Dr. Essam A. Makky
Associate Professor, Faculty of Industrial Sciences & Technology, University Malaysia Pahang (UMP), Malaysia.

ISBN 978-93-90768-58-5 (Print)
ISBN 978-93-90768-59-2 (eBook)
DOI: 10.9734/bpi/rpmb/v5

This book covers key areas of microbiology and biotechnology. The contributions by the authors include osteoarthritis, dietary supplement, cat’s claw, inflammation, flexibility, nutraceutical, natural product, maize lethal necrosis diseases, end stage renal disease, superficial fungal infections, dermatophytes, non-dermatophytes, heamodialysis, diabetic nephropathy, Imidacloprid, biodegradation, dipyridamole, lymphoid cells, IFN-synthesis, antiviral agent, tomato Indeterminate varietal entries, vegetative growth, organoleptic quality assessment, biochemical analysis, enterovirus infections, antiviral treatment, human pathology, latex agglutination tests, Clostridial toxins, industrial cultures, lectin domain, carbohydrate binding, intra-molecular relationships, medical biotechnology, microbiology, insecticide, xenobiotics, cervical cancer, exosomes, tetraspanins, phenolic compounds, fermentation, nutrients bioavailability, human nutrition. This book contains various materials suitable for students, researchers and academicians in the field of microbiology and biotechnology.


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Osteoarthritis is the most common form of joint disease and reflects a chronic inflammation that over time can result in joint destruction including the loss of articular cartilage and exposure of underlying bone. In a multi-center, randomised, placebo-controlled, double-blind clinical trial, a medicinal plant and L-leucine mixture (HLM) was studied in subjects with established osteoarthritis of the knee.

A total of 96 osteoarthritic subjects were enrolled and randomly assigned to either a placebo (n = 38) or an HLM treatment group (n = 38). The HLM group received a combination of Uncaria tomentosa (300 mg), Boswellia serrata (200 mg), Lepidium meyenii (1000 mg) and L-Leucine (700 mg) once a day. The placebo group received matching capsules with carboxymethylcellulose. The treatment lasted eight weeks, with assessments on days seven, fourteen, twenty-eight, and fifty six.The primary goal was to lower the total WOMAC score.VAS pain, tolerability, investigator assessments, use of rescue medication (acetominophen), and vital sign and laboratory assessments were all included.Subject randomization worked well in terms of age, gender, and disease severity. The trial was completed by 32/38 subjects in the placebo group and 35/38 in the HLM group. WOMAC scores (pain, stiffness, physical performance, and total) decreased steadily in both groups over the course of the 8-week study, but the magnitude was significantly greater in HLM (P 0.05). Total WOMAC was reduced 46.5% for HLM and 25.4% for placebo. VAS pain was reduced 21.8% in the placebo group (p < 0.002) but the changes were significantly greater (37.8% p < 0.03) with HLM treatment.

The investigator's overall rating of good-excellent for HLM was 24/35 (69%) and 14/32 (44%) for placebo (P = 0.05).The consumption and tolerability of rescue medications were comparable for HLM and placebo. These properties of HLM warranted a clinical investigation to further investigate its potential as a treatment for osteoarthritis, which is the focus of this report. There were no safety concerns with either group.Although a placebo effect was observed, HLM was clearly more effective in relieving osteoarthritis symptoms. This HLM represents a new, safe, and effective approach to managing osteoarthritis symptoms.

Brief Overview of Maize Lethal Necrosis and the Molecular Basis of Variability in Concentrations of the Causal Viruses in Co-infected Maize Plant

L. A. O. Awata, B. E. Ifie, P. Tongoona, E. Danquah, M. B. Jumbo, Manje Gowda, P. W. Marchelo-D’ragga, Chelang’at Sitonik, L. M. Suresh

