Advances and Trends in Biotechnology and Genetics Vol. 3

Thermophilic Bacteria: Novel Polymers for Biotechnological Applications

Palaniappan Ramasamy — 2019-12-21

Aims: We analyzed thermotolerant bacterial isolates of thermophiles from various regions in South East coasts of India. They were analyzed for Fatty Acid Methyl Ester (FAME) for identification and classification.

Study Design: A total of forty three (9.6%) thermotolerant bacterial isolates of thermophiles were secluded out of 444 isolates obtained from water samples from the Adyar River, Marina coast, Muttu Kadu Backwaters, Kovalam coast (India) and a prawn hatchery. In addition, a novel thermotolerant Gram positive Bacillus type bacterium from the pelletized feed of fish was examined. A total of 18 thermotolerant bacteria were analyzed for Fatty Acid Methyl Ester (FAME).

Results: Amongst 134 isolates that grew on E. coli FAGI agar, 23 (17%) were thermotolerant when tested at 15 psi for 30 min. Of the 72 isolates that grew on Staphylococcus Baird Parker agar 10 (13%) were thermotolerant, whilst among the 128 isolates that grew on Salmonella agar ÖNÖZ 10 (8%) were thermotolerant. In contrast, none of the 110 isolates that grew on TCBS agar (Vibrio sp.) exhibited thermotolerance, although they were isolated from the same regions as the other bacteria. The optimal temperature for the growth of most thermophiles was 45°C, except for 9 isolates. Of the latter, 7 grew at 50°C, one at 52°C and one at 55ºC, respectively.There were some extremely thermotolerant bacterial isolates that exhibited consistency in profuse growth at 15 psi (121°C) for 30 min. Other isolates showed inconsistency in growth after autoclaving the cultures at 15 psi (121°C) for 30 min, and these bacteria showed development of endospores. Isolates of thermotolerant bacteria that grew on Staphylococcus Baird Parker agar and Salmonella agar ÖNÖZ were predominantly antibiotic resistant when compared with those bacterial isolates that grew on E. coli FAGI agar. A major 43 kDa toxin-protein as well as 15 other minor protein subunits was detected in the thermotolerant Bacillus sp. isolate. The proteins were toxic and caused mortality to goldfish Cyprin macrophthalmus. The thermotolerant bacteria were identified as B. cereus, Bacillus subtilis, B. pumilus, B. laevolacticus), B. alcalophilus, Kurthiasibirica, Geobacillus stearothermophilus, Staphylococcus schleiferi and S. gallinarum,

Conclusions: This study documents 43 thermotolerant bacterial isolates from water samples and a Bacillus-type bacterium from fish feed in Chennai region, India and these bacterial isolates offer the potential for exploitation in biotechnological applications.

Effects of Moisture Barrier and Initial Moisture Content on the Storage Life of Some Horticultural Produce in Evaporative Coolant

Iwuagwu, Christian C. — 2019-12-21

Shelf-life of five horticultural produce were studied. These include three leafy vegetables: Telfairia occidentalis, Celosia argentea and Amaranthus cruentus and two fruit vegetables: Lycopersicum esculentum and Abelmuschus esculentus. The layout plan of the experiment was a 3x2 factorial in a completely randomized design and each treatment replicated three times. The two factors examined were moisture barrier at three levels namely: thick lining, thin lining and non-lining. The other factor included initial moisture content of the produce, namely, turgid and partially wilted. Partial wilting of the produce was achieved by exposing freshly harvested materials at ambient temperature to dry for 45 min. During this period, about 20% of the moisture content was lost. Eighteen (18) vegetable baskets which work on the principle of evaporative cooling system were used. Each type of produce was stored at a time inside the vegetable basket. Some quantity of each produce were kept on the laboratory benches to serve as controls. Data recorded includes length of storage of produce, severity of disease infection, visual quality, disease incidence, ambient temperature and relative humidity of the storage baskets. The result indicates that there was a significantly higher relative humidity (P< 0.05) in the lined baskets than in the non-lined baskets. Consequently, the shelf-life of produce in lined basket was prolonged. Turgid produce had better quality retention and stored much longer than partially wilted produce. Generally, the evaporative coolant baskets provided an average temperature of 3°C lower than the ambient condition. The shelf-life of leaf vegetables T. occidentalis, C. argentea and A. cruentus was extended appreciably for 7-8 days compared to the controls.

