2.3.2. Alpha-amylase
The α-amylase assay was performed according to Vega-Villasante
et al. (1993), using soluble starch (1%) as substrate. One unit
corresponded to the amount of enzyme required to increase by 0.01
units the absorbance at 540 nm per minute.
Specific and total enzyme activities in digestive extracts were
determined using the following equations:
(1) Units/mL=(Δabs reaction final volume (mL)) / (MEC·time
(min) extract volume (mL));
(2) Units/mg protein=Units per mL/mg of soluble protein
Δabs represents the increased absorbance at a determined
wavelength and MEC represents the molar extinction coefficient for
the product of the reaction (mL/μg/cm). The results are presented
using Eqs.(1) and (2), and all assays were carried out in triplicate.
2.4. Histology
Conventional histology and hematoxylin–eosin (H&E) staining
were performed on larvae fixed in 2% paraformaldehyde for 24 h at
4 °C, before being washed, dehydrated, cleared, and embedded in
paraffin. Sagittal sections (5 μm) were obtained with a conventional
microtome, placed on gelatin-coated slides, re-hydrated, and H&EMstained. Histological sections were viewed under a light microscope
and photographed with an Infinity digital camera and the PAXcam2
software (PAX-it version 6, MIS Inc., USA).
2.5. Statistics
Specific and total digestive enzyme activities in larval extracts
were expressed as the mean±SD. Homogeneity of variances and
normality tests were performed. The specific activity was calculated
using the one-way analysis of variance (ANOVA) procedure. Multiple
comparisons of enzymatic activity over time were obtained by a
Tukey comparison test. All statistics were conducted using Sigma-Plot
11.0 for Windows (Systat Software Inc.). A significance level of Pb0.05
was used.
2.3.2. Alpha-amylaseThe α-amylase assay was performed according to Vega-Villasanteet al. (1993), using soluble starch (1%) as substrate. One unitcorresponded to the amount of enzyme required to increase by 0.01units the absorbance at 540 nm per minute.Specific and total enzyme activities in digestive extracts weredetermined using the following equations:(1) Units/mL=(Δabs reaction final volume (mL)) / (MEC·time(min) extract volume (mL));(2) Units/mg protein=Units per mL/mg of soluble proteinΔabs represents the increased absorbance at a determinedwavelength and MEC represents the molar extinction coefficient forthe product of the reaction (mL/μg/cm). The results are presentedusing Eqs.(1) and (2), and all assays were carried out in triplicate.2.4. HistologyConventional histology and hematoxylin–eosin (H&E) stainingwere performed on larvae fixed in 2% paraformaldehyde for 24 h at4 °C, before being washed, dehydrated, cleared, and embedded inparaffin. Sagittal sections (5 μm) were obtained with a conventionalmicrotome, placed on gelatin-coated slides, re-hydrated, and H&EMstained. Histological sections were viewed under a light microscopeand photographed with an Infinity digital camera and the PAXcam2software (PAX-it version 6, MIS Inc., USA).2.5. StatisticsSpecific and total digestive enzyme activities in larval extractswere expressed as the mean±SD. Homogeneity of variances andnormality tests were performed. The specific activity was calculatedusing the one-way analysis of variance (ANOVA) procedure. Multiplecomparisons of enzymatic activity over time were obtained by aTukey comparison test. All statistics were conducted using Sigma-Plot11.0 for Windows (Systat Software Inc.). A significance level of Pb0.05was used.
การแปล กรุณารอสักครู่..