Bovine Biowaste Derived Nanocellulose for Pyrethrin Stabilization and Controlled Release

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Mugo, Weihao Lu This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5094131/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 12 Feb, 2025 Read the published version in Discover Materials → Version 1 posted 14 You are reading this latest preprint version Abstract Sustainable industrial and food production technologies are in demand with the heightened public environmental consciousness. For example, there is growing demand for organic agriculture where synthetic pesticides are replaced with biopesticides. While effective in the short term, biopesticides are unstable and decompose rapidly in nature, losing their pesticidal action. As such the use of biopesticides can be uneconomical. Pyrethrins are a good example of biopesticides that have found widespread application in both crop and animal husbandry. To improve pyrethrin stability, this article will demonstrate bovine biowaste derived nanocellulose (BBNC) as an effective support for stabilizing pyrethrins, and for controlled release for up to one month. The BBNC demonstrated functional similarities to commercial cellulose nanocrystals, hence the article points to a potential valorization technology potential for bovine biowaste. Bioallethrin Biopesticides controlled release Bovine biowaste nanocellulose Cellulose nanocrystals Figures Figure 1 Figure 2 Figure 3 1. Introduction To meet increased demand for food livestock farming trends has shifted to commercial large farms with concentrated feeding operations. The increase of livestock in a single location results in large manure production, and requires proper manure management. 1 Livestock is well known as a greenhouse gas contributor, particularly methane and nitrous oxide. 1 , 2 An excessive amount of manure can also pose an eutrophic concern to the surrounding environment through nitrogen and phosphorus leaching. 3 Present in animal feeds, hormones and antibiotics are present in manure, risking soil and surrounding groundwater contamination. 4 There has been a lot of interest towards investigating the valorization of livestock biowaste. 5 Some examples include anaerobic digestion of manure for smell reduction and generation of bioenergy (biogas), 3 , 6 and the production of biochar. 7 While livestock biowaste contains a large cellulosic component, the composition depends on the feedstock. 8 Generally, the composition of cattle manure consists of 25–30% cellulose, 15–25% hemicellulose and 5–15% lignin. 8,9 There is a need to investigate potential applications for animal processed fibers in the livestock manure valorization chain. The rumen is a bioreactor rich in microbial species that are responsible for numerous metabolic activities. 10 Microbes are responsible for the digestion and breakdown of cellulose from straw and animal feed into micro- and nanocellulose. 10 Additional cleaning and processing (i.e. removal of organics, bleaching and surface modification) can result in micro/nanocellulose utilizable in a variety of applications. 11 Cellulose nanomaterials have been used in many technologies and applications including paper electronics, 11 sensing and biosensing, 12 polymer reinforcement additives, 13 and water filtration. 14 Another example is the encapsulation and entrapment of pharmaceuticals, 15 essentials oils, and pesticides. 16 The purpose for encapsulation and entrapment is to stabilize and control the release of unstable molecules, i.e. pyrethroid insecticides. Pyrethrin is a natural insecticide produced by pyrethrum flower, Chrysanthemum cinerariaefolium , and is commonly cultivated in Australia, East Africa and Southern China. 17 Pyrethroids are synthetic compounds similar to pyrethrin that are widely used in households, agriculture and industry for insect control. 18 For example, insecticides have been applied as a textile finish to clothing as an insect repellant. 19 More specifically, bioallethrin is a potent allethrin that has been used in cotton finishing to combat houseflies, mosquitos, lice and cockroaches. 20 Unfortunately, pyrethroids are susceptible to microbial degradation, photodegradation, volatilization and hydrolysis in the environment. 18 There is a need to stabilize bioallethrin to prolong the insecticidal effects. Herein, we demonstrate cattle manure as a potential source of nanocellulose and is characterized in comparison to commercially available cellulose nanocrystals (CNC). Infrared spectroscopy (FTIR), zeta potential, material swelling, and electron microscopy were used to characterize the nanocellulose. Thin nanocellulose films were developed to entrap bioallethrin. The stability and slow-release of bioallethrin loaded nanocellulose films were studied over 31 days using gas chromatography mass spectrometry (GC/MS). 2. Material and Methods 2.1. Materials The cellulose nanocrystals (CNC) was donated by Alberta Innovates Technology Futures (AITF) and the cattle manure was obtained from a local farm in Alberta. The Clorox bleach was obtained from a local grocery store. 2,2,6,6-Tetramethyl-1-piperidinyloxy (TEMPO, 98%), ethanol (99%), bioallethrin, sodium bromide, acetone, toluene, dichloromethane (DCM), sodium hydroxide, sodium chloride, hydrogen peroxide (30 wt. %) were purchased from Sigma-Aldrich Canada (Oakville, ON). Fisherfinest Premium Cover Glass slides (25 mm x 25 mm) and MgSO 4 were purchased from Fisher Scientific, Canada. Centrifuge tubes (1.5 mL) were purchased from VWR International, Canada. Analytical grade deionized (DI) water was used for all experiments. 2.2. Extraction of bovine biowaste derived nanocellulose (BBNC) The cattle manure (355 g) was initially dried in the oven (80°C) to remove excess moisture. The cattle manure (14.30 g) was then rinsed with deionized water and filtered under vacuum using a Büchner funnel until the filtrate was colorless to remove excess organic material. Next, the manure was bleached by vigorously mixing with 10% H 2 O 2 solution for 8 hours at 80°C. Following bleaching the cellulose was oxidized via TEMPO oxidation method adapted from the literature. 21 The obtained cellulose (4 g) suspension was vigorously stirred with NaBr (20 % and TEMPO (4%). Clorox bleach (76 g, 10.3 % was slowly added to the solution and maintained at pH 10–11 using a NaOH. The oxidation was considered complete after no observable pH change. The reaction was terminated with the addition of ethanol (20 mL, 99 %, followed by 20 minutes of stirring. The oxidized nanocellulose was neutralized and washed using a Büchner funnel and vacuum filtration. The carboxyl content after TEMPO oxidation was determined using a conductometric titration. 21 , 22 The BBNC (~ 0.02 g) was dispersed in 100 mL of deionized water and 0.5 mL of 100 mM NaCl. The pH was adjusted to 2.8 using 0.1 M HCl and mixed at 400 rpm. The solution was titrated to a final pH of 10.5 using 0.01 M NaOH. The titration was performed in duplicates. 