Recent Progress in Microbiology and Biotechnology Vol. 5, 4 May 2021, Page 13-39

Maize lethal necrosis (MLN) disease is new to Africa, first report was in Kenya in 2012, since then the disease has rapidly spread to most parts of eastern and central Africa region including Tanzania, Burundi, DRC Congo, Rwanda, Uganda, Ethiopia and similar symptoms were observed in South Sudan. Elsewhere, the disease was caused by infection of Maize Chlorotic Mottle Virus (MCMV) in combination with any of the potyviruses namely, maize dwarf mosaic virus (MDMV) and sugarcane mosaic virus (SCMV), and tritimovirus wheat streak mosaic virus (WSMV). In Africa, the disease occurs due to combined infections of maize by MCMV and SCMV, leading to severe yield losses. Efforts to address the disease spread have been ongoing. Serological techniques including enzyme-linked immuno-sorbent assay (ELISA), polymerase chain reaction (PCR), genome-wide association (GWAS) mapping and next generation sequencing have been effectively used to detect and characterize MLN causative pathogens. Various management strategies have been adapted to control MLN including use of resistant varieties, phytosanitary measures and better cultural practices. The prevalence and survival of plant viruses in the tropics and subtropics are enhanced by the ideal tropical temperature conditions and relative humidity that encourage perpetuation of both the viruses and their insect vectors. This review looks at the current knowledge on MLN causative viruses, genetic architecture and molecular basis underlying their synergistic interactions. Lastly, some research gaps towards MLN management will be identified. The information gathered may be useful for developing strategies towards future MLN management and maize improvement in Africa.

Studies on Superficial Fungal Infections in ESRD Patients over the Past 16 Years

Upma Narain, Arvind Gupta

Recent Progress in Microbiology and Biotechnology Vol. 5, 4 May 2021, Page 40-47

Background: Mounting prevalence of end-stage renal disease is a major public health problem worldwide, resulting in millions of individuals affected. It is important to note that between 50% to 100% of such group have been found to have at least one or the other dermatological disorder. In the present study we have tried to analyze patients with end stage renal disease who had developed superficial fungal infections.

Methods and Findings: In between January 2000 to June 2016, analysis have been made on 2112 patients with end stage renal disease, who were undergoing haemodialysis at least thrice a week for a minimum of three months at our centre. Out of the above 2112 patients, 1500 suspected cases of superficial fungal infections we identified, 1040 cultures tested positive. The macroscopic examination of the scalp, skin and the nails revealed 0.2% Tinea capitis, 28.8% Pityriasis versicolor, 16.7% Tinea corporis, 5.3% Tinea mannum, 32.4% Onychomycosis, 13.5% Tinea cruris and 3.1% Tinea pedis. Culture examination highlighted 29.1% Trichyopyton rubrum, 22.9% Trychophyton mentagrophyte, 5.1% Trychophyton violaceum, 0.9% Trichophyton verrucosum, 0.1% Microsporum canis, 3.1% Epidermophyton flucossum, 3.1% Trychophyton tonsurans, 6.9% Scopulariopsis brevicaulis and 28.8% Malassezia. The predominant clinical abnormality observed was Onychomycosis and the prevalent fungal isolate was Trichophyton rubrum.

Conclusion: Superficial mycoses are prevalent worldwide. The current study illustrates that a notable number of patients with the end stage renal disease had a prevalence of superficial fungal infection; hence, a prompt recognition of skin lesion and identification of these superficial fungal infections alarm us to undertake early and judicious management in order to reduce the associated morbidity and thereby improve the quality of life in the said patients.

Purple Urine Bag Syndrome in a CKD Patient: A Case Report

Upma Narain, Arvind Gupta

Recent Progress in Microbiology and Biotechnology Vol. 5, 4 May 2021, Page 48-53

Purple urine bag syndrome is a rare disorder, in which the plastic disposable urinary catheter bag turns purple or blue following hours or days of urinary catheterization. This investigation reports the case of a 58 years woman was previously diagnosed with CKD resulting from diabetic nephropathy and also had a positive history of hypertension who presented with purple urine bag syndrome. It is a rare manifestation associated with urinary tract infection caused by bacteria that produce sulfatase or phosphatase.

Study on Imidacloprid Induced Intoxication and Its Biodegradation by Soil Isolate Bacillus weihenstephanensis

Arun Shetti, B. B. Kaliwal, R. B. Kaliwal

Recent Progress in Microbiology and Biotechnology Vol. 5, 4 May 2021, Page 54-66

Aims: The study was conducted to investigate the effect of imidacloprid on biochemical parameters and soil isolate growth. The soil isolate was also studied for imidacloprid degradation.

