Biosynthesis of raw starch degrading β-cyclodextrin glycosyltransferase by immobilized cells of <i>Bacillus licheniformis</i> using potato wastewater

The study was sought to enhance the synthesis of thermal stable β-cyclodextrin glycosyltransferase (β-CGTase) using potato wastewater as a low-cost medium and assess the degree to which it is efficient for industrial production of β-cyclodextrin (β-CD) from raw potato starch. Thermophilic bacteria producing β-CGTase was isolated from Saudi Arabia and the promising strain was identified as Bacillus licheniformis using phylogenetic analysis of the 16S rRNA gene. Alginate-encapsulated cultures exhibited twice-fold of β-CGTase production more than free cells. Scanning electron microscopy (SEM) of polymeric capsules indicated the potential for a longer shelf-life, which promotes the restoration of activity in bacterial cells across semi-continuous fermentation of β-CGTase production for 252 h. The optimal conditions for β-CGTase synthesis using potato wastewater medium were at 36 h, pH of 8.0, and 50°C with 0.4% potato starch and 0.6% yeast extract as carbon and nitrogen sources, respectively. The purified enzyme showed a specific activity of 63.90 U/mg with a molecular weight of ∼84.6 kDa as determined by SDS-PAGE analysis. The high enzyme activity was observed up to 60°C, and complete stability was achieved at 75°C. High levels of activity and stability were shown at pH 8.0, and the pH range from 7.0–10.0, respectively. The enzyme has an appreciable affinity for raw potato starch with a Km of 5.7 × 10⁻⁶ M and a Vmax of 87.71 µmoL/mL/min. β-CD production was effective against 25 U/g of raw potato starch. The outcomes demonstrated its feasibility to develop a fermentation process by integrating the cost-effective production of β-CGTase having distinctive properties for β-CD production with ecofriendly utilization of potato wastewater.     

Effect of <i>Lycopus lucidus</i> Turcz. supplementation on gut microflora and short chain fatty acid composition in Crj: CD-1 mice

We investigated the diversity and composition of microflora in feces of Lycopus lucidus Turcz.-fed mice. In addition, we evaluated the production of major cytokines (Interleukin-6 and -10) which are related to inflammation and fatty acid composition of several tissues. 16S ribosomal DNA sequencing-based microbiome taxonomic profiling analysis was performed utilizing the EzBioCloud data base. Male mice fed on L. lucidus showed a significantly reduced number of lactic acid bacteria and coliform in the feces compared with the control group (p < 0.05). 16S rDNA sequencing analysis of fecal samples showed that L. lucidus supplementation decreased the community of harmful microflora (Enterobacteriaceae including Escherichia coli and Bacteroides sp.) in feces compared with the control group (p < 0.05). There were no significant differences in mRNA expression of cytokine IL-6 and IL-10 between the control and L. lucidus fed groups. The fecal fatty acid composition in the L. lucidus group had percentages of 4:0, 6:0, 8:0 and 10:0 in the intestine but those short chain fatty acids were not detected in the control group. Our results showed that L. lucidus supplementation influenced gut environment by decreasing harmful microflora and increased the percentages of several short fatty acids.     

Enzyme-gold affinity labelling of cellulose

The enzyme-linked colloidal gold affinity labelling technique was tested as a method to localize cellulose on thin sections of plant cell walls and slime mold spores. Commercially available cellulase from cultures of Trichoderma reesei, the main components being cellobiohydrolase I and II (CBH I, CBH II) and endoglucanase (EG), was linked to colloidal gold by using standard techniques and applied as a dilute, buffered suspension to thin sections. After brief exposure, e.g., 15–30 minutes, cellulose exposed on the surface of sections was labelled with the enzyme-gold complex. Poststaining did not appear to have a deleterious effect on the labelled sections. The specificity of labelling was demonstrated by its complete inhibition when carboxymethylcellulose was incorporated in the labelling mixture, by lack of labelling of 1,4-β-mannans or 1,3-β-xylans in noncellulosic walls of marine algae, by lack of labelling of 1,4-β-glucans in chitin, by much lower labelling density when done at 4°C, and by lack of labelling when sections were predigested with cellulase. Labelling with the crude commercial cellulase was compared to labelling with purified CBH I-, CBH II-, and EG-linked colloidal gold, and the labelling pattern was similar. This method was found useful on conventionally fixed material and required no special preparation other than the use of inert (Ni or Au) grids and 0.5% gelatin to reduce nonspecific binding of the gold complex. Labelling was similar in the several embedding resins tested: LR White, Lowicryl K4M, Epon 812, and Spurr's. The cellulase-gold probe remained active for at least 4 weeks at 4°C and much longer when frozen at ?80°C in 20% glycerol. This technique should prove useful in studies of cellulose degradation and cellulose deposition and of the interaction of cellulose with other wall components.     

