Applied Microbiology Research Articles

Heat resistance differences are common between both vegetative cells and spores of Clostridium perfringens type F isolates carrying a chromosomal vs plasmid-borne enterotoxin gene.

Clostridium perfringens type F isolates utilize C. perfringens enterotoxin (CPE) to cause food poisoning (FP) and nonfoodborne gastrointestinal diseases. The enterotoxin gene (cpe) can be located on either the chromosome or plasmids, but most FP isolates carry a chromosomal cpe (c-cpe) gene. Our 2000 article in Applied and Environmental Microbiology (66:3234-3240, 2000, https://doi.org/10.1128/aem.66.8.3234-3240.2000https://doi.org/10.1128/AEM.66.8.3234-3240.2000) determined that vegetative cells and spores of c-cpe isolates are more heat resistant than those of plasmid cpe (p-cpe) isolates, which is favorable for their survival in improperly cooked or held food. However, that 2000 article was recently retracted (90:e00249-24, 2024, https://doi.org/10.1128/aem.00249-24). To our knowledge, the 2000 article remains the only study reporting that heat resistance differences are common between both vegetative cells and spores of type F c-cpe isolates vs type F p-cpe isolates. To confirm and preserve this information in the literature, the heat resistance portion of the 2000 study has been repeated. The 2024 results reproduced the 2000 results by indicating that, relative to the surveyed type F p-cpe isolates, the vegetative cells of surveyed type F c-cpe isolates are ~2-fold more heat resistant and the spores of most surveyed c-cpe isolates are ~30-fold more heat resistant. However, consistent with several reports since our 2000 paper, one surveyed type F c-cpe isolate (which did not appreciably sporulate in 2000 but sporulated in 2024) produced spores with intermediate heat sensitivity, confirming that spores of some type F c-cpe isolates lack exceptional heat resistance.IMPORTANCEClostridium perfringens type F food poisoning (FP), which is the second most common bacterial cause of FP, involves the production of C. perfringens enterotoxin. While the enterotoxin gene (cpe) can be located on either the chromosome or plasmids in type F isolates, most FP cases are caused by chromosomal cpe isolates. The current results support the conclusion that the vegetative cells and spores of type F chromosomal cpe isolates are often more heat resistant than vegetative cells and spores of type F plasmid cpe isolates. Greater heat resistance should favor the survival of the spores and vegetative cells of those chromosomal cpe isolates in temperature-abused food, which may help explain the strong association of type F chromosomal cpe strains with FP.

Electrokinetics with Surface-Enhanced Raman Spectroscopy for Bacterial Identification in Wastewater

Bacterial presence in wastewater serves as a crucial bioindicator for monitoring population-level infections and offering early disease outbreak warnings [1,2]. However, identifying bacteria in wastewater presents outstanding challenges; notably, current culturing or fluorescence-based methods [3] to identify bacteria are slow and expensive, unsuitable for high-throughput screening of diverse bacterial species, and may not work well in the complex wastewater matrix. Here, we harness the potential of surface chemistry and electrokinetics to introduce an innovative electro-optical approach that has the promise to detect a broad spectrum of pathogenic bacteria in wastewater.We combine surface-enhanced Raman spectroscopy (SERS) [4] with alternating current electric fields, complemented by machine learning models [5], to enable rapid and amplification-free bacteria detection in filtered wastewater, at cell concentrations as low as 104 cells/mL. First, we synthesize gold nanorods that can electrostatically bind to bacteria surfaces, allowing for SERS from cell surfaces. We collect SERS from bacteria spiked into filtered wastewater, including Staphylococcus aureus, Staphylococcus epidermidis, and Escherichia coli, spanning concentrations from 109 cells/mL to 104 cells/mL. Spectral clustering analysis shows that bacteria signals become more indistinguishable from wastewater as the concentrations decrease. Further, we incorporate electrokinetic effects into SERS by employing gold microelectrodes to apply electric fields, utilizing dielectrophoresis [6] to rapidly displace and concentrate bacteria based on their dielectric responses within minutes. The enrichment of bacteria with nanorods is directly visualized through microscopy, resulting in Raman signal intensities increased by up to tenfold under electrical fields for bacterial concentrations ranging from 106 down to 104 cells/mL. Such enhancement may enable the detection sensitivity to reach environmentally-relevant concentrations. Moreover, employing data science analysis, we identify biologically-relevant “fingerprint” Raman peaks characterizing proteins, nucleic acids, and lipids from bacteria surfaces, allowing for rapid identification of bacteria species in wastewater. This novel method exhibits potential for generalized pathogen detection and molecular recognition in complex liquid samples, such as wastewater, blood, and seawater.[1] Hellmér, et al. Applied and environmental microbiology. (2014).[2] Keshaviah, et al. The Lancet Global Health. (2023).[3] Jahn, et al. Nature Microbiology. (2022).[4] Tadesse, et al., Nano Lett. (2020).[5] Ho, et al., Nat. Comm. (2019).[6] Pethig. John Wiley & Sons (2010).

