Dr. Francisco Cruz Sosa,
Professor, Department of Biotechnology, Universidad Autónoma Metropolitana-Iztapalapa (UAM-Iztapalapa), Mexico.

ISBN 978-93-5547-696-8 (Print)
ISBN 978-93-5547-697-5 (eBook)
DOI: 10.9734/bpi/imb/v7

This book covers key areas of Microbiology and Biotechnology. The contributions by the authors include  methicilin resistant, methicillin susceptible, random amplified Polymorphic DNA, Antimicrobial activity,  zone of inhibition, matrix assisted laser desorption ionization time , flight mass spectrometry, Computer-based iterated prisoner’s dilemma scoring matrix, moran model, capillary feeder assay, prisoner's dilemma, clonal interference model, game theory method, Bacteria inherent biofilm, probiotics, genotypic methods, antibiotics, Syneresis, sodium caseinate, lacto-fermentation, starch, acid gel, non-dairy product, Pig fat, Lactic acid bacteria,  16S rDNA,  cancer, antioxidants,  rotavirus, diarrhoea, children, Bacillus subtilis subsp. subtilis 168, broth microdilution technique, and gas chromatography-mass spectrometry. This book contains various materials suitable for students, researchers and academicians in the field of Microbiology and Biotechnology.

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Molecular Identification of Methicillin-Resistant and Methicillin Susceptible Staphylococcus aureus Using RAPD Indicators

Suha M. Abed, Batol I. Dheeb, Akeel H. A. Assie, Basim M. Khashman, Anmar Sael Hussein

Innovations in Microbiology and Biotechnology Vol. 7, 25 August 2022, Page 1-16

The present study aimed to investigate the relatedness and genomic variability between strains of methicillin resistant and of methicillin susceptible Staphylococcus aureus. Staphylococcus aureus, a gram-positive coccoid bacterium, is a common human pathogen that has the ability to cause a wide array of severe hospital and community-acquired infections, such as pneumonia, bacteremia, sepsis and toxic shock syndrome. The study included 100 local isolates of S. aureus collected from different clinical cases of nonrelated patients. All sample isolates were grown at 37°C on manitol salt agar medium and were been identified at the species level using morphological characteristics and biochemical tests. DNA profile of the samples obtained were detected by inspecting the banding pattern visually. Depending on the primers, the amount of bands and banding pattern varied. The presence or absence of a band at any place on the gel was scored to create a two-dimensional binary matrix. The application of RAPD technique was preceded by extraction of genomic DNA using the boiling method. mec A gene was amplified to distinguish MRSA strains without the need to perform antibiotic susceptibility. Our data had demonstrated ability of RAPD analysis to differentiate strains at intra-specific level also it can be applied successfully to assess the genetic backgrounds among isolates.

Assay of Commenila benghalensis Linn. Leaf Extract on Selected Microbes

Urvashi Sinha

Innovations in Microbiology and Biotechnology Vol. 7, 25 August 2022, Page 17-22

The present study was designed to investigate the antibacterial activity of leaf extract of C. benghalensis L. in order to examine the pharmacological basis of the use of the plant in folk medicine for the treatment of infectious diseases. The existence of tannin, favonoid, phenol, carbohydrate, and volatile oil was discovered in the chloroform extract, which demonstrated the greatest activity against the pathogenic bacterial strains Escherechia coli, Staphylococcus aureus, Salmonella typhi, and Candida albicans, with inhibition zones of 7mm, 1mm, 3mm, and 4mm, respectively, when compared to the hexane and methanolic extracts. The Minimum Inhibition Concentration (MIC) of the chloroform extract was determined using the dilution susceptibility test method. The current findings support the use of Commelina benghalensis L leaf extract as a therapeutic agent against common pathogenic microorganisms.

