB)

Biotechnology and Bioengineering (BB)

pISSN 1017-7825 eISSN 1738-8872

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Most Influential Articles

Current Status of Epidemiology, Diagnosis, Therapeutics, and Vaccines for Novel Coronavirus Disease 2019 (COVID-19)

Abstract : Kombucha, a fermented beverage, is gaining popularity due to its numerous beneficial health effects. Various substrates such as herbs, fruits, flowers, and vegetables, have been used for kombucha fermentation in order to enhance the flavor, aroma, and nutritional composition. This study aims to investigate the potential suitability of cascara as a novel ingredient for kombucha production. Our findings suggested that cascara is a suitable substrate for kombucha production. Fermentation elevated the total phenolic and flavonoid content in cascara, which enhanced the antioxidant, antibacterial, and prebiotic characteristics of the product. Furthermore, the accumulation of acetic acid-induced the pH lowering reached 2.7 after 14 days of fermentation, which achieved the microbiological safety of the product. Moreover, 14 days of fermentation resulted in a balanced amalgamation of acidity, sweetness, and fragrance according to sensory evaluation. Our findings not only highlight the potential of cascara kombucha as a novel substrate for kombucha production but also contribute to repurposing coffee by-products, promoting environmentally friendly and sustainable agricultural development.

Dae-Gyun Ahn 1 , Hye-Jin Shin 1 , Mi-Hwa Kim 1, 2 , Sunhee Lee 1 , Hae-Soo Kim 1 , Jinjong Myoung 3 , Bum-Tae Kim 1* and Seong-Jun Kim 1*

2020; 30(3): 313-324 https://doi.org/10.4014/jmb.2003.03011

Deciphering Diversity Indices for a Better Understanding of Microbial Communities

Bo-Ra Kim 1 , Jiwon Shin 1 , Robin B. Guevarra 1 , Jun Hyung Lee 1 , Doo Wan Kim 2 , Kuk-Hwan Seol 2 , Ju-Hoon Lee 3 , Hyeun Bum Kim 1* and Richard E. Isaacson 4

2017; 27(12): 2089-2093 https://doi.org/10.4014/jmb.1709.09027

Advances in Rapid Detection Methods for Foodborne Pathogens

Xihong Zhao 1, 2, 3 , Chii-Wann Lin 3 , Jun Wang 2 and Deog Hwan Oh 2*

2014; 24(3): 297-312 https://doi.org/10.4014/jmb.1310.10013

A Brief Overview of Escherichia coli O157:H7 and Its Plasmid O157

Ji Youn Lim 1 , JangWon Yoon 2 and Carolyn J. Hovde 1*

2010; 20(1): 5-14 https://doi.org/10.4014/jmb.0908.08007

Isolation, Characterization, and Use for Plant Growth Promotion Under Salt Stress, of ACC Deaminase-Producing Halotolerant Bacteria Derived from Coastal Soil

Md. Ashaduzzaman Siddikee 1 , Puneet. S. Chauhan 1 , R. Anandham 2 , Gwang-Hyun Han 1 and Tongmin Sa 1*

2010; 20(11): 1577-1584 https://doi.org/10.4014/jmb.1007.07011

Production of Biosurfactant Lipopeptides Iturin A, Fengycin and Surfactin A from Bacillus subtilis CMB32 for Control of Colletotrichum gloeosporioides

Pyoung Il Kim 1 , Jaewon Ryu 2 , Young Hwan Kim 3 and Youn-Tae Chi 4*

2010; 20(1): 138-145 https://doi.org/10.4014/jmb.0905.05007

Extremozymes: A Potential Source for Industrial Applications

Kelly Dumorné 1* , David Camacho Córdova 2 , Marcia Astorga-Eló 3 and Prabhaharan Renganathan 4

2017; 27(4): 649-659 https://doi.org/10.4014/jmb.1611.11006

Role of Probiotics in Human Gut Microbiome-Associated Diseases

Seon-Kyun Kim 1 , Robin B. Guevarra 2 , You-Tae Kim 1 , Joongi Kwon 1 , Hyeri Kim 2 , Jae Hyoung Cho 2 , Hyeun Bum Kim 2* and Ju-Hoon Lee 1*

2019; 29(9): 1335-1340 https://doi.org/10.4014/jmb.1906.06064

Disruption of Established Bacterial and Fungal Biofilms by a Blend of Enzymes and Botanical Extracts

Gitte S. Jensen 1* , Dina Cruickshank 2 , and Debby E. Hamilton 3

2023; 33(6): 715-723 https://doi.org/10.4014/jmb.2212.12010

Gut Microbiome as a Possible Cause of Occurrence and Therapeutic Target in Chronic Obstructive Pulmonary Disease

Eun Yeong Lim 1 , Eun-Ji Song 1 , and Hee Soon Shin 1,2*

2023; 33(9): 1111-1118 https://doi.org/10.4014/jmb.2301.01033

Recent Advancements in Technologies to Detect Enterohaemorrhagic Escherichia coli Shiga Toxins

Jeongtae Kim 1† , Jun Bong Lee 2† , Jaewon Park 3 , Chiwan Koo 1* , and Moo-Seung Lee 4,5*

2023; 33(5): 559-573 https://doi.org/10.4014/jmb.2212.12025

Anti-Inflammatory Response in TNFα/IFNγ-Induced HaCaT Keratinocytes and Probiotic Properties of Lacticaseibacillus rhamnosus MG4644, Lacticaseibacillus paracasei MG4693, and Lactococcus lactis MG5474

Ji Yeon Lee, Jeong‐Yong Park, Yulah Jeong, and Chang‐Ho Kang *

2023; 33(8): 1039-1049 https://doi.org/10.4014/jmb.2301.01028

Anticancer Effects of Gut Microbiota-Derived Short-Chain Fatty Acids in Cancers

Mi-Young Son 1,2,3* and Hyun-Soo Cho 1,2,3*

2023; 33(7): 849-856 https://doi.org/10.4014/jmb.2301.01031

Current Status of Epidemiology, Diagnosis, Therapeutics, and Vaccines for the Re-Emerging Human Monkeypox Virus

Wooseong Lee, Yu-Jin Kim, Su Jin Lee, Dae-Gyun Ahn * , and Seong-Jun Kim *

2023; 33(8): 981-991 https://doi.org/10.4014/jmb.2306.06033

Lactobacillus rhamnosus JY02 Ameliorates Sarcopenia by Anti-Atrophic Effects in a Dexamethasone-Induced Cellular and Murine Model

Juyeon Lee 1† , Minkyoung Kang 1† , Jiseon Yoo 1 , Sujeong Lee 1 , Minji Kang 1 , Bohyun Yun 2 , Jong Nam Kim 3 , Hyoungsun Moon 4 , Yihyung Chung 5 , and Sangnam Oh 1*

2023; 33(7): 915-925 https://doi.org/10.4014/jmb.2303.03001

Probiotic Characteristics and Safety Assessment of Lacticaseibacillus casei KGC1201 Isolated from Panax ginseng

Yun-Seok Lee 1 , Hye-Young Yu 2 , Mijin Kwon 2 , Seung-Ho Lee 2 , Ji-In Park 4 , Jiho Seo 3* , and Sang-Kyu Kim 2*

2023; 33(4): 519-526 https://doi.org/10.4014/jmb.2211.11029

Antimicrobial Resistance of Seventy Lactic Acid Bacteria Isolated from Commercial Probiotics in Korea

Eunju Shin 1 , Jennifer Jaemin Paek 1,2 , and Yeonhee Lee 1*

2023; 33(4): 500-510 https://doi.org/10.4014/jmb.2210.10041

Journal of Microbiology and Biotechnology

Subject Area and Category

• Biotechnology
• Applied Microbiology and Biotechnology
• Medicine (miscellaneous)

Korean Society for Microbiolog and Biotechnology

Publication type

17388872, 10177825

1991, 1996-2022

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The set of journals have been ranked according to their SJR and divided into four equal groups, four quartiles. Q1 (green) comprises the quarter of the journals with the highest values, Q2 (yellow) the second highest values, Q3 (orange) the third highest values and Q4 (red) the lowest values.

The SJR is a size-independent prestige indicator that ranks journals by their 'average prestige per article'. It is based on the idea that 'all citations are not created equal'. SJR is a measure of scientific influence of journals that accounts for both the number of citations received by a journal and the importance or prestige of the journals where such citations come from It measures the scientific influence of the average article in a journal, it expresses how central to the global scientific discussion an average article of the journal is.

Evolution of the number of published documents. All types of documents are considered, including citable and non citable documents.

This indicator counts the number of citations received by documents from a journal and divides them by the total number of documents published in that journal. The chart shows the evolution of the average number of times documents published in a journal in the past two, three and four years have been cited in the current year. The two years line is equivalent to journal impact factor ™ (Thomson Reuters) metric.

Evolution of the total number of citations and journal's self-citations received by a journal's published documents during the three previous years. Journal Self-citation is defined as the number of citation from a journal citing article to articles published by the same journal.

