Assessment and Improvement of Nutritive Value of Agricultural Residue Feed for Caprine

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SHITTU, Biliksu ABDULKAREEM, Halimat B. ALADE, Jimoh O. TIJANI This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5564179/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The use of agricultural residues as caprine feed is essential for sustaining livestock, and ensuring food security. However, the presence of fungal communities in these residues poses a threat to the feed quality and animal health. This study aimed to determine the fungal diversity and optimize of selenium nanoparticle (SeNPs) biosynthesis for antifungal scavengers for the feeds. The isolation and identification of the fungal species were characterised through morphological, microscopic, and phylogenetic analyses. The optimization of SeNPs was done using sodium selenite concentration, pH, and incubation time. While characterized was carried out with ultraviolet-visible spectroscopy (UV-Vis), and ZETA sizer. The presence of fungal species such as Aspergillus flavus, Rhizopus species, Aspergillus niger, Trichophyton rubrum, Mucor sp., Macrosporium audounii, Epidermophyton floccosum, Verticillium tenesum, Tricophytonmetagrophi and Exophialajeanselmei was confirmed across the feeds. With fungal load of varying concentrations ranging from 1.76 x 10 4 cfu/g of bean husk (BH) to 6.08 x 10 4 cfu/g of groundnut husk (GOH) was also confirmed. The evolutionary classification of the fungi isolated showed a monophyletic pattern of Aspergillus flavus species clustered with a bootstrap of 59%. The best condition for synthesizing PT-SeNPs was confirmed has 7 days of incubation time, 2mM of PT concentration and pH 6. The PT-SeNPs with particle size of 26.30 nm at wavelength of 300 nm show antifungal activity against Aspergillus nidulans (24.25 ± 0.45 mm) at 3.0ml. Hence, the present of toxigenic fungi in agricultural residue feed for caprine requires proper safety measures and the potential of PT-SeNPs as an antifungal scavenger for Aspergillus contamination. Caprine feed Nutritional value and quality Microbial load Phylogenicity Molecular taxonomy Figures Figure 1 Figure 2 Figure 3 Figure 4 1.0 INTRODUCTION CAPRINE is a multi-functional animal that plays a significant role in the economy and nutrition of landless, small and marginal farmers in Nigeria. This is an enterprise, that has been practiced by a large section of the population in rural areas. Caprine can efficiently survive on available shrubs and trees in adverse harsh environments in low fertility lands where no other crop can be grown. Also, agricultural residues, such as crop by-products play a crucial role in sustaining both agricultural productivity and the global food supply chain of caprine farming (Perdana et al ., 2023). These low-quality residues available after crop harvest has become a viable practice worldwide for alleviating some of the feeding burdens (Kumar et al ., 2023). The increasing scarcity of free grazing areas and the rising cost of commercial feed contribute to the growing utilization of these by-products (Bhandari, 2019). Also, the decision to use these residues are driven by the need for roughage during feed scarcity or drought conditions. Most crop residues are inherently poor in nutrition and high in fiber content and feeding untreated residues may hinder animal acceptance and subsequently impact animal performance (Derara and Bekuma, 2021). However, the presence of microorganisms, including fungi, in these residue feeds raises concerns about the potential impact on both feed quality and animal health. Fungi are ubiquitous in the environment and have diverse effects on agricultural systems. Some fungi contribute to the degradation of organic matter, aiding in nutrient cycling, while others may produce mycotoxins that can be harmful to animals and humans (Devi et al ., 2020). A greater number of possible risks that call for understanding and awareness of mycotoxins have been brought about by the globalization of the trade in agricultural commodities (Mona et al ., 2016). According to Bhandari et al. (2023), nanotechnology has the potential to have a significant impact on the agricultural industry through a variety of means, including targeted and intelligent delivery of nutrients and bioactive compounds, Nanoparticle-mediated genetic material delivery for crop improvement, nano fertilizers, disease management, nanosensors for pathogen detection and soil monitoring, nanoencapsulation of seeds, nano pesticides, and nano herbicides, increased crop yield, and nutritional quality. Nanoparticles (NP) have been synthesized using a wide range of techniques, including chemical, physical, biological, and biogenic methods (Agbebati-Maleki et al ., 2009). On the other hand, it has been demonstrated that the biogenic reduction of metal precursors to produce the corresponding NPs is less costly, environmentally benign, and devoid of chemical pollutants. Natural goods that have been a good source of biogenic reduction of metallic particles to nanoparticles include plant extract, which is embedded with naturally occurring stabilizing, growth terminating, and capping chemicals. Selenium nanoparticles (SeNPs), a type of metallic nanoparticle generated through biogenic reduction, have demonstrated antibacterial activities against certain pathogenic organisms (Nadaroglu et al ., 2017). Because of its distinct characteristics, structure, and size, it finds use in the biomedical sciences for antibacterial therapy, drug transport, cancer treatment, medical diagnosis, and sensor fabrication (Nadaroglu et al ., 2017). The leguminous plant Piliostigma thonningii Schum . is a member of the 133-genus Caesalpinioideae subfamily of the Fabaceae family (Cyril et al ., 2021). It has been discovered that P. thonningii roots, bark, and leaves are used to cure haematochezia, stomach issues, and loss of appetite (Cyril et al ., 2021). There have been reports of antilipidemic, antibacterial, antihelminthic, and anti-inflammatory properties (Cyril et al ., 2021). Therefore, P. Thonningii can function as a biogenic reduction of selenium salt. Hence, this research aims to analyze and characterize fungal communities in caprine agricultural residue feeds and also optimize the synthesis of selenium nanoparticles (SeNPs) as fungi scavengers in agricultural residue caprine feed. 2.0 MATERIALS AND METHODS 2.1 MATERIALS 2.1.1 Sample Collection The samples; beans husk, Guinea corn husk, groundnut husk, and maize shaft were purchased from Major Market in Minna, Niger state. The natural habitat of P. thonningii, was found near Bosso Area in Minna, Niger state, where the fresh leaves were collected. The Federal University of Technology, Minna, Niger State's Department of Plant Biology performed the taxonomic authentication of the plant. 2.1.2 Reagents and Chemicals All chemicals and reagents used in this study were of analytical grade which include; Genomic Lysis Buffer, DNA Elution Buffer, DNA Prewash Buffer, Bashing Bead Buffer, g-DNA Wash Buffer, Potato dextrose agar (PDA), Chloramphenicol, Lactophenol cotton blue. sulfuric acid, ascorbic acid, and sodium selenite, all of which were produced by Sigma Chemical Co. 2.2 METHODS 2.2.1 Isolation and estimation of fungal species from caprine agricultural residue feed samples Fungal isolation was carried out using the pour plate technique with 1ml of the 10 -4 dilutions of each sample aseptically transferred into sterile petri dishes and molten potato dextrose agar medium was poured into respective petri dishes and allowed to solidify before incubating at 37°C for 48-72 hours. This process was carried out in duplicates for each sample. After incubation, the plates were then screened for the presence of discrete colonies and the actual numbers of fungi, Mold, and yeast were estimated in colony-forming units per gram (cfu/g), (Bird et al ., 2015). 2.2.2 Estimation of the microbial load in a sample The microbial concentration is expressed in colony forming unit (cfu/g) per gram of sample, which is an estimate of viable fungal cells in a sample. Colony colony-forming unit is calculated using the formula: Subculture of fungal species : Following incubation, each plate was examined and fungi colonies were sub-cultured onto fresh potato dextrose agar plates to obtain pure cultures which was stored on appropriate agar slants for further identification and analysis. 2.2.3 Identification of fungal species Isolated fungal species were identified based on colony morphological characteristics on the surface of the culture medium and microscopic features. The technique of Oyeleke and Manga (2008) was adopted for the identification of the isolated fungi using lactophenol cotton blue stain. Determination of Microscopic features was achieved by placing a drop of the lactophenol stain on a clean grease-free glass slide and a small portion of the aerial mycelia of the fungi culture placed in the drop of lactophenol stain and a cover slip was gently placed over it. The slide was then mounted and viewed under a light microscope at ×10 and ×40 objectives. 2.3 DNA Extraction and Molecular Characterization of Fungi Isolates Molecular identification was performed to confirm the identities of the fungal species recovered from samples as outlined by Samson et al ., (2010). Genomic DNA was extracted from the fungal cultures using the ZR fungal DNA kit (Zymo Research D6005, California, USA). After DNA extraction, Polymerase Chain Reaction (PCR) was performed to amplify the DNA of interest within the Internal Transcribed Spacer (ITS) region using Econo Tag Plus Master Mix (Lucigen), ITS 1 forward and ITS 4 reverse primers with sequences TCCGTAGGTGAACCTGCGG and TCCTCCGCTTA TTGATATGC. After amplification, the PCR products was run on a gel and the gel extracted using ZymoClean Gel DNA recovery clean-up kit (Zymo Research, D4001). The extracted fragments were then sequenced in the forward and reversed directions (Applied Biosystems, Thermofisher Scientific, Big Dye terminator kit v3.1, Carlsbad, California, USA) and purified using ZR-96 DNA sequencing clean-up kit (Zymo Research, D4050). The purified fragments were run on an ABI 3500 x L Genetic Analyser (Applied Biosystems, Thermofisher Scientific) for each reaction of every sample. CLC Bio Main Workbench v7.6 was used to analyze the data (.abi files) generated by the ABI 3500 xL Genetic Analyser (Applied Biosystems, Thermofisher Scientific). The similarities of the fragments with previously published sequence data were examined with the BLASTN 2.2.31+ version (Sadhasivam et al ., 2017). 2.4 Phylogenetic analysis A phylogenetic study was performed based on a data set of fungi isolated from the sampled PM and published sequence from Gen bank. Consensus sequences were obtained from forward and reverse sequences, which was aligned by clustalO in MEGA version 11. The evolutionary history was inferred using the neighbour-joining (NJ) method. The evolutionary distances were computed using the Maximum Composite Likelihood method (Tamura et al ., 2004) and was in the units of the number of base substitutions per site. The analysis involved 22 nucleotide sequences. All positions containing gaps and missing data were eliminated. Evolutionary analyses were conducted in Molecular Evolutionary Genetics Analysis App version 11 (MEGA 11) (Kumar et al ., 2016). 2.5 Sample Preparation and Extraction of Piliostigma Thonningii After gathering fresh P. thonningii leaves, they were cleaned using pure water and allowed to air dry for 15 days at room temperature to shield the plant's thermolabile constituents from the sun. The leaves were destalked and then ground into a coarse powder. Then, twenty-five (25 g) of the powdered P. thonningii leaves were weighed, combined with 500 ml of distilled water in a 1000 ml conical flask, allowed to boil for twenty-five minutes. Filter paper and muslin cloth were used to filter the aqueous extract (Whatman no. 1). To expedite the creation of selenium nanoparticles, the filtrate was maintained at a low temperature (Shittu and Ihebunna, 2017). 2.6 Phytochemical screening of the plant extracts 2.6.1 Qualitative phytochemical screening Standard techniques were used to screen extracts of P. thonningii for the presence of secondary metabolites before testing for flavonoids, alkaloids, saponins, tannins, and phenols (Sofowora, 2008). 2.6.2 Quantitative phytochemical screening of the crude extracts of P. Thonningii Oloyede, (2005) states that quantitative estimation of phytochemicals such as alkaloids and saponins was done. The method outlined by Singleton et al. , (1999) was utilized to estimate the total phenolic content, while Chang’s (2002). Aluminum Chloride Colorimetric Method was employed to assess the flavonoid levels. 2.6.2.1 Total flavonoid determination After mixing 0.5 ml of the plant extract with 0.1 ml of 10% aluminum chloride, 2.8 ml of distilled water, 0.1 ml of 1 M sodium acetate, and 1.5 ml of methanol, the mixture was left to stand at room temperature for 30 minutes. At 415 nm, the absorbance of the reaction mixture was measured with a spectrophotometer. 2.6.2.2 Determination of total phenol Using the method outlined by Singleton et al. (1999), the total phenol concentration of the crude extracts was determined. After 2.5 milliliters of 10% Folin-Ciocalteau's reagent (v/v) were used to oxidize two milliliters of the crude extract (0.5 ml), two milliliters of 7.5% sodium carbonate were used to neutralize the reaction. A spectrophotometer was used to measure the absorbance at 765 nm after the reaction mixture was incubated at 450C for 40 minutes. 2.6.2.3 Alkaloids determination After combining 20 milliliters of 96% ethanol and 20% H2SO4 in a 1:1 ratio, zero-point five grams (0.5 g) of the crude extract were filtered. They mixed five milliliters of 60% H2SO4 with one milliliter of the filtrate. After five minutes, the mixture was given a 5-ml formaldehyde solution at a 0.5% concentration, and it was left to stand for three hours. At 565 nm of absorbance, the reading was obtained (Oloyede, 2005). 2.6.2.4 Saponins determination 0.5 g of the crude extract and 20 milliliters of 1M HCl were combined and brought to a boil for a duration of four hours. 