Prof. Chen Chin Chang
Adjunct Professor, Hebei Normal University, China.

Short Biosketch

ISBN 978-81-19761-72-2 (Print)
ISBN 978-81-19761-14-2 (eBook)
DOI: 10.9734/bpi/ramb/v7

This book covers key areas of microbiology and biotechnology. The contributions by the authors include dissimilatory arsenate-reducing bacteria, arsenic contamination, arsenic toxicity, anthropogenic activity, fossil fuels, sulfide minerals, DNA immortalization, homemade ultra-rapid thermal cycler, homemade inexpensive transformation device, stable surveillance kit, DNA synthesis, rapid detection, DNA isolation, midgut binding activity, vector-borne diseases, mosquitocidal protoxin crystals, feathery mottle virus, chlorotic stunt virus, multistep oncogenesis, adult T-cell, carbon anhydrase ix, hypoxia responsive element, human T-cell leukemia viruses, T-cell immunoreceptor, c-c chemokine receptor type-4, phytoconstituents, oxidative pathophysiology, oxidative stress, malaria parasites, antimalarial drugs, bioactive compounds, novel markers, DNA polymorphisms, marker-assisted selection, gender determination. This book contains various materials suitable for students, researchers and academicians in the field of microbiology and biotechnology.


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Midgut Binding Activity of Mosquitocidal Extracellular Protein of Pseudomonas fluorescens Strain

B. Usharani , R. Venkateswari , P. Suganthi , M. Muthuraj

Research Advances in Microbiology and Biotechnology Vol. 7, 27 September 2023, Page 1-25

This chapter aimed to describe the preparation of purified midgut BBMV using whole larvae and pupae of Cx. quinquefasciatus as a starting material and investigated the in-vivo binding of the mosquitocidal protein of P. fluorescens Migula B426 to the mosquito larvae midgut through immunohistochemical methods to determine this protein's primary site of action.

Upon ingestion by a susceptible mosquito larva, the alkaline midgut environment promotes solubilization of crystalline inclusions releasing the protoxins. Subsequent cleavage by gut proteases results in formation of active toxins. The activated toxin fragments then bind to specific protein receptors on midgut epithelial cells, leading to membrane insertion and pore formation. The presence of 55kDa and 35kDa bands in western blot assay revealed the binding of mosquitocidal protein to the midgut of treated larvae and pupae of Cx. quinquefascistus. The mosquitocidal protein attaches preferentially to the midgut of larvae and pupae of the mosquito species Cx. quinquefasciatus, according to the immunofluorescence localization research. The present investigation confirmed that the binding of mosquitocidal protein to the gut regions of the pupae (non-feeding stage) by degrading the cuticle and overcoming the peritrophic membrane and thus found to be effective pupicidal activity. It has been reported that the binding of the protein to the midgut epithelium causes swelling of mitochondrial and endoplasmic reticulum and enlargement of vacuoles, followed by lysis of epithelial cells, midgut perforation and the death of the larvae. It was discovered that the treated larvae and pupae had lower alkaline phosphatase and amino peptidase activity than the untreated larvae and pupae. The findings showed that the enzymatic machinery of midgut cells was compromised and the specific activity of the enzymes was decreased as a result of protein binding to receptor molecules on Brush Border Membrane Vesicles of Cx. quinquefasciatus.

The objective of this review is to explain the multistep oncogenesis of adult T-cell leukemia/lymphoma (ATL). Human T-cell leukemia virus type 1 (HTLV-1) is a causative agent of adult ATL. Both ATL and HTLV-1-associated myelopathy (HAM) are caused by the type C RNA retrovirus known as the HTLV-1. It has remained a mystery as to why the HTLV-1-derived proteins may not have played significant roles in the conclusion of ATL's oncogenesis. Oncogenic processes of ATL are highly complicated, and there is an enigma that the HTLV-1-derived proteins Tax and HBZ may not play major roles in completion of its oncogenesis. Tax activates several targeted genes and molecules during the early polyclonal stage, but HBZ regulates and inhibits Tax and its actions during the intermediate stage. Additional oncogenic activities in host cells complete the oncogenesis of ATL at the monoclonal stage. In particular, the nuclear factor kappa B (NF-\(_K\)B)/hypoxia inducible factor (HIF)/carbon anhydrase IX (CA9) axis and the phosphatidylinositol 3-kinase (PI3K)/HIF/CA9 axis play essential roles in ATL oncogenesis. The in vitro experimental studies with CA9 inhibitors also present new therapeutic approaches for treating ATL.

