Meta-analysis of Genetic Diversity of the VP1 Gene Among the Circulating O, A, and SAT2 Serotypes and Vaccine Strains of FMD Virus in Egypt.

INTRODUCTION: Three strains of the FMD virus (A, O, and SAT 2) were recognised as causes of the FMD circulating in Egypt. The aims of this study were to trace the FMDV isolates from outbreaks in Egypt to understand their epidemiology and evolution and to understand the situation of the vaccine strains compared with the circulating serotypes. MATERIAL AND METHODS: A meta-analysis was carried out by using the data available for FMD outbreaks in Egypt from GenBank and the World Reference Laboratory for Foot-and-Mouth Disease (WRLFMD); a comparison was done with both data sets for the three serotypes. MEGA-X was used for the evolution analysis, through constructions of phylogenetic trees for all sequences recorded in GenBank for each serotype in different Egyptian outbreaks in different years and also within the same year. Additionally, nucleotide substitution rate, molecular clock, and mean evolutionary rates were estimated for the three serotypes to understand and compare their evolution. RESULTS: Absence of some records of certain serotype outbreaks from the WRLFMD database was noted as were subsequent missing appropriate vaccine programmes. Genetic variation was recorded among the virus isolates within the same years and also the vaccine strain was associated with up to 26 amino acid substitutions. The evolution rate of the SAT2 strain was the highest of the circulating strains. SAT2 had high amino acid substitution per year at an important immunogenic site (130-170), serotype A had less, and serotype O the least. CONCLUSION: The need for different strategies for vaccine serotype selection is indicated.

Serological evaluation for the current epidemic situation of foot and mouth disease among cattle and buffaloes in Egypt.

AIM: The present study was aimed to investigate the epidemic situation of foot-and-mouth disease (FMD) in Egypt from 2016 to 2018 based on the detection of FMD virus (FMDV) in carrier or previously infected animals, by determination of antibodies against non-structural protein (NSP), implementation a pilot study on circulating FMDV serotypes and assure the efficacy of locally produced inactivated trivalent vaccine. MATERIALS AND METHODS: A total of 1500 sera were collected from apparent healthy vaccinated cattle and buffaloes from three Egyptian geographical sectors, representing ten governorates. Determination of FMD antibodies against NSP was carried out using 3ABC enzyme-linked immunosorbent assay (ELISA) test. Serotyping of the circulating FMDV and assure the vaccine efficacy was performed using solid-phase competitive ELISA. RESULTS: The 3ABC ELISA test revealed 26.4% and 23.7% positive for FMDV-NSP antibodies in cattle and buffalo sera, respectively. The highest positivity was in Delta Sector among both cattle 42.3% and buffaloes 28.8%. Serotyping of FMDV-positive NSP sera in El-Qalyubia Governorate for the circulating FMDV serotypes O, A, and Southern African Territories (SAT) 2 was 52.2%, 17.4%, and 30.4% in cattle and 31.8%, 27.3%, and 40.9% in buffaloes, respectively. The overall protection level due to the vaccination program was 62.1 and 60.9% in cattle and buffaloes, respectively, while the protective level of the FMDV serotypes O, A, and SAT2 included in the inactivated trivalent vaccine was 73.9, 84.6, and 63.8% in cattle and 72.3, 82.3, and 63.5% in buffaloes, respectively. CONCLUSION: The present study recommended full determination for the immunogenic relationship between the vaccine strains and the field strains to attain maximum protection against the circulating viruses.

Melanomacrophage centers in Clarias gariepinus as an immunological biomarker for toxicity of silver nanoparticles.