50 milliliters of petroleum ether were added to the ether layer filtrate after it had cooled and been filtered, and the mixture was then allowed to evaporate until entirely dry. The residue was mixed with two milliliters of concentrated H2SO4, five milliliters of acetone and five milliliters of ethanol, and six milliliters of ferrous sulfate reagent per milliliter. The combination was homogenized, allowed to stand for 10 minutes, and then the absorbance at 490 nm was determined (Oloyede, 2005). 2.6.2.5 Tannin determination After adding 20 milliliters of 50% methanol and zero-point two grams (0.2 g) of the extract, a 50-milliliter beaker was sealed with parafilm, heated to 800 degrees Celsius for one hour, and then covered again. The contents were transferred into a 100 ml volumetric flask after the mixture had been well-shaken. Afterwards, 20 milliliters (20 ml) of water were used, along with 10 milliliters of 17% Na2CO3 and 2.5 milliliters of Folin-Denis reagent. Twenty minutes were spent after mixing in the mixture. At the very end of the 12.5–100 μg/ml range, the bluish-green hue was noticed. After the sample had developed its color, a spectrophotometer calibrated to 760 nm was used to measure the absorbance of the tannin and acid reference solution. 2.6.2.6 Terpenoids determination 100 mg (wi) of dried plant extract was obtained and let to soak for 24 hours in 9 milliliters of ethanol (Indumathi et al ., 2014). Following filtering, 10 mL of petroleum ether was used to extract the extract using a separating funnel. After being divided into glass vials that had been previously weighed, the ether extract was allowed to fully dry (wf). Using the formula total terpenoids = yield (%) of total terpenoids contents after ether was evaporated 2.6.2.7 Steroids determination 10 ml volumetric flasks were filled with 1 ml of the test extract of the steroid solution. After adding iron (III) chloride (0.5% w/v, 2 ml) and sulfuric acid (4N, 2 ml), potassium hexacyanoferrate (III) solution (0.5% w/v, 0.5 ml) was added. The combination was heated for thirty minutes, shaking occasionally, in a water bath kept at 70±20C. After that, it was diluted with distilled water to the appropriate level. At 780 nm, the absorbance was measured against the reagent blank. 2.7 Synthesis of P. thonningii Mediated Selenium Nanoparticles (PT-SeNPs) The approach previously described by Nadaroglu et al., (2017) was utilized to synthesize PT-SeNPs, with few changes. To summarise, at room temperature (25°C), 10 ml of PT (1 mg/ml) was continuously stirred at 500 rpm with an aqueous solution of sodium selenite (10 ml, 0.01 M). The mixture was then gradually mixed with a freshly made 10 ml ascorbic acid (0.04 M) solution, added dropwise. Using sodium hydroxide (1 M NaOH) and glacial acetic acid (1 M hac), the reaction system's pH was then brought to 7.5. Following the addition of 10 milliliters of ascorbic acid, the reaction mixture was continuously stirred magnetically at 500 rpm for two hours at room temperature. The production of orange-red color indicated the synthesis of PT-SeNPs. The remainder was eliminated by repeatedly centrifuging the pellet at 6000 rpm for 20 minutes, then re-suspending it in 100% ethanol (about three to four times) to eliminate any remaining Na 2 SeO 3 contaminants. After that, the SeNPs were immersed in a water bath at 80 degrees Celsius for six hours, yielding powdered PT-SeNPs (Figure 1). The resulting PT-SeNPs were gathered and kept for additional analysis. Figure 1: Schematic representation of the synthesis of SeNPs through biological routes 2.8 Optimization of parameters for the Synthesis of PT-SeNPs The parameters for the synthesis of P. thonningii mediated selenium nanoparticles (PT-SeNPs) were optimized using Taguchi design methodology. In the planned experiments, three (3) factors of PT concentration (2 and 8, 14 mM), pH (5, 6, and 7), and incubation time (7 days) at 3 different levels were studied. According to the designed experimental conditions, the prepared supernatant was then mixed in equal proportions with solutions comprising 2-, 8- and 14 mM PT, at 5, 6, and 7. The obtained solutions were incubated at 30°C for 7 days in an incubator shaker at 140 rpm. The nanoparticles produced by centrifugation were separated and purified at 5000 rpm for 15 min. The obtained solutions were then taken for further characterization (Ng, 2014). Table 1: Optimization of parameters for the synthesis of PT-SeNPs Run pH Concentration(mM) Incubation time (Days) 1 5 8 7 2 6 2 7 3 7 8 7 4 6 14 7 2.9 Characterization of Selenium Nanoparticles 2.9.1 UV-Vis Spectra Using a Shimadzu UV-1800 UV-VIS Spectrometer, the absorption maxima of the reaction mixtures between 200 and 1100 nm were monitored to verify the synthesis of the PT-SeNPs. Before and after adding SeNPs to the plant extract, the spectrum absorbance of the SeNPs salt was measured. 2.9.2 Particle Size Analysis The average particle size of artificial nanoparticles was assessed using dynamic light scattering (DLS), which is based on the laser diffraction approach with several scattering techniques. After being mixed with deionized water, the prepared sample was ultrasonically sonicated. The solution was then filtered, and the supernatant was collected after it was centrifuged for 15 minutes at 5000 rpm and 25 o C. A computer-controlled particle size analyzer (ZETA sizer Nano series, Malvern instrument Nano Zs) was used to examine the particle distribution in liquid after the supernatant had been diluted four or five times. 2.10. Adsorption Gradation and Screening of Fungi Resistance Three (3) distinct PT-SeNP concentrations (1.5, 2.0, 2.5, and 3.0 mm) were made. Ramar (2015) describes the agar well diffusion method that was used to screen for adsorption by resistance. Following the manufacturer's instructions, molten potato dextrose agar (PDA) was made. Aseptic conditions were then maintained, and a loopful of standardized fungi were inoculated into a solidified sterile PDA plate and spread with a sterile wire loop. A sterile cork-borer was used to punch a hole with a diameter of 6 mm, and 0.2 ml of PT-SeNPs was dispensed into the cork-bored holes in dishes seeded with the test isolates. This was done for seven days at a temperature of 27±2ºc. Using a meter rule to measure the diameter colony extension of each fungus on each plate, the resistance of each fungus to each treatment was ascertained on a daily basis, and the average diameter of resistance was noted for the study. 2.11 Statistical Analysis and Data Evaluation The study's data was displayed as mean ± Standard Error of Mean (S.E.M.). Analysis of Variance (ANOVA) was used to compare data from different groups. Using the Statistical Package for Social Sciences (SPSS) version 26, the Duncan Multiple Range Test (DMRT) was used to assess if there were any significant differences between the control and experimental groups. 3.0 RESULTS 3.1 Microscopic Identification of Isolated Fungi Species in Selected Caprine Agricultural Residue Feeds in Minna Metropolis The distribution of fungal species in selected agricultural residue cattle feed samples are presented in Table 1. The Beans husk residue group, BH1 and BH2 displayed specific morphological features indicative of Aspergillus flavus and Rhizopus species, respectively. BH1 exhibited a rapid transition from white to green, with powdery edges, suggesting Aspergillus flavus . On the other hand, BH2, characterized by colorless growth turning grey, non-septate hyphae, and umbrella-shaped sporangium, indicated the presence of a Rhizopus species. The Groundnut husk (GOH) group encompassed fungal isolates with a green appearance, white edges, and a powdery texture. GOH1 and GOH2 were identified as Aspergillus flavus and Aspergillus niger , respectively. Both displayed characteristic features such as green coloration, white edges, and powdery texture. Additionally, GOH3 showed characteristics typical of Trichophyton rubrum , including fluffy white colonies with pigmentation. The Maize shaft (MS) group exhibited diverse morphological traits. MS-3 resembled Aspergillus fumigatus with blue-green fluffy colonies. MS-4, MS-5, and MS-6 were indicative of Mucor species, displaying white, fluffy, and fast-growing colonies. MS-7 and MS-8 were identified as Epidermophyton floccosum and Verticillium tenesum , respectively, based on their distinctive characteristics. The guinea corn husk (GH) group included isolates with green and dark-colored colonies. GH2 displayed features consistent with Aspergillus nudulans , while GH3 exhibited traits characteristic of Trichophyton metagrophi . GH4, identified as Exophialajeanselmei , showed slow-growing colonies with green-black coloration. Table 1: Microscopic and morphological identification of isolated fungi species in selected caprine Agricultural residue feeds in Minna Metropolis Feed sample Morphological features Microscopic features Suspected organism BH 1 Begins white colour lies but rapidly develops into green has white edges and a powdery Dark conidia head and septate hyphae Aspergillus flavus BH 2 Colourless grow rapidly initially white turns grey with the time and Non septate hyphae umbrella shaped sporangium Rhizopus specie BH 4 Black fluffy colonies with white edges Smooth coloured conidiosphores and conidia, hyaline& septate hyphae Aspergillus niger GOH 1 Begins whites’ colonies but rapidly develops into green has white edges & powdery Dark conidia head & septate hyphae Aspergillus flavus GOH 2 Black fluffy colonies with dark edges Smooth coloured coniodioshores and conidia hyaline and septate hyphae Aspergillus niger GOH 3 Fluffy white, slow growing colonies with white and reverse pigmentation Septate hyphae with a smooth walled macroconidium Trichophyton rubrum GOH 4 White fluffy fast growing, this are known as lid further Non septate hyphae round sporangia filled with sporagiospore Mucor specie MS-3 Fluffy blue green with white edges Septate and hyaline hyphae columnar conidia heads Aspergillus fumigatus MS-4 White fluffy fast growing known as lid lifter fungi Non septate hyphae round sporangia filled with sporangium Mucor sp. MS-5 White fluffy fast growing known as lid lifter fungi Non septate hyphae sporangia filled with sporangium Mucor sp MS-6 Colonies have a bright golden yellow to brownish yellow reverse pigment Septate hyphae with a spindle shaped macroconidium Macrosporium audounii MS-7 Colonies are slow growing with a raised and folded centre Septate hyphae with smooth walled macroconidia and sparse microconidia Epidermophyton floccosum MS-8 Begins as white colonies to black with smooth edges Hyphae with formation of conidiophores bearing conidia Verticillium tenesum GH2 Dark green colonies wool like texture Septate and hyaline hyphae conidia heads are columnar Aspergillus nudulans GH3 White in colour with a powdery surface and smooth edges Septate hyphae and macronidia Tricophyton metagrophi GH4 Slow growing colonies that are green black in color Septate hyphae, conidiophores and dark small, spherical conidia Exophialajeanselmei KEY: BH- Beans husk, GH- Guinea corn husk, GOH- Groundnut husk, MS- Maize shaft 3.2 Fungal load in caprine agricultural residue feed samples From the result of the fungal load across all the samples, it is evident that Groundnut Husk (GOH) exhibits the highest fungal contamination, with a total mean of 6.08 x 10 4 cfu/g (Table 2). Following closely, Maize Shaft (MS) shows a moderate to high fungal load, with a total mean of 5.48 x 10 4 cfu/g. Beans Husk (BH) presents a moderate fungal load, with a total mean of 1.76 x 10 4 cfu/g. Guinea Corn Husk (GH) shows the lowest fungal load among the feed types, with a total mean of 4.05 x 10 3 cfu/g. The feed types can be ranked from the highest to the lowest fungal load as follows: GOH > MS > BH > GH. Table 2: Fungal load in caprine agricultural residue feed samples in Minna Feed sample Plate 1 (cfu/g) Plate 2 (cfu/g) Plate 3 (cfu/g) Total mean (cfu/g) BH 1.76 x 10 4 4.2 x 10 3 3.1 x 10 4 1.76 x 10 4 GOH 6.08 x 10 4 1.36 x 10 4 1.08 x 10 5 6.08 x 10 4 GH 4.05 x 10 3 1.1 x 10 3 7 x 10 3 4.05 x 10 3 MS 5.48 x 10 4 2.16 x 10 4 8.8 x 10 4 5.48 x 10 4 KEY: BH- Beans husk, GH- Guinea corn husk, GOH- Groundnut husk, MS- Maize shaft 3.3 Phylogenetic tree of isolated fungi from selected caprine feed Phylogenetic trees of fungal isolates revealed that the isolates were cluster of monophyletic patterns of close resemblance (Figure 2) The fungi species belonging to genus of Aspergillus , with Aspergillus flavus as the predominant species identified. Test of phylogeny was bootstrap of 1000 replications with matrix of 9 nucleotide sequences. All isolates of Aspergillus flavus species were clustered with bootstrap of 59%. Figure 2: Phylogenetic tree of the evolutionary relationship between isolated fungi from selected caprine -crop feed 3.4 Phytochemical components of Piliostigma thonningii aqueous extract The qualitative phytochemical components Piliostigma thonningii aqueous extract is shown in Table 3. The qualitative phytochemical composition of the P. thonningii aqueous extract shows that alkaloids, saponins, phenols, flavonoids, tannins, terpenoids, steroids and glycosides were found to be present in the aqueous extract of P. thonningii while, anthocyanin and phlobatannins were found to be absent. (Table 3). Table 3: Qualitative phytochemical constituents of Piliostigma thonningii aqueous extract Phytochemicals Inference Alkaloid + Saponins + Phenols + Flavonoids + Tannins + Terpenoids + Steroids + Anthocyanin - Phlobatannins - Glycosides + The quantitative phytochemical components of P. thonningii aqueous extract are presented in Table 4. The result showed that the phytochemicals components (tannins and steroids) recorded in the P. thonningii aqueous extract (2.67±0.01 and 4.34±0.06 mg/100g) were significantly lower than other phytochemical components in the extract. However, phytochemical contents (saponins and flavonoids) recorded in the P. thonningii aqueous extract (14.56±0.01 and 11.88±0.01 mg/100g) were significantly (p<0.05) the highest amongst all the phytochemical components present in the P. thonningii aqueous extract. Table 4: Quantitative phytochemical constituents of Piliostigma thonningii aqueous extract Phytochemicals Concentration(mg/100g) Alkaloid 9.66±0.01 b Saponins 14.56±0.01 d Phenols 8.36±0.04 b Flavonoids 11.88±0.01 c Tannins 2.67±0.01 a Terpenoids 6.13±0.02 ab Steroids 4.34±0.06 a Values are expressed as mean ± S.E.M (Standard error of Mean). At the 5% level, there is no significant difference (P~0.05) between the values of the experimental treatments inside the column that has the same superscript. 