Production of Cellulase and Xylanase Using an Amazon Fungus (Trametes sp.)

Rafael Lopes e Oliveira — 2019-12-21

In order to use the residue from the beneficiation of Brazil nuts (Bertholletia excelsa HBK) as substrate in solid-sate fermentation (SSF), in this work, the production of cellulase and xylanase by the fungus Trametes sp. was investigated, using the residue as a carbon source. Employing a 2^4-1 fractional experimental design, the influence of substrate moisture, nutrient addition and inoculum quantity on enzymatic activities was verified. Moisture was detected to be statistically significant for the production of both enzymes, and increasing the moisture leads to the improvement of cellulase and xylanase activities. Nitrogen and phosphate were also important for enzymes production by the Amazon Trametes sp. The use of this Amazon strain to obtain cellulase and xylanase via SSF of Brazil nut residue appears to be feasible when maintaining substrate moisture at 80%, nitrogen source at 0.9% and low inoculum concentrations.

Food Condiment Series 1: Fungal Contamination of Two Food Condiments (Brachystegia eurycoma and Detarium macrocarpum) Sold in Some Local Markets in Rivers State

Ikechi-Nwogu Chinyerum Gloria — 2019-12-21

This work is aimed at identifying these organisms in order to reduce the contamination of food condiments. A survey of seed-borne fungi associated with the seeds of Brachystegia eurycoma and Detarium macrocarpum (Whole and Ground) obtained from two different local markets in Rivers State namely; Etche and Bori was conducted. Fungi were isolated using Standard Blotter Method and pure cultures of Aspergillus flavus, Aspergillus niger, Aspergillus spp, Botrydiplodia theobromae, Fusarium solani, Rhizopus stolonifer, Penicillium spp, Penicillium notatum and Fusarium moniliforme, were obtained on Potatoes Dextrose Agar (PDA). Comparing the fungal contamination of the food condiments Detarium macrocarpum and Brachystegia eurycoma from 2 different markets in Rivers State, shows that the fungi obtained from Etche Market has the highest percentage incidence of 181.83% as compared to Bori Market (79.16%). It shows that these legumes are good substrate, for the growth of pathogenic fungi that produce toxins that are detrimental to human health because of the diseases they cause. Fungi produce toxins which cause harm to us thus, consumers are advised to buy the whole seeds of (Brachystegia eurycoma and Detarium macrocarpum) and use immediately. Preservation of food condiments should be a major concern in order to improve food security and safe guard the health of the consumers.

The Use of Streptomyces capoamus Isolated from Caatinga in the Production and Characterization of Microbial Endoglucanase

Rafael Lopes e Oliveira — 2019-12-21

Cellulases are hydrolases of great importance to industries, especially due to their ability to produce ethanol via hydrolysis of cellulolytic materials. Actinomycetes are the producers of these enzymes, particularly the genus Streptomyces sp. The present study is the first report on the production and characterization of cellulolytic complex secreted by Streptomyces capoamus, isolated from the rhizosphere soil of Caatinga. In selecting the microbial producers of cellulolytic complex in qualitative tests, 171 x micro organism showed the most expressive enzymatic index. Regarding the production time of the complex, fermentation was done for 7 days, with aliquots being taken every 24 h. Peak production was obtained during 48 h fermentation. It was done at 37°C and under an agitation of 180 rpm. It was noted also that the 171x micro-organism produced the enzyme in greater quantity. The experiment was done with the most significant actinomycetes (171x), optimal substrate concentration (carboximeticellulose), cultivation temperature and pH of initial output. The results showed that a higher cellulolytic complex was obtained with 2% substrate, 45°C temperature and initial pH 4.0. The microorganism was identified at genus level by microculture method; and with molecular identification method, it was identified as S. capoamus UFPEDA-3410. In optimal culture conditions, this strain produced 0.309 U/mL cellulose, a good production for a thermostable endoglucanase stable in a broad range of pH and stable temperature. It has potential applications in a wide range of industries. Industrial processes are generally carried out at elevated temperatures. Therefore enzymes with a high optima temperature and stability are desired for such applications.