Environmental microbiology explains the largest positive carbon isotope excursion in Earth history, the Lomagundi-Jatuli Event.

The geological record of stable carbon isotopes preserved in marine carbonate rocks spans nearly 4 billion years. Numerous perturbations mark this record, but one stands out for its magnitude, the Lomagundi-Jatuli Event, which spanned the transition of the Earth's surface from an anoxic to an oxic state. An Applied and Environmental Microbiology article by D. Y. Sumner (90:e00093-24, 2024, https://doi.org/10.1128/aem.00093-24) provides, for the first time, a biological explanation for its initiation, cessation, environmental specific restriction, and geological singularity.

In vivo Antiplasmodial Assessment of Phyllanthus odontadenius against Plasmodium berghei in NMRI Mice

Aims: Phyllanthus odontadenius is one of the genus Phyllanthus species, used for number diseases treatment including malaria. Malaria today poses a real public health problem for more than hundred countries, afflicted millions people and killed an estimated 405 000 in the World. The chemoresistance of Plasmodium falciparum to modern antimalarials which are either expensive, toxic or ineffective leads to the search for new antimalarials at lower cost, non-toxic and effective within plant biodiversity. Given the interest in various P. odontadenius crude extracts in vitro on P. falciparum, an in vivo study seems necessary in order to judge the extracts effectiveness of this plant. This study therefore aims to find justification for P. odontadenius secondary metabolites antiplasmodial activity which would have been revealed in the three samples from three different province sites. Study Design: Plants P. odontadenius samples from three province harvested and dried separately, plant phytochemistry screening realized, extracts preparation for antiplasmodial test, mice parasitization with P. berghei strain, orally administration drug; Parasiteamia determination using an immersion microscope. Place and Duration of Study: Department of Radiobiology, Applied Microbiology section, General Atomic Energy Commission, Regional Nuclear Studies Center of Kinshasa. MPI and pharmacognosy laboratories in the National Biomedical Research Institute (INRB). This work took place over the period from October 11, 2020 to March 12, 2021. Methodology: Phytochemical screening P. odontadenius samples was previously determined with the chemical reagent reactions and TLC. Then, P. odontadenius methanol extracts from aerial parts harvested in three sites (Kinshasa, Kasangulu and Kwango-bridge) were administered to test mice (12.5 mg/kg and 25 mg//kg bw) after infected mice with the Plasmodium berghei strain. DMSO 10% and quinine 10 mg/kg bw were also used as controls for comparison with the samples of P. odontadenius extracts. After 5 days, parasitemia of each test and controls mice was determined. Percent of parasitemia, parasite density and percent of inhibition were calculated. Finally, the effect dose 50 of each P. odontadenius specimen was finally determined. Results: Parasiteamia rates of negative control (DMSO 10%) was high (69.98±15.03%) comparing to positive control (27.43±11.46%) and tested mice with P. odontadenius extracts (12.5 and 25 mg/kg bw) which percent’s varied from 24.66±15.84% to 59.01±22.44%. Negative control presented high parasite density with 11,342 (±2,436) comparing to the positive control (4,447±1,857) and all P. odontadenius methanol extracts which varied from 3,995±2,343 for 25 mg/kg bw to 9,570±3,319 for 12.5 mg/kg bw. Parasiteamia reduction rates followed inversely parasite density, thus, Po3 25 mg/kg bw had high parasiteamia reduction rate (65.23%) comparing to positive control with 61.32% and to P. odontadenius methanol extracts. Po3 presented 2.44 mg/kg bw as effect dose 50 comparing to Po1 (2.93 mg/kg bw) and Po2 (2.68 mg/kg bw). Males mice were highly affected to P. berghei than the females. Conclusion: This study revealed that all specimens of P. odontadenius had good in vivo antiplasmodial activities on P. berghei. All P. odontadenius extracts showed good parasitaemia inhibition compared to negative control, but P. odontadenius from Kwango-brigde (Po3) presented good behavior concerning in vivo antiplasmodial activity in comparison to P. odontadenius from Kasangulu (Po2) and that from Kinshasa (Po1). However, further studies are necessary on the in vivo toxicity of the plant and on the medicinal form that could be applied.

