Dr. Emmanouil Magiorkinis
Department of Laboratory Haematology, General Hospital for Chest Diseases "Sotiria", Greece.

Short Biosketch

ISBN 978-81-967198-5-2 (Print)
ISBN 978-81-967198-4-5 (eBook)
DOI: 10.9734/bpi/ramb/v8

This book covers key areas of microbiology and biotechnology. The contributions by the authors include yeast fungus, denture stomatitis, microwave sterilization, mechanical brushing, polycyclic aromatic hydrocarbons, terrestrial ecosystems, soil microbial population, biodegradation, anthropogenic activities, antimicrobial resistance, bacterial strains, antibiotics, umbilical cord, biological association, placenta, hormonal changes, DNA profiling, anti-inflammatory activities, eutrophication, environmental degradation, bioactive compounds, DNA synthesis, sequence tagged microsatellite sites, gene identification, genetic polymorphism, amplified polymorphic DNA, human cytomegalovirus, natural compounds, medicinal mushrooms, cytotoxicity, anti-viral response, neutropenia, wound healing, extracellular matrix, inflammation, fibroblasts, collagen, hemostasis, cell proliferation, granulation, actin proteolysis, metalloproteinases, bacterial invasion, arboviruses, insecticides, chemical insecticides, human monkeypoxgoatpox virus, zoonosis, sporadic human infections, ola haemorrhagic fever, Hanta virus disease, Nipah virus disease, rift Valley fever, bacterin, antigenic potential, adaptive immune system, granulocyte colony-stimulating factor, cytokine. This book contains various materials suitable for students, researchers and academicians in the field of microbiology and biotechnology.


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Impact of Dentures’ All-night Immersion in Distilled Water on Candidal Colonization

Georges Aoun , Wissam Sharrouf , Pamela Aoun

Research Advances in Microbiology and Biotechnology Vol. 8, 17 November 2023, Page 1-9

Objective: In order to keep their dentures clean and properly stored, patients generally soak them in water at night. Candida albicans is a commensal yeast fungus that colonizes dentures, and in some conditions, it becomes an opportunistic pathogen and causes fungal infections known as candidiasis. This mycological study aimed to evaluate the effect of distilled water on Candida albicans colonizing dentures.

Methods: Twenty patients (9 men, 11 women; age range 40-75 years) with complete maxillary dentures infected by Candida albicans were included in this study. The dentures of these patients were soaked in distilled water for 4 days (8 hours at night). Swab samples from the dentures were collected before and after distilled water use and examined mycologically.

Results: The Candida albicans colony counts increased after soaking the dentures in distilled water for 8 hours for 4 days.

Conclusion: Patients should be dissuaded from soaking their dentures overnight in distilled water, as the result is a significant increase in fungal colonization.

The present investigation examined the potential of the filamentous fungus Penicillium sp. CHY-2, which was isolated from Antarctic soil, to biodegrade eight different aliphatic and aromatic hydrocarbons, including octane, decane, dodecane, ethylbenzene, butylbenzene, naphthalene, acenaphthene, and benzo[a]pyrene. The degree of polycyclic aromatic hydrocarbon contamination of environmental matrices has increased over the last several years due to an increase in industrial activities. Interest has surrounded the occurrence and distribution of polycyclic aromatic hydrocarbons for many decades because they pose a serious threat to the health of humans and ecosystems. Among all the compounds, CHY-2 showed the highest level of degradation for decane (49.0%), followed by butylbenzene (42.0%) and dodecane (33.0%), and lower levels of degradation for naphthalene (15.0%), acenaphthene (10.0%), octane (8.0%), ethylbenzene (4.0%), and benzo[a]pyrene (2.0%) at 20 \(^{\circ}\)C. The addition of carbon sources such as glucose (5 g L-1) and Tween-80 (5 g L-1) enhanced decane degradation by about 1.8-fold and 1.61-fold respectively at 20 \(^{\circ}\)C. The metabolites produced during the degradation of decane were identified by gas chromatography-mass spectrometry (GC-MS). Furthermore, the enzyme manganese peroxidase (MnP) from CHY-2 was purified. MnP was found to consist of monomers with a molecular mass of 36 kDa. The purified MnP had an optimum pH of 5.0 and temperature of 30 \(^{\circ}\)C. The Km and V max values of MnP towards Mn2+ were 1.31 mM and 185.19 mM min-1 respectively. These results indicated that the strain CHY-2 could be used for the degradation of hydrocarbons and could have promising applications in the treatment of hydrocarbon contaminated sites.

