Biomass concentration and productivity in continuous culture, in autotrophic and mixotrophic conditions (D = 0.03 h−1).
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More than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\\n\\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\\n\\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\\n\\nAdditionally, each book published by IntechOpen contains original content and research findings.
\\n\\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\\n\\n\\n\\n
\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'
Simba Information has released its Open Access Book Publishing 2020 - 2024 report and has again identified IntechOpen as the world’s largest Open Access book publisher by title count.
\n\nSimba Information is a leading provider for market intelligence and forecasts in the media and publishing industry. The report, published every year, provides an overview and financial outlook for the global professional e-book publishing market.
\n\nIntechOpen, De Gruyter, and Frontiers are the largest OA book publishers by title count, with IntechOpen coming in at first place with 5,101 OA books published, a good 1,782 titles ahead of the nearest competitor.
\n\nSince the first Open Access Book Publishing report published in 2016, IntechOpen has held the top stop each year.
\n\n\n\nMore than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\n\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\n\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\n\nAdditionally, each book published by IntechOpen contains original content and research findings.
\n\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\n\n\n\n
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Industrial reactors for microalgae cultivation have been generally constructed using channels with movement and adapted for a better gas exchange. One of the biggest problems in this culture system is the low density of microalgae cells; they are constructed between 15 and 30 cm deep along the canal, limiting therefore the available light in addition to increasing the potential for contamination. A system proposed to solve the problem of low density and pollution has been found in closed polyethylene pipe systems, having the geometric design of the reactor as its main objective. Some strategies addressing three aspects have been developed to improve cultivation of microalgae in photobioreactors and produce fine chemicals: (1) the culture medium design—it is necessary to fix the nutrient composition to provide the right source of carbon and energy depending on the microalgae strain and secondary metabolite to be produced; (2) reducing adverse conditions for culture, such as oxygen accumulation, CO2 efficient supply, and sufficient light distribution. For this purpose, studies on the photobioreactor prototype should be performed; (3) once that photobioreactor prototype works well, critical factor criteria for scale-up bioengineering process should be fixed [1, 2, 3].
\nBiomass and product productivity are significantly affected by the culture condition; energy and carbon supply impacts directly biomass and product concentration. In effect, different metabolic growth modes for microalgae have been recognized: (a) autotrophy, in which light is the sole source of energy and inorganic carbon is the sole source of carbon; (b) heterotrophy, in which energy and carbon are both obtained exclusively from an organic carbon source, such as glucose, glycerol, and acetate, and growth can proceed without light supply; (c) mixotrophy, in which the photosynthetic microorganisms obtain energy from light and organic carbon sources and carbon is obtained from organic and inorganic carbon sources [4, 5]; and (d) photoheterotrophy, in which carbon can be obtained from organic compound but strictly with a light supply [5]. Chojnacka and Noworyta designed an empirical mathematical model to describe mixotrophic growth; in this model heterotrophic and autotrophic cultures are fractions of mixotrophic growth, but the metabolic interaction of photosynthesis and heterotrophy is important to improve biomass density and consequently secondary metabolite productivity [6].
\nLight as a source of energy for photosynthetic organisms is the main limiting factor during cultivation process of these organisms. In light intensities above the light saturation point, photosynthesis rate is directly proportional to the incident light supplied. The photosynthetic system of many microalgae becomes saturated to a radiation close to 30% of the total solar irradiance, i.e., between 1700 and 2000 μEm−2 s−1. Some species of phytoplankton grow to optimal intensities of 50 μEm−2 s−1 and are photoinhibited at around 130 μEm−2 s−1. The culture limitation by photoinhibition is the most important problem for commercial cultivation of microalgae. A possible solution is to assume that the heterotrophic metabolism in photosynthetic cells occurs, replacing or supplementing energy and carbon requirements from organic sources. Some studies suggest that mixotrophic, autotrophic, and heterotrophic metabolic activities occur simultaneously during cell growth [7]. The relative contribution of autotrophy to biomass production increases by increasing the light supply coefficient (kJ kg m2 s−1) or with an increase in the supply of CO2 and a decrease of organic carbon source supply. For example, at a light supply coefficient of 0.5 at 0.03 and 10% of CO2 concentration, the ratio of contribution of autotrophy (heterotrophy/autotrophy) to the biomass production was of 98:2 and 70:30, respectively [8]. A respirometric procedure has been proposed to obtain half saturation constant values for several nutrients; it is useful for modeling bioprocess for photosynthetic microorganisms [9]. These methods can be useful to evaluate organic substrates to be used in cultures, in a practical way.
\nPrior works [8, 10] conducted a detailed analysis of the heterotrophic and autotrophic modes simultaneously in Euglena gracilis and Spirulina platensis, respectively, under strict control of culture conditions; mixotrophic biomass concentration and growth rates resulted in the sum of the biomass or growth rates of heterotrophic and autotrophic cells in growing culture in parallel. The yields produced approximately the same amount of biomass produced in mixotrophic conditions; a mathematical approach can be summarized in Eqs. (1)–(8) [8, 10]. Assuming that mixotrophic growth is derived from cells growing in autotrophic and heterotrophic conditions, it can be described mathematically by defining the contribution of both metabolic growth modes. This can be simplified in Eq. (1):
\nSupposing that α is the heterotrophic fraction in mixotrophic growth, XH (biomass from heterotrophy) may be expressed as follows:
\nCombining Eqs. (1) and (2), biomass in mixotrophic (XM) conditions can be expressed as follows:
\nin which XA is the biomass from autotrophy; the growth rate ratio can be expressed as follows:
\nThe α coefficient is low when the incident light reaches all cells in the photobioreactor, where cell density should be low enough to avoid hiding among cells. Once that cell density increases, heterotrophic fraction increases as well, in the presence of an organic substrate as carbon source. To estimate α at any time, the value of α can be represented by an αi which can be constant for a period of time, Δti.
\nThen, Eq. (4) can be expressed as follows:
\nBy replacing in Eq. (5) of autotrophic growth, the equation included incident light:
\nwhere \n
Integrating Eq. (6) results in
\nAnd then, αi at any time can be calculated:
\nWith Eq. (8), α value was 0.02 at the beginning of culture, independent of incident light, to 0.61 after several hours of cultivation. Carbon balance showed inorganic carbon and carbon from glucose were consumed simultaneously; CO2 produced from respiration was used as carbon source during autotrophic growth [11]. Ogbonna and McHenry observed similar behavior in Euglena gracilis. Heterotrophic and autotrophic growth occurred simultaneously and independently [8]; this work defined two fractions instead of one as follows:
\nThe mixotrophic growth rate dXM/dt was equal to the sum of the heterotrophic growth rate dXH/dt and the rate of autotrophic growth dXA/dt. However, when two metabolic activities interact and when the presence of an organic carbon source affects the autotrophic metabolism or when the light affects the heterotrophic metabolic activity, then the heterotrophic rate in the mathematical description can be modified in accordance with Eq. (10), where dXM/dt is the total mixotrophic growth rate and β and α are coefficients of the autotrophic and heterotrophic fractions of the total mixotrophic growth rate, respectively:
\nThe values of α and β can be calculated on the basis of dXA/dt autotrophic growth rate and dXH/dt heterotrophic growth rate, both during mixotrophic culture.
\nThe sum of the values of β and α is 1.0 when the growth proceeds independently and simultaneously; when the sum is more than 1.0, there is an effect of promotion; and when it is <1.0, there is an inhibitory effect. It is clear that when mixotrophic growth occurs, both growth modes, namely, autotrophic and heterotrophic growth, have contribution in the final biomass and metabolite production; Eqs. (1)–(12) may be used to describe mathematically the secondary metabolite formation. An important stoichiometric relationship exists on carbon metabolism, with the pH changes driven by consumption of carbon source. Bicarbonate consumption increases pH, whereas glucose consumption decreases the pH due to CO2 production, being the reason that pH is kept almost constant during mixotrophic growth, CO2 consumption by photosynthesis, this balance may reflect the type of predominant metabolism.