Selection and validation of reference genes for quantitative real-time polymerase chain reaction analyses of <i>Serratia ureilytica</i> DW2

Background: Serratia ureilytica DW2 is a highly efficient phosphate-solubilizing bacteria isolated from Codonopsis pilosula rhizosphere soil that can promote the growth of C. pilosula; nonetheless, until now, no validated reference genes from the genus Serratia have been reported that can be used for the normalization of quantitative real-time polymerase chain reaction (RT–qPCR) data. Methods: To screen stable reference genes of S. ureilytica DW2, the expression of its eight candidate reference genes (16S rRNA, ftsZ, ftsA, mreB, recA, slyD, thiC, and zipA) under different treatment conditions (pH, temperature, culture time, and salt content) was assayed by RT–qPCR. The expression stability of these genes was analyzed using different algorithms (geNorm, NormFinder, and BestKeeper). To verify the reliability of the data, the expression of the glucose dehydrogenase (gdh) gene under different soluble phosphate levels was quantified using the most stably expressed reference gene. Results: The results showed that the zipA and 16S rRNA genes were the most stable reference genes, and the least stable genes were thiC and recA. The expression of gdh was consistent with the phosphate solubilization ability on plates containing the National Botanical Research Institute phosphate growth medium. Conclusion: Therefore, this study provides a stable and reliable reference gene of Serratia for the accurate quantification of functional gene expression in future studies.     

Elemental sulfur upregulated testicular testosterone biosynthesis by associating with altered gut microbiota in mice

Elemental sulfur has been used as a traditional Chinese medicine to treat the late-onset hypogonadism and impotence without a clarified mechanism for many hundreds of years. In the present study, mice were received sulfur or distilled water for 35 days by daily intragastric gavage at a dose of 250 mg/kg body weight. Then, the serum testosterone level and genes associated with testicular testosterone biosynthesis (TTB) were detected. The gut microbiota was also analyzed by 16S rRNA gene sequencing. Serum testosterone level was significantly increased by 291.1% in sulfur-treated mice. The H₂S levels in serum and feces were significantly increased. The expression of genes associated with TTB including StAR, p450c17, 3β-HSD, and P450scc in testes were significantly upregulated by Sulfur and NaHS, suggesting that sulfur promotes TTB depending on H₂S. In addition, sulfur increased the diversity of gut microbiota and the abundance of several bacteria associated with sulfur metabolism, including genus Prevotella, which might be positively associated with serum level of testosterone in boys. Five pathways including bile secretion, carotenoid biosynthesis, lipid biosynthesis proteins, propanoate metabolism, and biosynthesis of type II polyketide products, were identified to associate with sulfur. Together, our results suggested that sulfur upregulated testicular testosterone biosynthesis via H₂S, which was associated with alteration of gut microbiota in mice. Our study highlights a mechanism for the treatment of late-onset hypogonadism and impotence by sulfur.     

Microscopic,molecular, and biochemical investigations to characterize a benthic cyanoprokaryote Leptolyngbya strainfrom Egypt

Microscopic, molecular, and biochemical investigations were conducted to describe a benthic mat‐forming Leptolyngbya isolate collected from wastewater canal in Helwan area, Egypt. Microscopic examination revealed that the isolate was filamentous, nonheterocystous, with obvious granular surface ornamentation. Electron microscopy was used to reveal the isolate's ultrastructure. Cross walls were thick with uneven deposition. Thylakoids were convoluted and irregularly distributed. Granular content differed from one cell to the other probably due to their physiological stages/position within the filaments and/or their age. Nycridial cells were present. Highly refractile gas vesicle‐like structures were detected and their identity as gas vesicles was confirmed by amplifying the gene coding for the gas vesicle protein GvpA. The presence of gas vesicles in benthic microorganisms is intriguing, and it is possible that those vesicles serve as a floating and dispersal mechanism as they increase in filaments that are about to break and release vacuolated hormogonia. To further confirm the isolate's identity, molecular analysis using 16S rRNA gene was performed. The sequence showed only 94% similarity to Leptolyngbya badia and less than 92% to other leptolyngbya. The phylogenetic analyses showed the coclustering of this strain with other Leptolyngbya strain. The fatty acid composition, used as a chemotaxonomic marker, revealed the presence of a considerable amount of polyunsaturated acids. Nevertheless, saturated fatty acids represented the highest proportion of the total fatty. Surprisingly, fatty acids of relatively limited occurrence within oscillatorian cyanobacteria such as saturated myristic fatty acid and polyunsaturated fatty acid C16:3 were found. Microsc. Res. Tech. 76:249–257, 2013. © 2013 Wiley Periodicals, Inc.     

