Abstract: Termites are eusocial insects that are found on all continents except Antarctica. Termites have serious destructive impact, damaging local huts and crops of poor subsistence. The annual cost of termite damage and its control is determined in the billions globally. In Egypt, most of these damages are due to the subterranean termite species especially the sand termite, P. hypostoma. Pyrethroids became the primary weapon for subterranean termite control, after the use of chlorpyrifos as a soil termiticide was banned. Despite the important role of pyrethroids in termite control, its extensive use in pest control led to the eventual rise of insecticide resistance which may make many of the pyrethroids ineffective. The ability to diagnose the precise mechanism of pyrethroid resistance in any insect species would be the key component of its management at specified location for a specific population. In the present study, detailed toxicological and biochemical studies was conducted on the mechanism of pyrethroid resistance in P. hypostoma. The susceptibility of field populations of P. hypostoma against deltamethrin, α-cypermethrin and ƛ-cyhalothrin was evaluated. The obtained results revealed that the workers of P. hypostoma have developed high resistance level against the tested pyrethroids. Studies carried out through estimation of detoxification enzyme activity indicated that enhanced esterase and cytochrome P450 activities were probably important mechanisms for pyrethroid resistance in field populations. Elevated esterase activity and also additional esterase isozyme were observed in the pyrethroid-resistant populations compared to the susceptible populations. Strong positive correlation between cytochrome P450 activity and pyrethroid resistance was also reported. |Deltamethrin could be recommended as a resistance-breaking pyrethroid that is active against resistant populations of P. hypostoma.
Abstract: The cotton leafworm, Spodoptera littoralis (Boisduval) is a major insect pest of vegetables and cotton crops in Egypt, and exhibits different levels of tolerance to certain insecticides. Chlorantraniliprole has been registered recently in Egypt for control this insect. The susceptibilities of three S. littoralis populations collected from El Behaira governorate, north Egypt to chlorantraniliprole were determined by leaf-dipping technique on 4th instar larvae. Obvious variation of toxicity was observed among the laboratory susceptible, and three field populations with LC50 values ranged between 1.53 µg/ml and 6.22 µg/ml. However, all the three field populations were less susceptible to chlorantraniliprole than a laboratory susceptible population. The most tolerant populations were sampled from El Delengat (ED) Province where S. littoralis had been frequently challenged by insecticides. Certain enzyme activity assays were carried out to be correlated with the mechanism of the observed field population tolerance. All field populations showed significantly enhanced activities of detoxification enzymes compared with the susceptible strain. The regression analysis between chlorantraniliprole toxicities and enzyme activities revealed that the highest correlation is between α-esterase or β-esterase (α-β-EST) activity and collected field strains susceptibility, otherwise this correlation is not significant (P > 0.05). Synergism assays showed the ED and susceptible strains could be synergized by known detoxification inhibitors such as piperonyl butoxide (PBO), triphenyl phosphate (TPP) and diethyl-maleate (DEM) at different levels (1.01-8.76-fold and 1.09-2.94 fold, respectively), TPP showed the maximum synergism in both strains. The results show that there is a correlation between the enzyme activity and tolerance, and carboxylic-esterase (Car-EST) is likely the main detoxification mechanism responsible for tolerance of S. littoralis to chlorantraniliprole.
Abstract: The use of probiotics engineered to express specific enzymes has been the subject of considerable attention in poultry industry because of increased nutrient availability and reduced cost of enzyme supplementation. Phytase enzyme is commonly added to poultry feed to improve digestibility and availability of phosphorus from plant sources. To construct a probiotic with potential of phytate degradation, phytase gene (appA) from E. coli was cloned and transformed into two probiotic bacteria Lactobacillus salivarius and Lactococcus lactis. L. salivarous showed plasmid instability, unable to express the gene. The expression of appA gene in L. lactis was analyzed by detecting specific RNA and zymography assay. Phytase enzyme was isolated from cellular extracts of recombinant L. lactis, showing a 46 kDa band upon the SDS-PAGE analysis. Zymogram also confirmed the phytase activity of the 46 kDa band corresponding to the enzyme. An enzyme activity of 4.9U/ml was obtained in cell extracts of L. lactis. The growth of native and recombinant L. lactis was similar in the presence of two concentrations of ox bile.
