Current Strategies in Biotechnology and Bioresource Technology Vol. 2

Investigation of Enzyme Activities during Composting of Soiled Cotton Waste: Recent Developments

A. Patnaik — 2020-07-14

Huge amount of soiled cotton waste is being produced by utilization of sanitary napkins by the women who have attained puberty. An attempt has been made to manage these wastes through composting and vermicomposting. Enzyme activities were studied during the composting and vermicomposting of soiled sanitary napkins. The highest protease (354.356 µg/g/hr), urease (0.104 µg/g/hr), amylase (119.08 µg/g/hr) and cellulase (170.13 µg/g/hr) activity was observed in vermicomposting sets. There was significant increase (p<0.001) in the enzyme activity in the vermicomposting and composting sets as compared to control.

Cervical Smear Analysis in Cross River State, Nigeria

Ayodele Omotoso — 2020-07-14

Background: Cervical cancer is one of the most common female genital tract malignancy and one of the leading causes of death among the female population. Cervical cancer continues to be with high prevalence among the populace in Africa. It is one of the cancers whose incidence has been drastically reduced due to continuous systematic screening.

Aim: To evaluate the cervical smear analysis in Cross river State, Nigeria.

Methodology: This is a cross sectional study of cervical smear of women in 5 out of the 18 local government areas of Cross River State (CRS), Nigeria. These local governments were randomly selected. These local government areas include, Akpabuyo, Biase, Calabar south, Municipality and Akamkpa local government areas. The primary health care centres in each of these five local governments were visited, Pap smear was done for the women who visited these primary health care centre and the results were analysed.

Results: A total of 114 cervical smears were analyzed. The age range of the subjects is between 20-69 years. The mean age in this study is 34.2 years. In all, 7.0% of the cervical smear show squamous intraepithelial lesions out of which 6.14% had low grade squamous intraepithelial lesion (LSIL) and 0.9% had high grade squamous intraepithelial lesion (HSIL). Cervical smear that were negative for intraepithelial lesion had a prevalence of 93.0%.

Conclusion: The prevalence of squamous intraepithelial lesion (SIL) is relatively high in semi-rural areas in Cross River State and there is urgent need for improvement in screening programs in order to reduce the incidence rate of the cervical cancer.

Impact of Bacillus and Pseudomonas sp. on Growth and Development of Pearl Millet and Ragi

B. Harinathan — 2020-07-14

In the present exploration, the effect of Bacillus sp. (C2) and Pseudomonas sp. (C7) isolated from Calotropis gigantea and Cyanodon dactylon from Shenbagathoppu hills, Srivilliputhur, Tamil Nadu, India on growth and development of pearl millet and ragi were considered. The outcome of these isolates was recorded for morphological characters like plant height, fresh and dry weight. In the present study, maximum fresh weight and dry weight of pearl millet and ragi were recorded when treated with isolate C2 compared to C7 on all schedules of 30 and 45 days. Plant height recorded firm in pearl millet on 45 days when treated with isolate C7 while ragi recorded inclined growth when treated with C2 isolate. Physiological characters like total chlorophyll, IAA, starch, fresh and dry weight were recorded on 45 day after sowing both in pearl millet and ragi treated with C2 and C7 isolates. It was found that, the isolate C2 treated plants recorded maximum total chlorophyll, IAA, starch, fresh and dry weight compared to C7 isolate treated plants both in pearl millet and ragi. Among the parameters tested, ragi plants recorded better total chlorophyll, IAA, starch, fresh and dry weight compared to pearl millet plants.