A Comparative Overview on Algal Inhibitory Efficiency of Secondary Metabolites of Tamarindus indica and Azadirachta indica

Mathews P. Raj , Anitha A. Abraham , Kasturi Banerjee , Mainak Chakrabarty , Lakshmi Sagar K.

Research Advances in Microbiology and Biotechnology Vol. 8, 17 November 2023, Page 28-38

This chapter explores the potential of Tamarindus indica extract to inhibit the proliferation of algae in an environmentally friendly way and compare its efficiency of inhibition with Azadirachta indica leaf extract. Tamarindus indica is a medicinal plant belonging to the family Fabaceae. It has been used as a medicinal plant for centuries; its fruits being the most valuable part. This study showed that Tamarindus indica extracts effectively inhibited the growth of algae. The secondary metabolites of Azadirachta indica are more effective than the secondary metabolites of Tamarindus indica in reducing the growth of algae, according to a comparative analysis of inhibitory efficacy between the extracts of both plants. The observed result suggests that fruit extract of Tamarindus indicacan be used as an alternative way to prevent algal bloom.

More efficiency of inhibition of algal growth might be achieved by scrutinizing the secondary metabolites of Tamarindus indica and administering them specifically.

This chapter aims to determine the prevalence and pattern of antibiotic resistance of Escherichia coli isolates from children admitted at selected health care facilities in Moshi municipality, Tanzania. Antimicrobial resistance (AMR) among Gram negative bacteria has become a major public health concern worldwide. The currently available information on the prevalence and patterns of AMR to different geographical regions has demonstrated that it is on rise. This descriptive cross sectional study was conducted during October 2017 to April 2018.

The study sought to answer the following questions:  What are the carriage rates of Escherichia coli among children admitted at selected health facilities in Moshi municipality, What is the resistance rates of Escherichia coli isolated from children against the commonly used antibiotics in Moshi Municipality and What is the prevalence of the Extended spectrum beta lactamase producing Escherichia coli isolates from children admitted at health facilities in Moshi municipality?

In this investigation, Escherichia coli was identified from all 282 cultured archived rectal swabs, and all isolates were sensitive to amikacin and meropenem. Extended spectrum beta lactamase producers made up 13.1% of the total, while non-extended spectrum beta lactamase producers made up 86.9%.  Antimicrobial resistance was detected in 282 isolates on the commonly used antimicrobial agents: ampicillin (83%), trimethoprim (75.9%), cefuroxime (32.6%), ceftriaxone (30.1%), ceftazidime (29.4%), ciprofloxacin (27.7%), amoxicillin/clavunic (24%), gentamicin (21%), chloramphenicol (16.3%) and piperacillin-tazobactam (8.1%). Ampicillin resistance was found in all Extended Spectrum beta lactamase generating isolates. Amikacin and meropenem are remain the antibiotics of choice for second-line therapy of E. coli infections. The majority of the isolates were resistant to ampicillin and trimethoprim. Both ESBL and non-ESBL producing Escherichia coli displayed resistance to routinely used antibiotics; therefore, more research is needed to understand the processes employed by non-ESBL producing Escherichia coli to develop resistance.

Non-Invasive Forensic Identification Methods of Neonates from Umbilical Cord and Cord Blood Sample

Soma Roy , Ishani Mitra , Dinesh D.

Research Advances in Microbiology and Biotechnology Vol. 8, 17 November 2023, Page 51-57

The most preferred forensic specimen collection technique for DNA Profiling in Justice delivery system is blood sample either in liquid form or in stain forms. But blood collections relating to newborn babies are always very difficult and painful invasive method. To prevent that issue, we have successfully isolated genomic DNA from a new born in a non-invasive way, by collecting the dried umbilical cord and the cord blood sample. It has given similar yield as of liquid or stained blood samples. Autosomal STR typing of isolated DNA from the cord blood and dried umbilical cord contributed satisfactory results in drawing conclusive opinion.