Evolution of the number of total citation per document and external citation per document (i.e. journal self-citations removed) received by a journal's published documents during the three previous years. External citations are calculated by subtracting the number of self-citations from the total number of citations received by the journal’s documents.

International Collaboration accounts for the articles that have been produced by researchers from several countries. The chart shows the ratio of a journal's documents signed by researchers from more than one country; that is including more than one country address.

Not every article in a journal is considered primary research and therefore "citable", this chart shows the ratio of a journal's articles including substantial research (research articles, conference papers and reviews) in three year windows vs. those documents other than research articles, reviews and conference papers.

Ratio of a journal's items, grouped in three years windows, that have been cited at least once vs. those not cited during the following year.

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Announcing JIMB 's New Editor-in-Chief: Yi Tang

The Society for Industrial Microbiology and Biotechnology is pleased to announce the journal's next Editor-in-Chief, Yi Tang, the Chancellor Professor at the University of California Los Angeles Department of Chemical and Biomolecular Engineering. 

"I am looking forward to working with scientists and engineers from both academia and industry to promote and communicate cutting-edge research activities in the areas of microbiology, synthetic biology and biotechnology." 

SIMB caught up with Yi Tang at the 2023 SIMB Annual Meeting for a short chat about publishing your research with the journal. 

Watch the video

Call for Papers: Applied Genomics and Systems Biotechnology

The Journal of Industrial Microbiology and Biotechnology is currently seeking papers for one of the core subjects that JIMB publishes on: Applied Genomics and Systems Biotechnology. 

Learn more here & view other articles on this subject

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Featured content.

Explore articles that have been hand-picked from Editor-in-Chief Yi Tang, and former Editor-in-Chief Ramon Gonzalez, as the best research from the  Journal of Industrial Microbiology and Biotechnology . 

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High-Impact Research Collection

Explore a collection of the most read and most cited articles making an impact in the Journal of Industrial Microbiology and Biotechnology from the past two years. 

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Special Collection: Articles from Industry

JIMB  is pleased to present a collection of articles highlighting recent, impactful research from authors in the microbiology and biotechnology industries. The collection covers various topics, including biofuels and biochemicals, vaccines, natural products, and biodegradation of plastics. The authors’ affiliations include ExxonMobil, BP, Petrobras, Braskem, DuPont, Dow, Merck, Sanofi, Pfizer, DSM, Chr. Hansen, Novozymes, Cargill, and others. 

All Special Issues and Collections from JIMB

In addition to its regular issues, the  Journal of Industrial Microbiology and Biotechnology  also actively contributes to the scientific community by releasing special issues and collections that encompass a wide range of research topics within the field of industrial microbiology and biotechnology.

Browse past special issues and collections

A journal for the industrial microbiology and biotechnology communities

The  Journal of Industrial Microbiology and Biotechnology (JIMB) publishes papers in all aspects of industrial microbiology and biotechnology and features a variety of benefits to publishing your research with the journal including: 

• Impact Factor & ranking in the field
• Rapid time to first decision
• Quality peer review
• Fully open access
• Listed in the DOAJ

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Authors interested in publishing in Journal of Industrial Microbiology and Biotechnology may be able to publish their paper Open Access using funds available through their institution’s agreement with OUP.

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JIMB has switched to a continuous publication model 

The Journal of Industrial Microbiology and Biotechnology has moved to a continuous publication model in 2023. This means that articles are published online as soon as they have completed the production process meaning researchers can access and cite the content faster than ever . 

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About the society

The Society for Industrial Microbiology and Biotechnology (SIMB) is a nonprofit, international association dedicated to the advancement of microbiological sciences, especially as they apply to industrial products, biotechnology, materials, and processes. Founded in 1949, SIMB promotes the exchange of scientific information through its meetings and publications, and serves as liaison among the specialized fields of microbiology. 

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The SIMB Annual Meeting and Exhibition will take place in Minneapolis, Minnesota from July 30 - August 2, 2023. This is an exciting time for science and technology development and a year of great advancement in microbial biotechnology. Topics will include: Biocatalysis, Cell Culture/Fermentation, Environmental Microbiology, Metabolic Engineering, Natural Products, and more. 

RAFT® 15 - Recent Advances in Fermentation Technology

The annual Recent Advances in Fermentation Technology meeting will take place in Naples, Florida from October 29 - November 1, 2023. RAFT is where leading industry, government, and academic stakeholders gather to discuss the latest developments and applications in industrial-scale fermentation technologies, and we hope to see you there! 

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Journal Metrics Reports 2022

Microbiology & medical microbiology, announcement of the latest impact factors from the journal citation reports.

Researchers consider a number of factors in deciding where to publish their research, such as journal reputation, readership and community, speed of publication, and citations. See how we share a whole range of information to help the research community decide which journal is the best home for their research as well as what the metrics can tell you about the performance of a journal and its articles.

Explore journal impact metrics

Indian Journal of Microbiology

Impact Factor 3.0 (2022)

5 Year Impact Factor 3.1 (2022)

Cite Score 6.1 (2022)

H5 Index 30 (2021)

Social Media Mentions 229 (2022)

Downloads 141,252 (2022)

Current Fungal Infection Reports

Impact Factor 1.4 (2022)

5 Year Impact Factor 1.3 (2022)

Cite Score 3.0 (2022)

H5 Index 13 (2021)

Social Media Mentions 316 (2022)

Downloads 53,651 (2022)

Current Clinical Microbiology Reports

Impact Factor 5.2 (2022)

5 Year Impact Factor 3.6 (2022)

Cite Score 7.3 (2022)

H5 Index 19 (2021)

Social Media Mentions 240 (2022)

Downloads 82,582 (2022)

VirusDisease

Cite Score 5.2 (2022)

H5 Index 22 (2021)

Social Media Mentions 680 (2022)

Downloads 161,476 (2022)

Acta Parasitologica

Impact Factor 1.5 (2022)

5 Year Impact Factor 1.5 (2022)

Cite Score 2.7 (2022)

H5 Index 18 (2021)

Social Media Mentions 198 (2022)

Downloads 109,539 (2022)

Food and Environmental Virology

Impact Factor 3.4 (2022)

Cite Score 6.4 (2022)

H5 Index 27 (2021)

Social Media Mentions 560 (2022)

Downloads 116,531 (2022)

Journal of Microbiology

5 Year Impact Factor 3.7 (2022)

H5 Index 35 (2021)

Social Media Mentions 869 (2022)

Downloads 125,977 (2022)

Biodegradation

Impact Factor 3.6 (2022)

Cite Score 5.7 (2022)

H5 Index 25 (2021)

Social Media Mentions 153 (2022)

Downloads 129,407 (2022)

Molecular Genetics, Microbiology and Virology

Impact Factor 0.5 (2022)

5 Year Impact Factor 0.4 (2022)

Cite Score 0.7 (2022)

H5 Index 9 (2021)

Downloads 13,356 (2022)

Brazilian Journal of Microbiology

Impact Factor 2.2 (2022)

5 Year Impact Factor 2.8 (2022)

Cite Score 3.8 (2022)

H5 Index 43 (2021)

Social Media Mentions 4,549 (2022)

Downloads 178,214 (2022)

Genes & Genomics

Impact Factor 2.1 (2022)

5 Year Impact Factor 1.9 (2022)

Cite Score 3.7 (2022)

Social Media Mentions 336 (2022)

Downloads 185,576 (2022)

Folia Microbiologica

Impact Factor 2.6 (2022)

5 Year Impact Factor 2.6 (2022)

Cite Score 5.3 (2022)

Social Media Mentions 496 (2022)

Downloads 199,030 (2022)

Virus Genes

Impact Factor 1.6 (2022)

5 Year Impact Factor 2.0 (2022)

Cite Score 4.4 (2022)

H5 Index 24 (2021)

Social Media Mentions 746 (2022)

Downloads 223,233 (2022)

Impact Factor 3.9 (2022)

5 Year Impact Factor 4.3 (2022)

Cite Score 7.5 (2022)

H5 Index 32 (2021)

Social Media Mentions 904 (2022)

Downloads 226,923 (2022)

Mycopathologia

Impact Factor 5.5 (2022)

Cite Score 6.2 (2022)

Social Media Mentions 1,604 (2022)

Downloads 236,912 (2022)

Current Genetics

Impact Factor 2.5 (2022)

5 Year Impact Factor 2.4 (2022)

Cite Score 6.7 (2022)

Social Media Mentions 905 (2022)

Downloads 269,834 (2022)

Medical Microbiology and Immunology

Impact Factor 5.4 (2022)

Cite Score 9.5 (2022)

H5 Index 28 (2021)

Social Media Mentions 2,118 (2022)

Downloads 297,822 (2022)

Applied Microbiology and Biotechnology

Impact Factor 5.0 (2022)

5 Year Impact Factor 5.2 (2022)

Cite Score 9.9 (2022)

H5 Index 79 (2021)

Downloads 3,456,479 (2022)