A Review on Algal Plasmid DNA

S. R. Sivakumar — 2019-12-21

Plasmid plays a crucial role in molecular biotechnology as vectors. Once it is believed that plasmids are found only in prokaryotes but the development in molecular Bio-technology proves that plasmids are also found in eukaryotes such as yeasts and some algae. Several researches are carried out to isolate the plasmid DNA from both micro and macro algae. This review gives an overview about the plasmid DNA of various Algae.

Identification of the Essential Oils from Annona senégalensis Pers. Sous-espèce Oulotricha Le Thomas (Annonaceae)

Nkounkou-Loumpangou Célestine — 2019-12-21

Annona senegalensis Pers., Oulotricha Le Thomas subspecies (Annonaceae), is an evergreen shrub that is used for ethnomedicinal purposes and as a source of food. The chemical composition of the essential oils from leaves, stems bark, roots bark, epicarp and mesocarp of these plant, growing in Brazzaville (Congo), were analyzed by CG and CG-MS. These oils essentially contain sesquiterpenic compounds (58.3-97.7%), dominated by oxygenated sesquiterpenes (21.8-88.3%), with elemol (13.2-35.0%), β-and γ-eudesmols (3.7-58.3%) as characteristic components. The essential oils from roots and stems bark is distinguished by its high content in diterpenes (17.1 and 11.9% of the total), while the seeds presents a significant amounts of monoterpene hydrocarbons (25.8%) with α-pinene (6.2%) and β-phellandrene (11.5%) as major components accompanied by two oxygenated monoterpenes : bornyle acetate (4.5%) and smallest of 1,8-cineole. However, in the epicarp oil, the presence of about 5.8% of oxygenated monoterpenes as terpinen-4-ol and bornyle acetate in comparable rates (1.7%) was noted. The mesocarp oil is exclusively rich in aliphatic fatty acids (35.8%) which is absent in the other organs, but represented by lauric acid (18.0%), hexadecanoïc acid (8.6%), myristic acid (7.2%) and oleic acid (2.0%). Results were compared with same species collected in the democratic republic of Congo and in Cameroon essentially dominated by monoterpenes (84.2 and 87.6%).

Biopreservation of a Cameroonian Indigenous Fermented Milk (Pendidam) Using Biosurfactants

Mbawala Augustin — 2019-12-21

Natural compounds with bio-preservative properties are gaining attention of researchers worldwide. It is the case of biosurfactants known as surface active compounds produced by microorganisms which deserved antimicrobial and antioxidant properties. The present study was aimed to improve the shelf life of a Cameroonian indigenous fermented milk “pendidam” using biosurfactants derived from that food matrix. Biosurfactants was extracted from pendidam, purified, characterized preliminarly and its in vitro antimicrobial activity against microorganisms isolated from pendidam was assessed. Then, the biosurfactants was applied in pendidam and the microbiological quality of pendidam was followed up during 4 days of preservation at room temperature. The results obtained show that the crude extract of biosurfactants exhibit emulsifying and surface activities which remained stable after heat treatment at 100°C for 15 min, NaCl concentration of 20% and pH values ranging from 2 to 12. Furthermore, the biosurfactants was microbicidal against Candida spp. and E. coli at a concentration of 100 mg/mL. Solution of Biosurfactants (50 mg/mL) introduced into pendidam at a ratio of 10% (v/v), reduced significantly (p<0.05) the proliferation of the total aerobic mesophilic flora, E. coli and Candida spp. during 4 days of preservation at room temperature while no adverse effects were noticed on the growth of lactic acid bacteria. The present study highlights the bio-preservative potential of biosurfactants and suggests its use in food industry as food preservatives.

