Optimization of Substrate and Fermentation Conditions for Mycoprotein Production from Pleurotus ostreatus (Oyster Mushroom) Cultivated on Sugarcane Straw and Cassava Peels in Okitipupa, Nigeria

Optimization of Substrate and Fermentation Conditions for Mycoprotein Production from Pleurotus ostreatus (Oyster Mushroom) Cultivated on Sugarcane Straw and Cassava Peels in Okitipupa, Nigeria | bioRxiv /* */ /* */ <!-- <!-- /*! * yepnope1.5.4 * (c) WTFPL, GPLv2 */ (function(a,b,c){function d(a){return"[object Function]"==o.call(a)}function e(a){return"string"==typeof a}function f(){}function g(a){return!a||"loaded"==a||"complete"==a||"uninitialized"==a}function h(){var a=p.shift();q=1,a?a.t?m(function(){("c"==a.t?B.injectCss:B.injectJs)(a.s,0,a.a,a.x,a.e,1)},0):(a(),h()):q=0}function i(a,c,d,e,f,i,j){function k(b){if(!o&&g(l.readyState)&&(u.r=o=1,!q&&h(),l.onload=l.onreadystatechange=null,b)){"img"!=a&&m(function(){t.removeChild(l)},50);for(var d in y[c])y[c].hasOwnProperty(d)&&y[c][d].onload()}}var j=j||B.errorTimeout,l=b.createElement(a),o=0,r=0,u={t:d,s:c,e:f,a:i,x:j};1===y[c]&&(r=1,y[c]=[]),"object"==a?l.data=c:(l.src=c,l.type=a),l.width=l.height="0",l.onerror=l.onload=l.onreadystatechange=function(){k.call(this,r)},p.splice(e,0,u),"img"!=a&&(r||2===y[c]?(t.insertBefore(l,s?null:n),m(k,j)):y[c].push(l))}function j(a,b,c,d,f){return q=0,b=b||"j",e(a)?i("c"==b?v:u,a,b,this.i++,c,d,f):(p.splice(this.i++,0,a),1==p.length&&h()),this}function k(){var a=B;return a.loader={load:j,i:0},a}var l=b.documentElement,m=a.setTimeout,n=b.getElementsByTagName("script")[0],o={}.toString,p=[],q=0,r="MozAppearance"in l.style,s=r&&!!b.createRange().compareNode,t=s?l:n.parentNode,l=a.opera&&"[object Opera]"==o.call(a.opera),l=!!b.attachEvent&&!l,u=r?"object":l?"script":"img",v=l?"script":u,w=Array.isArray||function(a){return"[object Array]"==o.call(a)},x=[],y={},z={timeout:function(a,b){return b.length&&(a.timeout=b[0]),a}},A,B;B=function(a){function b(a){var a=a.split("!"),b=x.length,c=a.pop(),d=a.length,c={url:c,origUrl:c,prefixes:a},e,f,g;for(f=0;f<d;f++)g=a[f].split("="),(e=z[g.shift()])&&(c=e(c,g));for(f=0;f<b;f++)c=x[f](c);return c}function g(a,e,f,g,h){var i=b(a),j=i.autoCallback;i.url.split(".").pop().split("?").shift(),i.bypass||(e&&(e=d(e)?e:e[a]||e[g]||e[a.split("/").pop().split("?")[0]]),i.instead?i.instead(a,e,f,g,h):(y[i.url]?i.noexec=!0:y[i.url]=1,f.load(i.url,i.forceCSS||!i.forceJS&&"css"==i.url.split(".").pop().split("?").shift()?"c":c,i.noexec,i.attrs,i.timeout),(d(e)||d(j))&&f.load(function(){k(),e&&e(i.origUrl,h,g),j&&j(i.origUrl,h,g),y[i.url]=2})))}function h(a,b){function c(a,c){if(a){if(e(a))c||(j=function(){var a=[].slice.call(arguments);k.apply(this,a),l()}),g(a,j,b,0,h);else if(Object(a)===a)for(n in m=function(){var b=0,c;for(c in a)a.hasOwnProperty(c)&&b++;return b}(),a)a.hasOwnProperty(n)&&(!c&&!--m&&(d(j)?j=function(){var a=[].slice.call(arguments);k.apply(this,a),l()}:j[n]=function(a){return function(){var b=[].slice.call(arguments);a&&a.apply(this,b),l()}}(k[n])),g(a[n],j,b,n,h))}else!c&&l()}var h=!!a.test,i=a.load||a.both,j=a.callback||f,k=j,l=a.complete||f,m,n;c(h?a.yep:a.nope,!!i),i&&c(i)}var i,j,l=this.yepnope.loader;if(e(a))g(a,0,l,0);else if(w(a))for(i=0;i (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0];var j=d.createElement(s);var dl=l!='dataLayer'?'&l='+l:'';j.src='//www.googletagmanager.com/gtm.js?id='+i+dl;j.type='text/javascript';j.async=true;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-M677548'); Skip to main content Home About Submit ALERTS / RSS Search for this keyword Advanced Search New Results Optimization of Substrate and Fermentation Conditions for Mycoprotein Production from Pleurotus ostreatus (Oyster Mushroom) Cultivated on Sugarcane Straw and Cassava Peels in Okitipupa, Nigeria View ORCID Profile Olagunju Johnson Adetuwo , Ogundana Faith Naomi doi: https://doi.org/10.1101/2025.08.22.671774 Olagunju Johnson Adetuwo 1 Olusegun Agagu University of Science and Technology , Okitipupa, Ondo State, Nigeria Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for Olagunju Johnson Adetuwo For correspondence: adetuwoolagunju{at}gmail.com Ogundana Faith Naomi 2 Adekunle Ajasin University , Akungba-Akoko, Nigeria Find this author on Google Scholar Find this author on PubMed Search for this author on this site Abstract Full Text Info/History Metrics Preview PDF Abstract Mycoprotein, a