Antonie van Leeuwenhoek

Cite Score 4.7 (2022)

Social Media Mentions 1,833 (2022)

Downloads 390,428 (2022)

Biotechnology Letters

Impact Factor 2.7 (2022)

5 Year Impact Factor 2.5 (2022)

Cite Score 5.1 (2022)

Social Media Mentions 2,115 (2022)

Downloads 570,619 (2022)

Microbiology

5 Year Impact Factor 1.4 (2022)

Cite Score 2.5 (2022)

Downloads 60,921 (2022)

Current Microbiology

Cite Score 3.9 (2022)

H5 Index 34 (2021)

Social Media Mentions 10,583 (2022)

Downloads 638,008 (2022)

Extremophiles

Impact Factor 2.9 (2022)

Cite Score 5.8 (2022)

H5 Index 29 (2021)

Social Media Mentions 543 (2022)

Downloads 715,058 (2022)

Archives of Microbiology

Impact Factor 2.8 (2022)

Cite Score 3.3 (2022)

H5 Index 33 (2021)

Social Media Mentions 11,332 (2022)

Downloads 719,004 (2022)

World Journal of Microbiology and Biotechnology

Impact Factor 4.1 (2022)

5 Year Impact Factor 4.2 (2022)

Social Media Mentions 1,312 (2022)

Downloads 744,263 (2022)

Parasitology Research

Impact Factor 2.0 (2022)

Cite Score 4.0 (2022)

H5 Index 36 (2021)

Social Media Mentions 2,304 (2022)

Downloads 800,825 (2022)

Mycotoxin Research

5 Year Impact Factor 3.9 (2022)

Cite Score 7.4 (2022)

H5 Index 26 (2021)

Social Media Mentions 121 (2022)

Downloads 85,447 (2022)

Microbial Ecology

Cite Score 8.3 (2022)

H5 Index 48 (2021)

Social Media Mentions 1,988 (2022)

Downloads 878,185 (2022)

International Microbiology

Impact Factor 3.1 (2022)

5 Year Impact Factor 3.2 (2022)

Cite Score 5.4 (2022)

H5 Index 21 (2021)

Social Media Mentions 382 (2022)

Downloads 90,583 (2022)

European Journal of Clinical Microbiology & Infectious Diseases

Impact Factor 4.5 (2022)

5 Year Impact Factor 3.8 (2022)

Cite Score 8.9 (2022)

H5 Index 54 (2021)

Social Media Mentions 11,229 (2022)

Downloads 917,043 (2022)

Archives of Virology

Cite Score 4.9 (2022)

H5 Index 47 (2021)

Social Media Mentions 8,244 (2022)

Downloads 954,375 (2022)

AMB Express

Impact Factor 3.7 (2022)

5 Year Impact Factor 4.0 (2022)

Cite Score 7.0 (2022)

H5 Index 42 (2021)

Social Media Mentions 490 (2022)

Downloads 956,597 (2022)

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Journal Metrics

This page provides information on peer review performance and citation metrics for Nature Microbiology . Our quick reference guide to journal metrics is also available for download.

2023 Peer Review Metrics

Submission to first editorial decision: the median time (in days) from when a submission is received to when a first editorial decision about whether the paper was sent out for formal review or not is sent to the authors.

Submission to Accept: the median time (in days) from the submission date to the final editorial acceptance date.

Submission to first editorial decision - 15

Submission to Accept - 198

2022 Journal Metrics

On this page you will find a suite of citation-based metrics for Nature Microbiology . Brief definitions for each of the metrics used to measure the influence of our journals are included below the journal metrics. Data has been produced by Clarivate Analytics.

For recently launched journals, metrics are calculated from available citation data. If a metric uses multiple years of data, new journals may have partial metrics.

While the metrics presented here are not intended to be a definitive list, we hope that they will prove to be informative. The page is updated on an annual basis.

2-year Impact Factor - 28.3

5-year Impact Factor - 22.9

Immediacy index - 5.1

Eigenfactor® score - 0.06382

Article Influence Score - 8.7

2023 Usage Metrics

Article-level metrics are also available on each article page, allowing readers to track the reach of individual papers.

2,976,242 Downloads

35,348 Altmetric mentions

Definitions

2-year impact factor.

The Journal Impact Factor is defined as all citations to the journal in the current JCR year to items published in the previous two years, divided by the total number of scholarly items (these comprise articles, reviews, and proceedings papers) published in the journal in the previous two years. (Courtesy of Clarivate Analytics )

5-year Impact Factor

The 5-year journal Impact Factor, available from 2007 onward, is the average number of times articles from the journal published in the past five years have been cited in the JCR year. It is calculated by dividing the number of citations in the JCR year by the total number of articles published in the five previous years. (Courtesy of Clarivate Analytics )

Immediacy index

The Immediacy Index is the average number of times an article is cited in the year it is published. The journal Immediacy Index indicates how quickly articles in a journal are cited. (Courtesy of Clarivate Analytics )

Eigenfactor® Score

The Eigenfactor Score calculation is based on the number of times articles from the journal published in the past five years have been cited in the JCR year, but it also considers which journals have contributed these citations so that highly cited journals will influence the network more than lesser cited journals. References from one article in a journal to another article from the same journal are removed, so that Eigenfactor Scores are not influenced by journal self-citation. (Courtesy of Clarivate Analytics )

Article Influence Score

The Article Influence Score determines the average influence of a journal's articles over the first five years after publication. It is calculated by multiplying the Eigenfactor Score by 0.01 and dividing by the number of articles in the journal, normalized as a fraction of all articles in all publications. This measure is roughly analogous to the 5-Year Journal Impact Factor in that it is a ratio of a journal's citation influence to the size of the journal's article contribution over a period of five years. (Courtesy of Clarivate Analytics )

Downloads reflect the number of times full text or PDF versions of articles are accessed directly from the journal website. Downloads are defined as HTML, LookInside, PDF and Epub clicks. Please note that this does not include article downloads from mirror databases such as PubMed Central.

Altmetric mentions

Total number of mentions (e.g. Twitter, Facebook, Reddit, Blogs, News articles, Policy documents and Faculty of 1000 reviews) for articles published in the specified timeframe, as provided by Altmetric .

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Applied Microbiology and Biotechnology

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Transcriptome analysis of kluyveromyces marxianus under succinic acid stress and development of robust strains.

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Pulchinenoside B4 ameliorates oral ulcers in rats by modulating gut microbiota and metabolites

Melanin depletion affects Aspergillus flavus conidial surface proteins, architecture, and virulence

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A candidate competitive ELISA based on monoclonal antibody 3A8 for diagnosis of contagious bovine pleuropneumonia

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Applied Microbiology and Biotechnology is now fully open access!

We are excited to announce that Applied Microbiology and Biotechnology has now become a fully open-access (OA) journal as of January 2024. This means that we will only be publishing articles as Open Access meaning content will be freely available to readers worldwide, enabling the widest possible dissemination and reuse.

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Journal of Microbiology and Biotechnology Research

URL: http://scholarsresearchlibrary.com/Journal-of-Microbiology-and-Biotechnology-Research/index.html

Keywords: Applied Biochemistry, Applied Microbiology Bacteriology, Bioactive Compounds and Chemical Biology, Biocatalysis Technology, Biochemistry, Biodiversity, Biomedical Sciences, Bioprocess Engineering, Botany and Plant Sciences, Cell Physiology, Cellular Microbiology, Clinical Microbiology, Endocrinology, Entomology, Environmental Biotechnology, Environmental Microbiology, Enzymology and Enzyme Engineering, Fermentation Technology, Food and Agricultural Technologies, Food Biotechnology, Food Microbiology, Genetics, Genomics and Proteomics, Immunology, Industrial Microbiology, Infection and Immunity, Medical Microbiology, Metabolic Engineering, Microbial Ecology, Microbial Structure and Function, Molecular and Cellular Biology, Molecular Biology, Molecular Microbiology, Mycology, Parasitology, Physiology and Metabolism, Plant and Animal Cell Cultures, Plant Pathology, Probiotics and Prebiotics, Protozoology, Soil and Environmental Sciences, Toxicology, Virology, Zoology

ISSN: 2231-3168

EISSN: 2231-3168

Subject: Biotechnology

Publisher: Dr H P Singh

Country: India

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Journal Of Microbiology And Biotechnology impact factor, indexing, ranking (2024)

Aim and Scope

The Journal Of Microbiology And Biotechnology is a research journal that publishes research related to Biochemistry, Genetics and Molecular Biology; Immunology and Microbiology; Medicine . This journal is published by the Korean Society for Microbiolog and Biotechnology. The ISSN of this journal is 17388872, 10177825 . Based on the Scopus data, the SCImago Journal Rank (SJR) of journal of microbiology and biotechnology is 0.563 .

Journal Of Microbiology And Biotechnology Ranking

The Impact Factor of Journal Of Microbiology And Biotechnology is 3.277.

The impact factor (IF) is a measure of the frequency with which the average article in a journal has been cited in a particular year. It is used to measure the importance or rank of a journal by calculating the times its articles are cited.