Pseudomonas oryzihabitans Bacteremia from North India in a Terminally Ill Patient: A Rare Case Report

Akanksha Dubey, Tasneem Siddiqui, Mitra Kar, Chinmoy Sahu, Sangram Singh Patel

Innovations in Microbiology and Biotechnology Vol. 7, 25 August 2022, Page 23-29

Pseudomonas oryzihabitans is a gram-negative bacillus that is commonly isolated from bacteremia, CNS infections, catheter-associated infections or any other device-related infections, sinusitis, wound infections, and skin infections in immunocompromised individuals in hospital settings. Only a few cases of bacteremia caused by this uncommon infection have been recorded. This is the first case report of a terminally ill patient with chronic liver disease from North India. The illness was resolved after proper isolation and early commencement of medications based on sensitivity patterns, but he died subsequently owing to other problems. This organism is rare but it can cause opportunistic infections in immunocompromised individuals. The need of automated methods for identification and sensitivity testing limits the reporting of this rare but important pathogen in hospital settings. By correct identification and antibiotic susceptibility testing we can easily treat infections due to this opportunistic pathogen.

Application in Bioremediation, Psychology, Forestry, and Biofilm Development

Harpal Vala

Innovations in Microbiology and Biotechnology Vol. 7, 25 August 2022, Page 30-44

When a living organism performs a procedure in a given situation, it begins to update itself based on that situation. In this hypothetical theoretical research work, game theory is used to provide a hypothetical theory. Two different species (assume hypothetical species A+ and B-) are transferred through the system of various five compartments, according to this theory. In the presence of a specific external environment element, species A + and B- travelling through the five compartment model system can only do so in one compartment (compartment E).
If compartment E has a limited amount of an essential carbon source for the bacteriological system, species A+ and B- are likely to compete at first, but continuous contact with each other in sequence will convert corporations into compartment E for restricted sucrose self-uptake by species A+. Competition will be decreased, allowing species B- to use it as well. By transferring adapted bacteria to drosophila, the Bacterial system's adaptation behaviour can be validated using a capillary feeder assay (CAFÉ Assay).

Biofilms a Rabble-rouser in the Environment Leading to Antibiotics Resistance

Harman Multani, Varsha A. Singh

Innovations in Microbiology and Biotechnology Vol. 7, 25 August 2022, Page 45-51

Background: Bacteria inherent biofilm resistance and it’s very difficult to overcome, even when appropriate antibiotic treatments are provided. They exhibit resistance to antibiotics by various methods like restricted penetration of antibiotic into biofilms, decreased growth rate and expression of resistance genes.
Diagnostic Methods: Phenotypic and Genotypic methods.
Treatment: For eradication of the biofilms concentrations of antibiotics in excess of the MIC are required. Probiotics (Good Bacteria) reduces upper respiratory tract infections.
Conclusion: Owing to injudicious use of topical antibiotics, Multi Drug Resistant Bacteria are becoming more resistant to most of the available antibiotics leading to Biofilm formation.

Syneresis Understanding by Modeling the Lacto-Fermentation of Sodium Caseinate

Soumaya El Bouchikhi, Philippe Pagès, Azeddine Ibrahimi, Yassir El Alaoui, Yahya Bensouda

Innovations in Microbiology and Biotechnology Vol. 7, 25 August 2022, Page 52-69

This study aimed to investigate the influence of sodium caseinate, starch, lactose, and lactic acid bacteria on syneresis in a mixed model system and to evaluate their impact on the acid gel formation throughout pH and zeta potential monitoring. The results showed a significant decrease of syneresis in all mixtures with sodium caseinate at 8% compared to 3%, ranging between 1.8%-20.6% and 22.2%-47.8%, respectively. The fermented mixed model's acidification profile and syneresis were considerably influenced by the addition of starch. Furthermore, the differences in acid gelation and syneresis have been better understood as a result of monitoring pH and zeta potential throughout the lacto-fermentation process. These findings indicate that syneresis varies very closely with sodium caseinate concentration, starch concentration, and their association, regardless of the concentrations of lactose and ferment. If a sodium caseinate-starch mixed system is used, syneresis could be decreased to an ideal level: If the concentration of starch is above 1%, less syneresis gels may be produced at sodium caseinate concentrations above 5%.