First Report of White Mucus Disease Caused by Fuligo gyrosa on Agrocybe chaxingu in China.

Agrocybe chaxingu is an edible and medicinal mushroom widely cultivated in China (Liu et al. 2021). Agrocybe chaxingu is extremely well-liked for the unique flavor and nutritional value. In May 2021, a serious white mucus disease was observed in the farms of A. chaxingu in the Ganxian district of Ganzhou City, Jiangxi Province, China, with an approximate disease incidence of 20%. In the years of 2022 and 2023, the same white mucus disease on A. chaxingu was observed in the farms in Nanchang City, Jiujiang City and Guangchang County, Jiangxi Province, China. The disease generally occurs on the media, stipe or pileus of A. chaxingu under condition of high humidity. The plasmodial slime molds migrated from the surface of culture media (78% hardwood sawdust, 15% wheat bran, 5% tea seed shell, 1% lime, and 1% gypsum) to the base of fruiting bodies, stipes and finally to pilei, showing as moist, sticky, and white reticulated structures. The infected fruiting bodies of A. chaxingu were completely covered by reticulated plasmodia, displaying a white or pale-yellow color. This resulted in the growth cessation, wilting and eventual death of fruiting body. Microscopic observation found that the plasmodia of slime mold enveloped the hyphae of A. chaxingu, resulting in the fragmentation of the hyphae. The disease can spread quickly, resulting in a 30% reduction in production. Slime mold cultures were isolated by transferring diseased fruiting bodies of A. chaxingu onto oat-agar medium (2% agar and 1% oatmeal) at 25 ℃. The isolates can be obtained after being subcultured for two to three generations. Purified plasmodia were placed on the semi-defined medium (1% tryptone, 1% glucose, 0.15% yeast extract, chick embryo extract and a balanced salt solution) to confirm the absence of bacteria (Daniel et al. 1964) and thus obtained the pure culture. Specimen of the voucher has been deposited in the Institute of Agricultural Applied Microbiology, Jiangxi Academy of Agricultural Sciences as number IAAM-W0002. The vegetative plasmodia have a large and well-developed scalloped structure that were white or milky white in colour. The white plasmodium became opaque pale yellow when exposed to light before fruiting. The veins merged and thickened. Fruiting bodies can be formed on the lid or side of the Petri dish under light condition. The fruiting bodies formed papillae with irregular shape, and then the color changed from translucent yellow to greyish black. Spores were usually spherical or subglobose, free, greyish brown in mass, purplish brown, 7-12 μm in diameter under light microscopy. These morphological characteristics were found to be consistent with those of Fuligo gyrosa (Synonym: Physarum gyrosum) (Kim et al. 2009; Shi et al. 2005; Jahn 1902). The identity of the isolates was further confirmed by sequence analysis of the 18S ribosomal RNA gene with primer SMNUR101/NS4 (Rusk et al. 1995; White et al. 1990). Using BLASTn searches, the sequence of 18S rRNA gene (GenBank accession number OR186216) matched the sequence of F. gyrosa (GenBank accession number LC744593) with the identity of 99.91% and coverage of 97%. A phylogenetic tree based on the 18S rRNA gene also demonstrated that the slime mold clustered with F. gyrosa. Over ten isolates have been obtained from the diseased A. chaxingu samples in different factories and identified as F. gyrosa. To test the pathogenicity of F. gyrosa, five healthy young fruiting bodies (three to five days of primordium) of A. chaxingu cultivated in mushroom-growing room were gently inoculated by a 12 mm diameter oat-agar medium with plasmodia at 24 ± 2 ℃ and then were kept with relative humidity of 90%-95%. Five fruiting bodies inoculated with a 12 mm oat-agar medium served as controls. After 5 days, white mucus characteristics and three fifths of death symptoms were observed on the fruiting bodies inoculated with the plasmodia, while the controls remained asymptomatic. The slime mold on the inoculated fruiting bodies was morphologically identical to F. gyrosa that was observed on the initial diseased fruiting bodies. It was also observed the envelopment A. chaxingu hyphae by the plasmodia of slime mold and fragmentation of the hyphae, and the fragmentation was not observed in the controls. Reisolations were prepared from the inoculated fruiting bodies and confirmed to be F. gyrosa based on morphological characteristics and 18S rRNA sequence, thus fulfilling Koch's postulates. Fuligo gyrosa has been reported to cause severe disease in oriental melon in Korea (Kim et al. 2009). This is the first report of F. gyrosa causing white mucus disease in cultivated A. chaxingu. The findings will provide important information on prevention and control of the disease, and be helpful for the development of A. chaxingu industry.