\nDespite certain variability in the shape of open and closed systems, technical designs for open systems are the type race track, moved by paddles, usually operating at depths of 15–20 cm. At this depth, the growth rate of microalgae can be 15 g m−2 d−1, with a lipid content of 25%. Similar designs in terms of operation are the circular ponds, which are commonly found in Asia and Ukraine [3]. The major disadvantages of open systems are the significant loss of water by evaporation, the loss of CO2 into the atmosphere, the pollution, and the need for considerable surface for cultivation. Since the 1990s, in certain parameters such as the selection of species with efficient incident light utilization, the path of the incident light through the photobioreactor (PBR), the thickness of the wall, the mixing regime, and release of O2 via degassing, CO2 supply, have been focus on several developments [12]. Closed or semi-closed PBRs, based on different design concepts, have been implemented and tested at a pilot level. The latest developments seem to be directed toward tubular or plate-type compact configurations as well as combinations of these major designs in the form of distributing light over an expanded surface [13].
\nMicroalgae need enough quality and quantity of light supply, and it should be taken into account as a primary critic factor to design proper PBR. Cell density can increase from 103 cells ml−1 to densities above 108 cells ml−1; it produces a reduction of the distance among cells over 250 times, and the cell size can reduce its size 10 times as well. By improving mix capabilities of the PBR, hydrodynamic shearing stress over the cells can be increased; also, it can reduce growth or even cell death at high stress conditions [14]. The temperature has a greater influence on respiration and photorespiration than photosynthesis; when CO2 or light is limiting for photosynthesis, the influence of temperature is negligible. In contrast, an increase in the temperature will increase significantly the respiration, but flow of carbon through the Calvin cycle increases marginally. In other words, the net efficiency of photosynthesis declines at high temperatures. This effect can worsen in culture suspension by the difference in the solubility of CO2 and O2 at high temperatures. Normal temperatures for the growth of microalgae ranged between 25 and 30°C; an increment in the temperature affects the lipid production; at higher temperatures saturated free fatty acids are produced, while low temperatures favor unsaturated free fatty acid formation [15]. High concentration of O2 can build up in closed PBR; if this happens photosynthesis can be damaged by decreasing microalgae growth, and an improvement in the PBR should be implemented as an effective gas exchange [16].
\nThe first generation of closed PBR finds limitations over 50–100 L of culture volume; this was not effective for light supply to produce higher biomass density. Several designs of light distribution over the PBR, mainly underwater lamps, optical fiber, and column-shaped photobioreactors, have been used to provide an efficient production system; however, not much success has been obtained [12]. This is the main challenge in the future to find the appropriate scaling criteria for a larger irradiate surface, mass transfer, and coupled steps upstream and downstream processes [17]. The difficulty to scale up PBRs is to establish the inherent relationship among physical parameters involved in the design and the physiology of the microalgae to be cultured. An important design rule is to define quantitatively parameters to describe the interactions between incident light, the light distribution in the PBR, cell growth, and secondary metabolite production.
\nTo encourage the use of microalgae, it is necessary to implement a step-by-step system at different levels. The first step is the bioprospecting for selecting the most promising strain to produce a specific secondary metabolite and is the interaction of various disciplines, such as the analytical chemistry, biochemistry, molecular biology, and microbiology. The second step is the development of the culture medium, applicable to the largest volume. The third step is the strategy to analyze the scaling-up; biochemical or bioprocess engineers play an important role at this point. Strain and medium selection is carried out at flask level; the type of metabolism for the desired metabolite production, namely, mixotrophic, heterotrophic, or autotrophic growth, is also defined in this step. Operation parameters are fixed at small PBR scale; once the critic factors are overcome, PBR is ready to apply a scale-up procedure, from pilot to industrial production [18]. At the same time, recovery and purification steps should be performed. The last step of scale-up process should be a feasibility economic and technological analysis, in which production costs are obtained [19]. Quinn et al. constructed and validated a scalable growth model with species-specific variables, such as light and temperature; it can be used with PBR dimensions to accurate growth modeling for life cycle analysis.
\nMany aspects should be considered to obtain high concentration of biomass and secondary metabolites. Microalgae need energy from light to drive photosynthesis and growth. However, many of these organisms are able to use organic compounds as a source of chemical energy from respiratory mechanism. Although the terms of mixotrophy, autotrophy, photoheterotrophy, and heterotrophy are not well-defined, the influence of organic carbon energy and incident light energy can be quantitatively described in terms of biomass and secondary metabolite production.
\nAssuming that autotrophy growth occurred in cells absorbing incident light on the irradiate surface of the reactor, growth depends on the specific energy yield (YkJA), in other words, the amount of energy required to produce an amount of biomass; it can be defined by the following equation for continuous cultures:
\nMoreover, microalgae growing in mixotrophic mode and energy from carbon source can be included in Eq. (13), as follows:
\nYield equations can be achieved in continuous cultivation, where D is the dilution rate (h−1). Energy efficiency in batch and continuous cultures for Spirulina was calculated in values of 5.0 × 10−3 g biomass kJ−1 [11] and 2.4–4.8 × 10−3 g biomass kJ−1 [20]. There are two different points of view concerning energy efficiency which should be mentioned: one recently, a photocolor spirometer has been used for direct measurements of photosynthesis (calorimetry) and oxygen evolution at different light intensities [21], and the other one uses a photobioreactor to measure the overall light and carbon energy necessary to produce biomass and secondary metabolites [5]. The first can be useful to provide a potential energetic yield measurement because it considers the metabolic energy flows in the cells; the second provides data necessary for bioengineering purposes, specifically for the photobioreactor design and scaling-up procedures to follow.
\nMixotrophic growth can be described according to Figure 1. On the left-hand side, in the photosynthetic growth by the consumption of CO2 from culture medium, in the presence of light, biomass is produced, and oxygen is produced as well. On the other side, biomass is also produced but from organic carbon source consumption. The photosynthesis and respiration rates depend on several factors, such as microalgae species, O2 and CO2 availability, light supply, organic carbon source availability, temperature, pH, etc., but the main factor is the ability of microalgae to use O2 and CO2 at the same time [5].
\nDrawing describing the interaction of heterotrophy and autotrophy during mixotrophic growth. A, biomass from autotrophy; H, biomass from heterotrophy, modified from [5].
The balance for energetic yield (YX/kJ) can be described as follows: (i) autotrophic growth; total energy comes from incident light, and the biomass is formed from inorganic carbon:
\nHeterotrophic growth, energy, and biomass are provided by an organic carbon source:
\nThen, in mixotrophic growth, energy is supplied by both incident light and chemical energy from the organic carbon source and biomass from both inorganic and organic carbon sources.
\nIn the mixotrophic growth mode, certain molecules are accumulated, and there is a need to elucidate which metabolite is able to accumulate under specific growth conditions, but in general mixotrophic growth, it seems to be an efficient way for secondary metabolite accumulation [15, 22].
\nSeveral high valuable products have been described to be produced by photosynthetic microorganisms: antitumor agent from Amphidinium sp., food supplements from Dunaliella and Isochrysis galbana, antioxidants from Phaeodactylum tricornutum, and elastase inhibitor from Oscillatoria agardhii [17]. Algae biomass can accumulate or produce (i) bioenergy-based products, such as ethanol, methanol, biodiesel, biohydrogen, biogas, and long-chain hydrocarbons; (ii) staple food and vitamins such as yellow-white proteins, β-carotene, and phycobiliproteins, such as phycocyanin; (iii) polyunsaturated fatty acids, such as linolenic acid and arachidonic acid, that is, omega-3 fatty acids [23, 24]; (iv) base compounds for cosmetic industry and plant growth regulators; and (v) compounds with anticancer, antimicrobial, and antiviral activities.