Molecular Identification of Lactic Acid Bacteria in Indigenous Fermented Pig Fat: A Case from Assam, India

Ranjita Yumkhaibam, Kimjolly Lhouvum

Innovations in Microbiology and Biotechnology Vol. 7, 25 August 2022, Page 70-86

The study aimed to identify probiotic bacteria in traditionally fermented pig fat (Sathu) obtained in Karbi Anglong, Assam. The objective of the study is to assess the presence of indigenous Lactic acid bacteria and in vitro characterization for probiotic characteristics by using standard techniques, with the aim to explore the importance of the food to the population. From the food sample, two types of bacteria (KJc8 C8 and KJR2 C9) were isolated. The isolates were discovered to be cocci and rod-shaped morphologically. The isolated strains were resistant to inhibitory compounds such as NaCl (1-10%) and bile salt (0.1-1%) and grew better in acidic conditions, which were important characteristics for bacteria to be probiotic. The isolates also demonstrated successfully in metabolising various sources of carbohydrates. Bacillus and Lactobacillus species, which are probiotic bacteria, were identified through molecular analysis utilising 16S rDNA gene sequencing. The experimental findings also showed that only Lactobacillus and Bacillus species predominated the food sample, with no additional spoilage bacteria found. This provides significant evidence that the food item may be a probiotic. Beyond its nutritional effects, further research on the food product will mark a significant step in the economic development of humanity.

Determination of Potential Aptitude of Four Olive Cultivars used as Anticancer and Antioxidant Agents

Shimaa A. Rashed, Tamannouha I. Saad, Salama M. El-Darier

Innovations in Microbiology and Biotechnology Vol. 7, 25 August 2022, Page 87-103

The present study has focused on the extraction and purification of oleuropein from leaves of different olive cultivars: Chemlali (CHM), Manzanilla (MAZ), Picaul (PIC) and Toffahi (TOF), to evaluate the antioxidant activity and therapeutic potential in cancer. A source of numerous bioactive chemicals with known advantages for both human health and technical applications is Olea europaea L. (olive, Oleaceae). Nowadays, there is growing evidence that DNA damage caused by reactive oxygen species may be the first step in the progress of malignant cells can be repressed by consuming olive leaf extracts. It occurred in leaf extracts of the four olive cultivars (Chemlali, Manzanilla, Picaul and Toffahi) as a source for some anticancer and antioxidant agents and their consequences on the action of Hordeum vulgare (barley). The total phenolic and flavonoid compounds were extracted from olive leaves by ethanol 95% then analyzed by high-performance liquid chromatography (HPLC). The study evaluates the anticancer activity of the ethanolic extract of olive leaves against breast and hepatocellular carcinoma cells showing high values. Also, When barley's antioxidant potential was evaluated using the diphenyl picryl hydrazyl method, the extract had a significant impact (DDPH). These findings open the door to using olive leaves as a natural anticancer and antioxidant source. 3. Application of CHM and MAZ olive cultivar crude powder in barley cultivations enhances the production of antioxidants, a biomarker for medicinal value of barley.

This study aim to evaluate some of the clinical signs, causes, and mortality associated with acute diarrhea in children under the age of five in a hospital environment in Abuja, Nigeria, for a year. The second greatest cause of death in children is diarrhea, and Nigeria has the highest mortality rate on the continent with limited knowledge of the particular cause and a high prevalence of Rotavirus and Cryptosporidium. The One-year cross-sectional study was carried out in Abuja, the Federal Capital Territory of Nigeria. Nigeria has a total population of 150 million people according to the 2006 population census, of which about 5 million people live in Abuja from June 2018 to May 2019.
Cryptosporidium and Rotavirus ELISA were done with commercially available kits.
Stool samples were collected from 1450 participants, of whom 1185 (81.7%) were ambulatory, 109 (7.5%) were hospitalised, and 156 (10.7%) were controls without diarrhoea. Cryptosporidium-ELISA was positive among 274 (21.1%) children with diarrhoea and 23 (1.7%) of children without diarrhoea, with August and September as peak months for infection. Rotavirus-ELISA was positive among 231 (17.8%) children with diarrhoea and 29 (2.2%) controls, with November, December, and January as peak months. Children of 12 to 17 months were most affected for both and Rotavirus (39.8%) and Cryptosporidium (37.2%).
Rotavirus and Cryptosporidium are important pediatric diseases, particularly in Abuja's unvaccinated youngsters for Rotavirus. To track the planned vaccine program, notably for Rotavirus, it will be necessary to develop local and national infrastructure for diarrhoeal illness monitoring and provide access to virological and parasite stool testing. The findings of this study will aid in the planning of the rotavirus vaccine's possible inclusion in the National Immunization Program and establish a baseline for assessing the vaccine's influence on prevalence if it is implemented in Nigeria's immunization program.