The impact factor was devised by Eugene Garfield, the founder of the Institute for Scientific Information (ISI) in Philadelphia. Impact factors began to be calculated yearly starting from 1975 for journals listed in the Journal Citation Reports (JCR). ISI was acquired by Thomson Scientific & Healthcare in 1992, and became known as Thomson ISI. In 2018, Thomson-Reuters spun off and sold ISI to Onex Corporation and Baring Private Equity Asia. They founded a new corporation, Clarivate , which is now the publisher of the JCR.

Important Metrics

Journal of microbiology and biotechnology indexing.

The journal of microbiology and biotechnology is indexed in:

• Web of Science (SCIE)

An indexed journal means that the journal has gone through and passed a review process of certain requirements done by a journal indexer.

The Web of Science Core Collection includes the Science Citation Index Expanded (SCIE), Social Sciences Citation Index (SSCI), Arts & Humanities Citation Index (AHCI), and Emerging Sources Citation Index (ESCI).

Journal Of Microbiology And Biotechnology Impact Factor 2024

The latest impact factor of journal of microbiology and biotechnology is 3.277 .

The impact factor (IF) is a measure of the frequency with which the average article in a journal has been cited in a particular year. It is used to measure the importance or rank of a journal by calculating the times it's articles are cited.

Note: Every year, The Clarivate releases the Journal Citation Report (JCR). The JCR provides information about academic journals including impact factor. The latest JCR was released in June, 2023. The JCR 2024 will be released in the June 2024.

Journal Of Microbiology And Biotechnology Quartile

The latest Quartile of journal of microbiology and biotechnology is Q2 .

Each subject category of journals is divided into four quartiles: Q1, Q2, Q3, Q4. Q1 is occupied by the top 25% of journals in the list; Q2 is occupied by journals in the 25 to 50% group; Q3 is occupied by journals in the 50 to 75% group and Q4 is occupied by journals in the 75 to 100% group.

Call for Papers

Visit to the official website of the journal/ conference to check the details about call for papers.

How to publish in Journal Of Microbiology And Biotechnology?

If your research is related to Biochemistry, Genetics and Molecular Biology; Immunology and Microbiology; Medicine, then visit the official website of journal of microbiology and biotechnology and send your manuscript.

Tips for publishing in Journal Of Microbiology And Biotechnology:

• Selection of research problem.
• Presenting a solution.
• Designing the paper.
• Make your manuscript publication worthy.
• Write an effective results section.
• Mind your references.

Acceptance Rate

Journal publication time.

The publication time may vary depending on factors such as the complexity of the research and the current workload of the editorial team. Journals typically request reviewers to submit their reviews within 3-4 weeks. However, some journals lack mechanisms to enforce this deadline, making it difficult to predict the duration of the peer review process.

The review time also depends upon the quality of the research paper.

Final Summary

• The impact factor of journal of microbiology and biotechnology is 3.277.
• The journal of microbiology and biotechnology is a reputed research journal.
• It is published by Korean Society for Microbiolog and Biotechnology .
• The journal is indexed in UGC CARE, Scopus, SCIE, PubMed .
• The (SJR) SCImago Journal Rank is 0.563 .

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Innate immunity, gene therapy, genes & genomics, genome research, international journal of genomics, mammalian genome, european cytokine network, journal of interferon and cytokine research, top research journals.

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ORIGINAL RESEARCH article

Plumbagin inhibits fungal growth, hmgb1/lox-1 pathway and inflammatory factors in a. fumigatus keratitis.

• Department of Ophthalmology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China

To investigate the anti-inflammatory and antifungal effects of plumbagin (PL) in Aspergillus fumigatus ( A. fumigatus ) keratitis, the minimum inhibitory concentration (MIC), time-killing curve, spore adhesion, crystal violet staining, calcium fluoride white staining, and Propidium Iodide (PI) staining were employed to assess the antifungal activity of PL in vitro against A. fumigatus . The cytotoxicity of PL was assessed using the Cell Counting Kit-8 (CCK8). The impact of PL on the expression of HMGB1, LOX-1, TNF-α, IL-1β, IL-6, IL-10 and ROS in A. fumigatus keratitis was investigated using RT-PCR, ELISA, Western blot, and Reactive oxygen species (ROS) assay. The therapeutic efficacy of PL against A. fumigatus keratitis was assessed through clinical scoring, plate counting, Immunofluorescence and Hematoxylin-Eosin (HE) staining. Finally, we found that PL inhibited the growth, spore adhesion, and biofilm formation of A. fumigatus and disrupted the integrity of its cell membrane and cell wall. PL decreased IL-6, TNF-α, and IL-1β levels while increasing IL-10 expression in fungi-infected mice corneas and peritoneal macrophages. Additionally, PL significantly attenuated the HMGB1/LOX-1 pathway while reversing the promoting effect of Boxb (an HMGB1 agonist) on HMGB1/LOX-1. Moreover, PL decreased the level of ROS. In vivo , clinical scores, neutrophil recruitment, and fungal burden were all significantly reduced in infected corneas treated with PL. In summary, the inflammatory process can be inhibited by PL through the regulation of the HMGB-1/LOX-1 pathway. Simultaneously, PL can exert antifungal effects by limiting fungal spore adhesion and biofilm formation, as well as causing destruction of cell membranes and walls.

1 Introduction

Fungal keratitis is a highly destructive infectious disease of the cornea. It occurs most commonly in the tropics and subtropics. The incidence of fungal keratitis is estimated to be over 1,000,000 cases per year, and about 10% of them will progress to corneal perforation or even enucleation ( Bongomin et al., 2017 ; Brown et al., 2021 ). The disease is mainly concentrated in agricultural and outdoor workers, long-term contact lens wearers, and patients with excessive use of antibiotics and glucocorticoids ( Bongomin et al., 2017 ). The most common pathogenic fungi are Aspergillus and Fusarium ( Liu et al., 2023 ). In addition to the invasion and colonization of fungal hyphae in the corneal stroma, the excessive inflammation is one of the reasons for accelerating the development of fungal keratitis, which can lead to stromal damage and corneal opacity ( Mills et al., 2021 ; Sha et al., 2021 ). The current treatment drugs are mainly natamycin, which is used in the form of suspension. It not only has poor dispersion and low solubility but also limits penetration into the corneal epithelium. Furthermore, natamycin is also associated with the occurrence of adverse reactions and the development of resistance ( Sahay et al., 2019 ; Ung et al., 2019 ). Therefore, it is necessary to find new methods for the treatment of fungal keratitis.

Plumbagin (5-hydroxy-2-methyl-1, 4-naphthoquinone, PL) is a potent naphthoquinone compound from the roots of traditional medicinal plants. It has a variety of pharmacological properties ( Jaradat et al., 2017 ) and has been proven to have anti-inflammatory, antimicrobial activity, anti-oxidative stress and anti-cancer effects ( Pan et al., 2022 ; Catalani et al., 2023 ; Lin et al., 2023 ; Xiong et al., 2024 ). PL has been utilized in various inflammatory models, such as the lipopolysaccharide (LPS)-induced glial cell inflammation model and rheumatoid arthritis, to downregulate the expression of inflammatory factors, including TNF-α, IL-6, and IL-1β ( Gupta et al., 2018 ; Shu et al., 2022 ). Moreover, in the sepsis model induced by LPS, PL has demonstrated significant efficacy in reducing HMGB1 levels and further suppressing proinflammatory cytokines ( Zhang Z. et al., 2016 ). The accumulation of ROS in cells is a contributing factor to the development of inflammatory diseases ( Kim and Kim, 2021 ). Multiple studies have demonstrated that PL can effectively attenuate cellular ROS levels, thereby inhibiting the release of HMGB1 and mitigating inflammatory damage ( Cao et al., 2022 ; Xiao et al., 2022 ). HMGB1 is a promising and significant therapeutic target for the management of bacterial and fungal keratitis, as it plays a crucial role in influencing inflammatory factors ( Wu, 2020 ; Yin et al., 2021 ). Previous research has demonstrated that HMGB1 triggers inflammation in A. fumigatus keratitis by upregulating LOX-1 levels, consequently elevating the levels of IL-1β, TNF-α, and other inflammatory factors ( Jia-Qian Jiang et al., 2019 ; Wu, 2020 ) Therefore, PL is anticipated to have a pivotal role in treating A. fumigatus keratitis based on previous studies highlighting its ability to inhibit HMGB1 levels.

Furthermore, PL displays robust and extensive antifungal activity against numerous fungi, including Fusarium graminearum and Candida albicans . It not only disrupts the cell and mitochondrial membranes of the fungi, as evidenced by a concentration-dependent increase in relative electrical conductivity but also demonstrates significant anti-biofilm activity through down-regulation of FAS1 and FAS2 expression ( Hassan et al., 2016 ; Shang et al., 2019 ; Qian et al., 2022 ; Bisso et al., 2023 ; Wang et al., 2023 ). These findings indicate that PL may also possess substantial antifungal effects in A. fumigatus keratitis.