2.3. Characterization of CNC and BBNC The surface functional groups were determined using a Bruker Platinum ATR-FTIR. The transmittance spectra were measured at a resolution of 4 cm - 1 , a scan time of 16 scans and a range of 600 to 4000 cm - 1 . The zeta (ζ) potential of the nanocellulose (2 mg/mL) solutions were determined using Malvern Zetasizer Nano ZSP (Malvern, UK). The instrument’s measurement principle is electrophoretic light scattering with a sample measurement range from 3.8 nm − 100 µm. Analysis was completed at ambient room temperature and at neutral pH. The % swelling of CNC and BBNC from water sorbency was measured gravimetrically. Dried nanocellulose (0.025 g) was hydrated in a centrifuge tube (1 mL) for 15, 30 and 60 minutes. Prior to incubation the samples were dispersed using sonication and vortexing. The samples were centrifuged at 14,000 rpm for 60 minutes and the supernatant was discarded. The amount of water retained can be calculated using Eq. 1 : 23 $$\:\%\:swelling=\:\frac{{m}_{s}-{m}_{d}}{{m}_{d}}*100\:\%$$ 1 where m s is the mass of the swelled nanocellulose (g) and m d is the mass of the dried nanocellulose (g). Scanning electron microscopy (SEM) was performed using a field emission scanning electron microscope. Samples were mounted and coated with a conductive gold coating using a Xenosput XE200. With a JEOL-6301F, the samples were imaged using secondary electron imaging (SEI) at 5 kV. Transmission electron microscopy (TEM) images were obtained on an ultrahigh-resolution transmission electron microscope (JEOL JEM-2010FEF) using an accelerating voltage of 200 kV. 2.4. Preparation, extraction and analysis of CNC/BBNC thin films loaded with bioallethrin using GC-MS A 10% ethanol stock solution of bioallethrin (952 ppm, 100 mL) was prepared for loading onto the nanocellulose films, extraction and analysis experiments. The bioallethrin solution (10 mL) was mixed with CNC/BBNC (0.9 wt. %) for 3 hours at room temperature (300 rpm). The solution was deposited into a Petri dish and left to dry at room temperature overnight. The theoretical bioallethrin loading on the nanocellulose film was 10.5% (g/g). The initial immobilization of bioallethrin in the nanocellulose thin films was determined via a DCM solvent extraction. From the initial mixture (before deposition), the nanocellulose is filtered from the solution using a pipet filter. The solution containing the non-immobilized bioallethrin (10% aqueous ethanol solution) was isolated by liquid-liquid extraction using DCM. The solution was mixed at 600 rpm for 30 minutes, then mixed with brine solution used to break the emulsion. Before analysis the bioallethrin extract solution was dried using anhydrous MgSO 4 . Following the immobilization of bioallethrin in nanocellulose films, the controlled release of bioallethrin was monitored over 2, 6, 7, 17 and 31 days. To determine the amount of bioallethrin present in the films, 0.01 g of bioallethrin loaded nanocellulose film was sonicated in 2 mL of DCM for 30 minutes at room temperature. The extracted bioallethrin solution (in DCM) was filtered to remove nanocellulose before analysis. The controlled release of bioallethrin from CNC/BBNC thin films was analyzed with an Agilent GC/MS system (GC/MS 5975C, Agilent Technologies, Santa Clara CA, USA). Data acquisition was completed with Agilent ChemStation software (version G1701EA). The GC separation column was a HP-5MS 5% Phenyl Methyl Siloxane (30 m x 0.25 mm x 0.25 µm) (Agilent Technologies, Santa Clara CA, USA). The mobile phase was helium at a flow rate of 1.0 mL/min. The sample injection volume was 1 µL via a splitless injector at 280°C. The initial oven temperature was 60°C and ramped at 20°C/minute until 280°C. The mass spectrometer scan range of m/z 50 to 500 was used. Determination of immobilized bioallethrin after various days used an external calibration curve using bioallethrin standards prepared in DCM. 3. Results and Discussion 3.1. Extraction and characterization of cellulose from cattle manure The composition of cattle manure is a combination of cellulose, lignin, hemicellulose, proteins and lipids. 24 The manure was initially dried in the oven at 80°C and the dry mass content was determined as 34.6 % Manure was rinsed with deionized water until the filtrate cleared, extracting water-soluble substances and isolating cellulosic material. Bleaching treatment then removed lignin and hemicellulose from the cellulosic component. 25 After filtration, the biomass was treated with hydrogen peroxide. The wt. % of remaining cellulose following the bleaching treatment was 40 %. his relates closely to what has been reported in the literature. 8 , 9 The BBNC was surface modified using TEMPO-oxidation, which selectivity converts C6 hydroxyl groups to C6 carboxylate groups. 26 FTIR spectroscopy was used to compare BBNC to commercially produced CNC and to determine whether lignin and hemicellulose has been removed. Figure 1 a) shows the FTIR spectra of CNC and BBNC. The band assignments for CNC and BBNC are found in Table 1 . The spectra of CNC and BBNC overlap closely which suggests that they are chemically similar in structure. There is a noticeable difference between the two spectra in the 1600 to 1750 cm − 1 range. The band at 1643 cm − 1 in CNC is associated with δ (O-H) from absorbed water. 27 – 30 In the literature, bands found between 1509–1609 cm − 1 and 1700–1740 cm − 1 have been attributed to the presence of lignin and hemicellulose. 27 , 29 However, the band found at 1604 cm − 1 in BBNC has been attributed to ν as (COO − ) 31,32 associated with the surface oxidation of C6 hydroxyls to carboxylate/carboxylic acid groups. The band at 1730 cm − 1 has been attributed to ν s (C = O) 33 likely due to minimal protonation of the carboxylate group, rather than the presence of lignin and hemicellulose. Furthermore, the absence of an absorbance band at 1255 cm − 1 indicates the removal of hemicellulose. 34 The carboxylic acid content was determined using a conductometric titration and quantified as 0.88 ± 0.04 mmol/gram of BBNC. The sulfate half-ester content following sulfuric acid hydrolysis is typically reported to be around 0.2 to 0.3 mmol/gram for CNC. 35 Table 1 Comparison of CNC and BBNC FTIR band assignments. CNC Wavenumber (cm − 1 ) BBNC Wavenumber (cm − 1 ) Band assignment Reference 3331 3334 ν (O-H) (Fahma et al., 2010; Johar et al., 2012; Mohamad Haafiz et al., 2013; Rosa et al., 2012) 2897 2918 ν (C-H) (Fahma et al., 2010; Johar et al., 2012; Mohamad Haafiz et al., 2013; Rosa et al., 2012) - 1730 ν s (C = O) (Follain et al., 2010) 1643 - δ (O-H) (Fahma et al., 2010; Johar et al., 2012; Mohamad Haafiz et al., 2013; Rosa et al., 2012) - 1604 ν as (COO − ) (Benkaddour et al., 2014; Soni et al., 2015) 1428 1408 δ (C-H 2 ) (Jiang et al., 2017; Soni et al., 2015) 1316 1315 π (C-H) (Soni et al., 2015) 1161 1157 ν as (C-O-C) (Mohamad Haafiz et al., 2013; Rosa et al., 2012) 1055 1053 ν (C-O) (Fahma et al., 2010) 898 901 ρ (C-H), cellulosic β-glycosidic linkage (Fahma et al., 2010), (Jiang et al., 2017) ν - stretching; δ - bending; π - wagging; ρ - rocking; s - symmetric; as - asymmetric ζ-potential is used to determine the surface potential and colloidal stability of suspensions. 36 In the literature, CNC has ζ-potential values commonly between − 20 to -50 mV. 36 The ζ-potential of CNC and BBNC were determined as -47.1 and − 47.8 mV, respectively. The negative ζ-potential is due to the anionic sulfate half-ester and carboxylate groups present on the CNC and BBNC surface. The surface potential is important in understanding the interaction between the nanocellulose and bioallethrin, which determines adsorption, retention, and release thermodynamic and kinetics. Figure 1 b) demonstrates the swelling behaviour of CNC and BBNC. The percent swelling of CNC and BBNC after 15, 30 and 60 minutes in deionized water was measured. The CNC and BBNC reached a percent swelling of approximately 700% and 1500% after 30 minutes. The percent swelling of BBNC is larger than CNC by about a factor of 2 and has been attributed to the difference in surface functionalization. While the surface potential is similar, the carboxylate content in BBNC is likely higher than the sulfate half-ester content in CNC. Assuming the sulfate half-ester content for CNC is near the maximum observed in the literature, the BBNC carboxylate content is about 2–3 times higher, which would promote increased swelling capability. The aspect ratio and sizes of nanocellulose will vary based on their source and the preparation procedure. 37 The length and width of CNC from wood sources have been reported as 100–200 nm and 3–5 nm, respectively. 37 , 38 Fig. 2 a) is a TEM image of commercial CNC. The length and width of these CNC is estimated to be 116 ± 24 nm and 5 ± 1 nm (n = 25), respectively. Figure 2 b) is a TEM image of BBNC. The BBNC is not as uniform in size as the commercial CNC. A range of lengths and width for BBNC can be observed. The length and width have been estimated to be 100–300 nm and 10–30 nm. Figure 2 c) is a SEM image of a dried BBNC aggregate. Nanocellulose will aggregate upon drying processes due to the intra- and intermolecular hydrogen bonding. 