Meeting Report: Microbes as Safeguards of the Environment

Abstract From 12 to 14 March, a gathering of microbiologists and biotechnologists convened at the International University of Andalucía (UNIA) in Baeza (Jaén, Spain) under the auspices of UNIA and the Applied Microbiology International (AMI) Society. The primary objective of this meeting was to analyse the pivotal role microbes play in sustaining our planet in a broader context—both from the descriptive (what is the state of affairs) and the prescriptive perspective (what to do and look for regarding activities of interest). The workshop focused on exploring the ecological aspects of microbes in soil and water, including some extreme environments, the potential of microorganisms as promoters of plant growth and biocontrol agents as well as active large-scale catalysts for environmental health. Various topics were examined in this context, encompassing the application of microbes as platforms for the biosynthesis of value-added chemicals, bioremediation technologies, the concept of the circular economy, the emergence of zoonotic concerns within a dynamically changing global environment and the role of microbes in precision nutrition and precision medicine. In alignment with the strategic plan of AMI, the discussion was focused on the background of the United Nations Sustainability Development Goals (UN SDGs). These targets were formulated at the beginning of the past decade to guide humanity towards a sustainable future. The UN SDGs aim to prevent the overstepping of planetary boundaries, which are endangering Earth's biodiversity and even human survival.

Assessment of Artificial Intelligence Platforms With Regard to Medical Microbiology Knowledge: An Analysis of ChatGPT and Gemini.

The performance of two artificial intelligence (AI) platforms, ChatGPT 3.5 (OpenAI, California, United States) and Gemini (Google AI, California, United States) was assessed by answering 200 questions of microbiology drawn from validated sources. The questions were selected from topics such as General Microbiology, Immunology, and Microbiology Applied to Infectious Diseases. The study was conducted from December 2023 to March 2024, and the responses of the different AI platforms were compared with an answer key. Statistical analysis was performed to assess accuracy. ChatGPT 3.5 and Gemini had comparable accuracy with correct response scores of 71% and 70.5%, respectively. Their performance varied across different sections. Gemini performed better in General Microbiology and Immunology, and ChatGPT 3.5 had a better score in the Applied Microbiology section. The study's findings highlight that AI platforms such as ChatGPT and Gemini can be utilized in microbiology and medical education.The evolution and continuous updating of AI platforms are required to improve their performance.

Antifungal potential of volatiles produced by Aspergillus neoniger F14 against Ganoderma boninense

Ganoderma boninense Pat. is a fungal pathogen that primarily afflicts oil palm, leading to decreased palm oil yield and significant economic losses. The recent focus on biocontrol agents producing acids and volatile compounds has gained momentum due to their straightforward synthesis and ease of interaction with the phytopathogen. This research is a component of a project aimed at exploring the local biodiversity of antagonistic fungi in the oil palm plantation environment, with a specific focus on volatile organic compounds (VOCs) detected using Gas Chromatography-Mass Spectrometry (GC-MS). Isolate F14, a collection in the Laboratory of Applied Microbiology, BRIN, Indonesia was identified as Aspergillus neoniger, a novel report as a soil-originating fungal community in Indonesian oil palm plantations. VOC profiling using GC-MS revealed five dominant compounds namely (Z)-9-Tricosene, 1,2,3,4,5-Pentamethylcyclopentane, Dihexyl carbonate, Pentadecane, and Tridecyl laurate. Based on aromatogram disc tests against G. boninense colonies, it was determined that VOCs from A. neoniger F14 inhibited growth by 48% on the fifth day. The findings of this study contribute supportive information for further investigations into VOCs produced by other local antagonistic isolates against G. boninense.

Evaluating the National Action Plan (NAP) on antimicrobial resistance, and recommendations for the next 5-year NAP: a roundtable discussion

Abstract The UK’s current 5-year National Action Plan (NAP) on tackling antimicrobial resistance (AMR), which runs from 2019 to 2024, is coming to an end. As such, the next 5-year NAP is currently under development, and progress made towards the UK’s commitments on AMR is being analysed. This analysis has involved the publication of an addendum to the current NAP, which includes some new and revised commitments. Applied Microbiology International (AMI) held a roundtable discussion in November 2022, inviting UK experts from across the human and animal sectors to discuss their thoughts on progress to date, as well as any recommendations and key considerations they thought should be taken forward. Overall, it was agreed that there is a major disparity between the increasing ambitions of the NAP, and the reducing financial support for achieving said ambitions. The gap between evidence generation and implementing recommendations into real-world settings also presents a key challenge to overcome, if the NAPs commitments are to be achieved. A lack of communication and funding appear to underpin many of the barriers that prevent the successful delivery of current NAP commitments and are therefore highlighted as key areas of concern for policymakers to consider when looking to tackle AMR. The focus should be on successfully implementing and delivering the current NAP commitments ahead of identifying future ambitions.