Study Design: The soil isolate was identified and used for toxicity testing. The isolate of Bacillus weihenstephanensis was further tested for its ability to degrade imidacloprid in minimal salt medium (MSM) and tryptic soya medium (TSB). The role of plasmid in imidacloprid degradation was established by curing experiments.

Place and Duration of Study: Department of Biotechnology and Microbiology, Karnatak University, Dharwad, India between June 2011 and December 2012.

Methodology: The soil isolate was identified by morphological, biochemical characters and 16s rDNA identification. Effect of imidacloprid on DNA, RNA, protein, glucose and growth in soil isolate was studied with 10-3 to 10-7 molar imidacloprid for 96 h. Imidacloprid degradation was determined in MSM and TSB for 28 days with samples taken on 7, 14, 21 and 28th day. The insecticide concentration was tested by HPLC. Plasmid curing was performed.

Results: The soil isolate was identified as Bacillus weihenstephanensis. The study involving soil isolate Bacillus weihenstephanensis with 10-3 to 10-7 molar imidacloprid showed significant (P<0.05) decrease in content of DNA, RNA, protein, glucose and growth. Bacillus weihenstephanensis in MSM and TSB showed 46 and 78% imidacloprid degradation in four weeks. The plasmid of Bacillus weihenstephanensis was cured in fourth generation. 18.80% and 75% degradation observed in cured and non cure cells of Bacillus weihenstephanensis in TSB.

Conclusion: Study showed that imidacloprid affects the biochemical contents and intern growth of soil isolate Bacillus weihenstephanensis. Study also revealed that Bacillus weihenstephanensis was able to degrade imidacloprid in MSM and TSB. Further plasmid curing revealed that the genes for imidacloprid degradation are located both in plasmid and chromosome. The soil isolate was able to utilize imidacloprid as sole carbon and nitrogen source as indicated by its growth in MSM.

Dipyridamole as an Antiviral Agent

Angel S. Galabov

Recent Progress in Microbiology and Biotechnology Vol. 5, 4 May 2021, Page 67-77

We described the pyrimido-pyrimidine derivative dipyridamole (DP) as an interferon (IFN) inducer in cultivated in vitro lymphoid cells, administrated orally in mice and in humans attaining high IFN titres within 24 hours. An IFN antagonistic period was recorded. Gut-associated lymphoid tissue was identified as the inducer target. Three double-blind placebo trials were carried out, involving more than 6000 participants. The results proved that DP acts as a prophylactic for influenza and viral acute respiratory diseases. Experimental evidence proved that the IFN-inducing activity of DP is due to its capacity to inhibit cAMP phosphodiesterase. Additionally, DP demonstrates a marked inhibitory effect on the replication of a broad spectrum of viruses belonging to various taxonomic groups. Contradictory results were obtained concerning DP’s IFN-synthesis in humans. A pharmacokinetic model and analysis convincingly explained the lack of DP-induced IFN production in humans. DP activities were followed and described in lupus erythematosus, common cold, chronic pulmonary diseases, liver diseases, ophthalmology, dermatology (herpesvirus and papillomavirus infections) and AIDS. Special attention was given to DP’s effect in blood aggregation.

The field experiment was carried out to study the performance evaluation of All India Co-Ordinated Vegetable Improvement Project (AICVIP) varietal trial (AVT-II) on growth, yield and quality of Tomato Indeterminate entries from 2014 to 2015 at the Department of Vegetable Crops, Horticultural College and Research Institute, Tamil Nadu Agricultural University, Coimbatore. The seeds of the tomato Indeterminate (AVT-II) Six entries were chosen for this study. Tomato cultivation can be conducted in open field conditions or in the greenhouse under environmentally regulated conditions. Among the Six entries tested (AVT-II), the highest fruit yield (345.0 q/ha) was recorded in 2012/ TOINDVAR-4 followed by ARKA VIKAS (C) (335.0 q/ha), whereas, the lowest fruit yield (291.0 q/ha) was recorded in 2012/ TOINDVAR-1.