3.5 Characterization of Selenium Nanoparticles (SeNPs) 3.5.1 Spectra of synthesized P. thonningii selenium nanoparticles (PT-SeNPs) The selenium nanoparticles synthesis with PT (2 and 8, 14 mM) at pH 5, 6, and 7, at 7-days incubation time was assessed by UV-Vis spectrophotometer (Shimadzu, UV-1800). The UV-Vis spectrum of the PT-SeNPs showed major absorption peaks at ~300 nm (Figure 3). Figure 3: UV–visible photometric spectra of PTSeNPs at Different pH, and Concentration, at 7 days incubations time 3.5.2 Particle size of synthesized P. thonningii selenium nanoparticles (PT-SeNPs) The particle size characterization of synthesized selenium nanoparticles of P. thonningii aqueous extract are presented in Figure 4. The particle sizes were determined under different conditions: 2mM at pH 6 resulted in 26.30 nm, 8mM at pH 5 showed 34.49 nm, 8mM at pH 7 exhibited 36.66 nm, and 14mM at pH 5 measured 80.30 nm (Fig 4). Figure 4: Average particle size distribution of synthesized selenium nanoparticles optimized at day seven (7). Key: 1= 2mM, pH 6 2= 8mM, pH 5 3 = 8mM, pH 7 4 = 14mM, pH 5 3.6 In vitro adsorption gradation and screening of fungi resistance activity of the P. Thonningii synthesized selenium nanoparticles (PT-SeNPs ) The in-vitro adsorption of the synthesized P. thonningii selenium nanoparticles (PT-SeNPs) of condition variable pH 6, 2mM and day 7 incubation time are presented in Table 5. All fungi isolate except Aspergillus nidulans shows no zone of inhibition. While PT-SeNPs show inhibition that is dose dependent compare to the control. 4.0 DISCUSSION The identification and quantification of fungal species in agricultural residues (Beans Husk (BH), Guinea Corn Husk (GH), Groundnut Husk (GOH), and Maize Shaft (MS), shed light on the presence of various identified fungi within each group and the potential consequences of their presence (Table 1 ). BH group exhibited the presence of Aspergillus flavus , a fungus known for producing mycotoxins, particularly aflatoxins. This is in corroboration with the study of Guluwa et al ., (2023) which evaluated fungal contamination in livestock feed ingredients of plant origin. The potential consequences of aflatoxin contamination in BH are significant, as aflatoxins are harmful to both animal health and the safety of derived products such as meat and milk Guinea Corn Husk (GH) group was found to harbour Aspergillus nudulans which is reported to be associated with various health concerns. This is agreement with the investigation of Nayak et al ., (2020) on roles of Aspergillus sp . in agricultural soil and environment. The presence of Aspergillus nudulans also raises potential issues related to mycotoxin production, although the specific mycotoxins associated with this species may vary. Groundnut Husk (GOH) group displayed the presence of Aspergillus flavus and Aspergillus niger ,in agreement with Bediako et al ., (2019) investigation on aflatoxin contamination of groundnut. Aspergillus flavus is a well-known mycotoxin producer, and its presence in the groundnut husk samples raises concerns about potential aflatoxin contamination. Aspergillus niger , while not typically associated with aflatoxin production, can impact feed quality and may lead to spoilage. The presence of Aspergillus fumigatus in the maize shaft (MS) group agrees with the study of Abdelaziz et al ., (2022). Aspergillus fumigatus is a fungus associated with respiratory issues in animals thus potentially unsafe for animals. Additionally, Mucor species were identified in agreement with the study of Muhammad et al ., (2019) which investigated mycoflora of maize in Niger state. Some members of this genus can produce mycotoxins. The potential consequences of these identified species include compromised respiratory health and the risk of mycotoxin contamination in the feed. The fungal load represented in Table 2 provides quantitative data on the abundance of fungi in each group, measured in colony-forming units per gram (cfu/g). The loads vary across the groups, with GOH displaying the highest total mean fungal load, followed by BH, MS, and GH. In the context of animal feed and fungal contamination, the presence of mold or yeast alone may not pose a significant health risk to animals. According to Golob (2007) and Tarr et al . (2006), Animals are typically thought to be safe from major dangers when exposed to non-mycotoxin generating mold species up to a maximum level of 106 CFU/g. But it's important to remember that this kind of contamination could cause an energy loss of 5–10%, and mycosis is only rarely caused by mold levels in feed that are significantly higher than what's considered safe (> 106 to 107 CFU/g). It emphasizes that the presence and concentration of particular mycotoxins in animal feed are the main concerns for safety control, with Aspergillus sp., Penicillium sp., and Fusarium sp. identified as the main mycotoxin-producing molds (Anthony et al ., 2021). Considering the fungal load values from this study, as observed in Beans Husk (BH), Guinea Corn Husk (GH), Groundnut Husk (GOH), and Maize Shaft (MS), it is evident that all groups have fungal loads below the acceptable level of 10 6 CFU/g, as specified by Golob (2007) and Tarr et al . (2006). The highest fungal load is observed in Groundnut Husk (GOH) at 6.08 x 10 4 CFU/g and despite this, it is below the threshold deemed potentially hazardous. The study aligns with the assertion that these fungal load levels are generally considered safe for farm animals, as noted in Canadian guidelines, where a Mold count in the feed of 10,000-500,000 (10 4 to 10 5 ) CFU/g is deemed safe, and even counts of 500,000–1,000,000 (5*10 5 to 10 6 ) CFU/g are still thought to be reasonably safe (Tarr, 1996). These levels are also considered safe in the United States (Adams et al ., 1993). The phylogenetic analysis result only identified fungal species that belonged to Aspergillus genera; as a consequence, this study has demonstrated that these genes should be employed as molecular markers for fungal species level identification. The majority of the plant extract's documented bioactivities are attributed to phytochemicals, which are secondary plant metabolites. It is well known that they have antibacterial, anti-inflammatory, anti-sickling, and antioxidant properties. Without a doubt, such metabolites indicate the plant extracts' potential medical value and suggest future medication candidates. Following the study of Babagana et al. , (2019), the qualitative phytochemical screening of P. thonningii revealed the presence of alkaloids, saponins, phenols, flavonoids, tannins, terpenoids, steroids, and glycosides (Table 3 ). Anthocyanin and phlobatannins were absent. These chemical classes have a history of being used to treat a wide range of ailments since they have been shown to have therapeutic action against some infections. A high concentration of flavonoids and saponins was found in the aqueous extracts of P. thonningii , according to the quantitative phytoconstituents analysis presented in this work (Table 3 .4). According to reports by (Ogbiko et al ., 2021), the plant extracts' antibacterial, antioxidant, and anti-inflammatory properties may be due to the presence of these phytonutrients. According to this study's findings, Kwaji et al. , (2010), Shittu and Ihebunna (2017), and Boualam et al. (2021) have all produced similar results. Additionally, some of these phytochemicals have been connected to the ability of plants utilized in the creation of nanoparticles to cap and stabilize themselves, according to research published in Shittu and Ihebunna (2017). Because of its many practical uses, nanotechnology is currently the most rapidly developing subject of study. The production of nanoparticles can be done in a variety of ways, including physical, chemical, and biological (Sani-e et al ., 2022). The utilization of biological methods, such as the synthesis of NPs using green plants, is preferred. The most advantageous method is green synthesis, which is also affordable, natural, safe, and environmentally friendly (Sani-e et al. , 2022). Furthermore, making SeNPs using medicinal plants may improve their advantageous qualities. Because they convert harmful oxyanions found in the environment into non-toxic elemental selenium, these biological catalysts are essential for the environment (Neha et al. , 2022). Prior research has demonstrated that the smaller-sized SeNPs were successful in preventing tumor cell growth via a process mediated by ROS (Natwar et al ., 2022), according to Fig. 3 . It was discovered that the ideal wavelength range for PT-SeNPs was 295.5–305.6 nm, and the ideal average size particles for the 7-day incubation period were 26.30, 34.49, 36.66, and 80.30 nm, respectively, at pH 2, 5, 6, and 7, concentrations of 2, 8, and 14 mM. After a seven-day incubation period, a pH of two, a PT concentration of eight milligrams, and a condition variable, the maximum wavelength was attained. These peaks verified the synthesis of SeNPs. The findings of Kirupagaran et al. , (2016), Vennila et al. , (2018), and Ban et al. , (2023) were consistent with this outcome, indicating that biosynthetic Se-NPs were absorbed between 250 and 400 nm. Vennila K. et al . (2018) provided support for these findings by confirming the importance of pH in regulating the form and size of metal nanoparticles. They found that nuclei congregate rather than form in acidic pH environments, while the largest number of particles form in alkaline medium at pH 9. Authors Chhabria and Desai (2016) also noted that by changing the pH from pH 5 to pH 7, big diameter nanospheres changed in size and shape to nanofibers with diameters of 50–75 nm. According to Natwar et al ., (2022) the data indicated that the presence of PT on the surface of SeNPs aids in the production and stabilization of SeNPs; in the absence of PT, it can aggregate into larger particles and produce unstable particles. Research has shown that P. thonningii (PT) phytochemical components are essential for the development and improvement of SeNPs stability (Kwaji et al ., 2010). Additionally, prior research has revealed that the phytochemical molecules bind with the SeNP surface by intermolecular hydrogen bonding (O-H-Se), giving the nanoparticles exceptional stability and dispersibility (Li et al. , 2019). Numerous studies have shown the importance of natural covering or capping agents, such as proteins, lipids, and phytochemicals, which can enhance the antimicrobial potential of selenium nanoparticles (SeNPs). These agents offer higher degree of stability and can be applied in various clinical treatments (Xu et al ., 2019; Shi et al ., 2021). Additionally, the dispersion and stability of the SeNPs formed are affected by the molecular weight and concentrations of the phytochemicals (Jia et al ., 2015). To prevent multidrug-resistant infections, SeNPs capped with phytochemicals have been identified as a unique possible antifungal agent that can be utilized instead of conventional medications (Natwar et al ., 2022). On the other hand, little research has been done on P. Thonningii's and selenium NPs' antifungal properties against fungus. However, according to Table 5, the synthesized PT-SeNPs exclusively showed antifungal efficacy against Aspergillus nidulans . By measuring the zone of inhibition (mm) against the studied pathogen, the antifungal activity of PT-SeNPs and the positive control were demonstrated. The results showed that PT-SeNPs had an inhibitory effect against Aspergillus nidulans that ranged from 19.11 to 24.25 mm and was greater than that produced by the control drug (Nystatine). The antifungal activity of PT-SeNPs against Aspergillus nidulans was found to increase as the concentration of the extracts increases from (1.5–3.0 mg/ml). The different extracts' ability to suppress the tested fungus showed a considerable difference. The research aligns with the findings of Perveen et al. (2020) concerning the investigation of green and sol-gel manufacturing of ZnO nanoparticles, as well as the assessment of their antibacterial and antifungal capabilities against active secondary metabolites of P. thonningii stem bark and phytochemicals. In addition, Aisha et al.'s work from 2023 evaluated the antifungal effectiveness of Piliostigma species against Aspergillus species that cause tomato fruit rot. However, PT-SeNPs' capacity to stop Aspergillus nidulans from growing suggests that they may have antifungal properties and be utilized to treat fungal diseases and cow feed. Declarations Ethical Statement: No ethical clearance is required Data Availability: All data that support all the experiment findings are in this article Conflict of Interest The authors have no conflict of interest to declare Consent to Participate declaration: Not applicable Acknowledgements The Authors are grateful to the Centre for Genetic Engineering and Biotechnology, Federal University of Technology, Minna for enabling direct access to the Centre's facilities. Funding Declaration This research was supported by the Tertiary Educational Trust Fund of Nigeria (grant number: TETF.ES.DR&D-CE/NRF2020/SETI/18/VOL.1). Author Contributions First Author (O.K.S.) performed DNA isolation, electrophoresis separation, polymerase chain reaction, and bioinformatics analysis and interpret the results. While, the second and third Authors (B. A and H.B.A) performed isolation, identification, and microbial load of the fungi present in the feeds under the supervision of O.K.S. 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Plates Plate 1 is available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files Plate1.docx Table5.docx Cite Share Download PDF Status: Posted Version 1 posted 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. 