Antidiabetic and Antiinflammatory Activity of Methanol and Aqueous Extracts of Pergularia daemia (Forssk.) Chiov.

A. Doss — 2019-12-21

The present study is aimed to investigate the antidiabetic and antiinflammatory activity of crude extracts (methanol and aqueous) of leaves of Pergularia daemia. The acute oral toxicity showed that the crude extracts of P. daemia leaf were safe until 2000 mg/kg body weight and no macroscopical organ abnormalities were observed in acute oral models. The results suggested methanol extract of leaves may contribute to the reduction of blood glucose levels and can be useful in the management of diabetes. The crude extracts were also assessed for its antiinflammatory activity by in-vitro methods. Out of the two extracts tested, the methanol extract exhibited significant activity. P. daemia extracts exhibited antidiabetic and antiinflammatory activities which may be attributed by the presence of active phytoconstituents

Control of Huanglongbing (HLB) Disease with Reference to Its Occurrence in Citrus Industry in the World

Hajivand Shokrollah — 2019-12-21

Huanglongbing (HLB) disease is one of the most severe diseases of citrus plantation all over the world. HLB has destroyed an estimated 60 million citrus trees in Africa and Asia and more than 40 countries were infected by HLB in Africa, Asia and USA, The HLB pathogens are highly fastidious phloem-inhabiting bacteria in the genus Candidatus liberibacter. The isolate from South Africa, Candidatus liberibacter africanus is considered heat-sensitive and only found in Africa regions. The isolate from Asia, Candidatus liberibacter asiaticus is the most severe strain and widespread. This strain is heat-tolerant and can show the disease symptoms in humid, cool and hot temperature (up to 35°C). The isolate from America has been named Candidatus liberibacter americanus and it was detected in Brazil and Florida. Current disease management strategies focus on vector control to avoid infection within the farm or management of infected trees to eliminate source of inoculum. For the Malaysia case, integrated pest management (IPM) is highly recommended to be adopted to control the disease. The successes of individual or combined approaches depend upon on the infestation level, knowledge and regulation. Basically, in the regions where disease incidence is low, the most common practices are removal of symptomatic trees, intensive monitoring, pesticides application and biological control of the vectors. Preventive and curative measures of infected trees includes enhanced nutrition and plant hormone application by foliar sprays, regulating soil pH, precision irrigation based on the needs of HLB-affected trees. In reality, effective disease control of HLB is still difficult, especially in the area where the disease is widespread, vectors are well established and the hosts are susceptible. In this article, possible HLB disease management approaches are discussed.

Effect of Lead (Pb) on in vitro Shoots Essential Oil Constituents of Jatropha curcas L.

Ravi Mahalakshmi — 2019-12-21

Essential oil chemical constituents of Jatropha curcas were analyzed for the effect of lead (Pb) on in vitro shoots. A total of 36 and 30 compounds obtained from in vitro and Pb treated shoots. The four compounds were α-copaene, β-bourbonene, T-muurolol and lauric acid increased parentage in Pb treated in vitro shoots. Lead treated in vitro shoots resulted in production of 21 new compounds and increased percentage of fatty acid, alkanes and aldehyde.

Assessment of Dental Pulp Stem Cell (DPSC) Biomarkers Following Induction with Bone Morphogenic Protein 2 (BMP-2)

Joseph Cinelli — 2019-12-21

Introduction: Tissue regeneration and biomedical engineering are the goals of modern research that have made tremendous strides in recent years. Dental pulp stem cells (DPSCs) have been demonstrated to exhibit functional multipotency, differentiating into neurons, adipocytes, and other cell types. The primary goal of this study was to investigate the ability of bone morphogenic protein (BMP-2) to induce proliferation and differentiation of DPSC isolates into mineral forming bone cell precursor lineages.

Study Design: This was a prospective study with the non-randomised experimental design.

Place and Duration of Study: This study was conducted at the University of Nevada, Las Vegas – School of Dental Medicine between May 2017 and August 2018.

Methodology: Eight previously isolated dental pulp stem cell (DPSC) isolates were grown in culture and treated with bone morphogenic protein (BMP-2) to evaluate any effects on growth, viability or biomarker expression.