Antibiotic Resistance Profiles of Salmonella and Escherichia coli Isolates Obtained from Garden Eggs Sold in Abakaliki Metropolis: Insights into Susceptibility Patterns

This investigation aimed to evaluate the antibiotic susceptibility profiles of Salmonella species and Escherichia coli isolated from 100 garden egg samples obtained from diverse vendors in Abakaliki. Samples were transported to the Department of Applied Microbiology Laboratory, Ebonyi State University, Abakaliki, for analysis. Organisms were isolated and identified using standard microbiological techniques, while their antibiotic susceptibility was assessed through the disc diffusion method. Results indicated a prevalence of 60% for Salmonella species and 80% for E. coli among the analyzed samples. The antibiotic susceptibility testing unveiled alarming resistance patterns: Salmonella species displayed complete resistance (100%) to meropenem, perfloxacin, colistin, aztreonam, neomycin, gentamycin, ciprofloxacin, chloramphenicol, and ofloxacin, with only 16.67% susceptibility to streptomycin. Similarly, E. coli isolates exhibited 87.5% susceptibility solely to ofloxacin and 12.5% to chloramphenicol, while showcasing absolute resistance (100%) to other antibiotics tested. This study underscores the role of garden egg sales and consumption in facilitating the widespread dissemination of multi-drug resistant bacterial pathogens within the region. It emphasizes the urgent need for stringent adherence to hygienic practices among garden egg and other fruit vendors as a crucial preventive measure to curb this escalating threat. Keywords: Garden eggs, Salmonella species, E. coli, antibiotic resistance

Bacillus thuringiensis pesticidal toxins: A global analysis based on a scientometric study (1980–2021)

Several studies have been conducted on Bacillus thuringiensis (Bt) pesticidal toxins due to their successful environmentally friendly biopesticide activity against various insect pest orders, protozoa, mites, and nematodes. However, no existing study has systematically examined the trends and evolution of research on Bt pesticidal toxins from a scientometric perspective. This study aimed to analyze the trends and hotspots of global research in this field. 5757 publications on Bt pesticidal toxins were extracted from the Web of Science Core Collection (WoS) from 1980 to 2021. Statistical and scientometric analyses were performed using Excel, CiteSpace, and VOSviewer visualization tools to evaluate research evolution, journal contribution and subject categories, contributing countries and institutions, highly influential references, and most used author keywords. The 5757 publications featured in 917 journals spanning 116 subject categories. The top 5 subject categories ranked as Entomology, Biotechnology & Applied Microbiology, Microbiology, Biochemistry & Molecular Biology, and Agriculture. Out of these publications, the USA contributed the most, with 1562 publications, 72,754 citations, and 46.58 average citations per paper (ACPP); however, Belgium had the highest (106.43) ACPP among the top 20 contributing countries. The Chinese Academy of Agricultural Sciences is the leading institution with 298 publications and 21.20 ACPP. The Pasteur Institute is ranked first (90.04) in terms of ACPP. Keywords analyses revealed that recent studies are inclined toward the evolution of insect resistance against Bt toxins. In future, studies related to the development of resistance mechanisms by insects against Bt pesticidal toxins and ways to overcome them will likely receive more attention. This study highlights the past and current situations and prospective directions of Bt pesticidal toxins-related research.

In vitro Cytoprotective Activity Study of Cocoa (Theobroma cacao L.) Pod Husk Ethanolic Extract against Blue Light Irradiation