This book review aims to introduce the current status of arsenic contamination in the environment and a biological arsenic treatment method. Arsenic is very toxic to living organism and cause high potential risk to human and animal health. Thus, the author conducted a comprehensive and critical review of the existing database on arsenic contamination worldwide in water and soil. Additionally, the characteristics of dissimilatory arsenate-reducing bacteria (DARB) as an example of biological arsenic treatment are summarized. DARB are known to contribute to the mobilization of arsenic and other elements from minerals. Despite this, metabolic capabilities of only a few DARB strains have been thoroughly investigated so far, and the influence of these bacteria on the bioavailability of arsenic in the environment is still a topic for discussion. In addition, evidence of microbial growth on arsenate is presented based on isolate analyses, after which a summary of the physiology of the following arsenate-respiring bacteria is provided: Chrysiogenes arsenatis strain BAL-1T, Sulfurospirillum barnesii, Desulfotomaculum strain Ben-RB, Desulfotomaculum auripigmentum strains OREX-4, GFAJ-1, Bacillus sp., Desulfitobacterium hafniense DCB-2T, strain SES-3, Citrobacter sp., Sulfurospirillum arsenophilum sp. nov., Shewanella sp., Chrysiogenes arsenatis BAL-lT, Deferribacter desulfuricans Aeromonas sp. O23A, Anaeromyxobacter sp. PSR-1, and Geobacter sp. OR1. Moreover, a brief explanation of arsenic extraction from a model soil artificially contaminated with As(V) using a novel DARB (Citrobacter sp. NC-1) is given in this review. The author concludes with a discussion of the importance of microbial arsenate reduction in the environment. The successful application and use of DARB should facilitate the effective bioremediation of arsenic-contaminated sites. This review can be readily comprehended by students and scientists and makes for a more informed selection of microbial reduction of arsenic for environmental cleanup.

Homemade Molecular Devices, Detections and Methods

Lahiri S. S. , Singh R. , Karothia B. S. , Giredhar M. , Chakraborty A.

Research Advances in Microbiology and Biotechnology Vol. 7, 27 September 2023, Page 98-120

Molecular methods are uniform, accurate and convenient procedures, but these need to be made inexpensive, ambient stable, simple and robust for operation under field conditions and operable by semi-skilled personnel. DNA being highly degradable, we have started with “DNA stabilization or immortalization”. The proposed eight homemade molecular devices, detection kits, and methods are a few steps in that direction.

We are reporting two inexpensive homemade instruments including an “ultra-rapid thermal cycler” for PCR detections and a “Portable Transformation device” costing only INR 1300 (15.63 USD) for putting any gene in the cells for expression, amplification or integration in the genome. Also reported are the development of two homemade “field Surveillance Kits” which has infinite shelf life under ambient conditions and maybe the least expensive in the world. It is most suitable for screening of large number of samples at the same time and cost, operable even under field conditions by the semi-skilled personnel. These are for infection detection in human and animals and for sex determination of large number of plants like “Banana” simultaneously at nursery stage. Also reported are a few simplified methods which include rapid identification of micro-organisms from clinical samples under field conditions, a method for inexpensive, efficient and easy DNA synthesis (used for generation of DNA probe, primer, translation, expression etc.), and a method for easy and quantitative isolation and preservation of large quantity of DNA.

Management of Sweet Potato Virus Disease Using Prophylactic Measure Strategy

Razean Haireen M. R. , Siti Noor Aishikin A. H. , Nur Zainih J. J. , Rawaida R. , Norma H. , Nurul Afza K. , Faizah S. A. R. , Mohd Nazri B. , Anuar A. , Mohd Aziz R. , Izyani R. , Nurul Ain A.

Research Advances in Microbiology and Biotechnology Vol. 7, 27 September 2023, Page 121-132