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SHITTU","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABEUlEQVRIiWNgGAWjYHADxocPKoAUG4THjFMdD4LJbGxwhsGANC1mEiAtMB5OLfbs7Q8/81QcTpw/I5mt4mDbn2g+6d6DHxgqrBMbpNsvYLWF54yxNM+Zw4kbbiSz3TjYZpDbJnMuWYLhTHpig8yZAqxaJHLYmHnbgFok8o/d/gjSIpFjIMEIFGmQyEnAriX9GTPvP4jDCg5CtBj/YPyHT0uCGTNvA1AB0GEMUC1mEowgEYn0A1i1nDljLDnnWLrxhjOPmSUOnDMGa7FIAIoAGVhDjL29/eGHNzXWsvPbkxk/HCiTy50/I8f4xgegSL9E+gOseiCgGY0P8gQbA48BFqUwUIfdCfhsGQWjYBSMgpEDAC4+YxU/B8PFAAAAAElFTkSuQmCC","orcid":"","institution":"Federal University of Technology","correspondingAuthor":true,"prefix":"","firstName":"Oluwatosin","middleName":"K.","lastName":"SHITTU","suffix":""},{"id":408154086,"identity":"79cf49e1-892b-4878-8ea5-90d5e6dc22ca","order_by":1,"name":"Biliksu ABDULKAREEM","email":"","orcid":"","institution":"Federal University of Technology","correspondingAuthor":false,"prefix":"","firstName":"Biliksu","middleName":"","lastName":"ABDULKAREEM","suffix":""},{"id":408154087,"identity":"1a084a8a-b796-4efa-9852-2fd2cc3808ec","order_by":2,"name":"Halimat B. ALADE","email":"","orcid":"","institution":"Federal University of Technology","correspondingAuthor":false,"prefix":"","firstName":"Halimat","middleName":"B.","lastName":"ALADE","suffix":""},{"id":408154088,"identity":"0c0b5c02-a15b-4472-8cbe-c9bfa1151d58","order_by":3,"name":"Jimoh O. TIJANI","email":"","orcid":"","institution":"Federal University of Technology","correspondingAuthor":false,"prefix":"","firstName":"Jimoh","middleName":"O.","lastName":"TIJANI","suffix":""}],"badges":[],"createdAt":"2024-12-02 11:38:20","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5564179/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5564179/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":75096698,"identity":"f0d6aa98-f721-436b-8a19-4e227240cb43","added_by":"auto","created_at":"2025-01-30 12:16:12","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":74935,"visible":true,"origin":"","legend":"\u003cp\u003eSchematic representation of the synthesis of SeNPs through biological routes\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5564179/v1/cc9f4c9fefa99ee0abff5a07.png"},{"id":75095920,"identity":"b3bfd3f8-63f4-408b-8f2f-76632382458b","added_by":"auto","created_at":"2025-01-30 12:08:11","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":57648,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree of the evolutionary relationship between isolated fungi from selected caprine-crop feed\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5564179/v1/9298564edcdd19a004a77287.png"},{"id":75095915,"identity":"5ae5c079-c567-42e2-9e16-92f1fcdd5d5f","added_by":"auto","created_at":"2025-01-30 12:08:11","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":54845,"visible":true,"origin":"","legend":"\u003cp\u003eUV–visible photometric spectra of PTSeNPs at Different pH, and Concentration, at 7 days incubations time\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5564179/v1/73b3d2a44499ac5c1b9c97d2.png"},{"id":75095922,"identity":"b6c41f49-b5ad-46cc-ae1a-30cb9f99b514","added_by":"auto","created_at":"2025-01-30 12:08:11","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":77166,"visible":true,"origin":"","legend":"\u003cp\u003eAverage particle size distribution of synthesized selenium nanoparticles optimized at day seven (7).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eKey: \u003c/strong\u003e1= 2mM, pH 6\u003c/p\u003e\n\u003cp\u003e2= 8mM, pH 5\u003c/p\u003e\n\u003cp\u003e3 = 8mM, pH 7\u003c/p\u003e\n\u003cp\u003e4 = 14mM, pH 5\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5564179/v1/0042dc50b80dea0581888e3e.png"},{"id":82360681,"identity":"5b2ba1e3-e543-4573-9c8f-220cfc85d18b","added_by":"auto","created_at":"2025-05-09 11:38:52","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1804343,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5564179/v1/a7bbf969-5082-4750-aa8a-50f57bbfa0e7.pdf"},{"id":75095892,"identity":"08e4bf72-0ec0-4c91-92a3-637bc3ede5ea","added_by":"auto","created_at":"2025-01-30 12:08:11","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":10417401,"visible":true,"origin":"","legend":"","description":"","filename":"Plate1.docx","url":"https://assets-eu.researchsquare.com/files/rs-5564179/v1/c700e59f97f5bd1f0e741d03.docx"},{"id":75096700,"identity":"527bd2a0-1296-402e-8e51-04df84dfff79","added_by":"auto","created_at":"2025-01-30 12:16:12","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":14917,"visible":true,"origin":"","legend":"","description":"","filename":"Table5.docx","url":"https://assets-eu.researchsquare.com/files/rs-5564179/v1/2ed2b05c7c0bd55df04fb2ee.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eAssessment and Improvement of Nutritive Value of Agricultural Residue Feed for Caprine\u003c/p\u003e","fulltext":[{"header":"1.0 INTRODUCTION","content":"\u003cp\u003eCAPRINE is a multi-functional animal that plays a significant role in the economy and nutrition of landless, small and marginal farmers in Nigeria. This is an enterprise, that has been practiced by a large section of the population in rural areas. Caprine can efficiently survive on available shrubs and trees in adverse harsh environments in low fertility lands where no other crop can be grown. Also, agricultural residues, such as crop by-products play a crucial role in sustaining both agricultural productivity and the global food supply chain of caprine farming (Perdana \u003cem\u003eet al\u003c/em\u003e., 2023). These low-quality residues available after crop harvest has become a viable practice worldwide for alleviating some of the feeding burdens (Kumar \u003cem\u003eet al\u003c/em\u003e., 2023). The increasing scarcity of free grazing areas and the rising cost of commercial feed contribute to the growing utilization of these by-products (Bhandari, 2019). Also, the decision to use these residues are driven by the need for roughage during feed scarcity or drought conditions. Most crop residues are inherently poor in nutrition and high in fiber content and feeding untreated residues may hinder animal acceptance and subsequently impact animal performance (Derara and Bekuma, 2021).\u003c/p\u003e \u003cp\u003eHowever, the presence of microorganisms, including fungi, in these residue feeds raises concerns about the potential impact on both feed quality and animal health. Fungi are ubiquitous in the environment and have diverse effects on agricultural systems. Some fungi contribute to the degradation of organic matter, aiding in nutrient cycling, while others may produce mycotoxins that can be harmful to animals and humans (Devi \u003cem\u003eet al\u003c/em\u003e., 2020). A greater number of possible risks that call for understanding and awareness of mycotoxins have been brought about by the globalization of the trade in agricultural commodities (Mona \u003cem\u003eet al\u003c/em\u003e., 2016).\u003c/p\u003e \u003cp\u003eAccording to Bhandari et al. (2023), nanotechnology has the potential to have a significant impact on the agricultural industry through a variety of means, including targeted and intelligent delivery of nutrients and bioactive compounds, Nanoparticle-mediated genetic material delivery for crop improvement, nano fertilizers, disease management, nanosensors for pathogen detection and soil monitoring, nanoencapsulation of seeds, nano pesticides, and nano herbicides, increased crop yield, and nutritional quality. Nanoparticles (NP) have been synthesized using a wide range of techniques, including chemical, physical, biological, and biogenic methods (Agbebati-Maleki \u003cem\u003eet al\u003c/em\u003e., 2009). On the other hand, it has been demonstrated that the biogenic reduction of metal precursors to produce the corresponding NPs is less costly, environmentally benign, and devoid of chemical pollutants. Natural goods that have been a good source of biogenic reduction of metallic particles to nanoparticles include plant extract, which is embedded with naturally occurring stabilizing, growth terminating, and capping chemicals. Selenium nanoparticles (SeNPs), a type of metallic nanoparticle generated through biogenic reduction, have demonstrated antibacterial activities against certain pathogenic organisms (Nadaroglu \u003cem\u003eet al\u003c/em\u003e., 2017). Because of its distinct characteristics, structure, and size, it finds use in the biomedical sciences for antibacterial therapy, drug transport, cancer treatment, medical diagnosis, and sensor fabrication (Nadaroglu \u003cem\u003eet al\u003c/em\u003e., 2017).\u003c/p\u003e \u003cp\u003eThe leguminous plant \u003cem\u003ePiliostigma thonningii Schum\u003c/em\u003e. is a member of the 133-genus Caesalpinioideae subfamily of the Fabaceae family (Cyril \u003cem\u003eet al\u003c/em\u003e., 2021). It has been discovered that \u003cem\u003eP. thonningii\u003c/em\u003e roots, bark, and leaves are used to cure haematochezia, stomach issues, and loss of appetite (Cyril \u003cem\u003eet al\u003c/em\u003e., 2021). There have been reports of antilipidemic, antibacterial, antihelminthic, and anti-inflammatory properties (Cyril \u003cem\u003eet al\u003c/em\u003e., 2021). Therefore, \u003cem\u003eP. Thonningii\u003c/em\u003e can function as a biogenic reduction of selenium salt. Hence, this research aims to analyze and characterize fungal communities in caprine agricultural residue feeds and also optimize the synthesis of selenium nanoparticles (SeNPs) as fungi scavengers in agricultural residue caprine feed.\u003c/p\u003e"},{"header":"2.0 MATERIALS AND METHODS","content":"\u003cp\u003e\u003cstrong\u003e2.1 MATERIALS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.1.1 Sample Collection\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe samples; beans husk, Guinea corn husk, groundnut husk, and maize shaft were purchased from Major Market in Minna, Niger state. The natural habitat of \u003cem\u003eP. thonningii,\u003c/em\u003e was found near Bosso Area in Minna, Niger state, where the fresh leaves were collected. The Federal University of Technology, Minna, Niger State\u0026apos;s Department of Plant Biology performed the taxonomic authentication of the plant.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.1.2 Reagents and Chemicals\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll chemicals and reagents used in this study were of analytical grade which include; Genomic Lysis Buffer, DNA Elution Buffer, DNA Prewash Buffer, Bashing Bead Buffer, g-DNA Wash Buffer, Potato dextrose agar (PDA), Chloramphenicol, Lactophenol cotton blue. sulfuric acid, ascorbic acid, and sodium selenite, all of which were produced by Sigma Chemical Co.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2 METHODS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2.1 Isolation and estimation of fungal species from caprine agricultural residue feed samples\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFungal isolation was carried out using the pour plate technique with 1ml of the 10\u003csup\u003e-4\u003c/sup\u003e dilutions of each sample aseptically transferred into sterile petri dishes and molten potato dextrose agar medium was poured into respective petri dishes and allowed to solidify before incubating at 37\u0026deg;C for 48-72 hours. This process was carried out in duplicates for each sample. After incubation, the plates were then screened for the presence of discrete colonies and the actual numbers of fungi, Mold, and yeast were estimated in colony-forming units per gram (cfu/g), (Bird \u003cem\u003eet al\u003c/em\u003e., 2015).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2.2 Estimation of the microbial load in a sample\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe microbial concentration is expressed in colony forming unit (cfu/g) per gram of sample, which is an estimate of viable fungal cells in a sample. Colony colony-forming unit is calculated using the formula:\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003cimg src=\"https://myfiles.space/user_files/127393_c7e80a1c9bb65875/127393_custom_files/img1738237918.png\"\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSubculture of fungal species\u003c/strong\u003e: Following incubation, each plate was examined and fungi colonies were sub-cultured onto fresh potato dextrose agar plates to obtain pure cultures which was stored on appropriate agar slants for further identification and analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2.3 Identification of fungal species\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIsolated fungal species were identified based on colony morphological characteristics on the surface of the culture medium and microscopic features. The technique of Oyeleke and Manga (2008) was adopted for the identification of the isolated fungi using lactophenol cotton blue stain. Determination of Microscopic features was achieved by placing a drop of the lactophenol stain on a clean grease-free glass slide and a small portion of the aerial mycelia of the fungi culture placed in the drop of lactophenol stain and a cover slip was gently placed over it. The slide was then mounted and viewed under a light microscope at \u0026times;10 and \u0026times;40 objectives.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.3 DNA Extraction and Molecular Characterization of Fungi Isolates\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMolecular identification was performed to confirm the identities of the fungal species recovered from samples as outlined by Samson \u003cem\u003eet al\u003c/em\u003e., (2010). Genomic DNA was extracted from the fungal cultures using the ZR fungal DNA kit (Zymo Research D6005, California, USA). After DNA extraction, Polymerase Chain Reaction (PCR) was performed to amplify the DNA of interest within the Internal Transcribed Spacer (ITS) region using Econo Tag Plus Master Mix (Lucigen), ITS 1 forward and ITS 4 reverse primers with sequences TCCGTAGGTGAACCTGCGG and TCCTCCGCTTA TTGATATGC. After amplification, the PCR products was run on a gel and the gel extracted using ZymoClean Gel DNA recovery clean-up kit (Zymo Research, D4001). The extracted fragments were then sequenced in the forward and reversed directions (Applied Biosystems, Thermofisher Scientific, Big Dye terminator kit v3.1, Carlsbad, California, USA) and purified using ZR-96 DNA sequencing clean-up kit (Zymo Research, D4050). The purified fragments were run on an ABI 3500 x L Genetic Analyser (Applied Biosystems, Thermofisher Scientific) for each reaction of every sample. CLC Bio Main Workbench v7.6 was used to analyze the data (.abi files) generated by the ABI 3500 xL Genetic Analyser (Applied Biosystems, Thermofisher Scientific). The similarities of the fragments with previously published sequence data were examined with the BLASTN 2.2.31+ version (Sadhasivam\u003cem\u003eet al\u003c/em\u003e., 2017).