39 However, the cellulose in this image is larger in size and is fibrous, rather than crystalline. This suggests that BBNC is a combination of micro- and nanocellulose. The difference in size and could be due to mild conditions of the ruminant digestive process compared to the traditional acid treatment used in commercial processes. Further treatment such as mechanical treatment and centrifugation could have also been used to limit the amount of microfibers and isolate the nanofibers. Figure 2 c) also shows evidence of porosity, which is a property relevant to the mass transfer kinetics for bioallethrin release. Figure 2 d) is a SEM image showing the cross-section of a dried BBNC mat. The mats are uniform with limited defects. Using the cross-sectional view, the mat thickness is estimated to be between ~ 5 µm. 3.2. Immobilization and controlled release of bioallethrin from nanocellulose films Bioallethrin was mixed and entrapped into CNC and BBNC films to stabilize and as a slow release platform. The amount of immobilized bioallethrin in CNC and BBNC was determined and quantified using the supernatant solution after filtering the nanocellulose. The bioallethrin was extracted from aqueous solution using DCM extraction. Figure 3 a) is a column chart showing the percent amount of bioallethrin in CNC and BBNC. The values have been normalized to the weight of sample (g). We determined a percent immobilization of 40% and 30% for CNC and BBNC, respectively. The immobilized bioallethrin is close in value and the difference could be related to the higher surface area of CNC and the size polydispersity in BBNC. The C2, C3 and C6 hydroxyls in nanocellulose present strong intra- and intermolecular hydrogen bonding capability. 39 The prevalent mode of interaction between the hydroxyl groups in CNC/BBNC and carbonyl groups in bioallethrin is hydrogen bonding. Figure 3 b) is a column chart monitoring the slow release of bioallethrin from CNC and BBNC over 2, 6, 7, 17 and 31 days. The inset image is a schematic of a chromatogram demonstrating the decrease in bioallethrin peak intensity over time. The bioallethrin was extracted from the nanocellulose and quantified using an external calibration curve of bioallethrin (R 2 = 0.9964). The initial immobilization into the CNC and BBNC mats is shown as day 0. While the CNC mats immobilized slightly more bioallethrin, the CNC released 62% of the bioallethrin after 2 days, while BBNC only released 39%. After 31 days the CNC released 91%, while BBNC released 77%. Pyrethrins are extremely sensitive to light and susceptible to breakdown within a few hours. 18 Moreover, Leng et al. performed a study to examine the exposure and elimination of pyrethrin in humans, and determined a half-life of about 4 hours. 40 The immobilization of bioallethrin into CNC and BBNC mats improved the stability and lifetime of the insecticide. The initial release of bioallethrin (within 48 hours) is much faster than the release after 48 hours. We suspect that this is from the initial rapid release of surface bound bioallethrin. Whereas the bioallethrin entrapped inside the CNC and BBNC mats demonstrate a much slower release rate. Comparatively, the BBNC mats released at a slower rate than the CNC mats. The physiochemical properties of the BBNC are thought to have a role in the slower release rate of bioallethrin. While the CNC are rod-like and crystalline in nature (Fig. 2 a), BBNC is a mixture of micro- and nanocellulose (Fig. 2 b and c). The crystalline CNC film and has less surface area for bioallethrin entrapment. In contrast, the more fibrous BBNC form physically crosslinked networks that easily entrap bioallethrin in micro-voids within the film. The physical absorption/adsorption interaction of the bioallethrin and the loading capacity in the nanocellulose is correlated with high fibrous content. 4. Conclusions In this work, we investigated potential applications for bovine biowaste nanocellulose. Nanocellulose was extracted from cattle manure and compared to commercially produced CNC. This article demonstrates the conventional and bovine biowaste derived nanocellulose are similar in structure and other physicochemical properties, making the latter a very sustainable valorization technology for bovine biowaste. Bioallethrin was entrapped into nanocellulose mats and its slow release was monitored over a month using GC/MS. Pyrethrins stabilization by nanocellulose closely mimics nature, where the biochemical pesticides (e.g. pyrethrins) are held in Chrysanthemum cinerariaefolium leaves until they are harvested. Films produced using commercial CNC released 91%, whereas BBNC released 77% after 31 days. In the future, this platform can be employed in diverse delivery systems of labile molecules, due to the lack of toxicity and the ease of functionalization to modify nanocellulose hydrophobicity. Declarations Author Contribution Weihao Lu was involved in carrying out most of the experiments in this work. Lu also wrote the draft manuscript. Samuel Mugo, the principal investigator designed the project and provided supervision and guidance on the project. Mugo extensively edited the draft manuscript to publishable quality. Acknowledgements Mugo research group acknowledges funding from NSERC and MacEwan University Research office. Data Availability The data used to generate the figures and tables in the manuscript is available on request. References Sefeedpari P, Vellinga T, Rafiee S, Sharifi M, Shine P, Pishgar-Komleh SH. J Clean Prod. 2019;233:857–68. Audsley E, Wilkinson M. J Clean Prod. 2014;73:263–8. Holm-Nielsen JB, Seadi T, Oleskowicz-Popiel P. Bioresour Technol. 2009;100:5478–84. Hill DN, Popova IE, Hammel JE, Morra MJ. J Environ Qual. 2019;48:47–56. Xu C, Nasrollahzadeh M, Selva M, Issaabadi Z, Luque R. Chem Soc Rev. 2019;48:4791–822. Amon T, Amon B, Kryvoruchko V, Zollitsch W, Mayer K, Gruber L. Agric Ecosyst Environ. 2007;118:173–82. Uzoma KC, Inoue M, Andry H, Fujimaki H, Zahoor A, Nishihara E. Soil Use Manag. 2011;27:205–12. Meyer S, Thiel V, Joergensen RG, Sundrum A. PLoS ONE. 2019;14:e0221266. Righi F, Simoni M, Visentin G, Manuelian CL, Currò S, Quarantelli A, de Marchi M. Livest Sci. 2017;206:105–8. Bickhart DM, Weimer PJ. J Dairy Sci. 2018;101:7680–9. Koga H, Nogi M, Komoda N, Nge TT, Sugahara T, Suganuma K. NPG Asia Mater. 2014;6:e93–93. Golmohammadi H, Morales-Narváez E, Naghdi T, Merkoçi A. Chem Mater. 2017;29:5426–46. Dufresne A. Curr Opin Colloid Interface Sci. 2017;29:1–8. Carpenter AW, de Lannoy CF, Wiesner MR. Environ Sci Technol. 2015;49:5277–87. Cao Y. Express Polym Lett. 2018;12:768–80. Tang C, Li Y, Pun J, Mohamed Osman AS, Tam KC. Colloids Surf A. 2019;570:403–13. Matsuo N. Proceedings of the Japan Academy Series B: Physical and Biological Sciences , 2019, 95, 378–400. Ullah S, Li Z, Zuberi A, Arifeen MZU, Baig MMFA. Environ Chem Lett. 2019;17:945–73. Chatha SAS, Asgher M, Asgher R, Hussain AI, Iqbal Y, Hussain SM, Bilal M, Saleem F, Iqbal HMN. Sci Total Environ. 2019;690:667–82. Hebeish A, Hamdy IA, EL–Sawy SM. Abdel–Mohdy. Res J Text Appar. 2009;13:24–33. Lin N, Bruzzese C, Dufresne A. ACS Appl Mater Interfaces. 2012;4:4948–59. Saito T, Isogai A. Biomacromolecules. 2004;5:1983–9. Mohaiyiddin MS, Ong HL, Othman MBH, Julkapli NM, Villagracia ARC, Md H, Akil. Polym Compos. 2018;39:E561–72. Liu Z, Zhang Y, Liu Z. Bioresour Technol. 2019;291:121855. Zeronian SH, Inglesby MK. Cellulose. 1995;2:265–72. Saito T, Kimura S, Nishiyama Y, Isogai A. Biomacromolecules. 2007;8:2485–91. Fahma F, Iwamoto S, Hori N, Iwata T, Takemura A. Cellulose. 2010;17:977–85. Johar N, Ahmad I, Dufresne A. Ind Crops Prod. 2012;37:93–9. Mohamad Haafiz MK, Eichhorn SJ, Hassan A, Jawaid M. Carbohydr Polym. 2013;93:628–34. Rosa SML, Rehman N, de Miranda MIG. Nachtigall and C. I. D. Bica. Carbohydr Polym. 2012;87:1131–8. Benkaddour A, Journoux-Lapp C, Jradi K, Robert S, Daneault C. J Mater Sci. 2014;49:2832–43. Soni B, Hassan EB, Mahmoud B. Carbohydr Polym. 2015;134:581–9. Follain N, Marais MF, Montanari S, Vignon MR. Polym (Guildf). 2010;51:5332–44. Jiang H, Wu Y, Han B, Zhang Y. Carbohydr Polym. 2017;174:291–8. Beck S, Méthot M, Bouchard J. Cellulose. 2015;22:101–16. Foster EJ, Moon RJ, Agarwal UP, Bortner MJ, Bras J, Camarero-Espinosa S, Chan KJ, Clift MJD, Cranston ED, Eichhorn SJ, Fox DM, Hamad WY, Heux L, Jean B, Korey M, Nieh W, Ong KJ, Reid MS, Renneckar S, Roberts R, Shatkin JA, Simonsen J, Stinson-Bagby K, Wanasekara N, Youngblood J. Chem Soc Rev. 2018;47:2609–79. Sacui IA, Nieuwendaal RC, Burnett DJ, Stranick SJ, Jorfi M, Weder C, Foster EJ, Olsson RT, Gilman JW. ACS Appl Mater Interfaces. 