The purpose of this study is to use plant components that have a pesticidal action to repel pests to control the pest vector. Sweet potato is one of the most economically important crops for addressing global food security and climate change issues, especially under conditions of extensive agriculture, such as those found in developing countries. However, osmotic stress negatively impacts the agronomic and economic productivity of sweet potato cultivation by inducing several morphological, physiological, and biochemical changes. Plants employ many signaling pathways to respond to water stress by modifying their growth patterns, activating antioxidants, accumulating suitable solutes and chaperones, and making stress proteins.  The sweet potato (Ipomoea batatas L.)  is a dicotyledonous plant that belongs to the bindweed or morning glory family, Convolvulaceae. Its large, starchy, sweet-tasting tuberous roots are used as a root vegetable. Sweet potato is well known as nutritious and healthy source of food that has been linked to viral infections all throughout the world, including Malaysia. When growing sweet potatoes, it's crucial to control the insect vectors that spread viral diseases. Aphid transmits Sweet potato feathery mottle virus (SPFMV) meanwhile whitefly transmits Sweet potato chlorotic stunt virus (SPCSV). SPFMV when singly or mix with other viruses had caused reduction in the quantity and quality of the sweet potato tubers. Planting chives (Allium tuberosum) in companion to sweet potato as a prophylactic measure was carried out for managing the virus pest vectors at farmer plot in Semenyih, Selangor. A total number of 192 bags of chives were planted companion to sweet potato in a ratio 1:2 as repellent crop. A continuity study which carried out to correlate between diseases severity in sweet potato and yield of harvest showed a significant interaction of negative correlation.  Sweet potato planted with chives was shown to have a lower mean number of aphid and whitefly when compared to mean number for aphid and whitefly in the control plants (sweet potato without chives). A lower virus incidence percentage was also significantly observed in sweet potato planted with chives (27.5%) compared to the control plants (41.2%). Furthermore, sweet potato grown with chives attracted more beneficial arthropod/ insects. The partial budgeting analysis showed that planting sweet potato with chives had a positive impact on the production with 13% increment of sweet potato yield compared to planting sweet potato alone. Farmer earned extra income from the sale of chives in addition to the existing sweet potato produce. More positive benefits were obtained with the value of in net income RM 344.57 when sweet potato was planted companion with chives.

Identification of Novel Genetic Markers Based on DNA Polymorphisms and Their Application in Animal Production

Hsiu-Lin Huang , I-Yen Huang , Chia-Yu Lin , Mu-Chiou Huang

Research Advances in Microbiology and Biotechnology Vol. 7, 27 September 2023, Page 133-150

This study offers practical methods for locating new and conventional markers based on DNA polymorphisms. In modern medical research, genetic markers are a helpful tool for predicting disease risk and medication response. They can be used in animal research for diseases evaluation, gender determination, species identification, and marker-assisted selection. Numerous unique genetic markers in candidate genes have been discovered by combining DNA microarray technology and SNP (single nucleotide polymorphism) genotyping. In comparison to conventional breeding techniques, these markers have additional benefits since they may be used to accurately select animals and to quickly boost reproductive performance. The investigation of DNA polymorphisms employed modified random amplified polymorphic DNA (RAPD) or random amplified microsatellite polymorphism (RAMPO) fingerprinting, and the outcomes showed that innovative and stable markers may be used for identifying species and determining the gender of animals.

Antiplasmodial Potentials and GC-MS Analysis of Bioactive Compounds in Methanol and Ethanol Leaf Extracts of Daniellia oliveri

Muhammed Muazu , Abdulsalami Mohammed Sani , Karderam Bukar Dikwa , Deborah Madi Dibal , Kokori Bajeh Tijani , Junaidu Yahaya

Research Advances in Microbiology and Biotechnology Vol. 7, 27 September 2023, Page 151-167

The goal of the study was to analyze D. oliveri utilizing qualitative, quantitative, and Gas Chromatography-Mass Spectroscopy (GC-MS) analytical methods to screen for phytochemical components and determine bioactive substances. The information regarding phytochemical compounds are not only supportive for discovery of therapeutic potential, but also have an active contribution towards discovery of new semi-synthetic and synthetic compounds. The identification of bioactive elements required for evaluating the antiplasmodial potentials of methanol and ethanol leaf extracts of Daniellia oliveri (D. oliveri) may be accomplished by qualitative, quantitative, and Gas Chromatography-Mass Spectrometry (GC-MS) analyses. The leaves were collected in Anyigba, Kogi state from which methanol and ethanol extracts were prepared, phytochemical components detected and bioactive compounds determined using GC-MS. Results showed the presence of alkaloid, tannin, reducing sugar, saponin, terpenoid, phenol, cardiac glycosides and flavonoid in the extracts. Phenol showed the highest concentration (46.14 and 43.09 mg/100g) while terpenoid showed the lowest concentration (10.63 and 9.97 mg/100g) in methanol and ethanol extracts respectively. GC-MS analysis revealed the presence of higher components (57) in methanol extract compared to ethanol extract (27). This study provides scientific evidence that methanol may be a better extraction solvent for GC-MS analysis of D. oliveri leaves meant to be used for the determination of antiplasmodial activity than ethanol due to higher components detected in methanol extract compared to ethanol extract.