Viability of endolithic micro-organisms in rocks from the McMurdo Dry Valleys of Antarctica established by confocal and fluorescence microscopy

The rocks of the McMurdo Dry Valleys desert in Antarctica harbour endolithic communities of micro‐organisms such as lichens, fungi, cyanobacteria and bacteria. Establishing the physiological status and viability of these microbial colonies in their natural microhabitat has far‐reaching implications for understanding the microbial ecology of the harsh environment of this polar desert. Here we describe the use of confocal microscopy and a specific fluorescent probe (FUN‐1) to evaluate the metabolic activity of fungal cells. Application of confocal microscopy also served to identify living and dead bacteria or cyanobacteria using the fluorescent assay reagents Live/Dead SYTO 9 and propidium iodide or SYTOX Green, respectively. In addition, through the use of epifluorescence microscopy, live/dead bacteria and cyanobacteria could be detected by estimating fluorescence from their cell components provoked by simultaneously staining with nucleic acids stains such as DAPI and SYTOX Green.     

Edematogenic and myotoxic activities of the duvernoy’s gland secretion of <i>Philodryas olfersii</i> from the north-east region of Argentina

Philodryas olfersii is found in South America, from Amazonas to Patagonia. It is important to characterize the venom of P. olfersii, who inhabits the North-East region of Argentina, since snake venoms are known to exhibit considerable variability in composition and biological activities. In this work, mice weighing 18-20 g (n = 4 for each experimental group) were used. For the edematogenic activity mice were injected s.c. in the right foot pad with 50 µl of solutions containing different amounts of venom, whereas the left foot pad was injected with 50 µl of PBS. Two hours after injection mice were killed by cervical dislocation and both feet were cut off and weighed individually. For the myotoxic activity mice were injected i.m. with 100 µl of solutions containing 40 µg of venom. Blood samples were extracted after 1, 3, 6, 8, 10, 12, 14, 16 and 24 h of venom injection to determinate serum CPK activity and mice were sacrificed at the same time intervals to obtain the inoculated gastrocnemius muscle. They were fixed with Bouin solution and stained with Hematoxylin-Eosin. Results showed that P. olfersii venom exhibits a high edematogenic activity (MED = 0.31 µg) and a moderate myotoxic activity. Myonecrosis reached its highest level after 12 h of venom injection as shown by plasmatic CPK levels (5,401 ± 330 IU/l) and microscopic assay. It demonstrates the potential toxicity of the venom of P. olfersii, who inhabits the North-East region of Argentina. It also reinforces the original warning concerning the potential danger of bites by colubrids.     

Antibacterial activity of lactose-binding lectins from <i>Bufo arenarum</i> skin

Amphibians respond to microbial infection through cellular and humoral defense mechanisms such as antimicrobial protein secretion. Most humoral defense proteins are synthetized in the skin. In this study we isolated two β-galactoside-binding lectins with molecular weights of 50 and 56 KDa from the skin of Bufo arenarum. These lectins have significant hemagglutination activity against trypsinized rabbit erythrocytes, which was inhibited by galactose-containing saccharides. They are water-soluble and independent of the presence of calcium. The antimicrobial analysis for each lectin was performed. At µmolar concentration lectins show strong bacteriostatic activity against Gram negative bacteria (Escherichia coli K12 4100 and wild strains of Escherichia coli and Proteus morganii) and Gram positive bacteria (Enterococcus faecalis). The antibacterial activity of these lectins may provide an effective defense against invading microbes in the amphibian Bufo arenarum.     