2014;6:6127–38. Habibi Y, Lucia LA, Rojas OJ. Chem Rev. 2010;110:3479–500. Peng Y, Gardner DJ, Han Y, Kiziltas A, Cai Z, Tshabalala MA. Cellulose. 2013;20:2379–92. Leng G, Gries W, Selim S. Toxicol Lett. 2006;162:195–201. Additional Declarations No competing interests reported. Supplementary Files image1.tiff.png Graphical Abstract Cite Share Download PDF Status: Published Journal Publication published 12 Feb, 2025 Read the published version in Discover Materials → Version 1 posted Editorial decision: Revision requested 14 Oct, 2024 Reviews received at journal 09 Oct, 2024 Reviews received at journal 07 Oct, 2024 Reviewers agreed at journal 04 Oct, 2024 Reviewers agreed at journal 04 Oct, 2024 Reviewers agreed at journal 04 Oct, 2024 Reviews received at journal 03 Oct, 2024 Reviews received at journal 30 Sep, 2024 Reviewers agreed at journal 27 Sep, 2024 Reviewers agreed at journal 26 Sep, 2024 Reviewers invited by journal 26 Sep, 2024 Editor assigned by journal 23 Sep, 2024 Submission checks completed at journal 23 Sep, 2024 First submitted to journal 15 Sep, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5094131","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":365639751,"identity":"1661bc8e-9e73-4b2b-a039-401d4c2fc45d","order_by":0,"name":"Samuel M. Mugo","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAuUlEQVRIiWNgGAWjYNCCCtK1nCFZB2MbKap1Z6RffPhz3jZ5/vYew88FDHbyBLWY3cgpNubddttwxpkzxtIzGJING4jQkibNuO024waJ3A3SPAwHGInSIvlzzm37DfJvN/8GarEnQkv6MQnehtuJGyR4t4FsSSSs5cwbZmOeY7eTZ5zJ/2bNY5CcTFjL8fSHD3/U3Lbtbz+WfJunws6WoBYGBh4DJI4BTmXIgP0BUcpGwSgYBaNgBAMAqhI+gUrGhfkAAAAASUVORK5CYII=","orcid":"","institution":"MacEwan University","correspondingAuthor":true,"prefix":"","firstName":"Samuel","middleName":"M.","lastName":"Mugo","suffix":""},{"id":365639752,"identity":"a02c5f12-cbee-4420-bdc9-898aef6d73d2","order_by":1,"name":"Weihao Lu","email":"","orcid":"","institution":"MacEwan University","correspondingAuthor":false,"prefix":"","firstName":"Weihao","middleName":"","lastName":"Lu","suffix":""}],"badges":[],"createdAt":"2024-09-15 20:30:52","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5094131/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5094131/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s43939-025-00207-9","type":"published","date":"2025-02-12T15:57:41+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":68348085,"identity":"b481ead6-44dc-43ab-ada3-4bbb23f25cb6","added_by":"auto","created_at":"2024-11-06 10:03:36","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":132095,"visible":true,"origin":"","legend":"\u003cp\u003ea)\u003cstrong\u003e \u003c/strong\u003eFTIR spectra of CNC and BBNC. b) Percent swelling of CNC and BBNC at 15, 30 and 60 minutes.\u003c/p\u003e","description":"","filename":"image2.tiff.png","url":"https://assets-eu.researchsquare.com/files/rs-5094131/v1/06dc9e24d671ef9f3282baf5.png"},{"id":68348087,"identity":"299798ee-de20-45fb-a39e-f73a6955191b","added_by":"auto","created_at":"2024-11-06 10:03:36","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1106476,"visible":true,"origin":"","legend":"\u003cp\u003ea) TEM image of commercial CNC. b) TEM and c) SEM image of BBNC after extraction from cattle manure and TEMPO oxidation. d) SEM image of the cross-sectional view of the BBNC mat.\u003c/p\u003e","description":"","filename":"image3.tiff.png","url":"https://assets-eu.researchsquare.com/files/rs-5094131/v1/5eb905370cabda70b42832fc.png"},{"id":68348086,"identity":"67da81f5-7646-4ec4-bcdb-7ec6cf818572","added_by":"auto","created_at":"2024-11-06 10:03:36","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1125669,"visible":true,"origin":"","legend":"\u003cp\u003ea) The percent immobilization of bioallethrin into CNC and BBNC. b) Column graph demonstrating the remaining immobilized bioallethrin in CNC and BBNC over 31 days. The percentages represent the amount of bioallethrin released. The inset image is a schematic representation of the decrease in signal intensity with the release of bioallethrin.\u003c/p\u003e","description":"","filename":"image4.tiff.png","url":"https://assets-eu.researchsquare.com/files/rs-5094131/v1/d5cb7e6b9936b2f6d9d5f404.png"},{"id":76488091,"identity":"f8f94bfc-27da-4a20-b642-44f4ac3d82ee","added_by":"auto","created_at":"2025-02-17 16:13:11","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3156705,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5094131/v1/86293114-25fa-4d5b-8945-22e6330adbc2.pdf"},{"id":68348088,"identity":"704dc482-6cd8-4c6d-8b9c-e5f4f146316e","added_by":"auto","created_at":"2024-11-06 10:03:36","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":562155,"visible":true,"origin":"","legend":"\u003cp\u003eGraphical Abstract\u003c/p\u003e","description":"","filename":"image1.tiff.png","url":"https://assets-eu.researchsquare.com/files/rs-5094131/v1/7e2e2f4918f93c9ea4e1642d.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Bovine Biowaste Derived Nanocellulose for Pyrethrin Stabilization and Controlled Release","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eTo meet increased demand for food livestock farming trends has shifted to commercial large farms with concentrated feeding operations. The increase of livestock in a single location results in large manure production, and requires proper manure management.\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e Livestock is well known as a greenhouse gas contributor, particularly methane and nitrous oxide.\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e,\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e An excessive amount of manure can also pose an eutrophic concern to the surrounding environment through nitrogen and phosphorus leaching.\u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e Present in animal feeds, hormones and antibiotics are present in manure, risking soil and surrounding groundwater contamination.\u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eThere has been a lot of interest towards investigating the valorization of livestock biowaste.\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e Some examples include anaerobic digestion of manure for smell reduction and generation of bioenergy (biogas),\u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e,\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e and the production of biochar.\u003csup\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e While livestock biowaste contains a large cellulosic component, the composition depends on the feedstock.\u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e Generally, the composition of cattle manure consists of 25\u0026ndash;30% cellulose, 15\u0026ndash;25% hemicellulose and 5\u0026ndash;15% lignin.\u003csup\u003e8,9\u003c/sup\u003e There is a need to investigate potential applications for animal processed fibers in the livestock manure valorization chain.\u003c/p\u003e \u003cp\u003eThe rumen is a bioreactor rich in microbial species that are responsible for numerous metabolic activities.\u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e Microbes are responsible for the digestion and breakdown of cellulose from straw and animal feed into micro- and nanocellulose.\u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e Additional cleaning and processing (i.e. removal of organics, bleaching and surface modification) can result in micro/nanocellulose utilizable in a variety of applications.\u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e Cellulose nanomaterials have been used in many technologies and applications including paper electronics,\u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e sensing and biosensing,\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e polymer reinforcement additives,\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e and water filtration.\u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e Another example is the encapsulation and entrapment of pharmaceuticals,\u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e essentials oils, and pesticides.