Kinetics of Methane Production from Co-digestion of Agricultural Wastes

Karlygash Korazbekova — 2020-07-14

Anaerobic digestion (AD) is widely used for treatment of organic waste for biogas production. Co-digestion of agricultural wastes with various substrate combinations is one of the ways to increase and stabilize biogas yield. The influence of co-substrates on kinetics of methane production in anaerobic fermentation of pig and manure slurry was studied by performing a series of laboratory experiments using food waste, biowaste and winemaking waste as co-substrates. Experiments were performed in the laboratory reactors of «Hohenheim» biogas testing system in batch mode at the temperature of 37ºC. Methane production rate was used to evaluate co-digestion of agricultural slurry. Research results showed that co-substrates gave significant effect to methane production kinetics. The kinetic parameters of methane production i.e. potential yield of methane (P), maximum methane yield rate (R_m) and duration of lag-phase (λ) were analyzed using modified Gompertz equation. Technical time to produce 95% of potential methane yield was investigated and efficient period of anaerobic digestion was calculated. The potential production of methane was reached in lag-phase duration (λ) of about 10-15 days. Maximal methane production rate was reduced 2-3 times in co-digestion than in mono-digestion of pig and cattle manure during anaerobic digestion for 35 days.

RNA Polymerase: The Master Executor

Vivek Pandey — 2020-07-14

RNA polymerases belong to a selected group of evolutionarily conserved proteins. Their role emerged with the coming of DNA molecules having more stability than the RNA molecules, which formed the RNA world before it. The chapter makes a detour through RNA polymerases working in a eukaryotic system and their prokaryotic counterpart—a special emphasis on their structural aspect and functional variation arising due to variation in structure. Further, the importance of various known transcription factors has been explained along with accessory transcription factors that serve as an essential regulator of RNAP activity. A mutation occurring in these proteins results in the aberration of polymerase activity that results in a plethora of diseases ranging from the developmental disorder, autoimmune diseases to cancers.

The Biology, Engineering and Applications of CRISPR/Cas9 System

Bilal Ahmad Lone — 2020-07-14

Microbes use diverse defense strategies that allow them to withstand exposure to a variety of genome invaders such as bacteriophages and plasmids. One such defense strategy is the use of RNA guided endonuclease called CRISPR-associated (Cas) 9 protein. The Cas9 protein, derived from type II CRISPR/Cas system, has been adapted as a versatile tool for genome targeting and engineering due to its simplicity and high efficiency over the earlier tools such as ZFNs and TALENs. With recent advancements, CRISPR/Cas9 technology has emerged as a revolutionary tool for modulating the genome in living cells and inspires innovative translational applications in different fields. Here, we review the developments and potential uses of CRISPR/Cas9 technology as well as recent advancements in genome engineering using CRISPR/Cas9.

Advanced Research on Malignant Hyperthermia: A Runaway Thermogenic Futile Cycle at the Sodium Channel Level

Charles H. Williams — 2020-07-14

Malignant Hyperthermia (“MH”)—the rapid onset of extremely high fever with muscle rigidity—is caused by a runaway heat production futile cycle mediated via the sodium channels at the myoneural receptor sites. MH is not triggered by non-depolarizing muscle relaxants; however, depolarizing muscle relaxants may trigger it [1]. Here we present a de novo hypothesis of how MH is triggered and develops. We believe that the acetylcholine receptor/sodium channels in the muscles of MH susceptible pigs initiate MH by allowing an increased flux of sodium ions when it is depolarized by acetylcholine or other depolarizing agents, such as succinylcholine and Halothane. Our theory is consistent with our observations of the effects of general anesthetics over twenty years. Succinylcholine is a depolarizing agent that is a potent MH trigger. Acetylcholine, the natural depolarizing muscle activator, may trigger MH if the susceptible patient or animal is exposed to sufficient stress, i.e., during strenuous activity, such as transport, fighting, breeding, etc. Halothane apparently destabilizes the myoneural sodium channels, which rapidly induces MH. The increased sodium channel activity releases heat with cascades that further releases of heat which results in the rapid onset of MH. MH susceptible pigs have increased action potential amplitudes at their myoneural junctions that are abnormally long in duration. This increased activity is thought to induce hypertrophy of muscle mass, increase metabolic rate, and cause other physical manifestations. When slaughtered, this increased metabolic activity causes the rapid post mortem release of heat in the muscles of MH susceptible pigs and, at the same time, the accumulation of low acidity, all of which denatures the muscle proteins to result in a pale, soft, exudative, pork meat considered to be of lesser quality for human consumption. The potency of inhalation anesthetics as a MH triggers varies widely. The inhalation anesthetic Halothane is a strong trigger of MH, causing MH within minutes of exposure. In contrast, the anesthetic Sevoflurane is a very weak trigger of MH, requiring several hours of inhalation exposure to trigger MH. Because of this, changing from Halothane to Sevoflurane as the general anesthetic of choice for surgeries in hospitals in the Greater Kansas City area during 1994 to 2006 led to an 11-fold decrease in the incidence of MH, from 1:50,000 to 1:550,000 [2]. One non-depolarizing muscle relaxant, Organon 9426 (“Rocuronium”) temporarily prevents MH in MH susceptible pigs when they are given sufficient dosages of it before being challenged with either Halothane or succinylcholine. Binding Rocuronium to the myoneural receptor sites apparently stabilizes them, thereby preventing increased sodium channel activity, and resulting MH. However, other non-depolarizing muscle relaxants do not have this protective effect— for examples Vecuronium, Arduan, and Organon 9616 do not. Uncoupling of mitochondria is not the source of accelerated heat production in MH susceptible pigs, as heart, liver, and skeletal muscle mitochondria isolated from MH susceptible pigs are all competent.