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.4 Phylogenetic analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA phylogenetic study was performed based on a data set of fungi isolated from the sampled PM and published sequence from Gen bank. Consensus sequences were obtained from forward and reverse sequences, which was aligned by clustalO in MEGA version 11. The evolutionary history was inferred using the neighbour-joining (NJ) method. The evolutionary distances were computed using the Maximum Composite Likelihood method (Tamura \u003cem\u003eet al\u003c/em\u003e., 2004) and was in the units of the number of base substitutions per site. The analysis involved 22 nucleotide sequences. All positions containing gaps and missing data were eliminated. Evolutionary analyses were conducted in Molecular Evolutionary Genetics Analysis App version 11 (MEGA 11) (Kumar \u003cem\u003eet al\u003c/em\u003e., 2016).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.5 Sample Preparation and Extraction of \u003cem\u003ePiliostigma Thonningii\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter gathering fresh \u003cem\u003eP. thonningii\u003c/em\u003e leaves, they were cleaned using pure water and allowed to air dry for 15 days at room temperature to shield the plant\u0026apos;s thermolabile constituents from the sun. The leaves were destalked and then ground into a coarse powder. Then, twenty-five (25 g) of the powdered \u003cem\u003eP. thonningii\u003c/em\u003e leaves were weighed, combined with 500 ml of distilled water in a 1000 ml conical flask, allowed to boil for twenty-five minutes. Filter paper and muslin cloth were used to filter the aqueous extract (Whatman no. 1). To expedite the creation of selenium nanoparticles, the filtrate was maintained at a low temperature (Shittu and Ihebunna, 2017).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6 Phytochemical screening of the plant extracts\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6.1 Qualitative phytochemical screening\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eStandard techniques were used to screen extracts of \u003cem\u003eP. thonningii\u003c/em\u003e for the presence of secondary metabolites before testing for flavonoids, alkaloids, saponins, tannins, and phenols (Sofowora, 2008).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6.2 Quantitative phytochemical screening of the crude extracts of P. Thonningii\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOloyede, (2005) states that quantitative estimation of phytochemicals such as alkaloids and saponins was done. The method outlined by Singleton \u003cem\u003eet al.\u003c/em\u003e, (1999) was utilized to estimate the total phenolic content, while Chang\u0026rsquo;s (2002). \u0026nbsp;Aluminum Chloride Colorimetric Method was employed to assess the flavonoid levels.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6.2.1 Total flavonoid determination\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter mixing 0.5 ml of the plant extract with 0.1 ml of 10% aluminum chloride, 2.8 ml of distilled water, 0.1 ml of 1 M sodium acetate, and 1.5 ml of methanol, the mixture was left to stand at room temperature for 30 minutes. At 415 nm, the absorbance of the reaction mixture was measured with a spectrophotometer.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6.2.2 Determination of total phenol\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eUsing the method outlined by Singleton et al. (1999), the total phenol concentration of the crude extracts was determined. After 2.5 milliliters of 10% Folin-Ciocalteau\u0026apos;s reagent (v/v) were used to oxidize two milliliters of the crude extract (0.5 ml), two milliliters of 7.5% sodium carbonate were used to neutralize the reaction. A spectrophotometer was used to measure the absorbance at 765 nm after the reaction mixture was incubated at 450C for 40 minutes.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6.2.3 Alkaloids determination\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter combining 20 milliliters of 96% ethanol and 20% H2SO4 in a 1:1 ratio, zero-point five grams (0.5 g) of the crude extract were filtered. They mixed five milliliters of 60% H2SO4 with one milliliter of the filtrate. After five minutes, the mixture was given a 5-ml formaldehyde solution at a 0.5% concentration, and it was left to stand for three hours. At 565 nm of absorbance, the reading was obtained (Oloyede, 2005).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6.2.4 Saponins determination\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e0.5 g of the crude extract and 20 milliliters of 1M HCl were combined and brought to a boil for a duration of four hours. 50 milliliters of petroleum ether were added to the ether layer filtrate after it had cooled and been filtered, and the mixture was then allowed to evaporate until entirely dry. The residue was mixed with two milliliters of concentrated H2SO4, five milliliters of acetone and five milliliters of ethanol, and six milliliters of ferrous sulfate reagent per milliliter. The combination was homogenized, allowed to stand for 10 minutes, and then the absorbance at 490 nm was determined (Oloyede, 2005).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6.2.5 Tannin determination\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter adding 20 milliliters of 50% methanol and zero-point two grams (0.2 g) of the extract, a 50-milliliter beaker was sealed with parafilm, heated to 800 degrees Celsius for one hour, and then covered again. The contents were transferred into a 100 ml volumetric flask after the mixture had been well-shaken. Afterwards, 20 milliliters (20 ml) of water were used, along with 10 milliliters of 17% Na2CO3 and 2.5 milliliters of Folin-Denis reagent. Twenty minutes were spent after mixing in the mixture. At the very end of the 12.5\u0026ndash;100 \u0026mu;g/ml range, the bluish-green hue was noticed. After the sample had developed its color, a spectrophotometer calibrated to 760 nm was used to measure the absorbance of the tannin and acid reference solution.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6.2.6 Terpenoids determination\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e100 mg (wi) of dried plant extract was obtained and let to soak for 24 hours in 9 milliliters of ethanol (Indumathi \u003cem\u003eet al\u003c/em\u003e., 2014). Following filtering, 10 mL of petroleum ether was used to extract the extract using a separating funnel. After being divided into glass vials that had been previously weighed, the ether extract was allowed to fully dry (wf). Using the formula total terpenoids = yield (%) of total terpenoids contents after ether was evaporated\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6.2.7 Steroids determination\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e10 ml volumetric flasks were filled with 1 ml of the test extract of the steroid solution. After adding iron (III) chloride (0.5% w/v, 2 ml) and sulfuric acid (4N, 2 ml), potassium hexacyanoferrate (III) solution (0.5% w/v, 0.5 ml) was added. The combination was heated for thirty minutes, shaking occasionally, in a water bath kept at 70\u0026plusmn;20C. After that, it was diluted with distilled water to the appropriate level. At 780 nm, the absorbance was measured against the reagent blank.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.7 Synthesis of \u003cem\u003eP. thonningii\u0026nbsp;\u003c/em\u003eMediated Selenium Nanoparticles (PT-SeNPs)\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe approach previously described by Nadaroglu \u003cem\u003eet al.,\u003c/em\u003e (2017) was utilized to synthesize PT-SeNPs, with few changes. To summarise, at room temperature (25\u0026deg;C), 10 ml of PT (1 mg/ml) was continuously stirred at 500 rpm with an aqueous solution of sodium selenite (10 ml, 0.01 M). The mixture was then gradually mixed with a freshly made 10 ml ascorbic acid (0.04 M) solution, added dropwise. Using sodium hydroxide (1 M NaOH) and glacial acetic acid (1 M hac), the reaction system\u0026apos;s pH was then brought to 7.5. Following the addition of 10 milliliters of ascorbic acid, the reaction mixture was continuously stirred magnetically at 500 rpm for two hours at room temperature. The production of orange-red color indicated the synthesis of PT-SeNPs. The remainder was eliminated by repeatedly centrifuging the pellet at 6000 rpm for 20 minutes, then re-suspending it in 100% ethanol (about three to four times) to eliminate any remaining Na\u003csub\u003e2\u003c/sub\u003eSeO\u003csub\u003e3\u003c/sub\u003e contaminants. After that, the SeNPs were immersed in a water bath at 80 degrees Celsius for six hours, yielding powdered PT-SeNPs (Figure 1). The resulting PT-SeNPs were gathered and kept for additional analysis.\u003c/p\u003e\n\u003cp\u003eFigure 1: Schematic representation of the synthesis of SeNPs through biological routes\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.8 Optimization of parameters for the Synthesis of PT-SeNPs\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe parameters for the synthesis of \u003cem\u003eP. thonningii\u0026nbsp;\u003c/em\u003emediated selenium nanoparticles (PT-SeNPs) were optimized using Taguchi design methodology. In the planned experiments, three (3) factors of PT concentration (2 and 8, 14 mM), pH (5, 6, and 7), and incubation time (7 days) at 3 different levels were studied. According to the designed experimental conditions, the prepared supernatant was then mixed in equal proportions with solutions comprising 2-, 8- and 14 mM PT, at 5, 6, and 7. The obtained solutions were incubated at 30\u0026deg;C for 7 days in an incubator shaker at 140 rpm. The nanoparticles produced by centrifugation were separated and purified at 5000 rpm for 15 min. The obtained solutions were then taken for further characterization (Ng, 2014).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1: \u0026nbsp;Optimization of parameters for the synthesis of PT-SeNPs\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 87px;\"\u003eRun\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 87px;\"\u003epH\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 20.1968%;\"\u003eConcentration(mM)\u003c/td\u003e\n \u003ctd style=\"width: 34.6012%;\"\u003eIncubation time (Days)\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003e1\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003e5\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.1968%;\"\u003e\n \u003cp\u003e8\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 34.6012%;\"\u003e\n \u003cp\u003e\u0026nbsp;7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003e2\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003e6\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.1968%;\"\u003e\n \u003cp\u003e2\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 34.6012%;\"\u003e\n \u003cp\u003e\u0026nbsp;7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003e3\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003e7\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.1968%;\"\u003e\n \u003cp\u003e8\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 34.6012%;\"\u003e\n \u003cp\u003e\u0026nbsp;7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003e4\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 87px;\"\u003e\n \u003cp\u003e6\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.1968%;\"\u003e\n \u003cp\u003e14\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 34.6012%;\"\u003e\n \u003cp\u003e\u0026nbsp;7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003e2.9 Characterization of Selenium Nanoparticles\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.9.1 UV-Vis Spectra\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eUsing a Shimadzu UV-1800 UV-VIS Spectrometer, the absorption maxima of the reaction mixtures between 200 and 1100 nm were monitored to verify the synthesis of the PT-SeNPs. Before and after adding SeNPs to the plant extract, the spectrum absorbance of the SeNPs salt was measured.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.9.2 Particle Size Analysis\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe average particle size of artificial nanoparticles was assessed using dynamic light scattering (DLS), which is based on the laser diffraction approach with several scattering techniques. After being mixed with deionized water, the prepared sample was ultrasonically sonicated. The solution was then filtered, and the supernatant was collected after it was centrifuged for 15 minutes at 5000 rpm and 25 \u003csup\u003eo\u003c/sup\u003eC. A computer-controlled particle size analyzer (ZETA sizer Nano series, Malvern instrument Nano Zs) was used to examine the particle distribution in liquid after the supernatant had been diluted four or five times.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.10. Adsorption Gradation and Screening of Fungi Resistance\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThree (3) distinct PT-SeNP concentrations (1.5, 2.0, 2.5, and 3.0 mm) were made. Ramar (2015) describes the agar well diffusion method that was used to screen for adsorption by resistance. Following the manufacturer\u0026apos;s instructions, molten potato dextrose agar (PDA) was made. Aseptic conditions were then maintained, and a loopful of standardized fungi were inoculated into a solidified sterile PDA plate and spread with a sterile wire loop. A sterile cork-borer was used to punch a hole with a diameter of 6 mm, and 0.2 ml of PT-SeNPs was dispensed into the cork-bored holes in dishes seeded with the test isolates. This was done for seven days at a temperature of 27\u0026plusmn;2\u0026ordm;c. Using a meter rule to measure the diameter colony extension of each fungus on each plate, the resistance of each fungus to each treatment was ascertained on a daily basis, and the average diameter of resistance was noted for the study.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.11\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eStatistical Analysis and Data Evaluation\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study\u0026apos;s data was displayed as mean \u0026plusmn; Standard Error of Mean (S.E.M.). Analysis of Variance (ANOVA) was used to compare data from different groups. Using the Statistical Package for Social Sciences (SPSS) version 26, the Duncan Multiple Range Test (DMRT) was used to assess if there were any significant differences between the control and experimental groups.