A field experiment was carried out to study the vegetative growth and yield performance of Bottle Gourds- All India Coordinated Vegetable Improvement Project (AICVIP) varietal trials (AVT-I) were conducted from 2013 to 2014 at the Department of Vegetable Crops, Horticultural College and Research Institute, Tamil Nadu Agricultural University, Coimbatore. The experiment was laid out in Randomized Block Design (RBD) with three replications, which included the seeds of the Bottle Gourds entries viz. 2012/BOGVAR-1, 2012/BOGVAR-2, 2012/BOGVAR-3, 2012/BOGVAR-4, 2012/BOGVAR-5, 2012/BOGVAR-6, 2012/BOGVAR-7, 2012/BOGVAR-8, NDBG-104 (C) and CO1 (LC) were chosen for this study. Lagenaria   siceraria,  known   as   bottle   gourd, is a member  of  the  Cucurbitaceae  family. Bottle  gourd, which  is  extremely  strong  climbing,  herbaceous  and monoecious, is    easily    distinguished    from    other members  of  the  Cucurbitaceous  family  with  its  large white flowers. The Bottle gourd genotypes were sown with care in the field during the year 2013 to 2014 at the spacing of 300 X 75cm with the plot size of 7.5 m X 3.0 m. Significant differences were observed among the genotypes for growth and yield parameters. Among the different bottle gourds genotypes tested (AVT-I), the highest fruit yield (253.6 q/ha) was recorded in 2012/BOGVAR-5 followed by 2012/BOGVAR-8 (240.3 q/ha). Whereas the checks, NDBG-104 (C) and CO1-(LC) recorded the yield of 225.5 and 226.4 q/ha respectively.

Chemo-Nutritional Analysis and Organoleptic Quality Characteristics of Smoked African Catfish (Clarias gariepinus) Treated with Scent Leaf (Ocimum gratissimum) Juice

Adeosun Olubunmi, Adegbola, Sunday Abiola, Bankole, Adebukola Folake, Olateju, Gbolahan Basit

Recent Progress in Microbiology and Biotechnology Vol. 5, 4 May 2021, Page 91-100

This study evaluated nutrient composition, biochemical and organoleptic quality characteristics of smoked Clarias gariepinus treated with scent leaf (Ocimum gratissimum) juice during five (5) weeks of storage at ambient temperature. Table-sized catfish were subjected to solution of O. gratissimum juice at 0.2%. 0.4%, 0.6%, 0.8% and 1.0%v/v for 5 minutes prior to smoking. 1.0%v/v brine was used as the treatment control. Proximate composition of fresh and smoked fish was done at week 0 and week 5. Biochemical analysis and organoleptic quality assessment were done weekly. Result of proximate composition showed that crude protein values obtained ranged between 64.42% and 66.80%. Biochemical analysis result revealed that the values of free fatty acids, peroxide value and total volatile nitrogen increased with storage time, the values remained within the limits (0.09% - 0.65%, 1.22 meq/kg – 1.77 meq/kg and 1.495 mg/100 g – 11.750 mg/100 g respectively). The organoleptic assessment revealed that sample treated with 0.2% juice recorded higher values for taste (7.43), aroma (7.46), texture (7.33), appearance (7.77) and general acceptability (7.57) by the taste panel compared to the other samples and control. This could be due to the effect of spicy property of scent leaf. At low concentration, the organoleptic characteristics of the smoked fish were best. The study also revealed that Ocimum gratissimum juice can be used to improve biochemical and organoleptic quality of stored smoked Clarias gariepinus.

On the Effective Antiviral Treatment of Enterovirus Infections

Angel S. Galabov, Adelina Stoyanova

Recent Progress in Microbiology and Biotechnology Vol. 5, 4 May 2021, Page 101-116