Detection of new antibiotic resistance gene profile in <i>Escherichia coli</i> associated with avian leukosis virus infection from broiler chickens

The Escherichia coli (E. coli) is prevailing worldwide, but the epidemiology of E. coli infections feature regional distribution characteristics to some extent. E. coli, as a zoonotic pathogen, can be transferred from animals to humans through food chain or via contact with wounds, causing a public health risk. We reported the swelling of proventriculus and tracheal bleeding following the death in two broiler chickens (Gallus gallus domesticus) from Beijing, China. To investigate whether a virus was involved in the infection, Madin Darby Bovine Kidney (MDCK) cells were co-cultured with supernatants of proventriculus, trachea and spleen homogenates. The avian leucosis virus was detected in the samples of proventriculus and trachea, but the avian influenza virus, the Newcastle disease virus and the avian infectious laryngotracheitis virus were not detected. E. coli isolates were resistant to almost all the antimicrobial as tested except for the combinations of amoxicillin/clavulanic acid and sulfamethoxazole/trimethoprim. PCR tests demonstrated the presence of antibiotic resistance genes in these E. coli isolates and further research revealed a novel gene profile with the presence of CTX-M-1, gyrA, gyrB, oqxA, oqxB, parC and Sul2 antibiotic resistance genes in a strain isolated from a proventriculus sample. These results demonstrated that the presence of antibiotic resistant E. coli would not necessarily cause outbreak of large-scale disease. However, when the bacteria carrying new antibiotic resistance genes enter the environment, it may result in the development of more virulent strains which will potentially impact human and animal health.     

Morphology visualization of irregular shape bacteria by electron holography and tomography

In the current work, irregular morphology of Staphylococcus aureus bacteria has been visualized by phase retrieval employing off‐axis electron holography (EH) and 3D reconstruction electron tomography using high‐angle annular dark field scanning transmission electron microscopy (HAADF‐STEM). Bacteria interacting with gold nanoparticles (AuNP) acquired a shrunken or irregular shape due to air dehydration processing. STEM imaging shows the attachment of AuNP on the surface of cells and suggests an irregular 3D morphology of the specimen. The phase reconstruction demonstrates that off‐axis electron holography can reveal with a single hologram the morphology of the specimen and the distribution of the functionalized AuNPs. In addition, EH reduces significantly the acquisition time and the cumulative radiation damage (in three orders of magnitude) over biological samples in comparison with multiple tilted electron expositions intrinsic to electron tomography, as well as the processing time and the reconstruction artifacts that may arise during tomogram reconstruction.     

Characterization of Talaromyces islandicus–mediated silver nanoparticles and evaluation of their antibacterial and anticancer potential

The biosynthesized silver nanoparticles (AgNPs) have been reported to possess several therapeutic applications. Silver is one of the important metals known for its bioactive properties not only as macromolecule but also as nanoparticle (NP). The current research focused on the eco-friendly synthesis of Talaromyces islandicus VSGF1(Lab code) –mediated AgNPs. The aqueous culture filtrate of T. islandicus VSGF1 was used as a reducing agent. The formation of AgNPs was confirmed by observing the color change from colorless to colloidal earthy-colored and a sharp absorption peak of ultraviolet–visible (UV-vis) spectroscopy at 400 nm. Fourier-transform infrared spectroscopy revealed the involvement of various functional groups for the formation and stabilization of AgNPs. The structure, size, and shape of mycosynthesized AgNPs were identified by X-ray diffraction (XRD), scanning electron microscopy, and atomic force microscopy (AFM) analysis. The XRD analysis exhibited crystalline nature of NPs whereas AFM analysis revealed the spherical shape of AgNP with average size range between 13 and 66 nm. The antibacterial activity of AgNPs (50 μg/ml) investigated against gram-positive and gram-negative bacteria revealed maximum zone of inhibition (ZOI) against drug-resistant Enterococcus faecalis MTCC439 (18.66 ± 0.57 mm) and Pseudomonas aeruginosa MTCC96 (16 ± 0 mm) followed by Staphylococcus aureus MTCC96 (15.33 ± 0.57 mm), Bacillus subtilis MTCC441 (14.66 ± 0.57 mm), and Escherichia coli MTCC45 (14.66 ± 0.57 mm). Further, the AgNPs evaluated for antitumor activity against human hepatocellular carcinoma (HepG2) cell line exhibited promising result with half-maximal inhibitory concentration (MIC) value at 38.17 μg/ml concentration through MTT (methylthiazolyl tetrazolium assay) assay. Apparently, this is the first report from T. islandicus to the best of our knowledge.     