Results: BMP-2 induced significant changes in cellular growth among a subset of DPSC with slow doubling times (sDT), which corresponded with similar increases in cellular viability. Also, BMP-2 was sufficient to induce mRNA expression of alkaline phosphatase (ALP) and other differentiation markers among the sDT isolates – although no significant changes were observed among the DPSC isolates with rapid or intermediate DTs (rDT, iDT).

Conclusions: This study may be the first to demonstrate not only the differential responsiveness of DPSC isolates to BMP-2, but also to identify the MSC biomarkers that may affect initial DPSC responsiveness to this stimulus. Although many studies have evaluated the role of the biomarkers NANOG, Sox-2 and Oct-4 in DPSC isolate, no other study of DPSC multipotency has evaluated the role of Nestin – which may be one of the key factors that potentiate or limits the responsiveness to BMP-2 and osteogenic potential among DPSCs.

Antibiotics Susceptibility Profile of Bacterial Isolates from Urinary Tract Infection among Honeymoon Women in Iraq

Nada Khazal Kadhim Hindi — 2019-12-21

Urinary Tract Infections (UTIs) is an infection caused by the presence and growth of microorganism anywhere in the urinary tract. This study aims at determining the most common microorganism that causes UTI in honeymoon women, detection the most effective antimicrobial agents that causative agents of UTI and select the best antibiotics for treated UTI. One hundred and thirty specimens were collected from women suspected with UTI {honeymoon (95) and other women (35)} in al-Qassim hospital; during the period (January 2012 to 30th September 2012). The bacterial isolates were identified tested for antibiotics sensitivity test6. Honeymoon women were showing higher rate of UTI than other women in (87 from 95) (91.5%) and (19 from 35) (54.3%) respectively. E. coli (46%) and Staphylococcus aureus (42%) were predominant isolate in honeymoon. Regarding Antibiotics sensitivity test, E. coli and Staphylococcus aureus showed highly sensitive to Nitrofurantoin (100%), (94.5%) followed by Amikacin (90%), (85%), Gentamicin (88.2%), (75%) and ciprofloxacin (80.7%) (88%) respectively, while Oxacillin were not effective against tested isolates. In summary, Honeymoon women were showing higher rate of UTI than other women. E. coli was isolated more than other bacteria that causing UTI in honeymoon women. Most bacterial isolates was showed highly sensitive to Nitrofurantoin and resistance to Oxacillin.

Morphological Characterization and Tissue Culture of Medicinal Plant Aloe vera L.

B. Chandra Sekhar Singh — 2019-12-21

The morphological characterization and micro propagation were carried out on Aloe vera (Aloe barbadensis Miller) accessions collected from different locations in India were discussed in this book chapter. The experiments were conducted on 12 Aloe accessions and the leaf length, width, thickness, stem length and thickness were measured using verniercalipers. The plantlets were chemically treated with 2.5% (w/v) of sodium hypochlorite which is an alternative safer surface sterilization method for replacement of mercury chloride by using shoot tip as explants. The sterilization results indicate that the highest number (93%) of viable explants (10) with the best regeneration potential was in Murashige and Skoog (MS) medium compared with other treatments. Multiple shoots/explants (35) were found in MS medium containing 6-benzylaminopurine (BAP, 1.5 mgL^-1; kinetin (0.5 mgL^-1), indole-3-butyric acid (IBA, 0.2 mg L^-1), indole-3-acetic acid (IAA, 0.1 mg L^-1), and naphthalene acetic acid (NAA, 0.2 mg L^-1). Based on the shoot proliferation, a maximum number (87.89%) of shoots obtained from the micro-shoot culture method and a maximum number (91.12%) of roots were obtained on MS medium supplemented with NAA (0.2 mg L^-1), IBA (0.2 mgL^-1) and IAA (0.1 mgL^-1) which was found to be the best. The healthy rooted plantlets were successfully transferred and obtained as the plants were of 100% survival. The regenerated plants were morphologically similar to the mother plant.