\nSpirulina platensis showed higher antioxidant activity than other microalgae tested; Nostoc muscorum and Oscillatoria sp., moreover, have an important increment of phycobiliproteins by increasing nitrogen to the culture medium. It produces an important increment of the antioxidant activity in aqueous extracts of these microalgae. These extracts exhibit anticancer activity as well; in the extracts phenolic compounds, terpenoids, and alkaloids have been detected which can be responsible for several biomedical activities [24]. Water extracts of S. platensis have shown vulvovaginal antifungal activity on Candida and antifungal activity on several strains of Candida sp.; this can be the basis for therapeutic treatments, where secondary effects seem to be absent [25].
\nDietary supplements have been produced from biomass of microalgae; they include pigments and colorants from Haematococcus pluvialis, Chlorella sp., Dunaliella, red algae, cyanobacteria, and S. platensis [23]. A profile of natural pigments in dietary supplements of Spirulina including 51 pigments has been found in commercial products [26]. Pre-column reaction with DPPH radical followed by fast UHPLC-PDA separation revealed different classes of pigments grouped among carotenes, xanthophylls, and chlorophylls. Diadinoxanthin, alloxanthin, canthaxanthin, diatoxanthin, zeaxanthin, and echinenone were found in powder and tablets as minor components, in addition to β-carotene as a major component of dietary supplements [26]. Astaxanthin from H. pluvialis, c-phycocyanin from Limnothrix sp., and phycoerythrin from Phormidium have been produced [27, 28, 29], respectively.
\nProduction of pigments is affected by the amount of light supplied, and in combination with mixotrophic growth mode, phycocyanin, chlorophyll-a, and carotenoid concentrations, increased as light intensity increased, the concentration increased at least 30% in S. platensis [4, 11]. Production of chlorophylls and carotenoids increases 1.5 fold in Chlorella vulgaris in stirred tank photobioreactor [4]. Carotenoid accumulation and composition seem to be induced by light intensity, nitrogen starving, and salt stress. Higher light and salt stresses active synergistically carotenogenesis, mechanism such as esterification of astaxanthin and adonixanthin in Scenedesmus sp. [30].
\nRed marine microalga has proven their ability to produce pigments and hydrocolloids, due to their diversity, and perhaps produce a diversity of high valuable compounds. Fine chemicals are used as cosmetics, nutraceutics, and therapeutic agents; some are used in the food industry, diagnostic, biomedical research, and biosensor. Carbon source such as glucose, sucrose, glycerol, or acetate in the culture medium can help for accumulation of β-carotene and zeaxanthin by red microalga [31].
\nMicroalgae produce a wide range of antioxidants, some of them involved in the scavenging machinery of photosynthesis, respiration, and oxidative protection mechanisms. Pigments (carotenoids, chlorophylls, phycobiliproteins) play an important role in the photosynthetic mechanism in tocopherols including α, δ, and γ tocopherols, and pigments are accumulated as secondary metabolites at different amounts. Tocopherols have been found in Nannochloropsis oculata, Tetraselmis suecica, Spirulina maxima, Chaetoceros sp., Synechococcus, and Porphyridium cruentum. These compounds are formed depending several factors, such as microalga specie, growth phase, nutrient availability, light supply, and oxygen concentration, but also their production is affected by the processes of extraction and purification [32, 33, 34].
\nLipid metabolism can be induced by a nitrogen-limiting condition; nitrogen obtained from amino acid catabolism is assimilated via the glutamate-glutamine pathway; then, it is stored as an amino acid. The excess of carbon obtained from photosynthesis or glycolysis is redistributed into carbon-containing compounds. Carbon enters lipid metabolism via gamma-aminobutyrate pathway, glycolysis, and the tricarboxylic acid cycle [35]; malonyl-CoA is formed via acetyl-CoA from respiration; then, lipogenesis proceeds [15]. Supplementing microalgae cultures with an organic carbon source increases the productivity of biomass, lipid, and carbohydrates, enhancing the production of biodiesel, ethanol, starch, and polyunsaturated fatty acids. However, organic carbon source addition has limitations, for example, the cost and the bacterial contamination during cultivation. Progress on biorefineries has been focused on mixotrophic cultivation to enhance either secondary metabolite accumulation or fine chemicals [36]. Triacylglycerol content in Neochloris oleoabundans, Dunaliella sp., and Botryococcus braunii is more abundant when glycerol was used as organic carbon source than with autotrophic cultures. Profile of free fatty acids is also different. Saturated free fatty acids increase significantly in the presence of glycerol, but unsaturated free fatty acids decrease in general [37]. Biomass productivity and also the lipid productivity increased with the addition of acetate, glucose, and glycerol; although lipid content is smaller than other cultures, light supply also affected the content of lipids [38]. In contrast, lipid concentration in Chlorella protothecoides was as high as 55%, four times those obtained in autotrophic growing cells. Microalgae metabolic pathways for lipid accumulation are influenced by nitrogen-limiting conditions and carbon metabolism, where distribution pathways contribute to lipid biosynthesis [39, 40]. Biomass is considered as a renewable fuel source and does not affect the overall balance of CO2 in the atmosphere. Algal biofuel production coupled to a biomass power plant waste can serve as a cost-effective process to enhance microalgae biomass and biofuel productivity by sequestration of the CO2 produced in the power plant [35]. Productivity of biodiesel from oily plant crops, in terms of produced oil by surface production, varies from 27.57 to 972 L per ha, whereas that from microalgae cultivation is 7688–23,067 L per ha [23].
\nBiofuels derived from algal biomass depend on algal species: for biodiesel, Cladophora fracta, C. protothecoides, and B. braunii; for biohydrogen, C. protothecoides, S. platensis, and Chlamydomonas reinhardtii; for bioethanol Palmaria, Porphyra, Ascophyllum, Ulva lactuca, Tetraselmis sp., and Chlorococum sp.; and for biogas C. reinhardtii, Chlorella kessleri, and Spirogyra neglecta [23, 41, 42, 43, 44]. To produce biodiesel from microalgae, it is very important to select strains with oil content over 50% to improve biodiesel yield. With respect to oil content, microalgae can be divided into low, medium, and high oil content strains [45].
\nSoluble proteins have been used as nutritional supplements and personal care products or insoluble proteins for animal feeds [36]. Protein production has been reported in S. platensis using beet vinasse-supplemented culture media, in tubular photobioreactor biomass, which reached to 6.5 g L−1 and 168 mg L−1 d−1 of protein productivity. Continuous cultivation was also suitable for protein production from S. platensis using a medium supplemented with beet vinasse [46].
\nIncorporation of carbon from an organic carbon source, the type of carbon source, the amount supplemented to the culture, and the specie of microalgae are important for lipid accumulation in the cells of microalgae. The content of protein mostly increases by the addition of an organic carbon source, but lipid content decreases, although productivity of biomass, protein, and lipids increases substantially in the presence of organic carbon source [38].
\nFigure 2 represents the secondary metabolite production along with biomass that should be included in balance equations. The main components are carbon, hydrogen, oxygen, and nitrogen; in other words, a secondary metabolite can be a fraction of the total biomass, and it can be defined as ΘΔX, whereө is the fraction corresponding to the secondary metabolite produced, for chlorophyll accumulation, and it depends on the availability of carbon and nitrogen sources [40].
\nDrawing describing the production of secondary metabolites under mixotrophy. P, metabolite from autotrophy and heterotrophy; A, biomass from autotrophy; H, biomass from heterotrophy. Modified from Ref. [5].
Mixotrophy is coupled with three metabolic mechanisms, glycolysis, Calvin-Benson-Bassham, and the tricarboxylic acid cycle, where ATP is formed in the tricarboxylic acid cycle helping to drive electron flux on the light reactions of the photosynthesis to generate NADH, which is needed in the tricarboxylic acid cycle. These mechanisms are focal point to perform metabolic engineering, which open new routes to enhance the synthesis of fine chemicals by microalgae [47].
\nIn the past, microalgae cultures were used as components of aquaculture feeds and human food supplements. Recently, new alternatives have been opened for the production of fine chemicals and biofuels. However, production costs have been a concern; several efforts have been made to reduce processing costs to construct a profitable process. In this context, Allen et al. propose an integration of biology, ecology, and engineering topics for a sustainable biofuel and bioproduct production from microalgae [48].