Abstract: Enzymes are the biocatalysts which catalyze the biochemical processes and thus have a wide variety of applications in the industrial sector. β-Galactosidase (E.C. 3.2.1.23) also known as lactase, is one of the prime enzymes, which has significant potential in the dairy and food processing industries. It has the capability to catalyze both the hydrolytic reaction for the production of lactose hydrolyzed milk and transgalactosylation reaction for the synthesis of prebiotics such as lactulose and galactooligosaccharides. These prebiotics have various nutritional and technological benefits. Although, the enzyme is naturally present in almonds, peaches, apricots and other variety of fruits and animals, the extraction of enzyme from these sources increases the cost of enzyme. Therefore, focus has been shifted towards the production of low cost enzyme from the microorganisms such as bacteria, yeast and fungi. As compared to yeast and bacteria, fungal β-galactosidase is generally preferred as being extracellular and thermostable in nature. Keeping the above in view, the present study was carried out for the isolation of the β-galactosidase producing fungal strain from the food as well as the agricultural wastes. A total of more than 100 fungal cultures were examined for their potential in enzyme production. All the fungal strains were screened using X-gal and IPTG as inducers in the modified Czapek Dox Agar medium. Among the various isolated fungal strains, the strain exhibiting the highest enzyme activity was chosen for further phenotypic and genotypic characterization. The strain was identified as Rhizomucor pusillus on the basis of 5.8s RNA gene sequencing data.
Abstract: Diagnostic enzymes like aspartate aminotransferase
(AST), alanine aminotransferase (ALT) and alkaline phosphatase
(ALP) were determined as indices of heavy metal pollution in Tilapia
guinensis. Three different sets of fishes treated with lead (Pb), iron
(Fe) and copper (Cu) were used for the study while a fourth group
with no heavy metal served as a control. Fishes in each of the groups
were exposed to 2.65mg/l of Pb, 0.85mg/l of Fe and 0.35 mg/l of Cu
in aerated aquaria for 96 hours. Tissue fractionation of the liver
tissues was carried out and the three diagnostic enzymes (AST, ALT,
and ALP) were estimated. Serum levels of the same diagnostic
enzymes were also measured. The mean values of the serum enzyme
activity for ALP in each experimental group were 19.5±1.62,
29.67±2.17 and 1.15±0.27 IU/L for Pb, Fe and Cu groups compared
with 9.99±1.34 IU/L enzyme activity in the control. This result
showed that Pb and Fe caused increased release of the enzyme into
the blood circulation indicating increased tissue damage while Cu
caused a reduction in the serum level as compared with the level in
the control group. The mean values of enzyme activity obtained in
the liver were 102.14±6.12, 140.17±2.06 and 168.23±3.52 IU/L for
Pb, Fe and Cu groups, respectively compared to 91.20±9.42 IU/L
enzyme activity for the control group. The serum and liver AST and
ALT activities obtained in Pb, Fe, Cu and control groups are
reported. It was generally noted that the presence of the heavy metal
caused liver tissues damage and consequent increased level of the
diagnostic enzymes in the serum.
Abstract: Beta-glucosidase, chitinase, leucine-aminopeptidase, acid phosphomonoesterase and acetate-esterase enzyme activities in
the soils under the impact of metallurgical industrial activity in Lori
marz (district) were investigated. The results of the study showed that
the activities of the investigated enzymes in the soils decreased with increasing distance from the Shamlugh copper mine, the Chochkan
tailings storage facility and the ore transportation road. Statistical
analysis revealed that the activities of the enzymes were positively
correlated (significant) to each other according to the observation
sites which indicated that enzyme activities were affected by the
same anthropogenic factor. The investigations showed that the soils
were polluted with heavy metals (Cu, Pb, As, Co, Ni, Zn) due to
copper mining activity in this territory. The results of Pearson
correlation analysis revealed a significant negative correlation
between heavy metal pollution degree (Nemerow integrated pollution
index) and soil enzyme activity. All of this indicated that copper
mining activity in this territory causing the heavy metal pollution of
the soils resulted in the inhabitation of the activities of the enzymes
which are considered as biological catalysts to decompose organic
materials and facilitate the cycling of nutrients.
Abstract: Lipases constitute one of the most important groups of
industrial enzymes that catalyze the hydrolysis of triacylglycerol to
glycerol and fatty acids. Muscarinic antagonist relieves smooth
muscle spasm of the gastrointestinal tract and effect on the
cardiovascular system. In this research the effect of a muscarinic
antagonist on the lipase activity of Pseudomonas aeruginosa was
studied. Lineweaver–Burk plot showed that the drug inhibited the
enzyme by competitive inhibition. The IC50 value (0.16 mM) and Ki
(0.03 mM) of the drug revealed the drug bound to enzyme with high
affinity. Determination of enzyme activity in various pH and
temperature showed that the maximum activity of lipase was at pH 8
and 60oC both in presence and absence of the drug.