\u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e The purpose for encapsulation and entrapment is to stabilize and control the release of unstable molecules, i.e. pyrethroid insecticides.\u003c/p\u003e \u003cp\u003ePyrethrin is a natural insecticide produced by pyrethrum flower, \u003cem\u003eChrysanthemum cinerariaefolium\u003c/em\u003e, and is commonly cultivated in Australia, East Africa and Southern China.\u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e Pyrethroids are synthetic compounds similar to pyrethrin that are widely used in households, agriculture and industry for insect control.\u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e For example, insecticides have been applied as a textile finish to clothing as an insect repellant.\u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e More specifically, bioallethrin is a potent allethrin that has been used in cotton finishing to combat houseflies, mosquitos, lice and cockroaches.\u003csup\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e Unfortunately, pyrethroids are susceptible to microbial degradation, photodegradation, volatilization and hydrolysis in the environment.\u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e There is a need to stabilize bioallethrin to prolong the insecticidal effects.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003cp\u003eHerein, we demonstrate cattle manure as a potential source of nanocellulose and is characterized in comparison to commercially available cellulose nanocrystals (CNC). Infrared spectroscopy (FTIR), zeta potential, material swelling, and electron microscopy were used to characterize the nanocellulose. Thin nanocellulose films were developed to entrap bioallethrin. The stability and slow-release of bioallethrin loaded nanocellulose films were studied over 31 days using gas chromatography mass spectrometry (GC/MS).\u003c/p\u003e"},{"header":"2. Material and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Materials\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eThe cellulose nanocrystals (CNC) was donated by Alberta Innovates Technology Futures (AITF) and the cattle manure was obtained from a local farm in Alberta. The Clorox bleach was obtained from a local grocery store. 2,2,6,6-Tetramethyl-1-piperidinyloxy (TEMPO, 98%), ethanol (99%), bioallethrin, sodium bromide, acetone, toluene, dichloromethane (DCM), sodium hydroxide, sodium chloride, hydrogen peroxide (30 wt. %) were purchased from Sigma-Aldrich Canada (Oakville, ON). Fisherfinest Premium Cover Glass slides (25 mm x 25 mm) and MgSO\u003csub\u003e4\u003c/sub\u003e were purchased from Fisher Scientific, Canada. Centrifuge tubes (1.5 mL) were purchased from VWR International, Canada. Analytical grade deionized (DI) water was used for all experiments.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Extraction of bovine biowaste derived nanocellulose (BBNC)\u003c/h2\u003e \u003cp\u003eThe cattle manure (355 g) was initially dried in the oven (80\u0026deg;C) to remove excess moisture. The cattle manure (14.30 g) was then rinsed with deionized water and filtered under vacuum using a B\u0026uuml;chner funnel until the filtrate was colorless to remove excess organic material. Next, the manure was bleached by vigorously mixing with 10% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e solution for 8 hours at 80\u0026deg;C. Following bleaching the cellulose was oxidized via TEMPO oxidation method adapted from the literature.\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e The obtained cellulose (4 g) suspension was vigorously stirred with NaBr (20 % and TEMPO (4%). Clorox bleach (76 g, 10.3 % was slowly added to the solution and maintained at pH 10\u0026ndash;11 using a NaOH. The oxidation was considered complete after no observable pH change. The reaction was terminated with the addition of ethanol (20 mL, 99 %, followed by 20 minutes of stirring. The oxidized nanocellulose was neutralized and washed using a B\u0026uuml;chner funnel and vacuum filtration.\u003c/p\u003e \u003cp\u003eThe carboxyl content after TEMPO oxidation was determined using a conductometric titration.\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e,\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e The BBNC (~\u0026thinsp;0.02 g) was dispersed in 100 mL of deionized water and 0.5 mL of 100 mM NaCl. The pH was adjusted to 2.8 using 0.1 M HCl and mixed at 400 rpm. The solution was titrated to a final pH of 10.5 using 0.01 M NaOH. The titration was performed in duplicates.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Characterization of CNC and BBNC\u003c/h2\u003e \u003cp\u003eThe surface functional groups were determined using a Bruker Platinum ATR-FTIR. The transmittance spectra were measured at a resolution of 4 cm\u003csup\u003e-\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e, a scan time of 16 scans and a range of 600 to 4000 cm\u003csup\u003e-\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe zeta (ζ) potential of the nanocellulose (2 mg/mL) solutions were determined using Malvern Zetasizer Nano ZSP (Malvern, UK). The instrument\u0026rsquo;s measurement principle is electrophoretic light scattering with a sample measurement range from 3.8 nm \u0026minus;\u0026thinsp;100 \u0026micro;m. Analysis was completed at ambient room temperature and at neutral pH.\u003c/p\u003e \u003cp\u003eThe % swelling of CNC and BBNC from water sorbency was measured gravimetrically. Dried nanocellulose (0.025 g) was hydrated in a centrifuge tube (1 mL) for 15, 30 and 60 minutes. Prior to incubation the samples were dispersed using sonication and vortexing. The samples were centrifuged at 14,000 rpm for 60 minutes and the supernatant was discarded. The amount of water retained can be calculated using Eq.\u0026nbsp;\u003cspan refid=\"Equ1\" class=\"InternalRef\"\u003e1\u003c/span\u003e:\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e\u003cdiv id=\"Equ1\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equ1\" name=\"EquationSource\"\u003e\n$$\\:\\%\\:swelling=\\:\\frac{{m}_{s}-{m}_{d}}{{m}_{d}}*100\\:\\%$$\u003c/div\u003e\u003cdiv class=\"EquationNumber\"\u003e1\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003ewhere \u003cem\u003em\u003c/em\u003e\u003csub\u003e\u003cem\u003es\u003c/em\u003e\u003c/sub\u003e is the mass of the swelled nanocellulose (g) and \u003cem\u003em\u003c/em\u003e\u003csub\u003e\u003cem\u003ed\u003c/em\u003e\u003c/sub\u003e is the mass of the dried nanocellulose (g).\u003c/p\u003e \u003cp\u003eScanning electron microscopy (SEM) was performed using a field emission scanning electron microscope. Samples were mounted and coated with a conductive gold coating using a Xenosput XE200. With a JEOL-6301F, the samples were imaged using secondary electron imaging (SEI) at 5 kV.\u003c/p\u003e \u003cp\u003eTransmission electron microscopy (TEM) images were obtained on an ultrahigh-resolution transmission electron microscope (JEOL JEM-2010FEF) using an accelerating voltage of 200 kV.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4. Preparation, extraction and analysis of CNC/BBNC thin films loaded with bioallethrin using GC-MS\u003c/h2\u003e \u003cp\u003eA 10% ethanol stock solution of bioallethrin (952 ppm, 100 mL) was prepared for loading onto the nanocellulose films, extraction and analysis experiments. The bioallethrin solution (10 mL) was mixed with CNC/BBNC (0.9 wt. %) for 3 hours at room temperature (300 rpm). The solution was deposited into a Petri dish and left to dry at room temperature overnight. The theoretical bioallethrin loading on the nanocellulose film was 10.5% (g/g).\u003c/p\u003e \u003cp\u003eThe initial immobilization of bioallethrin in the nanocellulose thin films was determined via a DCM solvent extraction. From the initial mixture (before deposition), the nanocellulose is filtered from the solution using a pipet filter. The solution containing the non-immobilized bioallethrin (10% aqueous ethanol solution) was isolated by liquid-liquid extraction using DCM. The solution was mixed at 600 rpm for 30 minutes, then mixed with brine solution used to break the emulsion. Before analysis the bioallethrin extract solution was dried using anhydrous MgSO\u003csub\u003e4\u003c/sub\u003e.