\nThe potential markets of value-added products from microalgae are nutraceuticals for human applications and nutraceutical with applications for animal and fish feed, bulk chemicals, and biofuels, with commercial costs of 100 €/kg biomass, 5–20 €/kg biomass, <5 €/kg biomass, and <0.4 €/kg biomass, with a volume market of 60 million, 3–4 billion, >50 billion, and >1 trillion €, respectively [49].
\nHigh value-added products such as antiviral, anticancer, and antioxidants are target products to be obtained from microalgae, since it is an alternative process that can be continuously cultivated of axenic cultures in a closed photobioreactor adapted with a special light source of irradiation, such as fiber-optic or halogen lamps. In this case, biomass increases as long as microalgae receive light and the broth hydrodynamic allows enough movement to reach the illuminated surface (see Table 1), in continuous cultivation. Once the light limitation occurred and due to the effect of washing out, biomass starts to decrease to a new dilution rate. When an organic carbon source has a positive effect on the growth, continuous cultivation can be used as well, to produce an increment in biomass density (Table 1) and secondary metabolite formation as well, producing an increment of biomass and in the metabolites. Productivity also has a substantial increment at same light intensity and same dilution rate (D, h−1). Productivity and biomass concentration have been obtained in semicontinuous cultivation with a biomass of 5.31 g L−1 and productivity of 1.32 g L−1 d−1 [50]. Therefore, semicontinuous cultivation seems to be a good strategy as well.
\nIo (J cm−2 h−1) | \nΔXA (g L−1) | \nΔXAD (g L−1 h−1 × 10−3) | \nΔXM (g L−1) | \nΔXMD (g L−1 h−1 × 10−3) | \n
---|---|---|---|---|
3.22 | \n0.050 | \n1.74 | \n0.079 | \n2.76 | \n
5.85 | \n0.092 | \n3.21 | \n0.136 | \n4.75 | \n
11.11 | \n0.175 | \n6.11 | \n0.241 | \n8.41 | \n
18.98 | \n0.301 | \n10.81 | \n0.405 | \n14.13 | \n
Biomass concentration and productivity in continuous culture, in autotrophic and mixotrophic conditions (D = 0.03 h−1).
Modified from Ref. [51].
Secondary metabolite production can be effectively improved, by three advantages,(i) using a continuous process (up- and downstream processes), (ii) implementing mixotrophic cultivation, and (iii) recycling broth medium at least three times (Figure 3).
\nSchematic representation of a series of photobioreactors to operate in continuous cultivation to produce fine chemicals.
The authors thank Mr. Mauricio Ramos for the drawing of figures and the Instituto Politécnico Nacional for funding this work.
\nThe authors declare that there is no known conflict of interest associated with this publication.
\nThanks to Lada Bozic for helping in an efficient communication with IntechOpen.
\nCoronaviruses are large, enveloped, single-stranded, positive-sense RNA viruses with a genome of approximately 30 kilobases in length. The genus Coronavirus belongs to the family Coronaviridae in the order Nidovirales. They are classified into three groups. Group 1 contains various mammalian viruses including porcine epidemic diarrhea virus, porcine transmissible gastroenteritis virus, and human coronaviruses 229E and NL63. Group 2 includes canine respiratory coronavirus among other mammalian viruses and human coronavirus OC43. Human severe acute respiratory syndrome coronavirus (SARS-CoV-1) is considered a distant relative of this group. Group 3 contains solely avian coronaviruses. Human coronaviruses (HCoVs) cause respiratory infections, mainly, but gastroenteritis and neurological disorders may also occur. So far, at least seven human coronaviruses have been described including SARS-CoV-2, which was just sequenced in 2020, and two of these coronaviruses (OC43 and 229E) are responsible for 10–30% of all common colds. HCoV-HKU1 is mostly associated with bronchiolitis and pneumonia [1, 2, 3].
The gross life cycle of the SARS-CoV-1 has been observed in Vero E6 cells (African green monkey kidney cells) following inoculation with the virus under an electron microscope. The SARS-CoV-1 enters the cells through membrane fusion. Then, the nucleocapsids are assembled in the rough endoplasmic reticulum (RER) and mature by budding into the smoothe vesicles derived from the Golgi apparatus. Finally, the smoothe vesicles fuse with the cell membrane and the mature virus particles are released [4]. SARS-CoV-2 displays a similar life cycle.
Recent molecular studies have revealed that in order to facilitate entry of the virus into a human cell, the “S” spike surface glycoprotein of SARS-CoV-2 binds to the angiotensin-converting enzyme 2 (ACE-2) cellular receptor. Binding of the virus occurs via the S1 subunit of the S protein to a receptor and entry requires S protein priming by the cellular serine protease in order to allow fusing together of viral and cell membranes, a process which is initiated by the S2 subunit [5]. Following the fusion of viral and plasma membranes, the virus RNA undergoes transcription and replication inside the cell cytoplasm. Viral proteins are synthesized and the new RNA genomes are assembled and packaged in the endoplasmic reticulum, in the Golgi apparatus, and in the endoplasmic reticulum-Golgi intermediate compartment prior to virion release in vesicles. In fact, the S protein of SARS-CoV-2 binds to ACE-2 receptors with an approximately 10–20 fold higher affinity than that of SARS-CoV-1 and this added feature may aid in the efficient spread of SARS-CoV-2 among human populations. However, SARS-CoV-2 does not employ the other usual CoV receptors such as aminopeptidase N and dipeptidyl peptidase 4 to enter human cells [6].
ACE-2 is a membrane-associated aminopeptidase that converts angiotensin II to angiotensin 1–7 and plays a role in the cleavage of peptides [3]. Expression of ACE-2 in human tissues correlates with known sites of SARS-CoV-1 infection including lungs (particularly airway epithelia), heart, kidneys, small intestine, testes, and vascular endothelia [7]. These same tissues also overlap with the sites of SARS-CoV-2 infection in humans due to ACE-2 receptor availability.
On a personal note, as a biochemist, I have been following every bit of new research on any chemical compound that might successfully combat the virus. Around January 6th, 2020, I developed a very bad flu while in India after meeting with a friend who had just travelled to Wuhan in China. Overnight, I got a sore-throat that lasted a few days followed by a severe head cold with sinus congestion and mucous and, finally, it developed into a dry cough. During this debilitating flu, I also had some loose bowel movements with mucous. In the aftermath of the flu that lasted around 14 days, I was plagued with dizziness and weakness for two more weeks.
Although we had heard of the novel coronavirus in China, there was no reason to believe that was what I had just experienced since there had been no unusual respiratory distress. So, it did not seem to overlap with the pneumonia-like symptoms of the new coronavirus from China. Moreover, the friend had returned at the beginning of January and, as far as we knew at that time, the virus had only appeared in December. Therefore, it seemed unlikely that the friend had been exposed to any infected individuals while in China. Furthermore, the traveller from China never became sick (although one other person who attended the same meeting as myself developed a very bad flu within two weeks of coming in contact with this person). At the same time, there are also many seasonal flus like swine flu (H1N1) that are endemic in India, so there was no reason to consider that it was a coronavirus infection. Finally, the only medicines I took initially were some herbal Ayurvedic cold remedies mainly with a licorice-root base (a potent anti-inflammatory), aspirin at night, and an electrolyte solution to prevent dehydration from diarrhea. When I had a relapse of the gastrointestinal symptoms in March including stomach pain after I returned to Canada, a course of azithromycin helped to resolve the symptoms.
However, it was only when the weakness and malaise persisted for 3–4 months after the initial illness and new data started to emerge about the differing patterns of COVID-19 infection, that I started to consider another possible cause. Firstly, all my symptoms were consistent with the disparate effects of the novel coronavirus including the lingering apathy. Secondly, it became apparent that the new coronavirus had appeared in Wuhan some time before December. Thirdly, unlike other flu viruses, the phenomenon of asymptomatic spreaders became widely known. So, now, even though I had not been tested for the new virus or COVID-19 antibodies, I started to suspect that I could have experienced a form of coronavirus infection.