Enteroviruses (EVs) as causative agents of diseases with varying clinical pictures occupy a significant place in human pathology. The great variability of these approximately 200 viruses is the basis of their comparatively quick development of drug resistance and thus the absence of effective antivirals for clinical use. Monotherapeutic applications of antivirals have been ineffective as anti-enteroviral chemotherapy. Therefore, to overcome the drug resistance barrier, we investigated EV replication inhibitors by looking at the effect of combinations of viral inhibitors in cell culture experiments. The next step was to test anti-EV combinations in vivo, in laboratory animals. The most successful achievement in this study was the development of the consecutive alternating administration (CAA) treatment course of triple combinations of antivirals with different modes of action. The CAA treatment scheme was applied in Coxsackievirus B1 and B3 infections by massive virus inocula (20 MLD50) in suckling albino mice. The CAA treatment scheme was used with three triple combinations, including five specific inhibitors of EV replication: disoxaril, pleconaril, guanidine.HCl, MDL-860 and oxoglaucine. Those experiments demonstrated a pronounced effectivity of the CAA approach expressed with a marked protective effect: decreased mortality and lengthening of the mean survival time. IC50 values (a phenotypic marker) of the virus progeny in virus samples isolated from target organs (brain, heart) of Coxsackievirus B infected mice treated with a triple combination of antivirals via the CAA scheme showed (a) a marked suppression in the development of drug resistance, and (b) in parallel, an unusual phenomenon of an increase in susceptibility to the partner compounds in the combinations. Sequence analysis of the genome (RNA) in samples of target isolates manifested the changes determining the increased susceptibility of the virus to the respective EV inhibitors. The data from these systematic studies show that the CAA treatment scheme with triple combinations of antivirals is an effective chemotherapy for EV infections.

Research on Semiquantification of Lethal Toxin at the Bottom of Reactors for Clostridium sordellii Culture

Diana Pérez-Etcheverry, Alberto Nieto-Cadenazzi, Iris Miraballes-Martínez

Recent Progress in Microbiology and Biotechnology Vol. 5, 4 May 2021, Page 117-129

Clostridium sordelli veterinary vaccine is made by cultivating the microorganism in large industrial bioreactors and purifying the toxin from culture supernatants. The harvesting time is currently established as a function of bacterial growth, but this time is not always associated with the maximum concentration of toxin present in the supernatant. As a result, it is simple to determine the total amount of toxin produced from batch to batch. Furthermore, the toxin concentration is currently measured using in vivo LD50 methods, and results can be obtained only after 72 hours, when the bioreactor has already been stopped. Moreover, the method is time-consuming and necessitates the use of a large number of experimental animals. The development and application of a latex agglutination reagent for semi-quantification of Clostridium sordelli lethal toxin (TcsL) in industrial bioreactors are described. The reagent was developed and characterised in our laboratory, with a detection limit of 8ng of toxin per ml of culture supernatant, and then validated in actual industrial conditions. The use of such a speedy (i.e., in minutes) and simple reagent will allow for real-time monitoring of the culture, standardising the optimal end-point for harvesting in terms of toxin quantity. As a result, the efficiency of TcsL industrial production may be improved immediately.  

Enzyme and Lectin Relationships in Cell Biology (Not Only Microbiology But Also Mammals and Plants)

Vladimir Lakhtin, Mikhail Lakhtin, Valeriy Davydkin, Vladimir Alyoshkin

Recent Progress in Microbiology and Biotechnology Vol. 5, 4 May 2021, Page 130-143

Current examples of inter- and intra-molecular relationships and directions in co-functioning lectins and enzymes are described and ordered. Regularities of such relationships are accented and used as additional criteria of the “true” lectins and lectinic enzymes. The data indicate importance and prospects of lectin-enzyme/ enzyme-lectin relationships (enzyme-biotics as lectins, lectin-biotics as enzymes, lectin-enzyme complexes and assemblies with regulated specificities) in molecular and clinical cell biology, microbiology, medical biotechnology and industry. Lectin-enzyme relationships give expanded platform/basis for further study and constructing advanced multi-functional agents influencing interactome of organism and supporting innovations in medicine and biotechnology.

Role of Cytochrome P450s in Insecticide Resistance in Anopheles gambiae (Diptera: Culicidae): The African Perspective

B. R. Mohammed, S. K. Malang, S. Mailafia, R. I. S. Agbede, R. D. Finn

Recent Progress in Microbiology and Biotechnology Vol. 5, 4 May 2021, Page 144-151