The present study validated the anti-inflammatory and antifungal effects of PL against A. fumigatus keratitis. The findings demonstrated that PL effectively inhibited the HMGB1/LOX-1 pathway, leading to the down-regulation of proinflammatory cytokine expression and attenuating intracellular ROS levels. Moreover, PL exhibited potent inhibition of spore adhesion and biofilm formation while also disrupting fungal cell membrane and cell wall. Consequently, PL holds great promise as a highly effective therapeutic agent for the treatment of fungal keratitis.

2 Materials and methods

2.1 pl preparation.

PL (HY-N1497, purity: 99.23%) was purchased from MedChemExpress (Shanghai, China). The concentration of PL was reduced to 100 mM by dissolving it in DMSO, and packaged and frozen at −80°C. The samples were taken in fractions and diluted to an appropriate concentration by phosphate-buffered saline (PBS) or corresponding cell culture medium with 1‰ DMSO.

2.2 A. fumigatus culture

A standard A. fumigatus strain (CPCC 3.0772), purchased from the China General Microbiological Culture Collection Center, was grown to mycelium after 2 days of cultivation in liquid Sabouraud medium at 37°C and 200 rpm. The mycelium was removed and ground in a sterile operating table, washed three times with PBS, and placed in 70% ethanol overnight to obtain inactivated mycelium, which was adjusted to a concentration of 3 × 10 8 CFU/mL. After culturing A. fumigatus strains for 2–3 days at 28°C (sabouraud agar medium), conidia on the surface of the medium were collected.

2.3 Cell culture and A. fumigatus stimulation

Human corneal epithelial cells (HCECs), provided by the laboratory of Xiamen University, Fujian, China, were cultured in Dulbecco's Modified Eagle Medium (96-well plate, 37°C, 5% CO 2 ) until reaching 80% confluence for CCK-8 and fungal spore adhesion experiments. RAW264.7 macrophages, purchased from the Chinese Academy of Sciences (Shanghai, China), were cultured in DMEM (light-protected 96-well plates, 37°C, 5% CO 2 ) and used for CCK-8 and ROS detection when the concentration of RAW264.7 reached 80%. For the extraction of peritoneal macrophages, mice were injected intraperitoneally with 1 mL of 3% thioglycollate medium and stimulated for 7 days. When the mice were sacrificed and the abdomen was wiped with 75% alcohol, 10 mL DMEM was injected intraperitoneally and collected. After centrifugation, purification, and suspension, the cells were then suspended and plated in either 6- or 12-well plates and stimulated with inactivated hyphae for 8 or 24 h for Reverse Transcription-PCR (RT-PCR), Western Blotting (WB), and Enzyme-Linked Immunosorbent Assay (ELISA). In order to validate the HMGB1/LOX-1 pathway, peritoneal macrophages were pre-treated with Boxb for 1 h, followed by stimulation with fungi and drugs. Subsequent experiments were conducted.

2.4 Mice and models of fungal keratitis

Eight-week-old healthy female C57BL/6 mice were obtained from Jinan Pengyue Laboratory Animal Co., Ltd. (Jinan, China). The study was carried out following the guidelines provided by ARVO for the ethical use of animals in research related to ophthalmology and vision. Additionally, it received approval from the Ethics Committee at Qingdao University's Affiliated Hospital (QYFY WZLL 28315). After the mice were anesthetized, A. fumigatus spores (3 μL, 0.5 × 10 5 μL −1 ) were injected into the corneal stroma of the right eye using a ×40 magnification stereoscopic microscope, while the left eye served as a blank control kept untreated. The experimental and control groups were treated with 4 μL PL (30 μM) and 1‰ DMSO drops on the ocular surface 4 times daily, respectively. Mouse corneas were examined daily using a slit-lamp microscope and photographed for clinical score at 1, 3, and 5 days post-infection (p.i.). After 5 days, corneas were collected for RT-PCR, ELISA, WB, and plate counting. HE staining and immunofluorescence staining were performed on infected eyeballs.

2.5 Cell counting kit-8 assay

After incubating HCECs with different concentrations of PL (0, 2, 3, 4, 5 and 6 μM) or 1‰ DMSO in a 96-well plate for 24 h, each well was supplemented with 10 μL of CCK-8 solution (Solarbio, Beijing, China) and further incubated for an additional period of 2 h at 37°C. The absorbance at a wavelength of 450 nm was measured using a microplate reader. RAW264.7 cells were subjected to toxicity testing using the same methodology mentioned above but with varying concentrations of PL (0, 2, 3, 4, 5, 6, 8, and 10 μM).

2.6 In vivo safety study

Chen et al. (2018) demonstrated that intravitreal injection of PL (0–50 μM) did not cause ocular toxicity in mice, which prompted this study to conduct in vivo toxicity tests. Mice were administered 4 μL PL (30, 40 μM) 4 times a day into the conjunctival sac of the right eye, while an equal amount of 1‰ DMSO was applied to the left eye at the same frequency. The sodium fluorescein staining of the ocular surface under cobalt blue light of the slit-lamp was observed and recorded at 1, 3, and 5 days. The ocular surface damage was assessed based on a comprehensive scoring system, which took into account corneal opacity and lesions, iris changes, as well as conjunctival congestion, bulbar chemosis, and secretion ( Gomez et al., 2020 ; Tian et al., 2022 ).

2.7 RT-PCR assay

The RNAiso Plus reagent (Takara, Dalian, China) was utilized to extract total RNA from corneal and peritoneal macrophages in mice. Spectrophotometry was employed for the quantification of the extracted RNA. For cDNA synthesis, HiScript III RT SuperMix (Vazyme, Nanjing, China) was used to process the RNA samples. Detailed RT-PCR methods were used, as described in a previous study ( Bernardino et al., 2021 ). Table 1 shows all primer sequences used for RT-PCR.

Table 1 . Primer list used for RT-PCR.

2.8 WB assay

The mouse corneal or peritoneal macrophages were fully lysed for 2 h using RIPA solution (Solarbio, Beijing, China) containing 1% each of protease inhibitors (PMSF) and phosphatase inhibitors. Protein concentrations were measured using the BCA kit (Solarbio). After the inclusion of SDS sample buffer (Elabscience, Wuhan, China), the samples were subjected to boiling, and protein separation was performed using a 12% SDS-PAGE gel. Subsequently, the separated proteins were transferred onto PVDF membranes (Solarbio). The membranes were then treated with a protein-free rapid-blocking buffer (1x, Yamei, Shanghai, China). Blocking was performed for 15 min, followed by overnight incubation with primary antibodies diluted in primary antibody diluent: HMGB1 antibody (1:1,000; Abcam), LOX-1 antibody (1:3,000; Proteintech), GADPH antibody (1:5,000; Elabscience), β-actin antibody (1:1,000; Elabscience). After a PBST shaken wash, the membranes were incubated with a secondary antibody diluted in secondary antibody diluent (1:5,000, Elabscience) for 60 min. Finally, the visualization of protein bands was achieved by employing an ECL chemiluminescence kit (Vazyme Biotech, Nanjing, China).

2.9 ELISA assay

The corneas of mice at 5 days p.i. were harvested and ground in PBS, or the cell culture medium from mouse peritoneal macrophages cultured for 24 h was collected, followed by centrifugation to obtain the supernatant. The protein levels of TNF-α, IL-6, and IL-1β were quantified using corresponding ELISA kits (BioLegend), and the absorbance was measured at 450 and 570 nm using a microplate reader.

2.10 Reactive oxygen species assay

The RAW264.7 cells were incubated with various concentrations of PL (0, 2, 3 μM) in a sterile and light-protected 96-well plate for 24 h. Subsequently, the cells were incubated in the dark for 30 min using the Reactive Oxygen Species Assay Kit (Beyotime). The absorbance was measured at a wavelength of 488 nm to determine ROS levels (%), which were calculated as follows: ROS level = (fluorescence value of experimental group/average fluorescence value of control group) × 100%.

2.11 Immunofluorescence staining

The mouse eyeballs were dissected and immersed in OCT (Sakura Tissue-Tek, Torrance, CA, USA) before being snap-frozen in liquid nitrogen. Subsequently, the corneas of the eyeballs were sectioned into 10 μm slices. These sections were then incubated with a goat anti-rat neutrophil monoclonal marker (NIMP-R14, 1:100, Santa Cruz, CA, USA) overnight. Following this primary antibody incubation step, secondary antibodies (1:500; Abcam) were applied for an additional hour. Finally, nuclear staining was performed using 4′,6-diamino-2-phenylindole dihydrochloride. Fluorescence microscopy images were captured after applying antifading reagents onto the sections.

2.12 HE staining

The eyeballs of mice from different experimental groups were extracted at day 5 p.i., fixed in a formalin fixative solution, embedded in paraffin, and sectioned into 5 μm slices. Subsequently, the cornea was stained with hematoxylin and eosin, followed by image acquisition.