\u003c/p\u003e"},{"header":"3.0 RESULTS","content":"\u003cp\u003e\u003cstrong\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e3.1 Microscopic Identification of Isolated Fungi Species in Selected Caprine Agricultural Residue Feeds in Minna Metropolis\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eThe distribution of fungal species in selected agricultural residue cattle feed samples are presented in Table 1. The Beans husk residue group, BH1 and BH2 displayed specific morphological features indicative of \u003cem\u003eAspergillus flavus\u003c/em\u003e and \u003cem\u003eRhizopus\u003c/em\u003e species, respectively. BH1 exhibited a rapid transition from white to green, with powdery edges, suggesting \u003cem\u003eAspergillus flavus\u003c/em\u003e. On the other hand, BH2, characterized by colorless growth turning grey, non-septate hyphae, and umbrella-shaped sporangium, indicated the presence of a \u003cem\u003eRhizopus\u003c/em\u003e species. The Groundnut husk (GOH) group encompassed fungal isolates with a green appearance, white edges, and a powdery texture. GOH1 and GOH2 were identified as \u003cem\u003eAspergillus flavus\u003c/em\u003e and \u003cem\u003eAspergillus niger\u003c/em\u003e, respectively. Both displayed characteristic features such as green coloration, white edges, and powdery texture. Additionally, GOH3 showed characteristics typical of \u003cem\u003eTrichophyton rubrum\u003c/em\u003e, including fluffy white colonies with pigmentation.\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eThe Maize shaft (MS) group exhibited diverse morphological traits. MS-3 resembled \u003cem\u003eAspergillus fumigatus\u003c/em\u003e with blue-green fluffy colonies. MS-4, MS-5, and MS-6 were indicative of \u003cem\u003eMucor\u003c/em\u003e species, displaying white, fluffy, and fast-growing colonies. MS-7 and MS-8 were identified as \u003cem\u003eEpidermophyton floccosum\u003c/em\u003e and \u003cem\u003eVerticillium tenesum\u003c/em\u003e, respectively, based on their distinctive characteristics. The guinea corn husk (GH) group included isolates with green and dark-colored colonies. GH2 displayed features consistent with \u003cem\u003eAspergillus nudulans\u003c/em\u003e, while GH3 exhibited traits characteristic of \u003cem\u003eTrichophyton metagrophi\u003c/em\u003e. GH4, identified as \u003cem\u003eExophialajeanselmei\u003c/em\u003e, showed slow-growing colonies with green-black coloration.\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003eTable 1: Microscopic and morphological identification of isolated fungi species in selected caprine Agricultural residue feeds in Minna Metropolis\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cdiv align=\"center\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003eFeed sample\u0026nbsp;\u003c/strong\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003eMorphological features\u003c/strong\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003eMicroscopic features\u003c/strong\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003eSuspected organism\u003c/strong\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eBH\u003csub\u003e1\u003c/sub\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eBegins white colour lies but rapidly develops into green has white edges and a powdery\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eDark conidia head and septate hyphae\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eAspergillus\u0026nbsp;\u003c/em\u003e\u003cbr\u003e\u003cem\u003eflavus\u0026nbsp;\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eBH\u003csub\u003e2\u003c/sub\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eColourless grow rapidly initially white turns grey with the time and \u0026nbsp; \u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eNon septate hyphae umbrella shaped sporangium \u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eRhizopus specie \u0026nbsp;\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eBH\u003csub\u003e4\u003c/sub\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eBlack fluffy colonies with white edges\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSmooth coloured conidiosphores and conidia, hyaline\u0026amp; septate hyphae\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eAspergillus niger\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eGOH\u003csub\u003e1\u003c/sub\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eBegins whites\u0026rsquo; colonies but rapidly develops into green has white edges \u0026amp; powdery\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eDark conidia head \u0026amp; septate hyphae \u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eAspergillus flavus\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eGOH\u003csub\u003e2\u003c/sub\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eBlack fluffy colonies with dark edges\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSmooth coloured coniodioshores and conidia hyaline and septate hyphae \u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eAspergillus niger\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eGOH\u003csub\u003e3\u003c/sub\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eFluffy white, slow growing colonies with white and reverse pigmentation\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSeptate hyphae with a smooth walled macroconidium\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eTrichophyton rubrum \u0026nbsp;\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eGOH\u003csub\u003e4\u003c/sub\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eWhite fluffy fast growing, this are known as lid further\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eNon septate hyphae round sporangia filled with sporagiospore\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eMucor specie\u0026nbsp;\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eMS-3\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eFluffy blue green with white edges\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSeptate and hyaline hyphae columnar conidia heads\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eAspergillus fumigatus\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eMS-4\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eWhite fluffy fast growing known as lid lifter fungi\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eNon septate hyphae round sporangia filled with sporangium\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eMucor sp.\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eMS-5\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eWhite fluffy fast growing known as lid lifter fungi\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eNon septate hyphae sporangia filled with sporangium\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eMucor sp\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eMS-6\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eColonies have a bright golden yellow to brownish yellow reverse pigment\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSeptate hyphae with a spindle shaped macroconidium\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eMacrosporium audounii\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eMS-7\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eColonies are slow growing with a raised and folded centre\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSeptate hyphae with smooth walled macroconidia and sparse microconidia\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eEpidermophyton floccosum\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eMS-8\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eBegins as white colonies to black with smooth edges\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eHyphae with formation of conidiophores bearing conidia\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eVerticillium tenesum\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eGH2\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eDark green colonies wool like texture\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSeptate and hyaline hyphae conidia heads are columnar\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eAspergillus nudulans\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eGH3\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eWhite in colour with a powdery surface and smooth edges\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSeptate hyphae and macronidia\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eTricophyton metagrophi\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eGH4\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSlow growing colonies that are green black in color\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSeptate hyphae, conidiophores and dark small, spherical conidia\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cem\u003eExophialajeanselmei\u003c/em\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eKEY: BH- Beans husk, GH- Guinea corn husk, GOH- Groundnut husk, MS- Maize shaft\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003e3.2 Fungal load in caprine agricultural residue feed samples\u0026nbsp;\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eFrom the result of the fungal load across all the samples, it is evident that Groundnut Husk (GOH) exhibits the highest fungal contamination, with a total mean of 6.08 x 10\u003csup\u003e4\u0026nbsp;\u003c/sup\u003ecfu/g (Table 2). Following closely, Maize Shaft (MS) shows a moderate to high fungal load, with a total mean of 5.48 x 10\u003csup\u003e4\u003c/sup\u003ecfu/g. Beans Husk (BH) presents a moderate fungal load, with a total mean of 1.76 x 10\u003csup\u003e4\u0026nbsp;\u003c/sup\u003ecfu/g. Guinea Corn Husk (GH) shows the lowest fungal load among the feed types, with a total mean of 4.05 x 10\u003csup\u003e3\u0026nbsp;\u003c/sup\u003ecfu/g. The feed types can be ranked from the highest to the lowest fungal load as follows: GOH \u0026gt; MS \u0026gt; BH \u0026gt; GH.\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e\u003cstrong\u003eTable 2: Fungal load in caprine agricultural residue feed samples in Minna\u0026nbsp;\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"560\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003eFeed sample\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003ePlate 1 (cfu/g)\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003ePlate 2 (cfu/g)\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003ePlate 3 (cfu/g)\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003eTotal mean (cfu/g)\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003eBH\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e1.76 x 10\u003csup\u003e4\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e4.2 x 10\u003csup\u003e3\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e3.1 x 10\u003csup\u003e4\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e1.76 x 10\u003csup\u003e4\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003eGOH\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e6.08 x 10\u003csup\u003e4\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e1.36 x 10\u003csup\u003e4\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e1.08 x 10\u003csup\u003e5\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e6.08 x 10\u003csup\u003e4\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003eGH\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e4.05 x 10\u003csup\u003e3\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e1.1 x 10\u003csup\u003e3\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e7 x 10\u003csup\u003e3\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e4.05 x 10\u003csup\u003e3\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003eMS\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e5.48 x 10\u003csup\u003e4\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e2.16 x 10\u003csup\u003e4\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e8.8 x 10\u003csup\u003e4\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cspan style='font-family: \"Times New Roman\", Times, serif;'\u003e5.48 x 10\u003csup\u003e4\u003c/sup\u003e\u003c/span\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eKEY: BH- Beans husk, GH- Guinea corn husk, GOH- Groundnut husk, MS- Maize shaft\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003e3.3 Phylogenetic\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003etree of isolated fungi from selected\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003ecaprine\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;feed\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003ePhylogenetic trees of fungal isolates revealed that the isolates were cluster of monophyletic patterns of close resemblance (Figure 2) The fungi species belonging to genus of \u003cem\u003eAspergillus\u003c/em\u003e, with \u003cem\u003eAspergillus flavus\u003c/em\u003e as the predominant species identified. Test of phylogeny was bootstrap of 1000 replications with matrix of 9 nucleotide sequences. All isolates of \u003cem\u003eAspergillus flavus\u003c/em\u003e species were clustered with bootstrap of 59%.\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eFigure 2: \u0026nbsp;Phylogenetic tree of the evolutionary relationship between isolated fungi from selected caprine -crop feed\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003e3.4 Phytochemical components of \u003cem\u003ePiliostigma thonningii\u0026nbsp;\u003c/em\u003eaqueous extract\u0026nbsp;\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eThe qualitative phytochemical components \u003cem\u003ePiliostigma thonningii\u0026nbsp;\u003c/em\u003eaqueous extract is shown in Table 3. The qualitative phytochemical composition of the \u003cem\u003eP. thonningii\u0026nbsp;\u003c/em\u003eaqueous extract shows that alkaloids, saponins, phenols, flavonoids, tannins, terpenoids, steroids and glycosides were found to be present in the aqueous extract of \u003cem\u003eP. thonningii\u0026nbsp;\u003c/em\u003ewhile, anthocyanin and phlobatannins were found to be absent. (Table 3).