Although there are many applications of silver nanoparticles (Ag-NPs) in human activities, there is still little known about their potential environmental toxicity, particularly to fish. In the present study, the effects of Ag-NPs on African catfish (Clarias gariepinus) were studied using melanomacrophage centers as immunohistological biomarkers. Fish were exposed to 25 mg/L, 50 mg/L and 75 mg/L 100-nm Ag-NPs. We studied the effects on the size and number of melanomacrophage centers in all target tissues. Many histopathological alterations in those tissues were observed. The histological changes were represented as dislocation of the epithelium, dilatation of central veins associated with inflammatory leukocytic infiltration, necrosis, and pyknotic nuclei of hepatocytes. There was shrinkage of Malpighian corpuscles, dislocation of nuclei of convoluted tubules, cellular degeneration, and dispersed infiltration of leukocytes in kidney tissue. Examination of spleen sections after exposure to Ag-NPs showed rupture within the red pulp and hemorrhage, dislocation of nuclei, accumulation of inflammatory leukocytes, and congestion in blood vessels. In conclusion, exposure to Ag-NPs induced alterations in tissues, suggesting a possible increase in oxidative stress in those tissues.

Supervised learning of perceptron and output feedback dynamic networks: a feedback analysis via the small gain theorem.

This paper provides a time-domain feedback analysis of the perceptron learning algorithm and of training schemes for dynamic networks with output feedback. It studies the robustness performance of the algorithms in the presence of uncertainties that might be due to noisy perturbations in the reference signals or due to modeling mismatch. In particular, bounds are established on the step-size parameters in order to guarantee that the resulting algorithms will behave as robust filters. The paper also establishes that an intrinsic feedback structure can be associated with the training schemes. The feedback configuration is motivated via energy arguments and is shown to consist of two major blocks: a time-variant lossless (i.e., energy preserving) feedforward path and a time-variant feedback path. The stability of the feedback structure is then analyzed via the small gain theorem, and choices for the step-size parameter in order to guarantee faster convergence are deduced by using the mean-value theorem. Simulation results are included to demonstrate the findings.

Serrapeptase and nattokinase intervention for relieving Alzheimer's disease pathophysiology in rat model.

Serrapeptase (SP) and nattokinase (NK) are proteolytic enzymes belonging to serine proteases. In this study, we hypothesized that SP and NK could modulate certain factors that are associated with Alzheimer's disease (AD) pathophysiology in the experimental model. Oral administration of aluminium chloride (AlCl3) in a dose of 17 mg/kg body weight (bw) daily for 45 days induced AD-like pathology in male rats with a significant increase in brain acetylcholinesterase (AchE) activity, transforming growth factor ß (TGF-ß), Fas and interleukin-6 (IL-6) levels. Meanwhile, AlCl3 supplementation produced significant decrease in brain-derived neurotrophic factor (BDNF) and insulin-like growth factor-1 (IGF-1) when compared with control values. Also, AlCl3 administration caused significant decline in the expression levels of disintegrin and metalloproteinase domain 9 (ADAM9) and a disintegrin and metalloproteinase domain 10 (ADAM10) genes in the brain. Histological investigation of brain tissue of rat model of AD showed neuronal degeneration in the hippocampus and focal hyalinosis with cellular as well as a cellular amyloid plaques formation. Oral administration of SP or NK in a rat model of AD daily for 45 days resulted in a significant decrease in brain AchE activity, TGF-ß, Fas and IL-6 levels. Also, the treatment with these enzymes produced significant increase in BDNF and IGF-1 levels when compared with the untreated AD-induced rats. Moreover, both SP and NK could markedly increase the expression levels of ADAM9 and ADAM10 genes in the brain tissue of the treated rats. These findings were well confirmed by the histological examination of the brain tissue of the treated rats. The present results support our hypothesis that the oral administration of proteolytitc enzymes, SP and/or NK, would have an effective role in modulating certain factors characterizing AD. Thus, these enzymes may have a therapeutic application in the treatment of AD.

The protective role of quince leaf extract against the adverse impacts of ultraviolet--a radiation on some tissues of Clarias gariepinus (Burchell, 1822).