Acute and Sub-chronic Toxicity Study of the Extract and Powder of Operculina macrocarpa (L.) Urb. in Mice: Recent Perspectives

Carpejane Ferreira da Silva — 2020-07-14

The aim of the study was to evaluate the experimental acute and sub-chronic toxicities of Operculina macrocarpa with plant extract and powder, respectively, in male and female mice. Phytochemical prospection was performed with extract and administration in single doses by intraperitoneal route to six groups and control at 1230, 970, 700, 350, 120 and 30 mg/kg doses and distilled water (0.1 ml/10 g), respectively. Sub-chronic doses of 1230, 700, 30 mg/kg/day and satellite (1230 mg/kg/day) were administered orally in feed. Major endpoints included alterations in the central and autonomic nervous system, water and food intake, body weight, hematological and biochemical parameters. Phytochemical screening identified compounds: Alkaloids, flavonoids, xanthones, leucoanthocyanidins and tannins condensate. In the acute study, mortality was observed with toxicity signs to the central nervous system (CNS) at LD₅₀ of 270 mg/kg. There were no significant changes in water and food intake, body weight, haematological and biochemical parameters, and histopathological examination in the sub-chronic study (p value). Results indicate that the oral administration of O. macrocarpa powder in feed is less toxic and relatively safer.

Antagonistic Efficacy of Lactic Acid Bacteria against the Human and Fish-borne Microbiota of Fresh Indian Mackerel Fish Chunks

Kannappan Sudalayandi — 2020-07-14

Various strains of lactic acid bacteria (LAB) namely Lactobacillus acidophilus NCIM 2287, Lactobacillus plantarum NCIM 2085, Lactobacillus helveticus NCIM 2126 and Lactococcus lactis NCIM 2114 were procured from the National Chemical Laboratory (NCL) Pune, India. These LAB cells were individually (10⁷cfu/ml) sprayed using a sterile syringe on the dressed fresh mackerel fish chunks and incubated at 37°C for 2 days. The growth pattern of each LAB and their antagonism against fish-borne spoilage bacteria namely, specific spoilage bacteria, halophilic bacteria, coliforms, lipolytic, proteolytic bacteria and total plate count were estimated for three days. Pathogenic Staphylococcus aureus was inhibited by Lb. acidophilus on the second day with 4.30 log difference as compared to control. The growth of specific spoilage bacteria was decreased by Lb. plantarum spray on the first day by 1.0 log difference. Lb. helveticus inhibited S. aureus on the third day by 3.5 log difference. Out of the four LABs tried, Lb. helveticus showed the best inhibitory effect against the fish-borne bacteria. All three LABs exhibited inhibition against the fish-borne spoilage bacteria, they may thus be potentially used as bio-preservative bacteria to preserve the whole fish meat or minced meat products etc. for a shorter duration.