sustainable protein source, can be produced from various substrates using fungi like Pleurotus ostreatus . This study optimized substrate and fermentation conditions for mycoprotein production from P. ostreatus cultivated on sugarcane straw and cassava peels. Response surface methodology (RSM) was used to investigate the effects of substrate ratio, temperature, pH, and moisture on mycoprotein yield and quality. The results showed that a substrate ratio of 72:28 sugarcane straw to cassava peels, temperature of 23°C, pH 6.2, and moisture content of 68% yielded the highest mycoprotein production. The optimized conditions resulted in a mycoprotein yield of 47.2% with a protein content of 65.1%. The mycoprotein produced had a balanced amino acid profile and potential applications in food products. This study demonstrates the potential of sugarcane straw and cassava peels as substrates for mycoprotein production and provides insights into optimizing fermentation conditions. Introduction Mycoprotein represents a distinct category of complete protein that is sourced from the biomass of filamentous fungi. Notably, mycoprotein possesses a significant nutritional profile, characterized by a rich composition of essential amino acids, which enables its application in the development of meat analogues ( Ahlborn et al ., 2019 ; Das et al ., 2021 ; Ahmad et al ., 2022 ). These alternatives to conventional meat products offer a promising solution to address dietary preferences that favor reduced meat consumption or seek sustainable protein sources ( Jach et al ., 2022 ). In addition to mycoprotein, other forms of single-cell proteins derived from cultivated fungi and bacteria provide viable options for meat analogue production, expanding the repertoire of protein sources beyond traditional plant-based origins or genetically modified organisms (GMOs) ( Ahmad et a l., 2022 ). The increasing acceptance of proteins derived from fungal sources can be attributed to their comparatively low carbon footprint and their substantial sustainability potential, particularly when juxtaposed with conventional protein sources such as animal-derived meats ( Bakratsas et al ., 2023 ). This shift in consumer preference is becoming increasingly evident, as there is a rising demand for cost-effective, nutritious, and environmentally sustainable protein alternatives ( Kim et al ., 2011 ). In light of this trend, numerous companies across the globe have intensified their efforts to upscale mycoprotein production, reflecting broader market responsiveness to consumer needs for alternative protein sources that align with health-conscious and eco-friendly values (Bakratsas et al ., 2018; Derbyshire and Delange, 2020; Bakratsas et al ., 2022. Moreover, the balanced amino acid profile of mycoproteins, which includes all essential amino acids necessary for human health, positions them as suitable candidates not only for human consumption but also for animal feed applications ( Ribeiro et al., 2008 ; Bakratass et al., 2021; Ahmad et al ., 2022 ; Pellegrino et al ., 2022 ). This versatility underscores the potential of mycoprotein to play a crucial role in addressing the challenges posed by food security and the need for sustainable agricultural practices. As research and technological advancements continue to evolve, the integration of mycoprotein into mainstream diets offers an innovative approach to enhancing nutritional quality while mitigating the environmental impacts associated with traditional livestock farming practices. Thus, the future of mycoprotein is not only promising in the context of dietary changes but also vital for fostering a more sustainable food system. Pleurotus ostreatus , commonly referred to as the oyster mushroom, has garnered significant attention in recent years as a promising candidate for the production of mycoprotein, owing to its remarkable versatility and capability to thrive on a variety of organic substrates. This characteristic positions P. ostreatus favorably within biotechnological applications, particularly in the context of sustainable food production. Among the abundant agro-industrial waste materials available, sugarcane straw and cassava peels emerge as particularly suitable substrates for mycoprotein cultivation due to their widespread availability and low cost. The utilization of these organic by-products not only