Studies on Genetic Diversity and Polymorphism of Acacia constricta and Acacia farnesiana

M. Balasubramanian , S. Janaki , M. Abinaya

Research Advances in Microbiology and Biotechnology Vol. 8, 17 November 2023, Page 58-77

In this study, the genetic diversity and polymorphism of the A. farnesiana and Acacia constricta species were examined using randomly amplified polymorphic DNA (RAPD). Several groups have reported on the utility of RAPD markers as a source of phylogenetic information.  The seeds of 10 varieties of A. constricta and A. farnesiana were randomly selected from five districts (Namakkal, Perambalur, Nagapattinam, Ramanathapuram and Pudukkottai) of Tamil Nadu, India. Analyses using RAPD data revealed 87.5% polymorphism.  The genetic closeness between varieties of both species ranged from 29 to 93%. Between the species, the GT-8 variety of A. constricta and GCT-2 of A. farnesiana had the lowest genetic similarity (29%), but the GT-8 and GT-6 varieties of A. constricta had the most genetic similarity (41%), with the variety GCT-3 of A. farnesiana.  Polymorphism was found in 51% of the fragments amplified with 22 SSR primers. The genetic similarity between cultivars of both species ranged from 20 to 91%.  The first cluster was exhibited by three A. constricta types, whereas the second cluster was revealed by seven A. farnesiana species with two sub clusters, and GCT-3 was linked separately to the main cluster.  The examination of genetic similarities and clustering patterns between RAPD and SSRs revealed almost 85% similarity.  SSR analysis has, the varieties Cocker-1, GCT-2 and GC-1 are grouped together with close genetic similarity when compared to the grouping pattern of varieties in RAPD analysis. In variants cultivated in Tami Nadu, isoenzyme analysis of the peroxidase and polyphenol oxidase enzymes revealed species-specific bands as well as bands specific to the Acacia genus. In analyzing the diversity of Acacia species, these important data may aid researchers in identifying duplicate accessions.

Culex quinquefasciatus Aedes aegypti and Anopheles stephensi responsible for deadly diseases to human beings and animals. Carassius auratus and Poecilia reticulata were used to evaluate for predatory efficiency against diseases causing mosquito larvae of Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi and remarkable larval consumption rate was observed that C. auratus feed all 50 larvae within 6 hours and P. reticulata feed all 50 at the end of 21st hours. Therefore, we suggest C. auratus and P. reticulata can be effectively used against mosquitoes for integrated vector control management programme.

Ethnomedicinal Approach against Human Cytomegalovirus Infection: A Search for Alternative Therapy

Debsopan Roy , Nilanjan Chakraborty

Research Advances in Microbiology and Biotechnology Vol. 8, 17 November 2023, Page 84-94

Human cytomegalovirus is a common opportunistic infection that can infect immunocompromised and immunosuppressed individuals as well as neonates. There are currently very few clinically authorized medications on the market for use in therapy. Although they are all incredibly effective against active HCMV infections but each of these medications has a certain amount of side effects which can leads to serious health complications to the patients. In search for the alternative approach to these synthetic drugs antiviral substances produced from natural resources are urgently needed as because due to less cytotoxic impact as their synthetic equivalents. The medicinal mushrooms have shown to be a really intriguing option in this regard. Utilizing these compounds might be a viable first line of defense against since traditional ethnomedicinal substances have been a rich source of bioactive molecules with different pharmacological actions, including antiviral activity. This chapter is hereby comprising with the investigative work of those promising ethnomedicinal resources to understand the new frontier of HCMV research.

The Extracellular Matrix (ECM) and Wound Healing: A Review

Robert B. Diller , Aaron J. Tabor

Research Advances in Microbiology and Biotechnology Vol. 8, 17 November 2023, Page 95-117

The purpose of this article is to summarize current and historical literature demonstrating the role of the Extracellular Matrix (ECM) in the various stages of the wound healing cascade. ECM is a three-dimensional framework that is found in all human tissues. It is made up of a variety of proteins, including collagens, elastin, and minor amounts of structural proteins. The ECM has been shown in studies to aid in cellular adhesion, tissue anchoring, cellular signaling, and cell recruitment. The ECM is affected at times of integumentary injury or damage, whether acute or chronic. The ECM is created and ideally returned to its native condition by a series of overlapping activities known as the wound healing phases: hemostasis, inflammation, proliferation, and remodeling. The authors believe that more study should be done to not only use the structural components of the ECM but also to imitate the physiological structure.

Human Mpox: A Deadliest Pox Virus till Date

Meenu Meena

Research Advances in Microbiology and Biotechnology Vol. 8, 17 November 2023, Page 118-126

The ongoing 2022 multicountry outbreak of Mpox (monkey pox) is an emerging and re-emerging Zoonotic viral disease. Mpox is a double-stranded DNA virus. It can be transmitted to humans through physical contact with someone who is infectious, with contaminated materials, or with infected animals. It draws world’s attention because of the increasing occurrence of human outbreaks recent years, beyond its traditional geographical location. The incubation period of the disease can be 5 to 21 days. The current 2022 outbreak of Mpox in newly reported countries primarily affects men (homosexual or bisexual) and genital skin lesions are a dominant symptom. Genome sequencing has revealed that the Clade IIb variant is responsible for the Mpox ourbreak. Healthcare professionals are at higher risk of becoming infected since they are in close contact with patients directly. Children, pregnant women & people with weak immune system are at risk for complication from Mpox.