2.13 MIC and time-killing curves

The broth microdilution protocol is performed in accordance with the guidelines provided by the Clinical and Laboratory Standards Institute (CLSI M27-A4) ( Albuquerque et al., 2018 ), the PL was diluted to concentrations of 0, 1.25, 2.5, 3, 6, 8, 10, 20, 30, and 40 μM using Sand's liquid medium. The DMSO content was quantified as a concentration of 1‰. Conidia were added to achieve a uniform concentration of spore suspension at a density of 2 × 10 5 CFU/mL. After a comprehensive blending process, the prepared mixture was subsequently transferred into a 96-well plate. The plates were then incubated for a duration ranging from 24 to 120 h and the absorbance was measured at 540 nm. Simultaneously, a control group consisting of drug-free medium without conidia was established as the negative control.

2.14 Anti-adhesion of A. fumigatus spores

In order to evaluate the anti-adhesion effect of PL on A. fumigatus conidia, HCECs were seeded into the four-chambered slides containing different concentrations of PL (2, 3 μM) or 1‰ DMSO, and the concentration of spores was set as 1 × 10 7 CFU/mL, referring to previous study ( Zhu et al., 2021 ). After a 3-h incubation period at a temperature of 37°C, the wells were rinsed with PBS in order to eliminate any non-adherent conidia. Subsequently, the cells were stained with hematoxylin and eosin, visualized under a microscope (magnification, 200×), and the average number of conidia adhered to the surface of each HCEC was calculated.

2.15 PI staining and calcofluor white staining

The A. fumigatus conidia (1 × 10 6 CFU/mL) were incubated in 6-well plates at 37°C for 24 h, followed by additional culture in liquid medium containing various concentrations of PL (2, 3, 8, 10, 20, and 30 μM) or 1‰ DMSO for another 24 h. After being washed 3 times with sterile PBS, DNA staining agent PI (50 μg/mL; Sorobio, Beijing, China) was applied for 15 min in the dark and then examined under fluorescence microscopy (magnification, 100×; Leica, Germany). The same concentration of conidia was incubated with PL (0, 2, 3, 8, and 30 mM) or DMSO (with a concentration of 1‰) in medium for 24 h (12-well plate, 37°C). Fluorescence images were captured at the same magnification after staining with calcofluor white (CFW, 2.5 μg/mL).

2.16 Biofilm formation inhibition test

The biofilm formation inhibition experiment was slightly modified based on a previous study ( Wang et al., 2022 ). In this experiment, 1 × 10 5 CFU/mL of A. fumigatus conidia were co-cultured with different concentrations of PL (0, 3, 6, 8, 10, 20, and 30 μM) or 1‰ DMSO in Sargasparin medium for a duration of 48 h. After fixing the biofilm with methanol, it was stained with crystal violet (0.01%) for 15 min. Subsequently, it was washed and dried before being treated with ethanol (95%) for 5 min. The supernatant was then removed and transferred to a new 96-well plate. Finally, the absorbance at 570 nm was measured.

2.17 Efficacy comparison and synergy experiment between PL and natamycin

After establishing A. fumigatus keratitis in mice, the left eye served as a blank control was kept untreated. Four microliter PL (30 μM), 5% NATA, and PBS were, respectively, dropped on ocular surface 4 times a day. Mouse corneas were examined using a slit-lamp microscope at 1 and 5 days p.i. We employed the checkerboard method to investigate the synergistic effect of PL and NATA ( Gaspar-Cordeiro et al., 2022 ). Different concentrations of PL and NATA were added to a 96-well plate. resulting the final drug concentrations of PL ranged from 1 to 32 μM and NATA concentrations ranged from 0 to 16 μM. The concentration of A. fumigatus in each well was maintained at 2 × 10 5 CFU/mL. After incubated at 37°C for 24 h, the absorbance was measured at 540 nm, and the inhibition rate was calculated accordingly. The fractional inhibitory concentration index (FICI) was calculated at MIC 70 . FICI = (MIC PL combined/MIC PL alone) + (MIC NATA combined/MIC NATA alone). Synergism between PL and NATA was indicated when FICI ≤ 0.5. Additionally, 0.5 < FICI ≤ 1 implies additivity, 1 < FICI ≤ 2 implies indifference, and FICI > 2 implies antagonism.

2.18 Statistical analysis

The statistical analysis was performed using GraphPad Prism 9.5 (San Diego, CA, USA). Student's t -test was applied to evaluate the statistical significance between two groups. The significance among three or more groups was determined by One-way ANOVA test. All experiments were conducted in a minimum of three biological replicates and are expressed as mean ± SEM. Statistical significance was determined at P < 0.05.

3.1 Evaluation of PL safety

To determine the appropriate concentration of PL for the treatment of fungal keratitis, experiments were carried out both in vitro and in vivo . The results from the CCK-8 assay demonstrated that PL concentrations below 3 μM exhibited no cytotoxic effects on HCECs within a 24-h timeframe ( Figure 1A ), while RAW264.7 cells showed no cytotoxicity at concentrations ≤ 4 μM ( Figure 1B ). Furthermore, the Draize test was performed to assess the potentially toxic effects of PL on mouse corneas. The findings revealed that neither 30 nor 40 μM concentrations of PL caused any damage to the mouse cornea ( Figure 1C ). The results suggested that 3 μM PL is safe for in vitro experiments, while 30 μM PL is safe for in vivo experiments.

Figure 1 . Safety evaluation of PL in vivo and in vitro . Cell viability of HCECs (A) treated with DMSO (1‰) and PL (2, 3, 4, 5, and 6 μM) and RAW 264.7 cells (B) treated with PL (2, 3, 4, 5, 6, 8, and 10 μM) for 24 h ( n = 6/group). Corneal fluorescein staining images (C) were captured after 0, 1, 3, and 5 days of PL treatment. All data were displayed as mean ± SEM (ns, no significant difference, ** P < 0.01, *** P < 0.001, **** P < 0.0001. N, normal cells without treatment; D, days of treatment).

3.2 PL alleviated the severity of mouse A. fumigatus keratitis

To investigate the therapeutic benefits of PL in treating keratitis caused by A. fumigatus , the symptomatic manifestations (including turbidity area, turbidity intensity, ulcer morphology) of mice corneas were recorded using a slit lamp at 1, 3, and 5 days p.i. ( Figure 2A ). Clinical scores based on symptoms were evaluated for different groups ( Figure 2B ), and the results demonstrated that PL treatment significantly improved symptoms and markedly reduced clinical scores compared to DMSO treatment. Plate counting experiments ( Figure 2D ) indicated that fungal load in the cornea was significantly lower following PL treatment than in the DMSO treatment group, with a statistically significant difference observed ( Figure 2C ). HE staining of corneal tissues ( Figure 2E ) and immunofluorescence experiment ( Figure 2F ) revealed that PL treatment effectively mitigated inflammatory cell infiltration and edema within the cornea. These findings suggested that PL exhibited efficacy in alleviating the inflammatory response and reducing the fungal load in the fungi-infected mouse cornea.

Figure 2 . PL alleviated A. fumigatus keratitis in mice. Corneal changed at 1, 3, and 5 days p.i. after treatment with DMSO or PL ( A , n = 6/group) and analysis of differences between the two groups (B) . Plate counts of fungal units (D) and quantities analysis (C) in mouse corneal homogenates at 5 days of treatment with DMSO and PL ( n = 6/group). After 5 days of corneal infection, HE staining ( E ; Magnification, 200×; Scale bar: 50 μm) and neutrophil immunofluorescence ( F ; Magnification, 200×; Scale bar: 100 μm) were conducted on the corneas of Normal, DMSO, and PL groups. All data were displayed as mean ± SEM (ns, no significance, ** P < 0.01, *** P < 0.01. D, days p.i.).

3.3 PL decreased the expression of inflammatory factors and HMGB1/LOX-1 in peritoneal macrophages, and also reduced the levels of ROS in RAW 264.7 cells

The anti-inflammatory effect of PL was investigated by evaluating the levels of inflammation-related factors and intracellular ROS content after treatment with PL in vitro . Following 8 h of stimulation with A. fumigatus , the mRNA levels of HMGB1 ( Figure 3A ), LOX-1 ( Figure 3B ), TNF-α ( Figure 3C ), IL-1β ( Figure 3D ), and IL-6 ( Figure 3E ) were significantly reduced in PL treatment group, while the mRNA expression of IL-10 ( Figure 3F ) was increased. Additionally, low-dose PL stimulation within a period of 24 h exhibited a concentration-dependent decrease in intracellular ROS levels ( Figure 3G ).

Figure 3 . PL inhibited the level of inflammation in peritoneal macrophages in vitro , down-regulate the expression of HMGB1/LOX-1, and showed anti-oxidation effect. PL treatment decreased the mRNA expression of HMGB1, LOX-1, TNF-α, IL-1β, and IL-6 in peritoneal macrophages (A–E) , while increased the mRNA level of IL-10 (F) . Co-culturing PL and RAW264.7 cells for 24 h resulted in a significant reduction of intracellular ROS levels (G) . All data were displayed as mean ± SEM ( n = 6/group, ns, no significance; * P < 0.05, ** P < 0.01, *** P < 0.001. N, normal peritoneal macrophages or normal RAW264.7 cells; AF, A. fumigatus ).