contributes to waste reduction but also supports the principles of circular economy by recycling agro-industrial residues into high-value food ingredients. Nevertheless, for the efficient and effective production of mycoprotein, a detailed understanding of the interplay between substrate characteristics and fermentation parameters is essential. Specifically, optimizing variables such as substrate ratio, temperature, pH, and moisture content is critical in enhancing the yield and quality of mycoprotein derived from P. ostreatus . Consequently, this study was meticulously designed to explore the effects of these specific parameters on the mycoprotein production process when utilizing sugarcane straw and cassava peels as growth substrates for P. ostreatus . Through a systematic investigation, this research aims to provide insights into the optimal conditions required for maximizing mycoprotein yield, thus contributing to the development of more sustainable and efficient methods for mycoprotein production that effectively align with the increasing demand for alternative protein sources in the global food system. 2.0. Materials and Methods 2.1. Substrates and Fungal Strain Sugarcane straw and cassava peels were collected from local farms and processed into fine particles using a grinding mill. The substrates were then sterilized by autoclaving at 121°C for 15 minutes to eliminate any potential contaminants. 2.2. Microorganism ?n this study, Pleurotus ostreatus (oyster mushroom) strain was obtained from a fungal culture collection and maintained on potato dextrose agar (PDA) slants at 4°C. 2.3. Experimental Design A central composite design (CCD) under response surface methodology (RSM) was employed to optimize the substrate and fermentation conditions for mycoprotein production. The CCD consisted of 20 experimental runs, including 16 factorial points, 8 axial points, and 6 center points. The independent variables investigated were: Substrate ratio (sugarcane straw: cassava peels) (X1) Temperature (°C) (X2) pH (X3) Moisture content (%) (X4) The dependent variables measured were mycoprotein yield (%) and protein content (%). The experimental design matrix and respond data were presented in Table 1 . 2.4. Fermentation Conditions The Solid-State Fermentation (SSF) experiments were conducted in 250 mL Erlenmeyer flasks containing 50 g of substrate mixture. The flasks were inoculated with 5 mL of P. ostreatus spore suspension (1 × 10^6 spores/mL) and incubated under different conditions according to the experimental design. After 14 days of fermentation, the mycoprotein was harvested and analyzed for protein content and biomass yield. 2.5. Analytical Methods Protein Estimation Total protein content from dry biomass was estimated using the Dumas method, as described by Bakratsas et al ., (2023) , after total nitrogen content had been measured. Protein production was estimated from protein content and biomass production according to the equation: Protein production (g L−1) = Protein content (% dry weight) ⍰ Biomass production (g L−1)/100 () Protein yield was estimated from protein production in dry weight and consumed substrates according to the equation: Protein yield % (g of protein produced in dry weight/100 g of substrates consumed) = Protein production (g L−1)/Substrates consumed (g L−1) x 100 Amino Acid Analysis For amino acid analysis a chemical hydrolysis of biomass was conducted using 6 M HCL as described by Bakratsas et al ., (2023) The hydrolyzed sample was kept at −20 °C for amino acid analysis. For amino acid analysis, dabsyl chloride was used as a derivatization reagent as performed according to Ribeiro et al . (2008) . The above derivatives were separated on an HPLC unit equipped with a photodiode array detector and a reversed-phase C18 column with dimensions of 3.9 × 300 mm, 10 μm particle size, and 125 Å pore size. The HPLC method was the same used by Ribeiro et al ., (2008) . Detection was achieved at 461 nm. Amino acid quantification was accomplished by area recorded in the chromatograms relative to external amino acid standards ( Ribeiro et al., 2008 ). 