\u003c/p\u003e \u003cp\u003eFollowing the immobilization of bioallethrin in nanocellulose films, the controlled release of bioallethrin was monitored over 2, 6, 7, 17 and 31 days. To determine the amount of bioallethrin present in the films, 0.01 g of bioallethrin loaded nanocellulose film was sonicated in 2 mL of DCM for 30 minutes at room temperature. The extracted bioallethrin solution (in DCM) was filtered to remove nanocellulose before analysis.\u003c/p\u003e \u003cp\u003eThe controlled release of bioallethrin from CNC/BBNC thin films was analyzed with an Agilent GC/MS system (GC/MS 5975C, Agilent Technologies, Santa Clara CA, USA). Data acquisition was completed with Agilent ChemStation software (version G1701EA). The GC separation column was a HP-5MS 5% Phenyl Methyl Siloxane (30 m x 0.25 mm x 0.25 \u0026micro;m) (Agilent Technologies, Santa Clara CA, USA). The mobile phase was helium at a flow rate of 1.0 mL/min. The sample injection volume was 1 \u0026micro;L via a splitless injector at 280\u0026deg;C. The initial oven temperature was 60\u0026deg;C and ramped at 20\u0026deg;C/minute until 280\u0026deg;C. The mass spectrometer scan range of m/z 50 to 500 was used. Determination of immobilized bioallethrin after various days used an external calibration curve using bioallethrin standards prepared in DCM.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results and Discussion","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Extraction and characterization of cellulose from cattle manure\u003c/h2\u003e \u003cp\u003eThe composition of cattle manure is a combination of cellulose, lignin, hemicellulose, proteins and lipids.\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e The manure was initially dried in the oven at 80\u0026deg;C and the dry mass content was determined as 34.6 % Manure was rinsed with deionized water until the filtrate cleared, extracting water-soluble substances and isolating cellulosic material. Bleaching treatment then removed lignin and hemicellulose from the cellulosic component.\u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e After filtration, the biomass was treated with hydrogen peroxide. The wt. % of remaining cellulose following the bleaching treatment was 40 %. his relates closely to what has been reported in the literature.\u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e,\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e The BBNC was surface modified using TEMPO-oxidation, which selectivity converts C6 hydroxyl groups to C6 carboxylate groups.\u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eFTIR spectroscopy was used to compare BBNC to commercially produced CNC and to determine whether lignin and hemicellulose has been removed. Figure\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea) shows the FTIR spectra of CNC and BBNC. The band assignments for CNC and BBNC are found in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The spectra of CNC and BBNC overlap closely which suggests that they are chemically similar in structure. There is a noticeable difference between the two spectra in the 1600 to 1750 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e range. The band at 1643 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e in CNC is associated with δ (O-H) from absorbed water.\u003csup\u003e\u003cspan additionalcitationids=\"CR28 CR29\" citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e In the literature, bands found between 1509\u0026ndash;1609 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and 1700\u0026ndash;1740 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e have been attributed to the presence of lignin and hemicellulose.\u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e,\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e However, the band found at 1604 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e in BBNC has been attributed to ν\u003csub\u003eas\u003c/sub\u003e (COO\u003csup\u003e\u0026minus;\u003c/sup\u003e)\u003csup\u003e31,32\u003c/sup\u003e associated with the surface oxidation of C6 hydroxyls to carboxylate/carboxylic acid groups. The band at 1730 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e has been attributed to ν\u003csub\u003es\u003c/sub\u003e (C\u0026thinsp;=\u0026thinsp;O)\u003csup\u003e33\u003c/sup\u003e likely due to minimal protonation of the carboxylate group, rather than the presence of lignin and hemicellulose. Furthermore, the absence of an absorbance band at 1255 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e indicates the removal of hemicellulose.\u003csup\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e The carboxylic acid content was determined using a conductometric titration and quantified as 0.88 \u0026plusmn; 0.04 mmol/gram of BBNC. The sulfate half-ester content following sulfuric acid hydrolysis is typically reported to be around 0.2 to 0.3 mmol/gram for CNC.\u003csup\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparison of CNC and BBNC FTIR band assignments.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCNC\u003c/p\u003e \u003cp\u003eWavenumber (cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBBNC\u003c/p\u003e \u003cp\u003eWavenumber (cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBand assignment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eReference\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3331\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3334\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eν (O-H)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(Fahma et al., 2010; Johar et al., 2012; Mohamad Haafiz et al., 2013; Rosa et al., 2012)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2897\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2918\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eν (C-H)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(Fahma et al., 2010; Johar et al., 2012; Mohamad Haafiz et al., 2013; Rosa et al., 2012)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1730\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eν\u003csub\u003es\u003c/sub\u003e (C\u0026thinsp;=\u0026thinsp;O)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(Follain et al., 2010)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1643\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eδ (O-H)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(Fahma et al., 2010; Johar et al., 2012; Mohamad Haafiz et al., 2013; Rosa et al., 2012)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1604\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eν\u003csub\u003eas\u003c/sub\u003e (COO\u003csup\u003e\u0026minus;\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(Benkaddour et al., 2014; Soni et al., 2015)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1428\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1408\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eδ (C-H\u003csub\u003e2\u003c/sub\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(Jiang et al., 2017; Soni et al., 2015)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1316\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1315\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eπ (C-H)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(Soni et al., 2015)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1161\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1157\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eν\u003csub\u003eas\u003c/sub\u003e (C-O-C)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(Mohamad Haafiz et al., 2013; Rosa et al., 2012)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1055\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1053\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eν (C-O)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(Fahma et al., 2010)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e898\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e901\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eρ (C-H), cellulosic β-glycosidic linkage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e(Fahma et al., 2010), (Jiang et al., 2017)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eν - stretching; δ - bending; π - wagging; ρ - rocking; s - symmetric; as - asymmetric\u003c/p\u003e \u003cp\u003eζ-potential is used to determine the surface potential and colloidal stability of suspensions.