Synthesis,characterization and use of iron oxide nano particles for antibacterial activity

In this decade, the use of nano particles (NPs) against bacterial growth is increasing day by day due to remarkable alternative properties compared to molecular antibiotics. Thus, the use of iron oxide nanoparticles (IONPs) has proven one of the most important transition metals oxide‐based remedy in nanotechnological advances and biological applications due to enriched biocompatibility of iron. In this study synthesis of IONPs was carried out via co‐precipitation method. The crystallographic morphology of the synthesized particles was studied via X‐ray diffraction which revealed cubic structure of the particles, whereas, the spinal shaped morphology of the prepared NPs was confirmed from scanning electron microscopy. Likewise, the presence of the major elements in the sample was determined through energy dispersive X‐ray analysis characterization. Bactericidal effect of the NPs was assessed at pre‐defined concentrations (50 and 100 μg/ml) against Gram +ve bacteria Staphylococcus aureus, Gram ?ve bacteria Shigella dysentry and Escherichia coli . Bacterial strains, which demonstrate the potential of NPs. The purpose of this study was assessing the structure of the synthesized NPs for protective effect against harmful bacterial activity.     

Microbiological examination of six industrial soluble oil emulsion samples

Soluble oil emulsion samples from machine tool sumps were examined for their total bacterial population and for potentially pathogenic enteric and haemolytic bacteria. All the samples contained a large bacterial population in which the most abundant organisms belonged to the genus Pseudomonas, and also a significant number of enteric and haemolytic bacteria. Very low numbers of anaerobic sulphide producing bacteria and no fungi were isolated. The samples were generally in poor condition with free oil and slime clearly visible. In only on instance was the oil/water ratio correct.     

Mass spectrometry for enzyme assays and inhibitor screening: an emerging application in pharmaceutical research

Robust methods that monitor enzyme activity and inhibitor potency are crucial to drug discovery and development. Over the past 20 years, mass spectrometric methods have increasingly been used to measure enzyme activity and kinetics. However, for rapid screening of inhibitory compounds, various forms of fluorescence and chemiluminscence readout have continued to dominate the market. As the sensitivity, speed, and miniaturization of mass spectrometry methods continue to advance, opportunities to couple mass spectrometry with screening will continue to come to the forefront. To appreciate the tremendous potential for MS‐based screening assays, it becomes necessary to understand the current state of capabilities in this arena. Thus, this review is intended to capture how mass spectrometry for studying enzymes activity has progressed from simple qualitative questions (i.e., is the product detected?) to quantitative measures of enzyme activity and kinetics and then as a tool for rapidly screening inhibitory compounds as an alternative to current methods of high throughput drug screening. © 2007 Wiley Periodicals, Inc., Mass Spec Rev     

Hydrogen peroxide staining to visualize intracellular bacterial infections of seedling root cells

Visualization of bacteria in living plant cells and tissues is often problematic due to lack of stains that pass through living plant cell membranes and selectively stain bacterial cells. In this article, we report the use of 3,3′‐diaminobenzidine tetrachloride (DAB) to stain hydrogen peroxide associated with bacterial invasion of eukaryotic cells. Tissues were counterstained with aniline blue/lactophenol to stain protein in bacterial cells. Using this staining method to visualize intracellular bacterial (Burkholderia gladioli) colonization of seedling roots of switch grass (Panicum virgatum), we compared bacterial free seedling roots and those inoculated with the bacterium. To further assess application of the technique in multiple species of vascular plants, we examined vascular plants for seedling root colonization by naturally occurring seed‐transmitted bacteria. Colonization by bacteria was only observed to occur within epidermal (including root hairs) and cortical cells of root tissues, suggesting that bacteria may not be penetrating deeply into root tissues. DAB/peroxidase with counter stain aniline blue/lactophenol was effective in penetration of root cells to selectively stain bacteria. Furthermore, this stain combination permitted the visualization of the bacterial lysis process. Before any evidence of H₂O₂ staining, intracellular bacteria were seen to stain blue for protein content with aniline blue/lactophenol. After H₂O₂ staining became evident, bacteria were often swollen, without internal staining by aniline blue/lactophenol; this suggests loss of protein content. This staining method was effective for seedling root tissues; however, it was not effective at staining bacteria in shoot tissues due to poor penetration. Microsc. Res. Tech. 77:566–573, 2014. © 2014 Wiley Periodicals, Inc.     