Finally, I had my COVID-19 test in August 2020 and, although it was negative, it did not preclude the possibility that I had the disease in January 2020 and that my body had formed and shed antibodies to the novel coronavirus (antibody testing was also negative). Since it is not known exactly how long antibodies persist following infection, even these may not be detected after a certain recovery period (there are recent reports antibodies decline after three months). Studies in rhesus monkeys show that re-infection does not occur in the recovered macaques up to 28 days after initial infection [8]. Nevertheless, prolonged inflammation and reports of re-infection in recovered humans are a surprising aspect of this virus. In my case, one additional negative C-reactive protein (inflammatory marker) test decisively clinched the matter.
Some scientists have opined that COVID-19 is highly contagious and highly lethal to a small subset of the population, while it produces milder symptoms in most people. Although the SARS-CoV-2 virus infects people of all ages, the World Health Organization (WHO) has determined that the evidence to date suggests that older adults and adults with underlying medical conditions are at a higher risk of developing severe COVID-19 disease [9].
One large study out of New York State seems to indicate that obesity, high blood pressure, and diabetes are strong risk factors for COVID-19 [10]. It has also been observed that cardiovascular disease and respiratory diseases could greatly affect the prognosis [11]. In fact, in an interesting German study involving autopsies on 12 COVID-19 patients the results revealed that coronary heart disease and asthma were common comorbid conditions in 50% of the deceased [12].
Other research suggests that cancer patients are more vulnerable to COVID-19 infection. A multicenter study showed that patients with cancer had higher risks in all severe outcomes of the disease tested. Hematologic cancer, lung cancer, or metastatic cancer (stage IV) cases experienced the highest frequency of severe events, while nonmetastatic cancer cases experienced similar frequencies to patients without cancer. Moreover, cancer patients who received surgery had higher risks of severe events than patients without cancer or those who underwent radiotherapy [13].
In addition, a surprising gender disparity appears to be present in relation to SARS-CoV-2 infection. Statistics from Australia, Belgium, Germany, Italy, the Netherlands, South Korea, Spain, the U.K and the US reveal that mortality rates from the virus are significantly higher in infected males than in infected females. In New York, approximately 60% of COVID-19-related deaths occurred in men. This may partly reflect biological characteristics since women produce stronger immune responses than men and are physically better at warding off viral and other types of infections. Nevertheless, biochemical differences in sex hormones are also likely to play a role in determining this dichotomy [14] and certain researchers have suggested it may be due to the presence of ACE-2 receptors in the testicles [15].
In the largest Chinese study to date assessing severity of coronavirus infection in smokers, it was found that higher percentages of current and former smokers needed ICU support or mechanical ventilation. Higher percentages of smokers among the severe cases also died [16]. Therefore, ultimately, the risk of any one individual is determined by the number of risk factors they display. For example, a ninety year old male smoker with diabetes and hypertension displaying five risk factors (age, gender, smoke inhalation, high blood pressure, and diabetes) would have an extremely high risk of contracting a terminal case of COVID-19.
However, genetic risk factors as a result of ethnic origin can only be considered once all these other significant risk factors have been taken into consideration. So far, despite attempts by various institutions to prove an ethnic link to COVID-19 infection, there is no compelling evidence to suggest that any one human group is genetically more susceptible to the novel coronavirus than any other beyond mitigating factors such as socioeconomic status or environmental conditions [17]. In order to establish a true genetic component, rigorous genetic testing must be undertaken to identify predisposing genes in susceptible ethnic groups. Prior to gene isolation and identification of a specific genetic polymorphism, a biochemical reaction resulting in a higher percentage of the disease is often demonstrated in a particular human population. As an example, the human sunburn cycle in response to UVA/B radiation only occurs in a minority of people with fair skin; however, most people simply tan when they are exposed to sunlight. In fact, these represent two separate physiological processes (burning and tanning). The former condition, scientific sunburn as a result of the human sunburn cycle, is mostly due to a genetic polymorphism involving the expression of very low levels of melanin in human skin since it can be corrected by wearing a sunscreen containing black sesame melanin [50 mg/ml] in a zinc oxide cream base [7.5%] [18, 19, 20]. It is also correlated with a high risk for skin cancer. Nonetheless, there may be other genetic factors like differences in DNA repair enzyme activity which can contribute to this unusual trait in certain individuals, as well [21].
Simultaneously, a surprising recent genetic association study has revealed that a major genetic risk factor for severe COVID-19 in humans may actually be inherited from Neanderthals. Outside the continent of Africa (0.3%), modern humans have inherited significantly more genetic material from other hominid species including Neanderthals (approximately 2%) and Denisovans [22]. Europeans and South Asians appear to have the greatest complement of Vindija Neanderthal genes from Croatia and a gene cluster on chromosome 3 inherited from this species has been identified as a risk locus for respiratory failure after infection with SARS-CoV-2. Among certain South Asian populations, up to 50% can carry at least one copy of this risk haplotype and the highest carrier frequency occurs in Bangladesh where 63% of the population carries it. In the UK it has been reported that individuals of Bangladeshi origin have roughly a two times higher risk of dying from COVID-19 than people of other nationalities [23].
Interestingly, there are high levels of air pollution in the two regions of China and Northern Italy that were hardest hit by the virus suggesting that environmental conditions can have an impact on the infectiousness of the disease [24]. Italian researchers have recently proposed an association between higher mortality rates in Northern Italy and peaks of particulate matter concentrations in this region. The most polluted northern provinces of Italy were found to have more infection cases than the less polluted southern provinces and this correlated well with ambient particulate matter concentrations that often exceeded the legal limit in these areas. All data for this study was collected prior to the lockdown [25]. Surprisingly, further research by the same group demonstrated that SARS-CoV-2 RNA was present on outdoor airborne particulate matter that was collected from an industrial site in Bergamo, Italy. This evidence suggests that, under the right atmospheric conditions, SARS-CoV-2 could create clusters with particulate matter and enhance persistence of the virus in the atmosphere by facilitating its capacity for diffusion. However, the vitality and virulence of the coronavirus diffused via this method remain to be confirmed [26].
This could have been a significant factor in the spread of the coronavirus in highly polluted and populated cities like Mumbai, India. Social conditions such as crowding in slums have also been considered contributory to dispersal of the virus in developing countries like Brazil and India. Proximity to infected individuals increases the risk of person-to-person transmission since the SARS-CoV-2 virus is spread mainly by respiratory droplets, but can be aerosolized, too [3].
No matter how healthy an individual may be, the more exposure they have to a particular virus, the greater risk they have of contracting the disease. The greater the number of particles of the virus one is exposed to, the greater the chance that they will overwhelm the body and immune responses. This is the reason that young doctors and other frontline healthcare workers are getting serious cases of COVID-19 and dying at a higher frequency than the general population.
View of Downtown Mumbai – December 2019
View of Mumbai Harbour – December 2019
In general, COVID-19 infection is associated with the increased production of pro-inflammatory cytokines, C-reactive protein, increased risk of pneumonia, sepsis, acute respiratory distress syndrome, and heart failure [24]. In fact, a cluster of unexplained pneumonia cases were first reported in Wuhan, China in late December 2019. A few days later, the cause of this pneumonia was identified as a new member of the coronavirus family. Since then, the virus has spread throughout China and precipitated a global pandemic [6].
Early reports from China suggested the most common symptoms of COVID-19 infection were fever (88%) and dry cough (67.7%). Rhinorrhea (4.9%) and gastrointestinal symptoms (diarrhea 4–14%) were less common. At the same time, a majority of patients (81%) had only mild symptoms (no pneumonia or mild pneumonia). Among patients with more pronounced symptoms, 14% experienced severe symptoms while 5% were critically ill with respiratory failure, septic shock, or multiorgan dysfunction or failure [3].