\u003csup\u003e\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u003c/sup\u003e In the literature, CNC has ζ-potential values commonly between \u0026minus;\u0026thinsp;20 to -50 mV.\u003csup\u003e\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u003c/sup\u003e The ζ-potential of CNC and BBNC were determined as -47.1 and \u0026minus;\u0026thinsp;47.8 mV, respectively. The negative ζ-potential is due to the anionic sulfate half-ester and carboxylate groups present on the CNC and BBNC surface. The surface potential is important in understanding the interaction between the nanocellulose and bioallethrin, which determines adsorption, retention, and release thermodynamic and kinetics.\u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb) demonstrates the swelling behaviour of CNC and BBNC. The percent swelling of CNC and BBNC after 15, 30 and 60 minutes in deionized water was measured. The CNC and BBNC reached a percent swelling of approximately 700% and 1500% after 30 minutes. The percent swelling of BBNC is larger than CNC by about a factor of 2 and has been attributed to the difference in surface functionalization. While the surface potential is similar, the carboxylate content in BBNC is likely higher than the sulfate half-ester content in CNC. Assuming the sulfate half-ester content for CNC is near the maximum observed in the literature, the BBNC carboxylate content is about 2\u0026ndash;3 times higher, which would promote increased swelling capability.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe aspect ratio and sizes of nanocellulose will vary based on their source and the preparation procedure.\u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u003c/sup\u003e The length and width of CNC from wood sources have been reported as 100\u0026ndash;200 nm and 3\u0026ndash;5 nm, respectively.\u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e,\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/sup\u003e Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ea) is a TEM image of commercial CNC. The length and width of these CNC is estimated to be 116\u0026thinsp;\u0026plusmn;\u0026thinsp;24 nm and 5\u0026thinsp;\u0026plusmn;\u0026thinsp;1 nm (n\u0026thinsp;=\u0026thinsp;25), respectively. Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb) is a TEM image of BBNC. The BBNC is not as uniform in size as the commercial CNC. A range of lengths and width for BBNC can be observed. The length and width have been estimated to be 100\u0026ndash;300 nm and 10\u0026ndash;30 nm. Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ec) is a SEM image of a dried BBNC aggregate. Nanocellulose will aggregate upon drying processes due to the intra- and intermolecular hydrogen bonding.\u003csup\u003e\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e However, the cellulose in this image is larger in size and is fibrous, rather than crystalline. This suggests that BBNC is a combination of micro- and nanocellulose. The difference in size and could be due to mild conditions of the ruminant digestive process compared to the traditional acid treatment used in commercial processes. Further treatment such as mechanical treatment and centrifugation could have also been used to limit the amount of microfibers and isolate the nanofibers.\u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ec) also shows evidence of porosity, which is a property relevant to the mass transfer kinetics for bioallethrin release. Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ed) is a SEM image showing the cross-section of a dried BBNC mat. The mats are uniform with limited defects. Using the cross-sectional view, the mat thickness is estimated to be between ~\u0026thinsp;5 \u0026micro;m.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Immobilization and controlled release of bioallethrin from nanocellulose films\u003c/h2\u003e \u003cp\u003eBioallethrin was mixed and entrapped into CNC and BBNC films to stabilize and as a slow release platform. The amount of immobilized bioallethrin in CNC and BBNC was determined and quantified using the supernatant solution after filtering the nanocellulose. The bioallethrin was extracted from aqueous solution using DCM extraction. Figure\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea) is a column chart showing the percent amount of bioallethrin in CNC and BBNC. The values have been normalized to the weight of sample (g). We determined a percent immobilization of 40% and 30% for CNC and BBNC, respectively. The immobilized bioallethrin is close in value and the difference could be related to the higher surface area of CNC and the size polydispersity in BBNC. The C2, C3 and C6 hydroxyls in nanocellulose present strong intra- and intermolecular hydrogen bonding capability.\u003csup\u003e\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e The prevalent mode of interaction between the hydroxyl groups in CNC/BBNC and carbonyl groups in bioallethrin is hydrogen bonding.\u003c/p\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb) is a column chart monitoring the slow release of bioallethrin from CNC and BBNC over 2, 6, 7, 17 and 31 days. The inset image is a schematic of a chromatogram demonstrating the decrease in bioallethrin peak intensity over time. The bioallethrin was extracted from the nanocellulose and quantified using an external calibration curve of bioallethrin (R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.9964). The initial immobilization into the CNC and BBNC mats is shown as day 0. While the CNC mats immobilized slightly more bioallethrin, the CNC released 62% of the bioallethrin after 2 days, while BBNC only released 39%. After 31 days the CNC released 91%, while BBNC released 77%. Pyrethrins are extremely sensitive to light and susceptible to breakdown within a few hours.\u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e Moreover, Leng et al. performed a study to examine the exposure and elimination of pyrethrin in humans, and determined a half-life of about 4 hours.\u003csup\u003e\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u003c/sup\u003e The immobilization of bioallethrin into CNC and BBNC mats improved the stability and lifetime of the insecticide. The initial release of bioallethrin (within 48 hours) is much faster than the release after 48 hours. We suspect that this is from the initial rapid release of surface bound bioallethrin. Whereas the bioallethrin entrapped inside the CNC and BBNC mats demonstrate a much slower release rate. Comparatively, the BBNC mats released at a slower rate than the CNC mats. The physiochemical properties of the BBNC are thought to have a role in the slower release rate of bioallethrin. While the CNC are rod-like and crystalline in nature (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ea), BBNC is a mixture of micro- and nanocellulose (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb and c). The crystalline CNC film and has less surface area for bioallethrin entrapment. In contrast, the more fibrous BBNC form physically crosslinked networks that easily entrap bioallethrin in micro-voids within the film. The physical absorption/adsorption interaction of the bioallethrin and the loading capacity in the nanocellulose is correlated with high fibrous content.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4. Conclusions","content":"\u003cp\u003eIn this work, we investigated potential applications for bovine biowaste nanocellulose. Nanocellulose was extracted from cattle manure and compared to commercially produced CNC. This article demonstrates the conventional and bovine biowaste derived nanocellulose are similar in structure and other physicochemical properties, making the latter a very sustainable valorization technology for bovine biowaste. Bioallethrin was entrapped into nanocellulose mats and its slow release was monitored over a month using GC/MS. Pyrethrins stabilization by nanocellulose closely mimics nature, where the biochemical pesticides (e.g. pyrethrins) are held in \u003cem\u003eChrysanthemum cinerariaefolium\u003c/em\u003e leaves until they are harvested. Films produced using commercial CNC released 91%, whereas BBNC released 77% after 31 days. In the future, this platform can be employed in diverse delivery systems of labile molecules, due to the lack of toxicity and the ease of functionalization to modify nanocellulose hydrophobicity.