Abstract: Heavy metals are one of the major groups of
contaminants in the environment and many of them are toxic even at
very low concentration in plants and animals. However, some metals
play important roles in the biological function of many enzymes in
living organisms. Metals such as zinc, iron, and cooper are important
for survival and activity of enzymes in plants, however heavy metals
can inhibit enzyme which is responsible for defense system of plants.
Polyphenol oxidase (PPO) is a copper-containing metalloenzyme
which is responsible for enzymatic browning reaction of plants.
Enzymatic browning is a major problem for the handling of
vegetables and fruits in food industry. It can be increased and
effected with many different futures such as metals in the nature and
ground. In the present work, PPO was isolated and characterized
from green leaves of red poppy plant (Papaverr hoeas). Then, the
effect of some known antibrowning agents which can form
complexes with metals and metals were investigated on the red poppy
PPO activity. The results showed that glutathione was the most
potent inhibitory effect on PPO activity. Cu(II) and Fe(II) metals
increased the enzyme activities however, Sn(II) had the maximum
inhibitory effect and Zn(II) and Pb(II) had no significant effect on the
enzyme activity. In order to reduce the effect of heavy metals, the
effects of metal-antibrowning agent complexes on the PPO activity
were determined. EDTA and metal complexes had no significant
effect on the enzyme. L-ascorbic acid and metal complexes decreased
but L-ascorbic acid-Cu(II)-complex had no effect. Glutathione–metal
complexes had the best inhibitory effect on Red poppy leaf PPO
activity.
Abstract: Enzyme activity was evaluated in the intestine of
juvenile dourado (Salminus brasiliensis) fed with diets containing 0,
10 or 20% of lyophilized bovine colostrum (LBC) inclusion for either
30 or 60 days. The intestinal enzymes acid and alkaline phosphatase
(ACP and ALP, respectively), non-specific esterase (NSE), lipase
(LIP), dipeptidyl aminopeptidase IV (DAP IV) and leucine
aminopeptidase (LAP) were studied using histochemistry in four
intestinal segments (S1, S2, S3 and posterior intestine). Weak
proteolitic activity was observed in all intestinal segments for DAP
IV and LAP. The activity of NSE and LIP was also weak in all
intestines, except for the moderate activity of NSE in the S2 of 20%
LBC group after 30 days and in the S1 of 0% LBC group after 60
days. The ACP was detected only in the S2 and S3 of the 10% LBC
group after 30 days. Moderate and strong staining was observed in
the first three intestinal segments for ALP and weak activity in the
posterior intestine. The activity of DAP IV, LAP and ALP were also
present in the cytoplasm of the enterocytes. In the present results,
bovine colostrum feeding did not cause alterations in activity of
intestinal enzymes.
Abstract: The hydrolysis of lactose using β-galactosidase is one of the most promising biotechnological applications, which has wide range of potential applications in food processing industries. However, due to intracellular location of the yeast enzyme, and expensive extraction methods, the industrial applications of enzymatic hydrolysis processes are being hampered. The use of permeabilization technique can help to overcome the problems associated with enzyme extraction and purification of yeast cells and to develop the economically viable process for the utilization of whole cell biocatalysts in food industries. In the present investigation, standardization of permeabilization process of novel yeast isolate was carried out using a statistical model approach known as Response Surface Methodology (RSM) to achieve maximal b-galactosidase activity. The optimum operating conditions for permeabilization process for optimal β-galactosidase activity obtained by RSM were 1:1 ratio of toluene (25%, v/v) and ethanol (50%, v/v), 25.0 oC temperature and treatment time of 12 min, which displayed enzyme activity of 1.71 IU /mg DW.
Abstract: Studies were carried out on the comparative study of the production of Avicelase enzyme using sugarcane bagasse-SCB in two different statuses (i.e. treated and untreated SCB) by thermophilic Geobacillus stearothermophilus at 50ºC. Only four thermophilic bacterial isolates were isolated and assayed for Avicelase production using UntSCB and TSCB. Only one isolate selected as most potent and identified as G. stearothermophilus used in this study. A specific endo-β-1,4-D-glucanase (Avicelase EC 3.2.1.91) was partially purified from a thermophilic bacterial strain was isolated from different soil samples when grown on cellulose enrichment SCB substrate as the sole carbon source. Results shown that G. stearothermophilus was the better Avicelase producer strain. Avicelase had an optimum pH and temperature 7.0 and 50ºC for both UntSCB and TSCB and exhibited good pH stability between "5-8" and "4-9", however, good temperature stability between (30-80ºC) for UntSCB and TSCB, respectively. Other factors affecting the production of Avicelase were compared (i.e. SCB concentration, inoculum size and different incubation periods), all results observed and obtained were revealed that the TSCB was exhibited maximal enzyme activity in comparison with the results obtained from UntSCB, so, the TSCB was enhancing the Avicelase production.