Although the novel coronavirus preferentially infects cells in the respiratory tract, autopsy results from Germany showed that it can be detected in multiple organs. The highest levels of the virus were detected in the lungs and the respiratory tract, while lower levels were usually present in the heart, liver, brain, kidneys, and spleen. This data suggests that SARS-CoV-2 may spread via the bloodstream and infect other organs. It also appears that COVID-19 may predispose patients to venous thromboembolism in several different ways including via endothelial dysfunction and promotion of a procoagulatory state by tissue factor pathway activation. High plasma levels of proinflammatory cytokines were observed in a small subset of patients with severe COVID-19 and, therefore, direct activation of the coagulation cascade by a cytokine storm is also plausible [12].
In one study, it was found that 22% of critically ill patients experienced myocardial injury from the infection [3]. In another study, the incidence of thrombotic complications in ICU patients with COVID-19 infections was reported to be 31%. It was concluded that COVID-19 may predispose to both venous and arterial thromboembolism due to excessive inflammation, hypoxia, immobilization, and diffuse intravascular coagulation [27].
In addition, the COVID-19 pandemic is surprisingly associated with neurological symptoms and complications including anosmia, hypogeusia, seizures, and stroke. Although statistics are not widely available at this point, the clinical course of COVID-19 is most severe in elderly male patients with comorbidities such as hypertension, diabetes, heart disease, and obesity which are all risk factors for stroke. There appears to be hypercoagulability associated with COVID-19 as a result of a “sepsis-induced coagulopathy” that may be a predisposing factor due to the formation of blood clots in the body [28]. COVID-19 complications in the brain can include delirium, inflammation, and encephalitis. A new study from UCL suggests that serious problems can occur even in individuals with mild cases of the virus [29].
A temporary loss of smell (anosmia) can be a consistent indicator of COVID-19 infection. An interesting finding is that the virus seems to change the sense of smell in patients by infecting and affecting the function of non-neural cells that support olfactory neurons. However, the neurons themselves do not appear to be infected as they do not express ACE-2 receptors [30].
Diabetes is already known to be a risk factor for COVID-19 and diabetics are more likely to die from the disease. Now, mounting evidence suggests that not only does diabetes make patients more vulnerable to the novel coronavirus, but the virus may actually trigger diabetes in some. Preliminary tissue studies indicate that the virus may act by damaging insulin-producing cells in the pancreas of affected individuals [31].
Even though, initially, children were thought to be unaffected by the novel coronavirus, a cluster of children with hyperinflammatory shock and features similar to Kawasaki disease and toxic shock syndrome was first reported in England. This hyperinflammatory condition could lead to severe illness, multiorgan failure, and even death in extreme cases. New reports out of the UK and US suggest that symptoms in young children (mainly toddler to elementary school age) can include inflammation of the blood vessels and coronary arteries. Almost all these pediatric cases had positive SARS-CoV-2 test results. As a result, this illness has been termed COVID-19-associated multisystem inflammatory syndrome [32].
Historically, there is no doubt that vaccines have provided a tremendous tool against infection for a variety of microbes including those causing small pox, tetanus, typhoid, cholera, and polio. Vaccines are an effective way for a population to achieve herd immunity (the concept that a pandemic will end once 60–70% of people become immune to any particular virus or microorganism). However, more recently, there are instances in which the production of viral vaccines has not been so successful as in the case of human immunodeficiency virus (HIV) and human coronaviruses (HCoVs) possibly due to their complex genomes. Virologists and immunologists maintain that it takes up to ten years to prepare a really good vaccine that has been properly tested. In fact, some of these specialists are skeptical about the race to find the first vaccine for the novel coronavirus within one year and often strike a cautionary note.
There are a number of things to consider in connection with a SARS-CoV-2 vaccine. Firstly, even if a safe and effective vaccine is made against the novel coronavirus, it may not be widely available in time to make a significant difference to the pandemic. Secondly, no successful vaccine against any coronavirus has been produced so far despite seventeen years of research. Moreover in March, the British Society for Immunology published an open letter stating that it is unknown whether this virus will induce long-term immunity in affected individuals as other related viruses do not [33]. Thirdly, certain vaccines can protect against a disease, but not against infection, so vaccinated individuals could potentially become asymptomatic carriers of SARS-CoV-2. Fourthly, some vaccines developed against SARS-CoV-1 (a close viral relative of SARS-CoV-2) actually exacerbated the disease in mice. Fifthly, although the easiest way to make a vaccine is to inactivate the pathogen, there are new vaccines in current trials based on RNA from coronaviruses or other RNA viruses that have never before been approved or tested in humans. Therefore, there could conceivably be unintended or irreversible consequences. Finally, at least one of the novel coronavirus vaccines approved for clinical trials so far has caused severe adverse events in three of eight healthy, young individuals that were tested [34] and other trials have been suspended. Unfortunately, the contamination of vaccines which are mass produced for a burgeoning human population also seems to be a potential problem and an ideal tool for rival countries to conduct biological warfare upon each other. Oral or nasal vaccines may be safer in this respect [35]. In addition, there is a physical limit to the number of vaccines a person can safely receive as new and deadlier viruses arise in the environment.
The action of UVB radiation striking and reacting thermally with 7-dehydrocholesterol in human skin results in the production of Vitamin D3 in the human body. This form of Vitamin D is converted to the hormonal metabolite, calcitriol, in a set of biochemical reactions in the liver, kidneys, and other organs as required. Then, calcitriol binds with the nuclear vitamin D receptor, which is a DNA binding protein, that interacts directly with regulatory sequences near target genes and affects their transcriptional output.
Vitamin D also enhances cellular innate immunity partly through the induction of antimicrobial peptides, including human cathelicidin, and, defensins. Cathelicidins exhibit direct antimicrobial activities against a spectrum of microbes including many types of bacteria, enveloped and nonenveloped viruses, and fungi. The main action of these host-derived peptides is to kill the invading pathogens by perturbing their cell membranes. Moreover, vitamin D is effective in reducing concentrations of pro-inflammatory cytokines that produce the inflammation that injures the lining of the lungs leading to pneumonia during viral infections like COVID-19 and increasing concentrations of anti-inflammatory cytokines [24].
According to a recent clinical study with a large sample size taken from different countries around the world, vitamin D supplements were found to protect against respiratory tract infections including colds and influenza. The most benefit was observed in patients who were very vitamin D deficient. This protective effect is likely provided by the capacity of vitamin D to boost levels of antimicrobial peptides in the lungs [36].
Vitamin D deficiency is a world-wide problem, but is particularly pronounced in the elderly, who are at greatest risk of contracting severe COVID-19 infection. The release of pro-inflammatory cytokines is one of the major causative factors in serious COVID-19 infections. However, vitamin D modulates their presence in the body by preventing macrophages from releasing too many inflammatory cytokines and chemokines. Calcitriol has also been found to exert an influence on ACE-2 receptors. Thus, it is not surprising that vitamin D deficiency has been correlated with COVID-19 cases and an increased risk of mortality in a European study [37].
RNA synthesis occurs in the life cycle of the SARS-CoV-1 virus in order to reproduce its genetic material and is catalyzed by an RNA-dependent RNA polymerase, which is the core enzyme of a multiprotein replication/transcription complex. In the case of SARS-CoV-1, an excess of intracellular zinc ions has been found to efficiently inhibit the RNA-synthesizing activity of this replication and transcription multiprotein. Enzymatic studies in vitro have revealed that zinc directly blocks the activity of the RNA polymerase by inhibiting elongation and reducing template binding. This RNA polymerase core, which is a central component of the coronaviral replication/transcription machinery, is well conserved among the members of the coronavirus family including SARS-CoV-2 [38, 39]. Therefore, it is quite possible that zinc treatment would have a similar biochemical effect on SARS-CoV-2 and interfere with its ability to replicate.