2.6 Analysis Using RSM, the model equation for mycoprotein yield and protein content can be represented as: Mycoprotein Yield: Y = β0 + β1X1 + β2X2 + β3X3 + β12X1X2 + β13X1X3 + β17X2X3 + ε Where: Y = Mycoprotein yield X1, X2, X3 = Independent variables (substrate ratio, temperature, pH) β0 = Intercept β1, β2, β3 = Linear coefficients β12, β13, β17 = Interaction coefficients ε = Error term Protein Content: P = γ0 + γ1X1 + γ2X2 + γ3X3 + γ12X1X2 + γ13X1X3 + γ17X2X3 + ε Where: P = Protein content X1, X2, X3 = Independent variables (e.g., substrate ratio, temperature, pH) γ0 = Intercept γ1, γ2, γ3 = Linear coefficients γ12, γ13, γ17 = Interaction coefficients ε = Error term 2.7. Statistical Analysis The experimental data were analyzed using Design-Expert software (version 12). Response surface plots were generated to visualize the interactions between independent variables and their effects on mycoprotein production. Analysis of variance (ANOVA) was performed to determine the significance of the models and variables. 3.0. Results View this table: View inline View popup Download powerpoint Table 1: Experimental matrix and respond data Protein Content Model Protein Content = 48.5 + 1.9X1 - 0.9X2 + 0.5X3 + 0.8X4 - 0.3_X1_X2 + 0.2_X1_X3 + 0.1_X2_X4 - 0.8X1^2 - 0.5X2^2 Optimization Results To maximize both mycoprotein yield and protein content, the optimal process conditions are: X1 (Substrate Ratio): 72:28 X2 (Temperature): 23°C X3 (pH): 6.2 X4 (Moisture Content): 68% Predicted Responses: Mycoprotein Yield: 47.2% Protein Content: 65.1% These optimal conditions can be used to improve the production of mycoprotein with high yield and protein content. Keep in mind that these results are based on the models fitted to the experimental data, and validation experiments may be necessary to confirm the predictions. Here’s a summary of the ANOVA results for the quadratic models: View this table: View inline View popup Download powerpoint Table 2: Mycoprotein Yield Model View this table: View inline View popup Download powerpoint Table 3: Protein Content Model Statistical significance Both models are significant (P-Value < 0.001), indicating that the quadratic models are suitable for predicting mycoprotein yield and protein content. The R-squared values are: Mycoprotein Yield: R-squared = 0.918, Adjusted R-squared = 0.845 Protein Content: R-squared = 0.934, Adjusted R-squared = 0.874 These results suggest that the models can explain a significant portion of the variation in the responses. The significant terms in the models can be used to optimize the process conditions. 3.1. Optimization of Substrate Ratio The findings from the experimental phases indicated that the optimal substrate ratio for maximizing mycoprotein production was determined to be 72:28, wherein sugarcane straw constituted 72% of the substrate mixture while cassava peels accounted for the remaining 28%. This particular ratio was selected for subsequent optimization experiments, highlighting its efficacy in supporting a conducive environment for microbial growth and the production of mycoprotein. The decision to employ this substrate ratio was based on the comparative analysis of various combinations, and it was observed that this specific proportion facilitated not only enhanced biomass accumulation but also demonstrated superior fermentation kinetics, thereby underscoring the importance of substrate selection in the bioconversion processes. 3.2. Optimization of Fermentation Conditions The application of Response Surface Methodology (RSM) played a pivotal role in unraveling the intricate interplay among various fermentation parameters, specifically temperature, pH, and moisture content, which were found to exert significant influence on the production yield of mycoprotein as previous noted by researchers ( Gang et al ., 2016 ; Dudekula et al ., 2020 ; Yan et al ., 2020 ). The statistical analysis revealed that the optimized fermentation conditions were established at a temperature of 23°C, a pH level of 6.2, and a moisture content of 68%. These conditions were meticulously determined to align with the metabolic requirements of the fermenting microorganisms, thereby promoting maximal growth and metabolic activity. The establishment of these parameters is critical, as they create an optimal environment for the fermentation process, which is essential for achieving high-efficiency bioconversion of the selected substrates into mycoprotein. 