\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003eTable 3: Qualitative phytochemical constituents of \u003cem\u003ePiliostigma thonningii\u0026nbsp;\u003c/em\u003eaqueous extract\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003ePhytochemicals \u0026nbsp;\u0026nbsp;\u003c/strong\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003eInference\u003c/strong\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eAlkaloid\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; +\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSaponins\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; +\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003ePhenols\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; +\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eFlavonoids\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; +\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eTannins\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; +\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eTerpenoids\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; +\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSteroids\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; +\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eAnthocyanin\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; -\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003ePhlobatannins\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; -\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eGlycosides\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 240px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; +\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eThe quantitative phytochemical components of \u003cem\u003eP. thonningii\u0026nbsp;\u003c/em\u003eaqueous extract are presented in Table 4. The result showed that the phytochemicals components (tannins and steroids) recorded in the \u003cem\u003eP. thonningii\u0026nbsp;\u003c/em\u003eaqueous extract (2.67\u0026plusmn;0.01 and 4.34\u0026plusmn;0.06 mg/100g) were significantly lower than other phytochemical components in the extract. However, phytochemical contents (saponins and flavonoids) recorded in the \u003cem\u003eP. thonningii\u0026nbsp;\u003c/em\u003eaqueous extract (14.56\u0026plusmn;0.01 and 11.88\u0026plusmn;0.01 mg/100g) were significantly (p\u0026lt;0.05) the highest amongst all the phytochemical components present in the \u003cem\u003eP. thonningii\u0026nbsp;\u003c/em\u003eaqueous extract.\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003eTable 4: Quantitative phytochemical constituents of \u003cem\u003ePiliostigma thonningii\u0026nbsp;\u003c/em\u003eaqueous extract\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003ePhytochemicals\u003c/strong\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003e\u0026nbsp;Concentration(mg/100g)\u003c/strong\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eAlkaloid\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e9.66\u0026plusmn;0.01\u003csup\u003eb\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSaponins\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e14.56\u0026plusmn;0.01\u003csup\u003ed\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003ePhenols\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e8.36\u0026plusmn;0.04\u0026nbsp;\u003csup\u003eb\u003c/sup\u003e\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eFlavonoids\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e11.88\u0026plusmn;0.01\u003csup\u003ec\u003c/sup\u003e\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eTannins\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e2.67\u0026plusmn;0.01\u0026nbsp;\u003csup\u003ea\u003c/sup\u003e\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eTerpenoids\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e6.13\u0026plusmn;0.02\u003csup\u003e\u0026nbsp;ab\u003c/sup\u003e\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eSteroids\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 189px;\"\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e4.34\u0026plusmn;0.06\u0026nbsp;\u003csup\u003ea\u003c/sup\u003e\u0026nbsp;\u003cbr\u003e\u003c/span\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eValues are expressed as mean \u0026plusmn; S.E.M (Standard error of Mean). At the 5% level, there is no significant difference (P~0.05) between the values of the experimental treatments inside the column that has the same superscript.\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003e3.5 Characterization of Selenium Nanoparticles (SeNPs)\u0026nbsp;\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003e3.5.1 Spectra of synthesized \u003cem\u003eP. thonningii\u003c/em\u003e selenium nanoparticles (PT-SeNPs)\u0026nbsp;\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eThe selenium nanoparticles synthesis with PT (2 and 8, 14 mM) at pH 5, 6, and 7, at 7-days incubation time was assessed by UV-Vis spectrophotometer (Shimadzu, UV-1800). The UV-Vis spectrum of the PT-SeNPs showed major absorption peaks at ~300 nm (Figure 3).\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eFigure 3: UV\u0026ndash;visible photometric spectra of PTSeNPs at Different pH, and Concentration, at 7 days incubations time\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003e3.5.2 Particle size of synthesized \u003cem\u003eP. thonningii\u003c/em\u003e selenium nanoparticles (PT-SeNPs)\u0026nbsp;\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eThe particle size characterization of synthesized selenium nanoparticles of \u003cem\u003eP. thonningii\u0026nbsp;\u003c/em\u003eaqueous extract are presented in Figure 4. The particle sizes were determined under different conditions: 2mM at pH 6 resulted in 26.30 nm, 8mM at pH 5 showed 34.49 nm, 8mM at pH 7 exhibited 36.66 nm, and 14mM at pH 5 measured 80.30 nm (Fig 4).\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003eFigure 4:\u0026nbsp;\u003c/strong\u003eAverage particle size distribution of synthesized selenium nanoparticles optimized at day seven (7).\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003eKey:\u0026nbsp;\u003c/strong\u003e1= 2mM, pH 6\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e2= 8mM, pH 5\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e3 = 8mM, pH 7\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e4 = 14mM, pH 5\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003e\u003cstrong\u003e3.6 In vitro adsorption gradation and screening of fungi resistance activity of the \u003cem\u003eP. Thonningii\u003c/em\u003e synthesized selenium nanoparticles (PT-SeNPs\u003c/strong\u003e\u003cstrong\u003e)\u003c/strong\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003e\u003cspan style=\"font-family: 'Times New Roman', Times, serif;\"\u003eThe \u003cem\u003ein-vitro\u0026nbsp;\u003c/em\u003eadsorption of the synthesized \u003cem\u003eP. thonningii\u0026nbsp;\u003c/em\u003eselenium nanoparticles (PT-SeNPs) of condition variable pH 6, 2mM and day 7 incubation time are presented in Table 5. All fungi isolate except \u003cem\u003eAspergillus nidulans\u0026nbsp;\u003c/em\u003eshows no zone of inhibition. While PT-SeNPs show inhibition that is dose dependent compare to the control.\u003c/span\u003e\u003c/p\u003e"},{"header":"4.0 DISCUSSION","content":"\u003cp\u003eThe identification and quantification of fungal species in agricultural residues (Beans Husk (BH), Guinea Corn Husk (GH), Groundnut Husk (GOH), and Maize Shaft (MS), shed light on the presence of various identified fungi within each group and the potential consequences of their presence (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e1\u003c/span\u003e). BH group exhibited the presence of \u003cem\u003eAspergillus flavus\u003c/em\u003e, a fungus known for producing mycotoxins, particularly aflatoxins. This is in corroboration with the study of Guluwa \u003cem\u003eet al\u003c/em\u003e., (2023) which evaluated fungal contamination in livestock feed ingredients of plant origin. The potential consequences of aflatoxin contamination in BH are significant, as aflatoxins are harmful to both animal health and the safety of derived products such as meat and milk\u003c/p\u003e \u003cp\u003eGuinea Corn Husk (GH) group was found to harbour \u003cem\u003eAspergillus nudulans\u003c/em\u003e which is reported to be associated with various health concerns. This is agreement with the investigation of Nayak \u003cem\u003eet al\u003c/em\u003e., (2020) on roles of \u003cem\u003eAspergillus sp\u003c/em\u003e. in agricultural soil and environment. The presence of \u003cem\u003eAspergillus nudulans\u003c/em\u003e also raises potential issues related to mycotoxin production, although the specific mycotoxins associated with this species may vary. Groundnut Husk (GOH) group displayed the presence of \u003cem\u003eAspergillus flavus\u003c/em\u003e and \u003cem\u003eAspergillus niger\u003c/em\u003e ,in agreement with Bediako\u003cem\u003eet al\u003c/em\u003e., (2019) investigation on aflatoxin contamination of groundnut. \u003cem\u003eAspergillus flavus\u003c/em\u003e is a well-known mycotoxin producer, and its presence in the groundnut husk samples raises concerns about potential aflatoxin contamination. \u003cem\u003eAspergillus niger\u003c/em\u003e, while not typically associated with aflatoxin production, can impact feed quality and may lead to spoilage.\u003c/p\u003e \u003cp\u003eThe presence of \u003cem\u003eAspergillus fumigatus\u003c/em\u003e in the maize shaft (MS) group agrees with the study of Abdelaziz \u003cem\u003eet al\u003c/em\u003e., (2022). \u003cem\u003eAspergillus fumigatus\u003c/em\u003e is a fungus associated with respiratory issues in animals thus potentially unsafe for animals. Additionally, \u003cem\u003eMucor species\u003c/em\u003e were identified in agreement with the study of Muhammad \u003cem\u003eet al\u003c/em\u003e., (2019) which investigated mycoflora of maize in Niger state. Some members of this genus can produce mycotoxins. The potential consequences of these identified species include compromised respiratory health and the risk of mycotoxin contamination in the feed.\u003c/p\u003e \u003cp\u003eThe fungal load represented in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e2\u003c/span\u003e provides quantitative data on the abundance of fungi in each group, measured in colony-forming units per gram (cfu/g). The loads vary across the groups, with GOH displaying the highest total mean fungal load, followed by BH, MS, and GH. In the context of animal feed and fungal contamination, the presence of mold or yeast alone may not pose a significant health risk to animals. According to Golob (2007) and Tarr\u003cem\u003eet al\u003c/em\u003e. (2006), Animals are typically thought to be safe from major dangers when exposed to non-mycotoxin generating mold species up to a maximum level of 106 CFU/g. But it's important to remember that this kind of contamination could cause an energy loss of 5\u0026ndash;10%, and mycosis is only rarely caused by mold levels in feed that are significantly higher than what's considered safe (\u0026gt;\u0026thinsp;106 to 107 CFU/g). It emphasizes that the presence and concentration of particular mycotoxins in animal feed are the main concerns for safety control, with \u003cem\u003eAspergillus sp., Penicillium sp., and Fusarium sp.\u003c/em\u003e identified as the main mycotoxin-producing molds (Anthony \u003cem\u003eet al\u003c/em\u003e., 2021). Considering the fungal load values from this study, as observed in Beans Husk (BH), Guinea Corn Husk (GH), Groundnut Husk (GOH), and Maize Shaft (MS), it is evident that all groups have fungal loads below the acceptable level of 10\u003csup\u003e6\u003c/sup\u003eCFU/g, as specified by Golob (2007) and Tarr \u003cem\u003eet al\u003c/em\u003e. (2006). The highest fungal load is observed in Groundnut Husk (GOH) at 6.08 x 10\u003csup\u003e4\u003c/sup\u003e CFU/g and despite this, it is below the threshold deemed potentially hazardous. The study aligns with the assertion that these fungal load levels are generally considered safe for farm animals, as noted in Canadian guidelines, where a Mold count in the feed of 10,000-500,000 (10\u003csup\u003e4\u003c/sup\u003e to 10\u003csup\u003e5\u003c/sup\u003e) CFU/g is deemed safe, and even counts of 500,000\u0026ndash;1,000,000 (5*10\u003csup\u003e5\u003c/sup\u003e to 10\u003csup\u003e6\u003c/sup\u003e) CFU/g are still thought to be reasonably safe (Tarr, 1996). These levels are also considered safe in the United States (Adams \u003cem\u003eet al\u003c/em\u003e., 1993). The phylogenetic analysis result only identified fungal species that belonged to Aspergillus genera; as a consequence, this study has demonstrated that these genes should be employed as molecular markers for fungal species level identification.\u003c/p\u003e \u003cp\u003eThe majority of the plant extract's documented bioactivities are attributed to phytochemicals, which are secondary plant metabolites. It is well known that they have antibacterial, anti-inflammatory, anti-sickling, and antioxidant properties. Without a doubt, such metabolites indicate the plant extracts' potential medical value and suggest future medication candidates. Following the study of Babagana \u003cem\u003eet al.\u003c/em\u003e, (2019), the qualitative phytochemical screening of \u003cem\u003eP. thonningii\u003c/em\u003e revealed the presence of alkaloids, saponins, phenols, flavonoids, tannins, terpenoids, steroids, and glycosides (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Anthocyanin and phlobatannins were absent. These chemical classes have a history of being used to treat a wide range of ailments since they have been shown to have therapeutic action against some infections. A high concentration of flavonoids and saponins was found in the aqueous extracts of \u003cem\u003eP. thonningii\u003c/em\u003e, according to the quantitative phytoconstituents analysis presented in this work (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e3\u003c/span\u003e.4). According to reports by (Ogbiko \u003cem\u003eet al\u003c/em\u003e., 2021), the plant extracts' antibacterial, antioxidant, and anti-inflammatory properties may be due to the presence of these phytonutrients. According to this study's findings, Kwaji \u003cem\u003eet al.\u003c/em\u003e, (2010), Shittu and Ihebunna (2017), and Boualam et al. (2021) have all produced similar results. Additionally, some of these phytochemicals have been connected to the ability of plants utilized in the creation of nanoparticles to cap and stabilize themselves, according to research published in Shittu and Ihebunna (2017).