The WHO declared, Mpox as a public health emergency of international concern on 25 July 2022. So, it is important to update knowledge of this zoonotic infection, including case finding, contact tracing, laboratory investigation, clinical management, isolation and implementation of infection prevention control measures and prophylaxis to understand the broader implication of the current outbreak. WHO published a strategic preparedness & response plan for Mpox.

CDC allows use of stockpiled Tecovirimat to Mpox during an outbreak. VIGIV is licenced by FDA for the treatment of complications due to vaccinia vaccination. A newer vaccine based on modified attenuated vaccinia virus (Ankara strain) was approved for the prevention of Mpox.

The Role of Actin-specific Bacterial Endoproteases Grimelysin and Protelysin in Serratia Invasion

Sofia Khaitlina , Ekaterina Bozhokina , Olga Tsaplina , Tatiana Efremova

Research Advances in Microbiology and Biotechnology Vol. 8, 17 November 2023, Page 127-148

This chapter describes the properties of the bacterial proteases grimelysin and protealysin, which are virulence factors in vitro and in vivo, and actin can be a target for these proteases upon their translocation into the host cell.

Bacterial invasion into eukaryotic cells is one of the major areas of research in infection biology, whereby they employ different strategies to invade the host cells. These interactions are highly complex, and the type of interaction depends on the bacterium, host, as well as environmental factors. The single actin polypeptide link, Gly42-Val43, is broken by grimelysin. No other proteases are able to break this link, which results in the reversible loss of actin polymerization. Similar characteristics were shared by protealysin, another bacterial protease. Grimelysin and protealysin are unique tools for studying the functional characteristics of actin because of their characteristics. Furthermore, bacteria Serratia grimesii and Serratia proteamaculans, producing grimelysin and protealysin, invade eukaryotic cells, and the recombinant Escherichia coli expressing the grimelysin or protealysins gene become invasive. Specifically, the grimelysin (ECP 32)-producing bacteria invaded the transformed epithelial and fibroblasts cells A431, HeLa and 3T3-SV40, but they were not found in embryonic fibroblasts, primary human keratinocytes and in cells of poorly transformed 3T3 cell lines. The invasion of eukaryotic cells by S. grimesii has been demonstrated to include the cellular c-Src and RhoA/ROCK signaling pathways, and it has also been proposed that E-cadherin plays a role in the invasion. Furthermore, it was discovered that S. grimesii produces membrane vesicles that contain grimelysin, enter eukaryotic cells, and promote bacterial invasion of these cells. These data indicate that the protease is a virulence factor, and actin can be a target for the protease upon its translocation into the host cell.

IL17 Responses in Rabbits Primed with E. coli-P. aeruginosa Bacterin Combinations

Ibrahim M. S. Shnawa, Ferial Abd

Research Advances in Microbiology and Biotechnology Vol. 8, 17 November 2023, Page 149-158

This chapter aimed for the developing prototype E. coli - P. aeruginosa heat killed bacterin balanced and unbalanced bacterin combinations, as well as monotypic E. coli and single P. aeruginosa bacterins. Microbial vaccines specific immune primed to different mammalian immune systems shown an array of IL17 responses as; increase, decrease, or variable from controls. The evolution of genetic engineering techniques has enabled the construction of recombinant microorganisms capable of expressing heterologous proteins in different cellular compartments, improving their antigenic potential for the production of vaccines against viruses, bacteria, and parasites. From a series of patients with urinary tract infections, a uropathic gram negative isolates were purified and identified by VITEK identification system as E. coli and P. aeruginosa. At the end of the specific immune-priming protocols, rabbits were bleed and sera were saved in 0.5 ml aliquots at -18C till testing due time. IL17 ELIZA assay was done on the test and control sera. In comparison to double balanced, unbalanced, and control bacterin combinations, a single strength balanced combined bacterin combination increased the mean IL17 concentrations. Inhibition in IL17 concentration means was observed in the other combinations compared to the control. The immunological interference correlated like follows: one bacterium amplifies the other, while another attenuates the other. Enhancement of IL17 response single strength combination might be a promise for the efficacy of such combination in regulation and protection against natural or experimental infectious challenges.