Abstract: The experimental design was 4 x 5 factorial with three
replications in fully controlled research greenhouse in Department of
Soil Sciences and Plant Nutrition, Faculty of Agriculture, University
of Selcuk in the year of 2009. Determination of tolerant chickpea
genotypes to drought was made in the research. Additionally,
sophisticated effects of drought on plant growth and development,
biochemical and physical properties or physical defense mechanisms
were presented. According to the results, the primary genotypes were
Ilgın YP (0.0063 g/gh) for leaf water capacity, 22235 70.44(%) for
relative water content, 22159 (82.47%) for real water content,
22159 (5.03 mg/l) for chlorophyll a+b, Ilgın YP (125.89 nmol
H2O2.dak-1/ mg protein-1) for peroxidase, Yunak YP (769.67
unit/ mg protein-1) for superoxide dismutase, Seydişehir YP
(16.74 μg.TA-1) for proline, Gökçe (80.01 nmol H2O2.dak-1/ mg
protein-1) for catalase. Consequently, all the genotypes
increased their enzyme activity depending on the increasing of
drought stress consider with the effects of drought stress on leaf
enzyme activity. Chickpea genotypes are increasing enzyme
activity against to drought stress.
Abstract: Cabbage seedlings grown in vitro were exposed to
excess levels of heavy metals, including Cd, Mo, and Zn. High metal
levels affected plant growth at cotyledonary stage. Seedlings under
Cd, Mo, and Zn treatments could not produce root hairs and true
leaves. Under stress conditions, seedlings accumulated a higher
amount of anthocyanins in their cotyledons than those in the control.
The pigments isolated from Cd and Zn stressed seedling cotyledons
appeared as pink, while under Mo stress, was dark pink or purple.
Moreover, excess Mo stress increased antioxidant enzyme activities
of APX, CAT, SOD. These results suggest that, under excess Mo
stress, the induced antioxidant enzyme activity of cabbage seedlings
may function as a protective mechanism to shield the plants from
toxicity and exacerbated growth.
Abstract: The purpose of the present work was to study the
production and process parameters optimization for the synthesis of
cellulase from Trichoderma viride in solid state fermentation (SSF)
using an agricultural wheat straw as substrates; as fungal conversion
of lignocellulosic biomass for cellulase production is one among the
major increasing demand for various biotechnological applications.
An optimization of process parameters is a necessary step to get
higher yield of product. Several kinetic parameters like pretreatment,
extraction solvent, substrate concentration, initial moisture content,
pH, incubation temperature and inoculum size were optimized for
enhanced production of third most demanded industrially important
cellulase. The maximum cellulase enzyme activity 398.10±2.43
μM/mL/min was achieved when proximally analyzed lignocellulosic
substrate wheat straw inocubated at 2% HCl as pretreatment tool
along with distilled water as extraction solvent, 3% substrate
concentration 40% moisture content with optimum pH 5.5 at 45°C
incubation temperature and 10% inoculum size.
Abstract: the objective of this study is to measure the levels of
cellulas activity of ostrich GI microorganisms, and comparing it with
the levels of cellulas activity of rumen-s microorganisms, and also to
estimate the probability of increasing enzyme activity with injecting
different dosages (30%, 50% and 70%) of pure anaerobic goat rumen
fungi. The experiment was conducted in laboratory and under a
complete anaerobic condition (in vitro condition). 40 ml of
“CaldWell" medium and 1.4g wheat straw were placed in incubator
for an hour. The cellulase activity of ostrich microorganisms was
compared with other treatments, and then different dosages (30%,
50% and 70%) of pure anaerobic goat rumen fungi were injected to
ostrich microorganism-s media. Due to the results, cattle and goat
with 2.13 and 2.08 I.U (international units) respectively showed the
highest activity and ostrich with 0.91 (I.U) had the lowest cellulose
activity (p < 0.05). Injecting 30% and 50% of anaerobic fungi had no
significant incensement in enzyme activity, but with injecting 70% of
rumen fungi to ostrich microorganisms culture a significant increase
was observed 1.48 I.U. (p < 0.05).