Pharmacognosy Research,2023,15,3,537-543.DOI:10.5530/pres.15.3.056Published:June 2023Type:Original Article Authors:Shakila Angjaya Adiyanto, Erika Chriscensia, Elizabeth Chrestella Wibowo, Joshua Nathanael, Audrey Amira Crystalia, Agus Budiawan Naro Putra, Gunawan Priadi, Fahrurrozi Fahrurrozi, and Pietradewi Hartrianti Author(s) affiliations:Shakila Angjaya Adiyanto1,#, Erika Chriscensia1,#, Elizabeth Chrestella Wibowo1, Joshua Nathanael1, Audrey Amira Crystalia1, Agus Budiawan Naro Putra2, Gunawan Priadi3, Fahrurrozi Fahrurrozi4, Pietradewi Hartrianti1,* 1Department of Pharmacy, School of Life Sciences, Indonesia International Institute for Life Sciences, Jakarta, INDONESIA. 2Research Center for Pharmaceutical Ingredients and Traditional Medicine, National Research and Innovation Agency, Cibinong, INDONESIA. 3Research Center for Applied Microbiology, National Research and Innovation Agency, Cibinong, INDONESIA. 4Research Center for Marine and Land Bioindustry, National Research and Innovation Agency, Mataram, INDONESIA. #These authors have contributed equally. Abstract:Background: Blue light exposure has been shown to induce ROS generation and subsequent inflammatory pathways that lead to cell death, in which antioxidants could counteract this effect. Although regarded as waste, cocoa (Theobroma cacao L.) Pod Husk (CPH) possesses a high polyphenolic contents, which could serve as antioxidants. Objectives: To characterize CPH ethanolic extract based on its antioxidant capacity and observe the cytoprotective ability of CPH in vitro upon blue light exposure. Materials and Methods: CPH ethanolic extract was characterized through total phenolic content, total flavonoid content, and three antioxidant assays, then treated on HaCaT cells, in which cell viability was measured through MTS assay. Results: CPH extract showed high phenolic and flavonoid contents and high antioxidant capacity through DPPH and FRAP assay. CPH extract started to exert cytotoxicity from concentrations of 400 μg/mL and above, while 100 μg/mL and above in AA. Furthermore, CPH extract showed significant cytoprotective effect at 50 μg/mL at 11.92 ± 0.83% cell viability increase, wherein AA failed to protect HaCaT cells at the same concentration at 15.79 ± 0.72% cell viability decrease. Conclusion: CPH could serve as an alternative as blue light protection agent as it was safe to be used at higher concentrations and was able to protect HaCaT cells from blue light irradiation better than AA. Keywords:Antioxidant, Blue light, Cell Culture, Cocoa pod husk, Cocoa pod husk cytoprotectivity against bluelight., HaCaTView:PDF (426.83 KB)

Evaluation of the applicability of immunochromatography to the identification of live plague vaccines and the tularaemia allergen (Tularin)

The regulatory standards require that the identification of live plague vaccines and the liquid tularaemia allergen (Tularin) should be performed by immunofluorescence. A major drawback of the recommended method is its labour intensive nature. However, immunochromatography represents an alternative method that offers a number of advantages, including rapid testing and easy result interpretation. The aim of the study was to assess the applicability of immunochromatography to the identification of live plague vaccines and the liquid tularaemia allergen (Tularin).Materials and methods. The authors performed identification tests using samples of the pharmacopoeia standard for live plague vaccines, three commercial batches of a live plague vaccine, and two batches of the liquid tularaemia allergen (Tularin). These samples were tested using immunochromatographic assay (ICA) reagent kits for rapid detection and identification of Yersinia pestis (ICA System for Y. pestis) and Francisella tularensis (ICA System for F. tularensis) manufactured by the State Scientific Center for Applied Microbiology and Biotechnology.Results. The findings show that immunochromatography is an effective, rapid, and species-specific method to confirm the presence of Y. pestis in a sample of a live plague vaccine or F. tularensis in a sample of the liquid tularaemia allergen (Tularin). To perform identification tests by immunochromatography, the authors recommend diluting live plague vaccine samples to a concentration of 109 bacterial cells/mL and using undiluted samples of the liquid tularaemia allergen (Tularin).Conclusions. The study results may support the inclusion of ICA into the regulatory standards for live plague vaccines and the liquid tularaemia allergen (Tularin) as an alternative identification method.

Comparative Analysis of Research Output of Tamil Nadu Agricultural University as Reflected in Scopus and Web of Science

The study presented in the paper has analyzed the research productivity of faculty members of Tamil Nadu Agricultural University for the period of 2001-2020. Total of 4372 publications in Scopus and 1980 publications in web of Science database were retrieved and compared the performance of the faculty members. This study analyzed the scientific parameters such as year-wise distribution of research productivity, most prolific authors, funding sponsors, Collaborating Institutions, Collaborating Countries and top-ranked sources preferred by the agricultural faculty members for publishing their research output. Tamil Nadu Agricultural University faculty members published more publications in Scopus 4372 compared to Web of Science 1980. Articles are the predominant publications by the faculty members of Tamil Nadu Agricultural University, Coimbatore 3928 in Scopus and 1822 in Web of Science Prof. Samiyappan R is the predominant and most impactful author of Tamil Nadu Agricultural University.USA is the most predominant country collaborated with Tamil Nadu Agricultural University publications about 187 in Scopus and 137 in Web of Science Legume Research is the most predominant journal with 69 publications in Web of Science and Electronic Journal of Plant Breeding 448 publications in Scopus. Agricultural College and Research Institute, Madurai is the most collaborating institution with 538 publications in Scopus and Kansas State University in Web of Science database.Plant Sciences (499), Agronomy (490), Biotechnology Applied Microbiology (190), Horticulture (159) and Environmental Science (129) are the predominant subject areas focused by the faculty members of Tamil Nadu Agricultural University in Web of Science and Agricultural and Biological Sciences subject category in Scopus 3317 publications. Indian Council of Agricultural Research is the predominant sponsor and supported for 104 research publications followed by Department of Biotechnology, Ministry of Science and Technology, India.