Antimicrobial Activity and GC-MS Based Analysis of the Extract of Bacillus subtilis subsp. subtilis 168 Isolated from a River Bank

Olasinbo Olumuyiwa Balogun, Sylvanus Chukwudi Ugoh, Peters Oluwale Oladosu

Innovations in Microbiology and Biotechnology Vol. 7, 25 August 2022, Page 126-145

Drug-resistant pathogens are becoming a greater threat, and they are becoming more adept at dodging traditional antibiotics. Therefore, it's crucial to find new antimicrobials to combat the problems we currently face. Our preliminary investigation identified Bacillus subtilis subsp. subtilis 168 isolated from soil sample sourced from a river bank in Abuja, Nigeria, as the most potent antibiotic-producing bacteria among the other identified producers. The objectives of the current study were to: (i) screen for the antimicrobial activity of the extract, and fractions of the candidate isolate by broth microdilution method (ii) identify the bioactive compounds by Gas Chromatography-Mass spectrometry (GC-MS) method. We calculated the minimum bactericidal concentration (MBC), minimum inhibitory concentration (MIC), and the MBC/MIC ratio. All the tested pathogens were susceptible to the ethyl-acetate extract (MIC between 28.70 mg/ml and 57.40 mg/ml). Proteus mirabilis was the least susceptible pathogen, and the extract demonstrated bactericidal efficacy against all tested pathogens (MBC/MIC between 1.00 and 2.00). Vacuum liquid chromatography was used to separate the extract, and pathogenic strains were challenged with the fractions after purification. The most effective fraction was fraction E, which was also the most bactericidal (MBC/MIC between 2.00 and 2.11) against all of the test microorganisms (MIC between 0.09 mg/ml and 0.75 mg/ml). 13 compounds with various retention times and peak regions were found by GC-MS analysis of the purified subfraction produced from fraction E. Among these were three major compounds which include: (i) bis(2-ethylhexyl) phthalate (ii) 1,4-epoxynaphthalene-1(2H)-methanol, 4,5,7-tris(1,1-dimethylethyl)-3,4-dihydro- (iii) D:B-Friedo-B':A'-neogammacer-5-en-3-ol, (3.beta.)-. According to our research, the locally isolated Bacillus subtilis subsp. subtilis 168 could be a rich source of lead compounds for use in biotechnology and medicine.

Study of Dengue Fever Cases at a Tertiary Care Hospital of India

R. J. Girish Babu, K. R. Madhuri

Innovations in Microbiology and Biotechnology Vol. 7, 25 August 2022, Page 146-154

Currently, dengue is the second most common vector-borne disease in the world. The objective is to study the clinical and laboratory profile of Dengue cases admitted at Sri Siddhartha Medical College and Hospital.

A hospital-based prospective study was conducted at Sri Siddhartha Medical College and Hospital from June to December 2015. Included were 150 seropositive patients who were hospitalised to the medical wards and ranged in age from 18 to 75. The patients' clinical profiles were noted. Out of 150 seropositive cases, 88 were males and 62 were females. Fever was the most prevalent initial symptom. Most of the time, thrombocytopenia measured between 50,000 and 100,000/cu.mm. IgM, IgG antibodies, and NS1 antigen were all found to be seropositive in 70%, 7%, and 3% of patients, respectively. The remaining 20% of cases had multiple seropositive markers.

Early diagnosis of dengue depends on a high degree of clinical suspicion, supported by laboratory evidence such thrombocytopenia and particular dengue serological testing.