Abstract: Glutathione S-transferase was purified from human
erythrocytes and effects of some polyphenols were investigated on
the enzyme activity. The purification procedure was performed on
Glutathione-Agarose affinity chromatography after preparation of
erythrocytes hemolysate with a yield of 81%. The purified enzyme
showed a single band on the SDS-PAGE. The effects of some
poliphenolic compounds such as catechin, dopa, dopamine, progallol
and catechol were examined on the in vitro GST activity. Catechin
was determined to be inhibitor for the enzyme, but others were not
effective on the enzyme as inhibitors or activators. IC50 value -the
concentration of inhibitor which reduces enzyme activity by 50%-
was estimated to be 10 mM. Ki constants were also calculated as 6.38
± 0,70 mM with GSH substrate, and 3.86 ± 0,78 mM with CDNB
substrate using the equations of graphs for the inhibitor, and its
inhibition type was determined as non-competitive.
Abstract: An experiment was conducted to study the effects of different types of probiotic on Sucrase enzyme activity of the small intestine mucosa in male broilers. The experimental design was arranged as randomized completely blocks in 4 × 2 factorial arrangement of treatment. 180 male broilers of Ross 308 commercial hybrid were designated into 4 groups. Three replicates of 15 birds were assigned to each treatment. Control treatments (diet contained no probiotic) were fed according to the NRC as base diet and three treatment groups were fed from the same diet plus three different types of probiotics. Birds were slaughtered after 21 and 42 days and different segments of small intestine (at 1,10,30,50,70 and 90% of total length the small intestine) were taken from each replicates (N=2) Sucrase enzyme activities were measured and recorded. Obtained data were analyzed by Spss (P
Abstract: The effect of phosphorus supplementation of ammoniated rice straw was studied. The in vitro experiment was carried out following the first stage of Tilley and Terry method. The treatments consisting of four diets were A = 50% ammoniated rice straw + 50% concentrate (control), B = A + 0.2% Phosphor (P) supplement, C = A + 0.4% Phosphor (P) supplement, and D = A + 0.6% Phosphor (P) supplement of dry matter. Completely randomized design was used as the experimental design with differences among treatment means were examined using Duncan multiple range test. Variables measured were total bacterial and cellulolytic bacterial population, cellulolytic enzyme activity, ammonia (NH3) and volatile fatty acid (VFA) concentrations, as fermentability indicators and synthesized microbial protein, as well as degradability indicators including dry matter (DM), organic matter (OM), neutral detergent fibre (NDF), acid detergent fibre (ADF) and cellulose. The results indicated that fermentability and degradability of diets consisting ammoniated rice straw with P supplementation were significantly higher than the control diet (P< 0.05). It is concluded that P supplementation is important to improve fermentability and degradability of rations containing ammoniated RS and concentrate. In terms of the most effective level of P supplementation occurred at a supplementation rate of 0.4% of dry matter.
Abstract: Malting is usually carried out on intact barley seed,
while hull is still attached to it. In this study, oat grain with and
without hull was subjected to controlled germination to optimize its
enzymes activity, in such a way that lipase has the lowest and α-
amylase and proteinase the highest activities. Since pH has a great
impact on the activity of the enzymes, the pH of germination media
was set up to 3 to 8. In dehulled oats, lipase and α-amylase had the
lowest and highest activities in pHs 3 and 6, respectively whereas the
highest proteinase activity was evidenced at pH 7 and 4 in the oats
with and without hull respectively. While measurements indicated
that the effect of hull on the enzyme activities particularly in lipase
and amylase at each level of the pH are significantly different, the
best results were obtained in those samples in which their hull had
been removed. However, since the similar lipase activity in
germinated dehulled oat were recorded at the pHs 4 and 5, therefore
it was concluded that pH 5 in dehulled oat seed may provide the
optimum enzyme activity for all the enzymes.
Abstract: Forty-five dairy cows were used to compare the
enzyme activity of alkaline phosphatase (ALP), lactate
dehydrogenase (LDH), α -amylase in the cervical mucus of cows
during spontaneous and induced estrus using progestagen or PGF2 α
and to determine whether these enzymes affect the fertility in cows
with induced estrus, at the time of Al. The animals were assigned to 3
groups (no treatment, a Crestar® for 12 days, a double im injection of
PGF2 α). The cows were artificially inseminated (AI). Cervical
mucus samples were collected from all cows 3 to 5 min before the
AI. The results are summarized as follows: ALP and α -amylase
activity for spontaneous estrus were similar to those for induced
estrus (P>0.05) . LDH activity levels during spontaneous and PGF2 α
induced estrus was significantly lower (P < 0.001) than that in
progestagene induced estrus groups. While no difference was found
between the first and the third groups. Our result showed a significant
difference in LDH activity levels between cows conceived with 2 or
more AI and those conceived with 1 AI. The result of this study
showed that the enzyme activity in cervical mucus is helpful for
detection of ovulation and time of AI.