Recent Advances in Applied Microbiology and Food Sciences, Volume II

The food industry is constantly evolving due to environmental, social, and geopolitical factors [...]

The shifting research landscape for PAH bioremediation in water environment: a bibliometric analysis on three decades of development.

Polycyclic aromatic hydrocarbons (PAHs) with their carcinogenic, teratogenic, and mutagenic effects can cause great damage to the ecosystem and public health when present in water. With bioremediation, PAH contamination in water environment can be greatly reduced in an eco-friendly manner. It has thus become the research focus for many environmental scientists. In this study, a bibliometric analysis on three-decade (1990-2022) development of PAH bioremediation in water environment was conducted from temporal and spatial dimensions using CiteSpace. A total of 2480 publications, obtained from Web of Science core collection database, were used to explore the basic characteristics, hotspots, and prospects of the research area. The results showed that (1) bioremediation/biodegradation of PAHs in water environment has been getting researchers' attention since 1990, and is gaining even more traction as time goes on. (2) In terms of countries, China and the USA were the major contributors in this research area, while at the institutional level, the Chinese Academy of Sciences has produced the most research results. However, international cooperation across regions was lacking in the field. (3) Environment Science and Technology, Chemosphere, Applied and Environment Microbiology, Journal of Hazardous Materials, and Environment Pollution were the 5 most cited journals in this field. (4) There were three major stages the field has gone through, each with distinct research hotspots, including initial stage (1990-1994), mechanism investigation (1995-2000), and application exploration (2001-2010; 2011-2022). Finally, research perspectives were proposed, covering three directions, namely, bioavailability, immobilization, and viable but nonculturable (VBNC) bacteria.

First Report of Neopestalotiopsis Disease in Ohio caused by an emerging and novel species of Neopestalotiopsis on Strawberry.

In October 2021, strawberry (Fragaria x ananassa) plants (cv. Ruby June) that had dark brown lesions with a diffuse black halo and light brown center and / or dark brown V-shaped necrotic areas often starting from the edge of the leaves were observed in a commercial planting in Washington County. The grower reported 50% incidence in the field when the sample was first submitted and two weeks later reported 80% incidence. The morphology of conidia present on symptomatic leaf tissue was consistent with species of Neopestalotiopsis (Maharachchikumbura et al. 2014). The conidia were ellipsoid to fusiform, five-celled, with three light brown colored median cells and one hyaline apical and basal cell. The apical cells had two to four flexuous appendages and the basal cell had one non-flexuous appendage. Average (N=30) conidia length, not including the appendages, and width was 24.1 ± 2.7 and 6.5 ± 1.4 µm respectively. Two isolates (MLI267-21 and MLI268-21) were purified on potato dextrose agar, producing a dense white mycelial mat with undulate margins. The underside color of the mycelial mat was pinkish-orange. Conidiomata formed randomly in the colonies and extruded black gelatinous spores. To confirm the identity of these isolates the genome of MLI267-21 was sequenced using the NextSeq 2000 Illumina platform and Nextera DNA CD indexes (OSU Applied Microbiology Service Laboratory, Columbus, OH). Partial internal transcribed spacer (ITS) region, β-tubulin (TUB), and translation elongation factor 1-alpha (TEF-1α) gene sequences (Accession numbers: OM649904, OM649905, and OM649906 respectively) were extracted from the MLI267-21 genome, concatenated, and aligned to published reference sequences. These same genes were amplified and sequenced from MLI268-21. Maximum likelihood phylogenetic analysis performed in IQ-TREE (Minh et al. 2020, Kalyaanamoorthy et al. 2017, Chernomor et al. 2016) with the aligned sequences revealed the clustering of MLI267-21 and MLI268-21 with seven other Neopestalotiopsis isolates, from strawberry (17-43L; Baggio et al. 2021) and pomegranate (GEV3426 to GEV3431; Xavier et al. 2021) leaves in Florida, which form a unique and emerging species group. The ITS, TUB, and TEF-1α sequences from both Ohio isolates were 100% similar to the same sequences from 17-43L and GEV3426 - GEV3431. Pathogenicity tests were performed using MLI267-21 by spray inoculating (~104 spore/ml) four-week-old 'Cabrillo' strawberry plants (n=4) and placing three drops (10µl each) of spore suspension (~104 spore/ml) on the calix area of detached fruit (n=4). Non-inoculated plants and fruit (n= 4 each) served as negative controls. The plants were covered with transparent plastic bags and maintained at 25 °C for 72 hours before the bags were removed (Baggio et al. 2021). Five days post-inoculation, dark brown circular spots on the leaves and petioles were observed on all four inoculated plants and acervuli were observed within the necrotic spots after an additional 72 hours in a moist chamber. Fruits were incubated in a moist chamber at 25 °C and after 72 hours orange-brown lesions formed on the fruit. After five days, fruit were mushy and covered with white mycelia, acervuli, and conidiomata. Neopestalotiopsis disease has been reported on strawberry in Florida (Baggio et al. 2021) and in several South American (Obregon et al. 2018, Hidrobo et al. 2021) and European (Chamorro et al. 2016, Gilardi et al. 2019) countries. The disease can cause rapid plant death when conditions are warm and wet. Research to investigate host susceptibility and to identify effective chemical and biological control has been initiated in Ohio to establish preventative management programs for commercial field operations.