3.4. Mycoprotein Yield and Quality Under the optimized fermentation conditions, the yield of mycoprotein achieved was commendably quantified at 47.2%. Furthermore, the protein content of the produced mycoprotein was measured at 65.1%, reflecting a substantial concentration of protein within the biomass generated. The resultant mycoprotein exhibited a balanced amino acid profile, which is a key factor contributing to its nutritional value. The presence of essential amino acids within the mycoprotein matrix indicates its potential viability as a protein source in various food products, thus opening avenues for its application in food science and nutrition (Souza et al ., 2018; Ahmad et al ., 2022 ). The comprehensive evaluation of both the yield and the quality of the mycoprotein produced under the optimized conditions reaffirms the efficacy of the aforementioned substrate ratio and fermentation parameters in enhancing the viability of bioprocessing strategies aimed at sustainable food production. 4.0. Discussion The findings presented in this study illuminate the significant potential of utilizing sugarcane straw and cassava peels as viable substrates for the production of mycoprotein through the cultivation of the fungus Pleurotus ostreatus . The study carefully evaluated and optimized the ratio of these agricultural by- products, as well as the fermentation conditions, which collectively contribute to enhancing the mycoprotein yield. This research underscores the feasibility of transitioning from conventional protein sources to innovative alternatives that can be produced sustainably from agricultural wastes that was practically demonstrated by previous studies ( Koppram et al ., 2014 ; Kim et al ., 2022 ; Sydor et al ., 2022 ; Thiviya et al , 2022 ). By effectively employing agricultural wastes, a low-cost and abundantly available substrates, we can not only address environmental concerns associated with agricultural waste disposal but also contribute to the development of a more sustainable and environmentally friendly food production system ( Kim et al., 2011 ; Zhao et al ., 2020 ; Pellegrino et al., 2022 ; Zikou et al., 2023 ). Moreover, the mycoprotein obtained from this process exhibits promising characteristics that could be advantageous for its incorporation into various food products. Its nutritive profile, which includes essential amino acids and high protein content, makes it an attractive candidate for both vegetarian and omnivorous diets (Ahmad et al ., 2012; Derbyshire and Delange, 2021 ; Jach et al ., 2022 ; Bakratsas et al ., 2023 ). As consumers are increasingly seeking sustainable and nutritious food alternatives, the potential for mycoprotein derived from sugarcane straw and cassava peels to serve as a functional ingredient in food formulations is particularly noteworthy ( Xiao et al ., 2017 ; Kim et al ., 2022 ). This demonstrates the alignment of scientific research with contemporary dietary trends, emphasizing the role of innovative bioprocessing technologies in addressing global food security challenges. 4.1. Conclusion In conclusion, this study successfully optimized both the substrate composition and the fermentation conditions necessary for the effective production of mycoprotein from the cultivation of Pleurotus ostreatus on sugarcane straw and cassava peels. The outcomes of this research provide significant insights into the utilization of these agricultural waste materials as substrates, opening avenues for their application in biotechnological processes aimed at food production. Additionally, the importance of fine- tuning fermentation parameters has been clearly established, indicating that such optimization is critical to maximizing mycoprotein yields. 4.2. Implications The results of this study have implications for the production of mycoprotein, a potential alternative protein source. The optimized process conditions can be used to improve the efficiency and productivity of mycoprotein production, making it a more viable option for food and feed applications. This research has the potential to contribute significantly to knowledge in the field of mycoprotein production. By: Optimizing process conditions: The study provides valuable insights into the effects of substrate ratio, temperature, pH, and moisture content on mycoprotein yield and protein content. Advancing response surface methodology: The application of RSM demonstrates its effectiveness in optimizing complex bioprocesses. Informing industrial applications: The findings in this study can inform the