Cyclical Changes in the Histology of the Gonads (Ovary and Testes) of African pike, Hepsetus odoe: An Overview

E. O. Idowu — 2020-07-14

Aims: To study the Morphology, stages of maturation and histology of gonads of African pike, Hepsetus odoe from Ado Ekiti Reservoir in order to establish its reproductive biology.

Study Design: Ecological and laboratory analysis were used in this study.

Place and Duration of Study: Hepsetus odoe samples were collected from Ado Ekiti Reservoir. The study was carried out between August 2014 and July 2015 at the Department of Zoology and Environmental Biology Laboratory, Ekiti State University, Ado Ekiti.

Methodology: Sex of each specimen collected was determined by examination of the gonads after dissection and the ratio of male to female calculated. The egg diameter was measured using an ocular micrometer in a binocular microscope. Gonad maturity stages were assessed and classified. Stage I, Immature; II, developing; III, mature; IV, ripe and running; V, spent. The fish were dissected and the gonads at each stage of development were removed. They were prepared according to standard histological procedures. Conclusive staging of the gonads were made on microscopic examination of the stained sections and photomicrographs taken.

Results: Out of 685 specimens examined 354 were males and 331 were females giving a sex ratio of 1:1.1 (female:male). There was no significant difference (P>0.05) in the sex ratio. The gonads were in pairs. All the gonad maturity stages were encountered in both male and female gonads. These stages were; immature, developing, mature (ripening), ripe and running, and spent. Histology of female ovary showed that six stages were encountered in the oogenesis, namely, oogonium, primary oocyte, primary vitellogenic oocyte, secondary vitellogenic oocyte, tertiary vitellogenic oocyte and hyaline oocyte while in male spermatogenesis five stages were present which are; spermatogonia, primary spermatocytes, secondary spermatocytes, spermatids and spermatozoa The oocyte diameter of H. odoe is higher when compared with other freshwater fishes (the range was from 0.25 to 2.07 mean 1.74 ± 0.16 mm).

Conclusion: All the five maturation stages occurred every month throughout the study period except the spent stage (stage V) which occurred only in October and ripe and running stage (Stage IV) which was absent only in December. This indicates that H. odoe is a multiple spawner in the reservoir.

Indian Tinospora Species: As Antibacterial Agents and Phytochemical Evaluation

Nikita Patel — 2020-07-14

Tinospora cordifolia (Family: Minispermaceae), mainly found at tropical regions of India. It is widely used as a medicine in the Folk and Ayurvedic Systems. It possesses general adatogenic, rejuvenating and immunomodulatory activity and therefore, it is widely used in Ayurveda. There are mainly three species, namely Tinospora cordifolia, Tinospora malabarica (sinensis) and Tinospora crispa, widely found to grow in India. Of these three species, only T. cordifolia is recommended in Ayurveda and used by Indian plant based Industries for manufacture of herbal medicines. Stem is the official commercial part used for preparation of various medicines. In view of importance of Indian Tinospora species in the Indian System of Medicine (ISM), the present study has been focused to evaluate the extracts of three Tinospora species for antibacterial activities and phytochemical evaluation in order to understand and recommend these species as an Antibacterial Agents to herbal medicine manufacturing Industries.

Experimental Investigation on Physico-chemical Properties of Bio-ethanol/Gasoline Blends and the Qualitative Effect of Different Blends on Gasoline Quality and Engine Performance

J. K. Tangka — 2020-07-14

Physico-chemical and operational properties of various gasoline bio-ethanol blends were evaluated. Bio-ethanol was obtained through distillation from maize (Zea mays), sugar cane (Saccharum L), raffia (Raffia vinefera) wine, and palm wine and then purified using a rotavapor. Engine trails involved combinations of various ratios of gasoline/bio-ethanol as fuel in a small unmodified gasoline engine connected to a dynamometer. The vapour pressure, octane number, flash point, specific gravity, and energy density of various compositions of the blends were evaluated. Sugar cane gave the highest yield of alcohol 97.99 g per kg of produce while the lowest amount of alcohol of 10.5 ml per kg of produce was obtained from palm wine. Engine power decreased from 0.400 kW with 100% gasoline as fuel to 0.108 kW with a gasoline ethanol ratio of 1: 10. The octane number increased from 93 at E10 to 106 at E90. The energy density decreased from 33.180 MJ/l at E10 to 23.600 MJ/l at E90. Other physical observations suggest that to successfully run a gasoline engine with bio ethanol/gasoline blends some modifications would have to be done on the engine, including advancing of ignition timing, provision of air tight fuel conduit network, and modification of piston heads to improve pre-combustion fuel homogenisation.