Ultrastructural and electron spectroscopic analyses of cyanobacteria and bacteria

The extracellular sheath material and some intracellular cell components of cyanobacteria and phosphate-accumulating sewage bacteria were analysed by electron spectroscopic imaging (ESI) and electron energy-loss spectroscopy (EELS). The specimens were embedded in water-soluble Nanoplast resin without any previous fixation and ultrathin sections were examined in a Zeiss CEM 902 microscope. A high sulphur content was detected in the inner sheath of the cyanobacterium Gloeothece. The elemental composition of some cell components and inclusion bodies, such as carboxysomes and cyanophycin, was determined by ESI and EELS. In addition, the phosphate content in specific granules of phosphate-accumulating sewage bacteria was estimated by EELS and nuclear magnetic resonance spectroscopy.     

Morphological and mechanical imaging of Bacillus cereus spore formation at the nanoscale

Bacteria from the genus Bacillus are able to transform into metabolically dormant states called (endo) spores in response to nutrient deprivation and other harsh conditions. These morphologically distinct spores are fascinating constructs, amongst the most durable cells in nature, and have attracted attention owing to their relevance in food‐related illnesses and bioterrorism. Observing the course of bacterial spore formation (sporulation) spatially, temporally and mechanically, from the vegetative cell to a mature spore, is critical for a better understanding of this process. Here, we present a fast and versatile strategy for monitoring both the morphological and mechanical changes of Bacillus cereus bacteria at the nanoscale using atomic force microscopy. Through a strategy of imaging and nanomechanical mapping, we show the morphogenesis of the endospore and released mature endospore. Finally, we investigate individual spores to characterize their surface mechanically. The progression in elasticity coupled with a similarity of characteristic distributions between the incipient endospores and the formed spores show these distinct stages. Taken together, our data demonstrates the power of atomic force microscopy applied in microbiology for probing this important biological process at the single cell scale.     

Bioactive Sheath/Core nanofibers containing olive leaf extract

This study aimed at producing silk fibroin (SF)/hyaluronic acid (HA) and olive leaf extract (OLE) nanofibers with sheath/core morphology by coaxial electrospinning method, determining their antimicrobial properties, and examining release profiles of OLE from these coaxial nanofibers. Optimum electrospinning process and solution parameters were determined to obtain uniform and bead‐free coaxial nanofibers. Scanning electron microscopy and transmission electron microscopy (TEM) were used to characterize the morphology of the nanofibers. The antimicrobial activities of nanofibers were tested according to AATCC test method 100. Total phenolic content and total antioxidant activity were tested using in vitro batch release system. The quality and quantity of released components of OLE were determined by high‐performance liquid chromatography. The changes in nanofibers were examined by Fourier‐transform infrared spectroscopy. Uniform and bead‐free nanofibers were produced successfully. TEM images confirmed the coaxial structure. OLE‐loaded nanofibers demonstrated almost perfect antibacterial activities against both of gram‐negative and gram‐positive bacteria. Antifungal activity against C. albicans was rather poor. After a release period of 1 month, it was observed that ～70–95% of the OLE was released from nanofibers and it was still bioactive. Overall results indicate that the resultant shell/core nanofibers have a great potential to be used as biomaterials. Microsc. Res. Tech. 79:38–49, 2016. © 2015 Wiley Periodicals, Inc.     

Leaf and stem micromorphology of Byrsonima sericea DC. by light and scanning electron microscopy

Micromorphological studies were carried out using multiple microscopic techniques on the leaves and stem bark of Byrsonima sericea DC. (Malpighiaceae), a species popularly known as “murici” and used medicinally, in order to identify both qualitative and quantitative features of leaf and stem anatomy and histochemistry as differential parameters to support both the quality control of its ethnodrugs and the taxonomy of the genus. The study was conducted using traditional techniques of plant anatomy, histochemical tests, and the stomatal index (SI). Byrsonima sericea has hypostomatic leaves, anomocytic stomata, and its epidermal walls are anticlinal and straight on the adaxial and curved on the abaxial faces. T‐shaped trichomes were observed mainly on the abaxial surface. The leaf epidermis showed waxes syntopism on both surfaces, with the occurrence of different crystalloid forms on a single phylloplane. The mesophyll is dorsiventral, with 3‐4 collateral vascular bundles. Phenolic compounds, starch, and proteins were identified in the petiole and stem. The SI was 14.5 ± 0.53% (p < .05), but did not showed significant variations. A set of characters were found to be distinctive for the studied species, however, constituting parameters that could be used to separate B. sericea from other species of the genus.