Cytochrome P450s are known to be critical for the detoxification and/or activation of xenobiotics such as insecticides in all living organisms including Anopheles gambiae. Many studies have demonstrated the role of P450s in insecticide resistance in An. gambiae. However, little is known about the impact of distribution in the African sub-continent. In this paper therefore, we analyse the P450 clans, the CYP6 family, localisation and function of An. gambiae CYPs, their insecticide substrates, regional distribution in the African continent and their role in insecticide resistance. This investigation from published data revealed that in the Central region; CYP6Z3, CYP6Z1, CYP12F2, CYP6P4, CYP6GA1, CYP6Z3 (Yaoundé, Cameroun) have bendiocarb, DDT and pyrethroids as substrates; in the Eastern region: CYP314A1 and CYP12F1 (Tanzania and Zanzibar) have DDT as a substrate, CYP32A3, CYP6Z1 and CYP6Z2 (Western Kenya) have DDT and carbaryl and permethrin; whilst in the Western region: CYP6AG1, CYP6Z2, CYP6Z3, CY6P3, CYP6P4, CYP6M2 (Ghana), CYP6M2, CYP6P3 (Benin), CYP325A3, CYP6P3 and CYP6M2 (Nigeria) all have DDT, carbaryl, permethrin, trans-and cis-permethrin, deltamethrin, bendiocarb as substrates. Additionally, CYP6M2, CYP6P3, CYP6Z3 (Côte d’Ivoire), CYP6P3, CYP6Z2 and CYP9J5 (Burkina Faso) have bendiocarb, DDT plus pyrethroids and only pyrethroids as substrates respectively. Interestingly, CYP6P3 is observed to metabolize all the available insecticides (DDT, pyrethroid, trans- and cis-permethrin, deltamethrin and bendiocarb), indicating possible insecticide cross resistance across all the three regions of Africa. A more detailed understanding of the substrate specificities of various P450s and the geographical distribution of insecticide resistance in Africa is quintessential for an effective resistance management.

Impact of Alcohol Exposure on the Composition of HeLa-Derived Extracellular Vesicles

Leandra B. Jones, Sanjay Kumar, Aliyah J. Curry, Jayde S. Price, Alexandre Krendelchtchikov, Brennetta J. Crenshaw, Courtnee’ R. Bell, Sparkle D. Williams, Tambre A. Tolliver, Sabita N. Saldanha, Brian Sims, Qiana L. Matthews

Recent Progress in Microbiology and Biotechnology Vol. 5, 4 May 2021, Page 152-170

Extracellular vesicles are nanosized vesicles that are under intense investigation for their role in intercellular communication. Extracellular vesicles have begun to be examined for their role in disease protection and their role as disease biomarkers and/or vaccine agents. With the high mortality rate of cervical cancers, more information is needed regarding the detection and treatment of the diseased. In this regard, several investigators have begun to examine extracellular vesicles and their role in disease protection. The goal of this study was to investigate the effects of alcohol exposure on the biogenesis and composition of extracellular vesicles derived from the cervical cancer line, HeLa. The HeLa cells were cultured in exosome-free media and were either mock-treated (control) or treated with 50 mM or 100 mM of alcohol for 24 h and 48 h. Our results demonstrated that alcohol significantly impacts HeLa cell viability and exosome biogenesis/composition. Importantly, our studies demonstrate the critical role of alcohol on HeLa cells, as well as HeLa-derived extracellular vesicle biogenesis and composition. Specifically, these results indicate that alcohol alters extracellular vesicles’ packaging of heat shock proteins and apoptotic proteins. Extracellular vesicles serve as communicators for HeLa cells, as well as biomarkers for the initiation and progression of the disease. HeLa cells used the primary model in this study, the research findings may have substantial implications on diagnostics and therapy for a variety of cell types and organ systems. Further investigation is needed to elucidate the mechanism(s) involved in these processes.

Solid-state Fermentation in the Bioavailability of Nutrients in Rice Bran

Anelise Christ Ribeiro

Recent Progress in Microbiology and Biotechnology Vol. 5, 4 May 2021, Page 171-177

Rice bran, a co-product from grain processing, is rich in nutrients and bioactive compounds. However, rice bran has some limitations for its application, such as the activity of hydrolytic and oxidative enzymes, median digestibility and antinutritional factors. However, fermentative processes are promising to minimize these effects efficiently. The purpose of this chapter is to evaluate the advantages that fermentation has on the composition of nutrients and benefits of rice bran for human consumption. With recent studies in the literature, the benefits of fermentation are evident in increasing the bioavailability of nutrients, mitigating antinutritional factors, increasing digestibility making biomass a promising alternative in human nutrition.