Endocrine Disrupting Compounds (EDCs): The Risks for Aquatic Fauna

Aniruddha Banerji — 2020-07-14

Many natural and synthetic compounds like fungicides, plasticizers, phytoestrogens and xenoestrogens released into the aquatic environment from sewage discharges, agricultural runoffs and industrial effluents can lead to endocrine disruption in exposed species of fish. These endocrine disrupting compounds (EDCs) exert adverse effects on fish by disrupting oestrogen, androgen and aromatase functions, by causing impairment of gonad development, by endocrine disruption and by promoting behavioural changes. Thus, the chances of survival of affected species are compromised. As fish constitute an important source of food for humans, elevated concentrations of EDCs can also exert adverse effects on human health. It is thus essential to control and reduce the levels of EDCs and to combat their effects upon fish species to prevent further damage to the Earth’s marine and freshwater ecosystems.

All-trans Retinoic Acid (ATRA), a Potential Inhibitor of Matrix Metalloproteinase-2 (MMP-2) and Tumour Invasion in Melanomas

Aniruddha Banerji — 2020-07-14

Increased expression and activity of matrix metalloproteinase-2 (MMP-2) has been correlated with an increase in invasive potential and a worse prognosis in many cancers including melanomas. The anti-tumorigenic potential of all-trans retinoic acid (ATRA) in melanomas was studied using the highly metastatic murine melanoma cell line B16F10 as a model. Treatment of B16F10 cells with 20 µM/ml ATRA for 24 hrs caused a significant downregulation of MMP-2 expression and activity. Inhibition of MMP-2 activity appeared to be mediated via a significant decrease in expression of membrane type-1 matrix metalloproteinase (MT1-MMP), a crucial component of the MMP-2 activation complex, and extracellular matrix metalloproteinase inducer (EMMPRIN) and a significant increase in expression of tissue inhibitor of metalloproteinases-2 (TIMP-2). Downregulation of MMP-2 expression and activity would appreciably lower the invasive potential of melanoma cells. Inhibition of phosphorylation of focal adhesion kinase (FAK) upon ATRA treatment could inhibit intracellular signalling cascades and also downregulate MMP-2 levels and cell migration. The effectiveness of ATRA in inhibiting the migratory capabilities of B16F10 cells was confirmed by wound healing assay. Our findings thus elucidate important aspects about the molecular mechanisms of the effect of ATRA treatment on MMP-2 in melanomas and also indicate the anti-tumorigenic potential of ATRA which could have therapeutic value in clinical management of invasive melanomas.

Agricultural Production and Climate Change: The Scope for Innovation in the Post-COVID 19 Scenario

Aniruddha Banerji — 2020-07-14

The adverse effects of climatic changes on agriculture have always been an issue of global concern. In the post COVID-19 crisis scenario, such issue related to food security will gain more importance, particularly after the lockdown period which has disrupted many economic activities including agricultural production. This article focuses on some of the effects of climatic changes (changes in temperature, precipitation and CO₂ levels) on agricultural production and their influence on various biological parameters related to agricultural yields. Along with scientific advances, proper planning and micro level road maps need to be developed to retain the beneficial effects of climatic changes (for example reduction of greenhouse gas emissions) resultant from the lockdown due to the crisis and to endorse sustainable means of agriculture for the future. Techniques like regenerative agriculture and crop rotation could gain renewed importance and the application of payment for ecosystem services (PES) approach to agricultural practices would allow the balanced co-existence of environment and anthropogenic activities as the human race re-orients itself to the altered situation in the post COVID-19 scenario.