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eWeihao Lu was involved in carrying out most of the experiments in this work. Lu also wrote the draft manuscript. Samuel Mugo, the principal investigator designed the project and provided supervision and guidance on the project. Mugo extensively edited the draft manuscript to publishable quality.\u003c/p\u003e\u003ch2\u003eAcknowledgements\u003c/h2\u003e \u003cp\u003eMugo research group acknowledges funding from NSERC and MacEwan University Research office.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe data used to generate the figures and tables in the manuscript is available on request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eSefeedpari P, Vellinga T, Rafiee S, Sharifi M, Shine P, Pishgar-Komleh SH. J Clean Prod. 2019;233:857\u0026ndash;68.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAudsley E, Wilkinson M. J Clean Prod. 2014;73:263\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHolm-Nielsen JB, Seadi T, Oleskowicz-Popiel P. Bioresour Technol. 2009;100:5478\u0026ndash;84.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHill DN, Popova IE, Hammel JE, Morra MJ. J Environ Qual. 2019;48:47\u0026ndash;56.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eXu C, Nasrollahzadeh M, Selva M, Issaabadi Z, Luque R. Chem Soc Rev. 2019;48:4791\u0026ndash;822.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAmon T, Amon B, Kryvoruchko V, Zollitsch W, Mayer K, Gruber L. Agric Ecosyst Environ. 2007;118:173\u0026ndash;82.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eUzoma KC, Inoue M, Andry H, Fujimaki H, Zahoor A, Nishihara E. Soil Use Manag. 2011;27:205\u0026ndash;12.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMeyer S, Thiel V, Joergensen RG, Sundrum A. PLoS ONE. 2019;14:e0221266.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRighi F, Simoni M, Visentin G, Manuelian CL, Curr\u0026ograve; S, Quarantelli A, de Marchi M. Livest Sci. 2017;206:105\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBickhart DM, Weimer PJ. J Dairy Sci. 2018;101:7680\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKoga H, Nogi M, Komoda N, Nge TT, Sugahara T, Suganuma K. NPG Asia Mater. 2014;6:e93\u0026ndash;93.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGolmohammadi H, Morales-Narv\u0026aacute;ez E, Naghdi T, Merko\u0026ccedil;i A. Chem Mater. 2017;29:5426\u0026ndash;46.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDufresne A. Curr Opin Colloid Interface Sci. 2017;29:1\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCarpenter AW, de Lannoy CF, Wiesner MR. Environ Sci Technol. 2015;49:5277\u0026ndash;87.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCao Y. Express Polym Lett. 2018;12:768\u0026ndash;80.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTang C, Li Y, Pun J, Mohamed Osman AS, Tam KC. Colloids Surf A. 2019;570:403\u0026ndash;13.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMatsuo N. \u003cem\u003eProceedings of the Japan Academy Series B: Physical and Biological Sciences\u003c/em\u003e, 2019, 95, 378\u0026ndash;400.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eUllah S, Li Z, Zuberi A, Arifeen MZU, Baig MMFA. Environ Chem Lett. 2019;17:945\u0026ndash;73.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChatha SAS, Asgher M, Asgher R, Hussain AI, Iqbal Y, Hussain SM, Bilal M, Saleem F, Iqbal HMN. Sci Total Environ. 2019;690:667\u0026ndash;82.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHebeish A, Hamdy IA, EL\u0026ndash;Sawy SM. Abdel\u0026ndash;Mohdy. Res J Text Appar. 2009;13:24\u0026ndash;33.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLin N, Bruzzese C, Dufresne A. ACS Appl Mater Interfaces. 2012;4:4948\u0026ndash;59.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaito T, Isogai A. Biomacromolecules. 2004;5:1983\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMohaiyiddin MS, Ong HL, Othman MBH, Julkapli NM, Villagracia ARC, Md H, Akil. Polym Compos. 2018;39:E561\u0026ndash;72.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu Z, Zhang Y, Liu Z. Bioresour Technol. 2019;291:121855.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZeronian SH, Inglesby MK. Cellulose. 1995;2:265\u0026ndash;72.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaito T, Kimura S, Nishiyama Y, Isogai A. Biomacromolecules. 2007;8:2485\u0026ndash;91.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFahma F, Iwamoto S, Hori N, Iwata T, Takemura A. Cellulose. 2010;17:977\u0026ndash;85.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJohar N, Ahmad I, Dufresne A. Ind Crops Prod. 2012;37:93\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMohamad Haafiz MK, Eichhorn SJ, Hassan A, Jawaid M. Carbohydr Polym. 2013;93:628\u0026ndash;34.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRosa SML, Rehman N, de Miranda MIG. Nachtigall and C. I. D. Bica. Carbohydr Polym. 2012;87:1131\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBenkaddour A, Journoux-Lapp C, Jradi K, Robert S, Daneault C. J Mater Sci. 2014;49:2832\u0026ndash;43.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSoni B, Hassan EB, Mahmoud B. Carbohydr Polym. 2015;134:581\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFollain N, Marais MF, Montanari S, Vignon MR. Polym (Guildf). 2010;51:5332\u0026ndash;44.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJiang H, Wu Y, Han B, Zhang Y. Carbohydr Polym. 2017;174:291\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBeck S, M\u0026eacute;thot M, Bouchard J. Cellulose. 2015;22:101\u0026ndash;16.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFoster EJ, Moon RJ, Agarwal UP, Bortner MJ, Bras J, Camarero-Espinosa S, Chan KJ, Clift MJD, Cranston ED, Eichhorn SJ, Fox DM, Hamad WY, Heux L, Jean B, Korey M, Nieh W, Ong KJ, Reid MS, Renneckar S, Roberts R, Shatkin JA, Simonsen J, Stinson-Bagby K, Wanasekara N, Youngblood J. Chem Soc Rev. 2018;47:2609\u0026ndash;79.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSacui IA, Nieuwendaal RC, Burnett DJ, Stranick SJ, Jorfi M, Weder C, Foster EJ, Olsson RT, Gilman JW. ACS Appl Mater Interfaces. 2014;6:6127\u0026ndash;38.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHabibi Y, Lucia LA, Rojas OJ. Chem Rev. 2010;110:3479\u0026ndash;500.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePeng Y, Gardner DJ, Han Y, Kiziltas A, Cai Z, Tshabalala MA. Cellulose. 2013;20:2379\u0026ndash;92.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLeng G, Gries W, Selim S. Toxicol Lett. 2006;162:195\u0026ndash;201.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"discover-materials","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"dime","sideBox":"Learn more about [Discover Materials](https://www.springer.com/journal/43939)","snPcode":"","submissionUrl":"","title":"Discover Materials","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Bioallethrin, Biopesticides controlled release, Bovine biowaste nanocellulose, Cellulose nanocrystals","lastPublishedDoi":"10.21203/rs.3.rs-5094131/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5094131/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eSustainable industrial and food production technologies are in demand with the heightened public environmental consciousness. For example, there is growing demand for organic agriculture where synthetic pesticides are replaced with biopesticides. While effective in the short term, biopesticides are unstable and decompose rapidly in nature, losing their pesticidal action. As such the use of biopesticides can be uneconomical. Pyrethrins are a good example of biopesticides that have found widespread application in both crop and animal husbandry. To improve pyrethrin stability, this article will demonstrate bovine biowaste derived nanocellulose (BBNC) as an effective support for stabilizing pyrethrins, and for controlled release for up to one month. 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