Fungal Production of Cellulase Using Fruit Waste as Economical Growth Substrates

Background: Vegetables and fruits are playing an important role in human life and diet. For eco-friendly usage the fruit peel wastes can be used as nutrients for microorganisms and animals as they consist of various growth promoting factors. Cellulolytic enzymes convert cellulose into simpler sugars. Due to suitable cellulase titers and rapid growth bacteria and fungi are becoming choice of interest. Aim: Production of fungal cellulase by using different types of fruit wastes. Method:This descriptive study was conducted by collecting the Fruit wastes (mango peels, melon peels, orange peels and watermelon rind) from fruit processing shops and transported to Applied and Environmental Microbiology laboratory.Inoculum was prepared by using basal medium and fruit peel powder.Descriptive statistical analysis of data will be performed. One way Analysis of Variance (ANOVA) will be applied for comparing mean values among species using Minitab 16. P < 0.05 will be considered significant. Result: The statistical analysis showed that in solid state fermentation the enzyme summary within 10 days was (181.32±0.00 μg/ml/min) after every 24 hour was (9.40±1.22μg/ml/min) and (0.39±0.05μg/ml/min) after every 1 hour and (0.01±0.00μg/ml/min) at 1 minute. The statistical analysis showed that in sub-merged fermentation the enzyme summary within 10 days was 191.83±4.96 after every 24 hour was (7.12±0.76μg/ml/min) and (0.30±0.03μg/ml/min) after every 1 hour and (0.0.00±0.00 μg/ml/min) at 1 minute. Conclusion: The present study clearly indicates the potential of Rhizopussp as the best producer of cellulolytic enzymes. Regarding the substrate fruit peels (mango peels, melon peels, orange peels, and watermelon rind) can be used for the efficient production of cellulase. Keywords: Rhizopus, cellulolytic enzymes, fruit peels, fungal production.

Acknowledgment to the Reviewers of Applied Microbiology in 2022

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Innovations and developments in single cell protein: Bibliometric review and patents analysis.

Global demand for food products derived from alternative proteins and produced through sustainable technological routes is increasing. Evaluation of research progress, main trends and developments in the field are valuable to identify evolutionary nuances. In this study, a bibliometric analysis and search of patents on alternative proteins from fermentation processes was carried out using the Web of Science and Derwent World Patents Index™ databases, using the keywords and Boolean operators "fermentation" AND "single cell protein" OR "single-cell protein." The dataset was processed and graphics generated using the bibliometric software VOSviewer and OriginPro 8.1. The analysis performed recovered a total of 360 articles, of which 271 were research articles, 49 literature review articles and 40 publications distributed in different categories, such as reprint, proceedings paper, meeting abstract among others. In addition, 397 patents related to the field were identified, with China being the country with the largest number of publications and patents deposits. While this topic is largely interdisciplinary, the majority of work is in the area of Biotechnology Applied Microbiology, which boasts the largest number of publications. The area with the most patent filings is the food sector, with particular emphasis on the fields of biochemistry, beverages, microbiology, enzymology and genetic engineering. Among these patents, 110 are active, with industries or companies being the largest depositors. Keyword analysis revealed that the area of study involving single cell protein has included investigation into types of microorganisms, fermentation, and substrates (showing a strong trend in the use of agro-industrial by-products) as well as optimization of production processes. This bibliometric analysis provided important information, challenges, and trends on this relevant subject.