In the present study the protective role of quince leaf extract against the adverse impacts of ultraviolet radiation-A (UVA) on some tissues of Clarias gariepinus (Burchell, 1822) was considered. Fishes were classified into four groups: control, UVR-treated group (for 3days/for 3h/day), UVR-treated group (for 3days/for 3h/day) with adding 10ml of quince extract, and UVR-treated group (for 3days/for 3h/day) with adding 20ml of quince leaf extract. Blood smears and sections of the liver, and skin were processed routinely for H & E paraffin embedding technique. Some UVA-induced malformations were recorded in the red blood cells including crenated cells (Cr), Acanthocytes (Ac), tear drop-like cells (Tr) and sickle cells (Sk). Also, UVA-induced disorganization of the normal architecture of hepatic tissues with lipidosis was evident. Hypertrophy and vacuolated club cells were recorded in skin exposed to UVA. In conclusion, quince leaf extract has a valuable antioxidant protective role to prevent and/or repair the histopathological changes induced by UVA.

Production of L-phenylacetyl carbinol by immobilized yeast cells: I. Batch fermentation.

Immobilization of Saccharomyces cerevisiae ATCC 834 within alginate beads enhances microbiological conversion of benzaldehyde to L-phenylacetyl carbinol (L-PAC), a precursor employed for synthesis of L-ephedrine. Yields of 90% L-PAC on benzaldehyde (initially 0.6% in medium) were obtained with immobilized cells, in contrast to about 10% with free cells which tend to form pellets in the presence of benzaldehyde. The predominant favorable action of immobilization appears to be a reduction in the toxic or inhibitory effects of benzaldehyde. With an initial benzaldehyde concentration of about 0.6% in the medium the optimum cell mass concentration was observed to be about 28 g cell mass (immobilized) per liter of medium.

Production of L-phenylacetyl carbinol by immobilized yeast cells: II. Semicontinuous fermentation.

The cyclic, semicontinuous production of L-phenylacetyl carbinol (L-PAC) from a benzaldehyde substrate by Saccharomyces cerevisiae ATCC 834 immobilized in calcium alginate beads was substantially enhanced to about 4.5 g/L in a second cycle by reactivation in fresh medium for 24 h, following an earlier 24-h period of production from substrate. Intermittent feeding of benzaldehyde was employed (four doses in 3 h). In subsequent similar cycles, however, the production returned to that produced in the first cycle, viz. L-PAC concentration of 2-3 g/L in the medium. Production of L-PAC was also increased by adaptation of the cells over 200 h of exposure to the benzaldehyde substrate (compared to wild-type cells) and by continuous (as compared to intermittent) feeding of the substrate. A liter as great as 10 g/L was obtained with wild-type cells by continuous feeding of benzaldehyde over 6 h. Immobilization not only protected the cells from toxic effects of substrate but also permitted them to be used during 7 cycles of semicontinuous operation over more than 200 h.

Comparative study of production of dextransucrase and dextran by cells of Leuconostoc mesenteroides immobilized on Celite and in calcium alginate beads.

In fed-batch fermentation, cells of L. mesenteroides immobilized on three types of Celite were used to produce dextransucrase (DS) followed by production of dextran. A layer of calcium alginate on the porous Celite R630 particles improved their mechanical stability, increased the amount of soluble DS produced and decreased the cell leakage from the highly porous support. Enzyme production with the immobilized cell cultures was significantly affected by both pore and particle size. Immobilized cultures using Celite R648 (average particle radius of 200 microm and pore size of 0.14 microm) produced the highest total enzymatic activity, followed by Celite R633, alginate-coated Celite R630, Celite R630, and then calcium alginate beads. Culture of free cells produced about 18% more total enzymatic activity than immobilized cells in calcium alginate beads, but about 64% less than immobilized cells on Celite R630. It is expected that larger amounts of enzymatic activity than measured are immobilized inside the alginate-coated Celite R630 and calcium alginate beads due to the mass transfer limitation conferred by the dextran product formed therein. The dextran yield from conversion of sucrose to dextran and fructose with all such enzyme-enriched, immobilized-cell cultures was higher than that obtained from free-cell culture under similar conditions.

Effect of beta-cyclodextrin on production of L-phenylacetyl carbinol by immobilized cells of Saccharomyces cerevisiae.