3.4 PL inhibited the levels of proinflammatory cytokines and down-regulated the expression of HMGB1/LOX-1 in mouse models of A. fumigatus keratitis

In addition to conducting in vitro experiments, we also conducted an assessment of the anti-inflammatory effect of PL on mouse corneas. Treatment with PL resulted in a significant down-regulation of proinflammatory cytokines TNF-α ( Figures 4A , B ) and IL-6 ( Figures 4C , D ) at both mRNA and protein levels compared to the DMSO treatment group at 5 days p.i. Furthermore, we investigated whether the mechanism underlying PL's efficacy in treating fungal keratitis is associated with HMGB1/LOX-1 by examining changes in mRNA and protein expression levels in the cornea. Our findings revealed a significant decrease in HMGB1/LOX-1 expression within the cornea of the PL treatment group compared to the DMSO treatment group ( Figures 4E – J ).

Figure 4 . PL inhibited the inflammatory progression of A. fumigatus keratitis in mice. Compared with DMSO treatment group, the mRNA and protein levels of TNF-α (A, B) and IL-6 (C, D) in the cornea of PL treatment group were significantly decreased, and the expression of HMGB1/LOX-1 (E–J) was significantly down-regulated. All data were displayed as mean ± SEM ( n = 6/group, ns, no significance; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Con, control, corneas in control group were not infected with A. fumigatus ; D, days p.i.).

3.5 PL exerted an anti-inflammatory effect by inhibiting HMGB1/LOX-1 signaling pathway

In order to further elucidate the mechanism of action of PL treatment on A. fumigatus keratitis, we employed pre-treatment with Boxb in mouse peritoneal macrophages. The results demonstrated that following A. fumigatus infection, PL treatment significantly reduced the protein levels of HMGB1 and LOX-1. Boxb stimulation increased the expression of HMGB1 and LOX-1 in cells; however, PL treatment could notably decrease Boxb-induced high expression of HMGB1 and LOX-1 ( Figures 5E – J ). Similar results were observed for TNF-α ( Figures 5A , B ) and IL-1β ( Figures 5C , D ). These findings suggested that PL could down-regulate LOX-1 expression by inhibiting HMGB1 activity, subsequently reducing IL-1β and TNF-α levels and exerting an anti-inflammatory effect.

Figure 5 . The inhibitory effect of PL on the inflammatory response was mediated through the inhibition of the HMGB1/LOX-1 pathway. Pre-treatment with Boxb significantly enhanced the mRNA and protein expression levels of HMGB1 (E–G) , LOX-1 (H–J) , TNF-α (A, B) , and IL-1β (C, D) . Furthermore, treatment with PL effectively attenuated the inflammatory amplification induced by Boxb ( n = 6/group). All data were displayed as mean ± SEM (ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001. N , normal peritoneal macrophages; AF, A. fumigatus ).

3.6 The antifungal efficacy of PL in vitro

When PL was cocultured with conidia for 24 h, the growth of fungi was concentration-dependently inhibited from 1.25 μM ( Figure 6A ). It was observed that the inhibitory rate of PL at 3 μM could reach 37%, and 30 μM PL inhibited fungal growth by 77%. Although the inhibitory effect of PL decreased over time, even at 120 h, a concentration of 30 μM still exhibited effectiveness of 45% ( Figure 6B ). Our study demonstrated that PL could inhibit fungal biofilm formation, concentrations ≥ 8 μM were found to significantly impede biofilm development ( Figure 6C ). Furthermore, we confirmed that PL exhibited a significant reduction in spore adhesion to HCECs compared to DMSO treatment, with statistical significance observed at concentrations of 2 and 3 μM ( Figures 6D , E ). PL also exhibited its ability to disrupt the cell membrane and cell wall of A. fumigatus . PI staining images revealed a positive correlation between the fluorescence intensity of mycelia and PL concentration, indicating that PL caused slight damage to the integrity of the fungal membrane starting from 2 μM, with a pronounced damaging effect observed at 30 μM ( Figure 6F ). Calcium fluoride white fluorescence staining demonstrated that treatment with PL at concentrations ≥ 10 μM resulted in obviously thinner and shorter mycelial morphology in a concentration-dependent manner ( Figure 6G ). These findings underscored the remarkable antifungal activity possessed by PL.

Figure 6 . PL exhibited remarkable antifungal efficacy. The inhibitory effect of PL on A. fumigatus was assessed at different concentrations within 24 h (A) , and the change in inhibitory rate was monitored over a period of 120 h (B) . Crystal violet chromogenic assay confirmed the inhibitory effect of PL on biofilm formation (C) . PL inhibited spore adhesion in vitro ( D, E ; Magnification, 200×, bar: 200 μm). The antifungal effects of PL treatment at various concentrations were evaluated through PI staining (F) and calcofluor white staining (G) (magnification, 100×, bar: 200 μm). All data were displayed as mean ± SEM (ns, no significance; *** P < 0.001, **** P < 0.0001).

3.7 Both PL and NATA attenuated A. fumigatus keratitis and had synergistic effects

In order to compare the effects of PL and NATA on A. fumigatus keratitis, we treated mice with 30 μM PL, 5% NATA, and PBS. At 5 days p.i., significant reductions of corneal opacity area were observed in both the PL and NATA treatment groups, accompanied by alleviated edema and improved visibility of the iris ( Figure 7A ). The differences were statistically significant ( Figure 7B ). Additionally, synergy experiments were conducted using the checkerboard method ( Figure 7C ). Our findings demonstrated that 16 μM NATA and 32 μM PL can individually achieve MIC 70 . When used in combination, the concentrations of PL and NATA at MIC 70 were both 4 μM. Therefore, the PL-NATA combination had a FICI value of 0.375 indicating a synergistic effect between them. The results from using either PL or NATA alone or in combination are presented in Table 2 .

Figure 7 . Comparison of efficacy and synergistic effect between PL and NATA. Corneal changed at 1 and 5 days p.i. after treatment with PBS, NATA or PL ( A , n = 6/group) and analysis of differences between the three groups (B) . The synergistic effect of different concentrations of NATA (0–16 μM) and PL (1–32 μM) treated for 24 h was analyzed by checkerboard method, and the inhibition rate was calculated after reading OD (C) . All data were displayed as mean ± SEM (ns, no significance; * P < 0.05. D, days p.i.). ** P < 0.01.

Table 2 . FICI values for PL-NATA combination.

4 Discussion

The incidence of fungal keratitis is progressively increasing over the years ( Oliveira dos Santos et al., 2020 ); 5% natamycin is considered the “gold standard” for treating fungal keratitis. Nevertheless, due to the poor dispersibility and low permeability of natamycin, 25% of cases still progress to corneal perforation following treatment ( Hoffman et al., 2022 ).

The major active component of various plants, including Plumbago indica L ., is PL, which has been demonstrated to possess a broad spectrum of biological and pharmacological activities. In vitro studies have shown that low doses of PL are non-toxic or exhibit low toxicity toward normal cutin cells and lens epithelial cells ( Oh et al., 2017 ; Zhang et al., 2021 ). Furthermore, the safety of intravitreal injection of high doses of PL has been confirmed by previous research ( Chen et al., 2018 ). These findings align with our study results and provide evidence for the safety of PL in A. fumigatus keratitis.

It is well-established that following fungal infection of the cornea, hyphae invade the corneal stroma and induce aggregation and infiltration of inflammatory cells. The growth of hyphae in the corneal stroma, along with protease secretion from neutrophil primary granules, lead to degradation of the corneal stroma, resulting in loss of corneal transparency ( Ratitong and Pearlman, 2021 ). Through clinical scoring, plate counting, HE staining, and immunofluorescence, we observed that treatment with PL significantly ameliorated corneal edema, reduced ulcer area, and inhibited fungal growth and inflammatory cell infiltration, thereby improving corneal transparency, and the therapeutic efficacy of NATA in treating A. fumigatus keratitis is consistent with that of PL. Furthermore, previous studies have shown that PL effectively downregulates IL-1β expression in a mouse model of osteoarthritis ( Wenhao Zheng et al., 2017 ), as well as inhibits TNF-α and IL-6 levels in a rat model of chronic periodontitis ( Zheng et al., 2017 ), which aligns with our observations. We observed a significant inhibitory effect of PL on proinflammatory cytokines IL-1β, IL-6, and TNF-α expression in peritoneal macrophages while increasing anti-inflammatory cytokine IL-10 expression. These results suggest that PL exerts its anti-inflammatory effects by altering levels of inflammatory factors in A. fumigatus keratitis. In addition, neutrophils constitute over 90% of the infiltrating inflammatory cells in fungal keratitis ( Ratitong and Pearlman, 2021 ), and previous studies have demonstrated that neutrophils are the primary source of IL-1β during infection ( Peiró et al., 2018 ; Sun et al., 2018 ). The findings of our study demonstrate that PL effectively attenuated neutrophil infiltration in A. fumigatus keratitis. This observation is consistent with the fact that PL reduces neutrophil infiltration in liver tissue during fulminant liver failure ( Wang et al., 2016 ). Therefore, it can be further inferred that the down-regulation of IL-1β levels by PL may also be attributed to its capacity for reducing neutrophil recruitment. In addition, PL had been reported to have anti-inflammatory effects in clinical trials, for example in allergic and COVID-19/uterine corpus endometrial carcinoma patients ( Kohli et al., 2011 ; Li et al., 2021 ). Collectively, these results suggest that PL could potentially enhance the prognosis of fungal keratitis by inhibiting fungal growth and mitigating corneal inflammation in mice.