\u003c/p\u003e \u003cp\u003eBecause of its many practical uses, nanotechnology is currently the most rapidly developing subject of study. The production of nanoparticles can be done in a variety of ways, including physical, chemical, and biological (Sani-e \u003cem\u003eet al\u003c/em\u003e., 2022). The utilization of biological methods, such as the synthesis of NPs using green plants, is preferred. The most advantageous method is green synthesis, which is also affordable, natural, safe, and environmentally friendly (Sani-e \u003cem\u003eet al.\u003c/em\u003e, 2022). Furthermore, making SeNPs using medicinal plants may improve their advantageous qualities. Because they convert harmful oxyanions found in the environment into non-toxic elemental selenium, these biological catalysts are essential for the environment (Neha \u003cem\u003eet al.\u003c/em\u003e, 2022).\u003c/p\u003e \u003cp\u003ePrior research has demonstrated that the smaller-sized SeNPs were successful in preventing tumor cell growth via a process mediated by ROS (Natwar \u003cem\u003eet al\u003c/em\u003e., 2022), according to Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. It was discovered that the ideal wavelength range for PT-SeNPs was 295.5\u0026ndash;305.6 nm, and the ideal average size particles for the 7-day incubation period were 26.30, 34.49, 36.66, and 80.30 nm, respectively, at pH 2, 5, 6, and 7, concentrations of 2, 8, and 14 mM. After a seven-day incubation period, a pH of two, a PT concentration of eight milligrams, and a condition variable, the maximum wavelength was attained. These peaks verified the synthesis of SeNPs. The findings of Kirupagaran \u003cem\u003eet al.\u003c/em\u003e, (2016), Vennila \u003cem\u003eet al.\u003c/em\u003e, (2018), and Ban \u003cem\u003eet al.\u003c/em\u003e, (2023) were consistent with this outcome, indicating that biosynthetic Se-NPs were absorbed between 250 and 400 nm.\u003c/p\u003e \u003cp\u003eVennila K. \u003cem\u003eet al\u003c/em\u003e. (2018) provided support for these findings by confirming the importance of pH in regulating the form and size of metal nanoparticles. They found that nuclei congregate rather than form in acidic pH environments, while the largest number of particles form in alkaline medium at pH 9. Authors Chhabria and Desai (2016) also noted that by changing the pH from pH 5 to pH 7, big diameter nanospheres changed in size and shape to nanofibers with diameters of 50\u0026ndash;75 nm. According to Natwar \u003cem\u003eet al\u003c/em\u003e., (2022) the data indicated that the presence of PT on the surface of SeNPs aids in the production and stabilization of SeNPs; in the absence of PT, it can aggregate into larger particles and produce unstable particles.\u003c/p\u003e \u003cp\u003eResearch has shown that \u003cem\u003eP. thonningii\u003c/em\u003e (PT) phytochemical components are essential for the development and improvement of SeNPs stability (Kwaji \u003cem\u003eet al\u003c/em\u003e., 2010). Additionally, prior research has revealed that the phytochemical molecules bind with the SeNP surface by intermolecular hydrogen bonding (O-H-Se), giving the nanoparticles exceptional stability and dispersibility (Li \u003cem\u003eet al.\u003c/em\u003e, 2019). Numerous studies have shown the importance of natural covering or capping agents, such as proteins, lipids, and phytochemicals, which can enhance the antimicrobial potential of selenium nanoparticles (SeNPs). These agents offer higher degree of stability and can be applied in various clinical treatments (Xu \u003cem\u003eet al\u003c/em\u003e., 2019; Shi \u003cem\u003eet al\u003c/em\u003e., 2021). Additionally, the dispersion and stability of the SeNPs formed are affected by the molecular weight and concentrations of the phytochemicals (Jia \u003cem\u003eet al\u003c/em\u003e., 2015).\u003c/p\u003e \u003cp\u003eTo prevent multidrug-resistant infections, SeNPs capped with phytochemicals have been identified as a unique possible antifungal agent that can be utilized instead of conventional medications (Natwar \u003cem\u003eet al\u003c/em\u003e., 2022). On the other hand, little research has been done on \u003cem\u003eP. Thonningii's\u003c/em\u003e and selenium NPs' antifungal properties against fungus. However, according to Table\u0026nbsp;5, the synthesized PT-SeNPs exclusively showed antifungal efficacy against \u003cem\u003eAspergillus nidulans\u003c/em\u003e. By measuring the zone of inhibition (mm) against the studied pathogen, the antifungal activity of PT-SeNPs and the positive control were demonstrated.\u003c/p\u003e \u003cp\u003eThe results showed that PT-SeNPs had an inhibitory effect against \u003cem\u003eAspergillus nidulans\u003c/em\u003e that ranged from 19.11 to 24.25 mm and was greater than that produced by the control drug (Nystatine). The antifungal activity of PT-SeNPs against \u003cem\u003eAspergillus nidulans\u003c/em\u003e was found to increase as the concentration of the extracts increases from (1.5\u0026ndash;3.0 mg/ml). The different extracts' ability to suppress the tested fungus showed a considerable difference. The research aligns with the findings of Perveen \u003cem\u003eet al.\u003c/em\u003e (2020) concerning the investigation of green and sol-gel manufacturing of ZnO nanoparticles, as well as the assessment of their antibacterial and antifungal capabilities against active secondary metabolites of \u003cem\u003eP. thonningii\u003c/em\u003e stem bark and phytochemicals. In addition, Aisha et al.'s work from 2023 evaluated the antifungal effectiveness of \u003cem\u003ePiliostigma\u003c/em\u003e species against \u003cem\u003eAspergillus\u003c/em\u003e species that cause tomato fruit rot. However, PT-SeNPs' capacity to stop Aspergillus nidulans from growing suggests that they may have antifungal properties and be utilized to treat fungal diseases and cow feed.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthical Statement:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo ethical clearance is required\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data that support all the experiment findings are in this article\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have no conflict of interest to declare\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Participate declaration:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe Authors are grateful to the Centre for Genetic Engineering and Biotechnology, Federal University of Technology, Minna for enabling direct access to the Centre\u0026apos;s facilities.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding Declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was supported by the Tertiary Educational Trust Fund of Nigeria (grant number: TETF.ES.DR\u0026amp;D-CE/NRF2020/SETI/18/VOL.1).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFirst Author (O.K.S.) performed DNA isolation, electrophoresis separation, polymerase chain reaction, and bioinformatics analysis and interpret the results. While, the second and third Authors (B. A and H.B.A) performed isolation, identification, and microbial load of the fungi present in the feeds under the supervision of O.K.S. Also, the Fourth Author (J.O.T) optimized and characterized the selenium nanoparticles. All authors contributed to interpreting the results and the final manuscript\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAbdelaziz, A. M., El-Wakil, D. A., Attia, M. S., Ali, O. M., AbdElgawad, H., \u0026amp; Hashem, A. H. (2022). Inhibition of Aspergillus flavus growth and aflatoxin production in Zea mays L. using endophytic Aspergillus fumigatus. \u003cem\u003eJournal of Fungi\u003c/em\u003e, \u003cem\u003e8\u003c/em\u003e(5), 482.\u003c/li\u003e\n \u003cli\u003eAdams, R. S., Kephart, K. B., Ishler, V. A., Hutchinson, L. J., \u0026amp; Roth, G. W. (1993). Mold and mycotoxin problems in livestock feeding. \u003cem\u003eDept of Dairy and Animal Science, Extension Publ. 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Circular supply chain governance for sustainable fresh agricultural products: Minimizing food loss and utilizing agricultural waste. \u003cem\u003eSustainable Production and Consumption\u003c/em\u003e, \u003cem\u003e41\u003c/em\u003e, 391\u0026ndash;403.\u003c/li\u003e\n \u003cli\u003ePerveen, R., Shujaat, S., Qureshi, Z., Nawaz, S., Khan, M. I., \u0026amp; Iqbal, M. (2020). Green versus sol-gel synthesis of ZnO nanoparticles and antimicrobial activity evaluation against panel of pathogens. \u003cem\u003eJournal of Materials Research and Technology\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e(4), 7817\u0026ndash;7827.\u003c/li\u003e\n \u003cli\u003eRamar, M., Manikandan, B., Marimuthu, P. N., Raman, T., Mahalingam, A., Subramanian, P., ... \u0026amp; Munusamy, A. (2015). Synthesis of silver nanoparticles using Solanum trilobatum fruits extract and its antibacterial, cytotoxic activity against human breast cancer cell line MCF 7. \u003cem\u003eSpectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy\u003c/em\u003e, \u003cem\u003e140\u003c/em\u003e, 223\u0026ndash;228.\u003c/li\u003e\n \u003cli\u003eSadhasivam, S., Britzi, M., Zakin, V., Kostyukovsky, M., Trostanetsky, A., Quinn, E., \u0026amp; Sionov, E. (2017). Rapid detection and identification of mycotoxigenic fungi and mycotoxins in stored wheat grain. \u003cem\u003eToxins\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e(10), 302.\u003c/li\u003e\n \u003cli\u003eSamson, R. A., Hoekstra, E. S., \u0026amp; Frisvad, J. C. (2004). \u003cem\u003eIntroduction to food-and airborne fungi\u003c/em\u003e (No. Ed. 7). Centraalbureau voor Schimmelcultures (CBS).\u003c/li\u003e\n \u003cli\u003eShi, X. D., Tian, Y. Q., Wu, J. L., \u0026amp; Wang, S. Y. (2021). 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Ca, Last updated: July\u003c/em\u003e, \u003cem\u003e22\u003c/em\u003e, 2002.\u003c/li\u003e\n \u003cli\u003eTarr, B. (2006). Managing the effects of molds and mycotoxins in ruminants. \u003cem\u003eShur-Gain, Nutrecko Canada Inc\u003c/em\u003e.\u003c/li\u003e\n \u003cli\u003eUsman, H., Abdulrahman, F. I., \u0026amp; Usman, A. (2009). Qualitative phytochemical screening and in vitro antimicrobial effects of methanol stem bark extract of Ficus thonningii (Moraceae). \u003cem\u003eAfrican Journal of Traditional, Complementary and Alternative Medicines\u003c/em\u003e, \u003cem\u003e6\u003c/em\u003e(3).\u003c/li\u003e\n \u003cli\u003eVennila, K., Chitra, L., Balagurunathan, R., \u0026amp; Palvannan, T. (2018). Comparison of biological activities of selenium and silver nanoparticles attached with bioactive phytoconstituents: green synthesized using Spermacoce hispida extract. \u003cem\u003eAdvances in Natural Sciences: Nanoscience and Nanotechnology\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e(1), 015005.\u003c/li\u003e\n \u003cli\u003eXu, C., Qiao, L., Ma, L., Yan, S., Guo, Y., Dou, X., ... \u0026amp; Roman, A. (2019). Biosynthesis of polysaccharides-capped selenium nanoparticles using Lactococcus lactis NZ9000 and their antioxidant and anti-inflammatory activities. \u003cem\u003eFrontiers in microbiology\u003c/em\u003e, \u003cem\u003e10\u003c/em\u003e, 1632.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTable 5 is available in the Supplementary Files section.\u003c/p\u003e"},{"header":"Plates","content":"\u003cp\u003ePlate 1 is available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Caprine feed, Nutritional value and quality, Microbial load, Phylogenicity, Molecular taxonomy","lastPublishedDoi":"10.21203/rs.3.rs-5564179/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5564179/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe use of agricultural residues as caprine feed is essential for sustaining livestock, and ensuring food security. However, the presence of fungal communities in these residues poses a threat to the feed quality and animal health. This study aimed to determine the fungal diversity and optimize of selenium nanoparticle (SeNPs) biosynthesis for antifungal scavengers for the feeds. The isolation and identification of the fungal species were characterised through morphological, microscopic, and phylogenetic analyses. The optimization of SeNPs was done using sodium selenite concentration, pH, and incubation time. While characterized was carried out with ultraviolet-visible spectroscopy (UV-Vis), and ZETA sizer. The presence of fungal species such \u003cem\u003eas Aspergillus flavus, Rhizopus species, Aspergillus niger, Trichophyton rubrum, Mucor sp., Macrosporium audounii, Epidermophyton floccosum, Verticillium tenesum, Tricophytonmetagrophi\u003c/em\u003e and \u003cem\u003eExophialajeanselmei\u003c/em\u003e was confirmed across the feeds. With fungal load of varying concentrations ranging from 1.76 x 10\u003csup\u003e4\u003c/sup\u003ecfu/g of bean husk (BH) to 6.08 x 10\u003csup\u003e4\u003c/sup\u003ecfu/g of groundnut husk (GOH) was also confirmed. The evolutionary classification of the fungi isolated showed a monophyletic pattern of \u003cem\u003eAspergillus flavus\u003c/em\u003e species clustered with a bootstrap of 59%. The best condition for synthesizing PT-SeNPs was confirmed has 7 days of incubation time, 2mM of PT concentration and pH 6. The PT-SeNPs with particle size of 26.30 nm at wavelength of 300 nm show antifungal activity against \u003cem\u003eAspergillus nidulans\u003c/em\u003e (24.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45 mm) at 3.0ml. Hence, the present of toxigenic fungi in agricultural residue feed for caprine requires proper safety measures and the potential of PT-SeNPs as an antifungal scavenger for \u003cem\u003eAspergillus\u003c/em\u003e contamination.\u003c/p\u003e","manuscriptTitle":"Assessment and Improvement of Nutritive Value of Agricultural Residue Feed for Caprine","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-01-30 12:08:03","doi":"10.21203/rs.3.rs-5564179/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"995485c9-e8c0-4a31-85bf-39256dc2ec11","owner":[],"postedDate":"January 30th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-05-09T11:38:21+00:00","versionOfRecord":[],"versionCreatedAt":"2025-01-30 12:08:03","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5564179","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5564179","identity":"rs-5564179","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}