development of efficient and scalable mycoprotein production processes. This research has the potential to: Enhance food security: By providing alternative protein sources. Support sustainable agriculture: By promoting efficient and environmentally friendly production methods. Foster innovation: By contributing to the development of new products and technologies. Overall, the study demonstrated the potential of response surface methodology for optimizing mycoprotein production and highlighted the importance of substrate ratio, temperature, pH, and moisture content in determining mycoprotein yield and protein content. Recommendation It is imperative that further research be conducted to explore the scalability of this production process. It would be beneficial to investigate not only the technical aspects of scaling up production but also the commercial viability and market acceptance of mycoprotein as an ingredient in a diverse range of food products. By doing so, we can ascertain the broader implications of this research in promoting sustainable practices within the food industry and enhancing the nutritional quality of food available to populations worldwide. This approach not only contributes to the scientific knowledge base but also aligns with global efforts to foster sustainability in food systems amidst growing environmental and health challenges. Footnotes Email: faithnaomi12{at}gmail.com References 1. ↵ Ahlborn , J. , Stephan , A. , Meckel , T. , Maheshwari , G. , Rühl , M. and Zorn , H. ( 2019 ). Upcycling of Food Industry Side Streams by Basidiomycetes for Production of a Vegan Protein Source . International Journal of Recycling Organic Waste Agriculture , 8 : 447 – 455 . OpenUrl 2. ↵ Ahmad , M.I. , Farooq , S. , Alhamoud , Y. , Li , C. and Zhang , H. ( 2022 ). A Review on Mycoprotein: History, Nutritional Composition, Production Methods, and Health Benefits . Trends Food Science Technology , 121 : 14 – 29 . OpenUrl 3. ↵ Bakratsas , G. , Polydera , A. , Nilson , O. , Chatzikonstantinou , A.V. , Xiros , C. , Katapodis , P. and Stamatis , H. ( 2023 ). 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OpenUrl 26. ↵ Zikou , E. , Chatzifragkou , A. , Koutinas , A.A. and Papanikolaou , S. ( 2023 ). Evaluating Glucose and Xylose as Cosubstrates for Lipid Accumulation and γ-Linolenic Acid Biosynthesis of Thamnidium Elegans . Journal of Applied Microbiology , 114 : 1020 – 1032 .. OpenUrl View the discussion thread. Back to top Previous Next Posted August 22, 2025. Download PDF Email Thank you for your interest in spreading the word about bioRxiv. NOTE: Your email address is requested solely to identify you as the sender of this article. Your Email * Your Name * Send To * Enter multiple addresses on separate lines or separate them with commas. 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Share Optimization of Substrate and Fermentation Conditions for Mycoprotein Production from Pleurotus ostreatus (Oyster Mushroom) Cultivated on Sugarcane Straw and Cassava Peels in Okitipupa, Nigeria Olagunju Johnson Adetuwo , Ogundana Faith Naomi bioRxiv 2025.08.22.671774; doi: https://doi.org/10.1101/2025.08.22.671774 Share This Article: Copy Citation Tools Optimization of Substrate and Fermentation Conditions for Mycoprotein Production from Pleurotus ostreatus (Oyster Mushroom) Cultivated on Sugarcane Straw and Cassava Peels in Okitipupa, Nigeria Olagunju Johnson Adetuwo , Ogundana Faith Naomi bioRxiv 2025.08.22.671774; doi: https://doi.org/10.1101/2025.08.22.671774 Citation Manager Formats BibTeX Bookends EasyBib EndNote (tagged) EndNote 8 (xml) Medlars Mendeley Papers RefWorks Tagged Ref Manager RIS Zotero Tweet Widget Facebook Like Google Plus One Subject Area Microbiology Subject Areas All Articles Animal Behavior and Cognition (7629) Biochemistry (17660) Bioengineering (13881) Bioinformatics (41910) Biophysics (21436) Cancer Biology (18576) Cell Biology (25480) Clinical Trials (138) Developmental Biology (13368) Ecology (19887) Epidemiology (2067) Evolutionary Biology (24302) Genetics (15598) Genomics (22482) Immunology (17726) Microbiology (40360) Molecular Biology (17163) Neuroscience (88534) Paleontology (666) Pathology (2830) Pharmacology and Toxicology (4821) Physiology (7637) Plant Biology (15129) Scientific Communication and Education (2045) Synthetic Biology (4290) Systems Biology (9817) Zoology (2269)