The rate and extent of microbial transformation of higher concentrations of benzaldehyde substrate to L-phenylacetyl carbinol (L-PAC) by immobilized cells of Saccharomyces cerevisiae ATCC 834 was markedly stimulated by addition of different concentrations of beta-cyclodextrin (BCD) to the fermentation medium. With 0.5, 1.0, and 1.5% BCD in the fermentation medium and cumulative doses of benzaldehyde of 12 and 14 g/L, significantly higher yields of L-PAC were obtained, about one- to twofold that of the yields of the control experiments. The favorable effects of BCD were evident in spite of its presence in stoichiometric concentrations significantly lower than those of benzaldehyde. The presence of BCD also appeared to stimulate microbial growth slightly. Enhanced cellular activity was reflected by faster D-glucose consumption and faster benzaldehyde utilization in the presence of BCD.

Production of dextransucrase by Leuconostoc mesenteroides immobilized in calcium-alginate beads: I. Batch and fed-batch fermentations.

In batch fermentation Leuconostoc mesenteroides immobilized in calcium alginate beads produced a total dextransucrase activity equal to about 93% of that by free, suspended bacterial cells under comparable conditions in a bubble column reactor. Continuous sucrose feeding (5 g/L h) to the immobilized-cell culture in the airlift bioreactor increased production of enzymatic activity by about 107% compared with ordinary batch operation of this reactor. About 14% of the enzymatic activity produced by the immobilized cells appears as soluble activity in the cell-free broth compared with about 40% in case of free cells. In an airlift bioreactor, both the soluble and the intact (sorbed and entrapped) enzymatic activity produced by the immobilized bacterial cells was about 34% greater under automatic pH control, compared to that produced in a bubble column reactor with only manual pH control. During formation of dextran by intact enzyme within cells and beads, declines are observed in apparent enzymatic activity.

Production of dextransucrase and dextran by Leuconostoc mesenteroides immobilized in calcium-alginate beads: II. Semicontinuous fed-batch fermentations.

Cells of Leuconostoc mesenteroides immobilized in calcium alginate beads were used to produce dextransucrase (DS) in three sequential cycles of semicontinuous fed-batch fermentations. Each cycle consisted of a fed-batch DS production period of 24 h followed by a batch dextran production period for another 24 h. Free, suspended cells were used in only one cycle of fed-batch DS production followed by a dextran production period. It was impractically tedious to separate and reuse free cells. Increasing sucrose feed rate from 5 to 10 g/L h led to increases of the total enzymatic activity by about 88% with immobilized cells and by about 100% with free cells. In DS fed-batch semicontinuous fermentation, total enzymatic activity produced by immobilized cells was 1.35 and 1.56 times greater than that produced by free cells with respective sucrose feeding rates of 10 and 5 g/L h. These increases in enzyme productivity with immobilized cells, however, required total overall operating times three times longer (three cycles) than with free cells (one cycle). Growing the microorganism at optimum conditions for DS production also increased the dextran yield and shortened the time of conversion of sucrose to dextran, regardless of whether the cells were free or immobilized. Moreover, during three cycles of semicontinuous operation (144 h) immobilized cells produced more than three times as much dextran as free cells during one cycle (24 h).

Investigation of production of dextran and dextransucrase by Leuconostoc mesenteroides immobilized within porous stainless steel.

Cells of Leuconostoc mesenteroides were immobilized within porus, stainless-steel (SS) supports and used for dextransucrase (DS) and dextran production. The pore size of the support significantly affected the dextran yields, which were greatest with average pore sizes of 2-5 microm. All immobilized-cell biocatalysts in porous stainless steel produced higher yields than free cells, with the exception of cells confined in submicrometer pores (0.5 microm). Coating supports of larger pore size (40 and 100 microm) with calcium alginate enhanced the cell-loading capacity of the supports and increased dextran and fructose yields in the cell-free broth. Controlled, fed-batch, DS production (activation), as a step preliminary to dextran production, significantly improved the subsequent dextran and fructose yields and shortened the time required to attain the maximum such yields. Scanning electron microscopy (SEM) of immobilized L. mesenteroides in stainless steel shows an irregular pattern of the microorganism inside the pores of the solid supports. Coating the porous solid supports with a cell-free calcium alginate layer led to an increase in the cell density inside the support. Cell growth inside the coated, porous stainless steel had no distinct growth form.