Since current research indicates that the mineral, zinc, can inhibit the replication of coronavirus and a variety of other RNA viruses in cell culture, it has become a potentially important and interesting supplement to study at this time. In the human body, zinc performs a variety of vital antioxidant functions and is required for maintaining good health. Inside the cell, the harmful effects of free radicals are balanced by the action of antioxidant enzymes (such as copper-zinc superoxide dismutase) and non-enzymatic antioxidants (such as metallothioneins). As zinc cannot pass easily through membranes, zinc-transporting proteins, ZIPs (Zrt-Irt-like protein or Zinc Iron permease) and ZnTs (Zinc transporters) help to facilitate this process. Metallothionein also aids in the regulation of zinc levels and the distribution of this metal in the extracellular space. The presence of zinc within the cell causes an increase in metallothionein, which is the major zinc-binding protein, and together they form a thermodynamically stable complex [40, 41]. Thus, low risk ways of increasing zinc bioavailability in the body can be safely considered.
In rats, rice fortified with zinc oxide or zinc carbonate is a feasible vehicle for zinc absorption, although zinc oxide displays lower bioavailability than zinc carbonate [42]. In young adults, zinc absorption from supplemental zinc citrate is comparable with that from zinc gluconate, but higher than from zinc oxide [43]. It is already known that zinc can be absorbed from topical (non-nano) zinc oxide by human skin in small quantities (nano forms of zinc oxide are not associated with significant zinc absorption) [44]. One of our recent studies suggests that zinc is absorbed by the human body from our suncare products (all with the same basic formula containing a medicinal form of zinc oxide) in sufficiently large quantities with regular use [45].
So, recently, when our company received an inquiry from Health Canada regarding any innovations that may benefit Canadian health workers at this critical time during the novel coronavirus pandemic, the answer was that we do have a product that may be useful to medical professionals and health workers in the field. It is a natural, award-winning suncare product specially formulated to block apoptotic sunburn (Skin Protector Plus). Its active ingredient is a non-nano, medicinal form of zinc oxide. The novel thing about this product is that it appears to be an efficient delivery system for boosting zinc levels in the whole body in a relatively short period of time. There is no toxicity associated with this product due to the use of high grade zinc oxide and natural ingredients. Since it is so safe and contains no harsh chemicals (already tested on human volunteers), no pre-clinical trials would be required to test its efficacy in protecting subjects from COVID-19 in a clinical study. The objective of such a comprehensive study would be to test and confirm the hypothesis outlined above, in vivo; namely, if maximum zinc levels are maintained in the human body via percutaneous zinc absorption from a topically applied zinc oxide cream, then it may provide one suitable defense against SARS-CoV-2 infection. Although oral supplementation is also an option, this type of topical application on the surface of the skin may be a faster method of ensuring even zinc distribution throughout the body and delivery to the various potential points of viral entry. Moreover, it may actually provide a physical barrier or blockade against entrance of the virus into the body by allowing suffusion and accumulation of zinc pools directly beneath the skin.
Quinine, an alkaloid derived from the bark of the cinchona tree, is most commonly found in South America, Central America, the islands of the Caribbean, and parts of the western coast of Africa. It is an important antimalarial drug and a synthetic form with a similar mode of action is known as chloroquine [46]. Chloroquine has been reported to inhibit the SARS-CoV-1 virus in infected cell cultures in vitro at doses equivalent to those used in the treatment of acute malaria in humans. Its antiviral effect appears to depend on the fact that chloroquine is a weak base that increases the pH of acidic vesicles when added extracellularly. The nonprotonated portion of chloroquine enters the cell where it becomes protonated and concentrated in acidic, low-pH organelles such as endosomes, Golgi vesicles, and lysosomes. The subsequent antiviral activity of the chloroquine depends partly on the extent to which a particular virus utilizes endosomes for entry into the cell [47]. In addition, this drug appears to interfere with terminal glycosylation of the angiotensin-converting enzyme 2 (ACE-2) cellular receptor, which is engaged by the virus for extracellular binding. This step may have a negative effect on the ability of the virus to gain entry into the host cell and, therefore, to initiate its replication cycle. Thus, infection may be deterred at clinically admissible concentrations [48]. Chloroquine also displays an immunomodulatory activity by suppressing the production and release of tumour necrosis factor alpha and interleukin 6 [49].
Furthermore, chloroquine was demonstrated to have strong antiviral activity against HCoV-OC43 in vitro. The anticoronaviral properties of chloroquine were also tested against HCoV-OC43 infection in newborn mice in vivo. Treatment with daily doses of chloroquine were found to have a long-lasting protective effect against lethal coronavirus OC43 infection in the newborn mice [1].
These favourable results suggest that chloroquine may be considered for use at antimalarial doses in the prevention of infections caused by coronaviruses, particularly SARS-CoV-2, which utilizes ACE-2 receptors in order to gain entry into host cells like its close relative, SARS-CoV-1.
Licorice root has been a commonly used ingredient in both Ayurvedic and traditional Chinese medicine for centuries, particularly in cough and cold remedies. Twenty triterpenoids and nearly three hundred flavonoids have been isolated from this herb. Scientific studies have shown that these metabolites possess many pharmacological activities including antiviral, antimicrobial, anti-inflammatory, and anti-tumour properties. However, glycyrrhizic acid or glycyrrhizin (GL), 18β-glycyrrhetinic acid (GA), liquiritigenin (LTG), licochalcone A (LCA), licochalcone E (LCE) and glabridin (GLD) are the main active components which possess antiviral and antimicrobial activities [50].
It has been known for some time that glycyrrhizic acid extracted from licorice (Glycyrrhiza glabra) root is active against viruses. This chemical is able to disrupt the growth and cytopathology of several unrelated DNA and RNA viruses without harming the host cell or its ability to replicate. Glycyrrhizic acid has also been demonstrated to inactivate herpes simplex virus particles irreversibly [51].
In a more recent study, the anti-SARSCoV activity of 15 glycyrrhizic acid derivatives was tested. Glycyrrhizin was shown to inhibit SARS-CoV-1 replication in vitro [52]. GL has also been reported to act by inhibiting viral gene expression and replication, reducing adhesion force and stress, and reducing High mobility group box 1 protein (HMGB1) binding to DNA. In addition, GL can enhance host cell activity by blocking the degradation of IκB, activating T lymphocyte proliferation and/or suppressing host cell apoptosis [50]. Thus, the potential for this licorice root component (GL) against SARS-CoV-2 infection is plausible.
Isoflavones and their related flavonoid compounds, particularly genistein, exert antiviral properties against a wide range of DNA and RNA viruses in vitro and in vivo [53]. The biological properties of the flavonoids are well studied, but the mechanisms of action underlying their antiviral properties are not fully understood. Isoflavones appear to have a combination of negative effects on viruses including affecting virus binding, entry, replication, viral protein translation and formation of certain viral envelope glycoprotein complexes. A variety of host cell signalling processes can also be affected by isoflavones including induction of gene transcription factors and secretion of cytokines. All these effects are dependent on dose, frequency of administration, and different combinations of isoflavones employed in bioassays in vitro. Genistein may be able to mimic the action of 17-beta-estradiol [E2] due to its similar structure or to act as an E2 antagonist and its activity as a broad-spectrum tyrosine kinase inhibitor may contribute to its ability to influence estrogen receptor-independent mechanisms [54]. Despite their unique effect on immune function and anti-inflammatory activity, there is still a lack of data confirming the antiviral efficacy of such soy isoflavones in vivo against coronaviruses and other viruses thereby forming a worthwhile subject for biochemical study.
At least seven human coronaviruses have been described to date including SARS-CoV-2, which is closely related to and resembles SARS-CoV-1 in many respects. Both viruses bind to ACE-2 receptors on human cells. ACE2 is a membrane-associated aminopeptidase that converts angiotensin II to angiotensin 1–7 and plays a general role in the cleavage of peptides. Expression of ACE2 in human tissues correlates with known sites of SARS-CoV-1 infection including lungs (particularly airway epithelia), heart, kidneys, small intestine, testes, and vascular endothelia. These same tissues overlap with known sites of SARS-CoV-2 infection in humans.