The extracellular form of HMGB1, known as damage-associated molecular pattern (DAMP), exhibits cytokine and chemotactic activity and plays a proinflammatory role in various disease models ( Kang et al., 2014 ; Tang et al., 2023 ). Numerous studies have demonstrated the effectiveness of targeting HMGB1 as an anti-inflammatory strategy, showing that inhibiting HMGB1 can significantly reduce inflammatory responses in myocarditis, sepsis, periodontitis, and other diseases by down-regulating inflammatory cytokines such as IL-1β, TNF-α, and IL-6 ( Yang H. et al., 2019 ; Jiang et al., 2021 ; Luo et al., 2022 ; Yang et al., 2022 ). Previous research on Pseudomonas aeruginosa keratitis has validated the potential of targeting HMGB1 as an anti-inflammatory approach ( Ekanayaka et al., 2018 ; Hazlett et al., 2021 ). Meanwhile, PL has been shown to effectively inhibit HMGB1 in different disease models, including cholestatic liver injury, hepatic ischemic re-injury, and sepsis ( Zhang Z. et al., 2016 ; Zaki et al., 2018 ; Pan et al., 2022 ). This research prompted us to conduct related experiments on corneal and peritoneal macrophages in mice. We observed that PL effectively suppressed the expression of HMGB1 both in vitro and in vivo , leading to a significant down-regulation of TNF-α, IL-6, and IL-1β. These results confirmed that PL may play an anti-inflammatory role in A. fumigatus keratitis by inhibiting HMGB1. It is well-known that LOX-1 plays a role in increasing the expression of proinflammatory cytokines during A. fumigatus keratitis ( Che et al., 2018 ; Sun et al., 2019 ). It has been reported that HMGB1 is positively associated with changes in LOX-1, IL-1β, and TNF-α in murine models of COPD and acute lung injury ( Liu et al., 2018 ; Xu et al., 2020 ). However, to date, no study has demonstrated a significant correlation between PL and LOX-1. To further investigate the mechanism of action of PL in fungal keratitis, relevant experiments were conducted. We observed that PL treatment not only suppressed the expression of HMGB1 but also attenuated the expression of LOX-1, while concurrently downregulating the levels of inflammatory factors such as IL-1β. Additionally, LOX-1 has the ability to recruit neutrophils ( Yin et al., 2022 ). We have confirmed the efficacy of PL in reducing neutrophil infiltration, indicating a potential correlation between decreased LOX-1 expression and the inhibition of neutrophil infiltration by PL. We further validated the anti-inflammatory mechanism of PL by using an HMGB1 agonist, Boxb. Our findings demonstrated that PL significantly attenuated the enhanced expression of HMGB1 and LOX-1 mediated by Boxb, as well as the elevation of inflammatory cytokines induced by HMGB1. These results suggest that the anti-inflammatory effect of PL is achieved through the inhibition of HMGB1/LOX-1. In addition, excessive intracellular accumulation of ROS can impact multiple cellular signaling pathways, thereby triggering various pathological processes, including inflammatory responses ( Zhang J. et al., 2016 ; Yang G.-G. et al., 2019 ). It is well-established that down-regulating the expression of HMGB1 and LOX-1 can inhibit intracellular ROS levels ( Gao et al., 2016 ; Zhou et al., 2021 ), and in models of liver injury and upper respiratory tract inflammation, ROS can stimulate the release of HMGB1, which subsequently triggers an inflammatory immune response and generates more ROS ( Bystrom et al., 2017 ; Min et al., 2017 ). Therefore, we hypothesize that there might be a cycling of ROS-HMGB1 during the inflammatory response. Previous studies have demonstrated the capacity of PL to attenuate intracellular ROS levels ( Chen et al., 2019 ; Zhang et al., 2020 ). To validate this, we conducted pertinent in vitro experiments and substantiated the reduction of intracellular ROS levels by PL. Consequently, it can be further inferred that the anti-inflammatory mechanism of PL may also involve inhibiting ROS production and disrupting the ROS-HMGB1 cycle.

The initial stage of fungal infection involves the adhesion of spore to the corneal surface ( Niu et al., 2020 ). We conducted experiments on this aspect and discovered that PL effectively inhibited the adhesion of A. fumigatus spores. Previous research has demonstrated that PL can disrupt the structural integrity of bacterial and fungal cell membranes and walls, leading to increased permeability and subsequent cellular demise ( Periasamy et al., 2019 ; Shang et al., 2019 ; Qian et al., 2022 ; Wang et al., 2022 ). A similar effect of PL on A. fumigatus keratitis was also confirmed in our study, where the destruction of the cell membrane and cell wall was enhanced in a concentration-dependent manner. Additionally, during the growth of A. fumigatus , the biofilm produced exhibits protective and adhesive properties, leading to increased drug resistance ( Di Somma et al., 2020 ). However, previous reports have indicated that PL can inhibit biofilm formation ( Qian et al., 2022 ; Wang et al., 2022 ). Therefore, we conducted experiments related to biofilm formation and found that PL significantly inhibited its development. More significantly, PL demonstrated notable antifungal effects against clinical isolates of Candida albicans ( Hassan et al., 2016 ), indicating the potential for clinical application in treating fungal keratitis. These findings suggest that PL possesses multifaceted antifungal effects, which are concentration-dependent, and exhibits promising therapeutic efficacy against A. fumigatus keratitis.

Additionally, the potential synergistic interaction between PL and NATA was investigated. Previous studies have reported a synergistic effect of PL with tetracyclines ( Xu et al., 2022 ). By employing the checkerboard method, our results showed that PL could exhibit synergy effect with NATA against A. fumigatus , thereby reducing the required drug concentrations and improving the prognosis of A. fumigatus keratitis. These findings suggest that PL has the potential to serve as a potent adjunctive therapeutic agent, highlighting its significant prospects for treating A. fumigatus keratitis.

The potential of PL in alleviating A. fumigatus keratitis lies in its ability to inhibit the growth of A. fumigatus , suppress inflammation by regulating the HMGB1/LOX-1 signaling pathway and inhibiting ROS production. Therefore, PL holds promise for the remission and treatment of A. fumigatus keratitis.

Data availability statement

The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.

Ethics statement

Ethical approval was not required for the studies on humans in accordance with the local legislation and institutional requirements because only commercially available established cell lines were used. The animal study was approved by the Ethics Committee at Qingdao University's Affiliated Hospital. The study was conducted in accordance with the local legislation and institutional requirements.

Author contributions

FC: Conceptualization, Methodology, Writing – original draft. LG: Supervision, Writing – review & editing. JL: Data curation, Validation, Writing – review & editing. GL: Data curation, Software, Writing – review & editing. QW: Formal analysis, Supervision, Writing – review & editing. LZ: Data curation, Software, Writing – review & editing. MC: Investigation, Methodology, Writing – review & editing. QX: Funding acquisition, Software, Writing – review & editing. GZ: Funding acquisition, Supervision, Visualization, Writing – review & editing. CL: Funding acquisition, Project administration, Resources, Writing – review & editing.

The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. This work was supported by the National Natural Science Foundation of China (Nos. 81870632, 82171029, and 81800800), the Taishan Scholars Program (No. tsqn202103188), and Natural Science Foundation of Shandong Province (No. ZR2020QH141).

Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Publisher's note

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Keywords: Aspergillus fumigatus keratitis, plumbagin, HMGB1, LOX-1, antifungal

Citation: Cong F, Gu L, Lin J, Liu G, Wang Q, Zhang L, Chi M, Xu Q, Zhao G and Li C (2024) Plumbagin inhibits fungal growth, HMGB1/LOX-1 pathway and inflammatory factors in A. fumigatus keratitis. Front. Microbiol. 15:1383509. doi: 10.3389/fmicb.2024.1383509

Received: 07 February 2024; Accepted: 26 March 2024; Published: 09 April 2024.

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Copyright © 2024 Cong, Gu, Lin, Liu, Wang, Zhang, Chi, Xu, Zhao and Li. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) . The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

*Correspondence: Cui Li, yankelicui@126.com ; Guiqiu Zhao, zhaoguiqiu_good@126.com

† These authors have contributed equally to this work and share first authorship

Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.