A cluster of unexplained pneumonia cases were first reported in Wuhan, China and, a few days later, the cause of this pneumonia was identified as a new member of the coronavirus family. SARS-CoV-2 infection appears to be associated with a puzzling array of symptoms and complications. The major symptoms noted in China were fever (88%) and dry cough (67.7%), while rhinorrhea (4.9%) and gastrointestinal symptoms (diarrhea 4–14%) were less common. A majority of patients (81%) had only mild symptoms (no pneumonia or mild pneumonia). Among patients with more pronounced symptoms, 14% experienced severe symptoms while 5% were critically ill with respiratory failure, septic shock, or multiorgan dysfunction or failure.
New data suggests that SARS-CoV-2 may spread via the bloodstream to infect other organs. In addition to the lungs, other target organs can include the heart, liver, brain, kidneys, and spleen. It also appears that COVID-19 may predispose patients to venous thromboembolism in several different ways including via endothelial dysfunction and promotion of a procoagulatory state. In fact, it was found that a significant percent of critically ill patients experienced myocardial injury from the infection and it has been concluded that COVID-19 may predispose to both venous and arterial thromboembolism due to excessive inflammation, hypoxia, immobilization, and diffuse intravascular coagulation. The COVID-19 pandemic is associated with various neurological symptoms and complications including anosmia, hypogeusia, seizures, and stroke, as well. COVID-19 complications in the brain can include delirium, inflammation, and encephalitis. Despite initial reports that children were unaffected by the novel coronavirus, it has emerged that pediatric patients are susceptible to a COVID-19-associated multisystem inflammatory syndrome that can cause serious inflammation of the blood vessels.
Several internal risk factors have been identified for SARS-CoV-2 infection. The main ones include age (older adults are more vulnerable to serious infection by the virus), gender (the virus is significantly more deadly in men than in women), obesity, heart disease, diabetes, cancer status, and smoking. However, there is no convincing evidence to date that any particular ethnic group displays a stronger genetic susceptibility to the virus (although, there may be a possible link to an inherited Neanderthal gene locus). Nevertheless, specific genetic variants such as those for the gene that encodes a protein that interacts with the ACE-2 receptor may be involved in determining individual patient responses to the disease. Simultaneously, external risk factors like environmental pollution, social conditions such as crowding, and frequency of exposure to infected persons also seem to play an important role.
Reports of re-infection in recovered humans is a surprising aspect of this virus. Recently, a team from the University of Hong Kong reported the first case of re-infection of COVID-19 within a period of approximately four and a half months. Genomic analyses confirmed that the patient had re-infection instead of persistent viral shedding from first infection. Moreover, there was a difference of 24 nucleotides between both viruses that infected the patient suggesting two different viral strains were involved [55].
Even though the virus is associated with positive COVID-19/COVID-19 antibody and high C-reactive protein test results, antibody levels may decline soon after infection. Consequently, it is quite possible that a lasting resistance to the virus will not be achievable. In the event that long-term immunity cannot be induced to the novel coronavirus by a vaccine, an annual, bi-annual, or even tri-annual inoculation may be required (current data suggests that antibodies begin to decrease or disappear three months after infection). This means that other modes of protection and prevention like supplementation may be more relevant in this case. Some candidates include Vitamin D, zinc, chloroquine/quinine, glycyrrhizic acid, and genistein due to anti-viral properties such as the ability to inhibit replication and reproduction of coronaviruses.
Scientists have concluded that drastic social distancing, quick detection and isolation of infected individuals and travel restrictions were the most effective steps for containment of COVID-19 in China. Genome sequencing has also helped to track and control COVID-19 infections quickly. However, if people do not continue to be careful, certain places may become vulnerable to further rounds of this disease. WHO recently reported that coronavirus infections among younger populations were skyrocketing. The proportion of cases in teens and young adults increased six-fold, while the proportion in young children and babies increased seven-fold by August. This may be attributable in part to the resurgence of large parties and social gatherings attended by young people following the relaxation of restrictions during the summer. Therefore, it seems very likely that the denouement of the COVID-19 story will be largely dictated by our social habits and ability to adapt to a new set of societal norms and conditions. This will include wearing face masks in public places, possibly, with a thin zinc coating along with a special zinc oxide crème formulation applied to the skin underneath [56].
IntechOpen implements a robust policy to minimize and deal with instances of fraud or misconduct. As part of our general commitment to transparency and openness, and in order to maintain high scientific standards, we have a well-defined editorial policy regarding Retractions and Corrections.
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\\n\\nA Statement of Concern detailing alleged misconduct will be issued by the Academic Editor or publisher following a 3rd party report of scientific misconduct when:
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\\n\\nA Correction will be issued by the Academic Editor when:
\\n\\n3.1. ERRATUM
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\\n\\nA published Erratum will adhere to the Retraction Notice publishing guidelines outlined above.
\\n\\n3.2. CORRIGENDUM
\\n\\nA Corrigendum will be issued by the Academic Editor when it is determined that a mistake in a Chapter is a result of an Author’s miscalculation or oversight. A published Corrigendum will adhere to the Retraction Notice publishing guidelines outlined above.
\\n\\n4. FINAL REMARKS
\\n\\nIntechOpen wishes to emphasize that the final decision on whether a Retraction, Statement of Concern, or a Correction will be issued rests with the Academic Editor. The publisher is obliged to act upon any reports of scientific misconduct in its publications and to make a reasonable effort to facilitate any subsequent investigation of such claims.
\\n\\nIn the case of Retraction or removal of the Work, the publisher will be under no obligation to refund the APC.
\\n\\nThe general principles set out above apply to Retractions and Corrections issued in all IntechOpen publications.
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\\n\\nPolicy last updated: 2017-09-11
\\n"}]'},components:[{type:"htmlEditorComponent",content:'IntechOpen’s Retraction and Correction Policy has been developed in accordance with the Committee on Publication Ethics (COPE) publication guidelines relating to scientific misconduct and research ethics:
\n\n1. RETRACTIONS
\n\nA Retraction of a Chapter will be issued by the Academic Editor, either following an Author’s request to do so or when there is a 3rd party report of scientific misconduct. Upon receipt of a report by a 3rd party, the Academic Editor will investigate any allegations of scientific misconduct, working in cooperation with the Author(s) and their institution(s).
\n\nA formal Retraction will be issued when there is clear and conclusive evidence of any of the following:
\n\nPublishing of a Retraction Notice will adhere to the following guidelines:
\n\n1.2. REMOVALS AND CANCELLATIONS
\n\n2. STATEMENTS OF CONCERN
\n\nA Statement of Concern detailing alleged misconduct will be issued by the Academic Editor or publisher following a 3rd party report of scientific misconduct when:
\n\nIntechOpen believes that the number of occasions on which a Statement of Concern is issued will be very few in number. In all cases when such a decision has been taken by the Academic Editor the decision will be reviewed by another editor to whom the author can make representations.
\n\n3. CORRECTIONS
\n\nA Correction will be issued by the Academic Editor when:
\n\n3.1. ERRATUM
\n\nAn Erratum will be issued by the Academic Editor when it is determined that a mistake in a Chapter originates from the production process handled by the publisher.
\n\nA published Erratum will adhere to the Retraction Notice publishing guidelines outlined above.
\n\n3.2. CORRIGENDUM
\n\nA Corrigendum will be issued by the Academic Editor when it is determined that a mistake in a Chapter is a result of an Author’s miscalculation or oversight. A published Corrigendum will adhere to the Retraction Notice publishing guidelines outlined above.
\n\n4. FINAL REMARKS
\n\nIntechOpen wishes to emphasize that the final decision on whether a Retraction, Statement of Concern, or a Correction will be issued rests with the Academic Editor. The publisher is obliged to act upon any reports of scientific misconduct in its publications and to make a reasonable effort to facilitate any subsequent investigation of such claims.
\n\nIn the case of Retraction or removal of the Work, the publisher will be under no obligation to refund the APC.
\n\nThe general principles set out above apply to Retractions and Corrections issued in all IntechOpen publications.
\n\nAny suggestions or comments on this Policy are welcome and may be sent to permissions@intechopen.com.
\n\nPolicy last updated: 2017-09-11
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