Background Honey bee venom contains various enzymes with wide medical and pharmaceutical applications. Results The phospholipase A2 (PLA2) has been apparently purified from the venom of Egyptian honey bee (Apis mellifera lamarckii) 8.9-fold to a very high specific activity of 6033 U/mg protein using DEAE–cellulose and Sephacryl S-300 columns. The purified bee venom PLA2 is monomeric 16 kDa protein and has isoelectric point (pI) of 5.9. The optimal activity of bee venom PLA2 was attained at pH 8 and 45 °C. Cu2+, Ni2+, Fe2+, Ca2+, and Co2+ exhibited a complete activating effect on it, while Zn2+, Mn2+, NaN3, PMSF, N-Methylmaleimide, and EDTA have inhibitory effect. Conclusions The purified bee venom PLA2 exhibited anti-platelet aggregation and anti-coagulation activities which makes it promising agent for developing novel anti-clot formation drugs in future.

Background: Cystic echinococcosis (CE), a zoonotic disease that affects animal and human health, is of increasing economic importance due to high morbidity rates and high economic losses in the livestock industry. Aim: The present study was conducted to purify the antigen from hydatid cyst fluid (HCF) with high diagnostic efficacy of camel hydatidosis using indirect enzyme-linked immunosorbent assay (ELISA). Materials and Methods: The HCF antigen was purified using Sephacryl S-300 column chromatography. Characterization of fractions was performed using reducing and non-reducing sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. Further, antibodies against Echinococcus granulosus cysts in camel serum were detected using indirect ELISA. Results: The purification process resulted in three fractions of antigens: FI, FII, and FIII. Indirect ELISA showed that higher diagnostic efficacy was observed in FI than in FII and FIII. Indirect ELISA, in which FI was utilized, showed 88% sensitivity and 91.7% specificity. Non-reducing SDS-PAGE showed that FI had two bands of molecular weights 120 and 60 kDa. Western blot analysis of FI demonstrated that 60, 38, and 22 kDa were antigenic bands when reacted with naturally infected camel sera with E. granulosus cysts. Using indirect ELISA, F1 recorded an infection percentage of 81.7% in randomly collected camel serum samples. Conclusion: FI is a promising antigen for accurate diagnosis of camel CE using indirect ELISA.

The effects of two probiotics on NH3 degradation, as well as the magnetic field (21.56 m tesla) on the germination and proliferation of Bacillus spores, were studied in-vitro. Additionally, the effect of these probiotics on water quality maintenance in Litopenaeus vannamei holding ponds was investigated. For 180 min, NH3 degradation was assessed as follows: Set 1: ammonia-free tap water with NH3; Probiotic A (5 × 1010 viable Bacillus spores/g) with NH3; Probiotic B (multi spp. 2 × 109 CFU/g) with NH3; and Set 2: same as set 1 with 30 mg L−1 OM. The magnetic field was tested on Probiotic A (3.5 × 107 CFU) for 36 h in triplicate. In the presence of organic matter, both probiotics degrade NH3. The viable Bacillus count increased within 6 h of being exposed to the magnetic field, reaching its peak after 36 h. Firstly, fifteen ponds (250,000 PL/acre) were investigated, then 360 water samples were collected from the same corresponding pond for 8 weeks, and subjected to T1: control; T2: Probiotic A (0.007 g/m3/2 weeks); T3: Probiotic B (0.03 g/m3/2 weeks). Both probiotics with TVC and NH3 demonstrated a negative correlation, on the other hand, they showed a significant (P ≤ 0.01) improvement in DO and pH. Overall, both probiotics were able to degrade NH3 and the magnetic field (21.56 m tesla) was efficient to improve the germination and proliferation of Bacillus spores in-vitro. Probiotics were also effective for reducing TVC and NH3 levels by increasing dissolved oxygen and pH in pond water.

No abstract

Schistosoma haematobium soluble egg antigens (SH SEAs) induce intense granulomas in human hosts that often culminate in severe disease. In an attempt to identify the SH SEA fractions that are responsible for pathology, we combined T-cell Western blotting and an in vitro model of granuloma formation. Whole SH SEAs were dotted onto nitrocellulose pieces or were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrotransferred onto nitrocellulose paper. Horizontal strips bearing the separated antigens were solubilized in dimethylsulfoxide and precipitated in carbonate/bicarbonate buffer. Antigen-free and antigen-bearing particles were used to stimulate peripheral blood mononuclear cells (PBMCs) obtained from S. haematobium-infected patients and sex- and age-matched healthy controls to form granulomas in vitro. Whole SH SEA-bearing nitrocellulose particles elicited in vitro formation of granulomas by PBMCs from infected donors. The response was similar in sensitivity, specificity, and reproducibility to that evoked by SH SEA-bound polyacrylamide beads. The results obtained in samples form 30 patients and 10 controls tested with SH SEA-separated fractions revealed that SEA bands of 84,000, 63,000, 57,000, 55,000, 40,000, 30,000, and 28,000 Da elicited in vitro granuloma reactions by PBMCs of almost all infected patients. Conversely, separated soluble adult-worm antigens failed to stimulate PBMCs of infected patients to form granulomas. This study is the first to identify the SH SEA fractions that evoke in vitro granuloma formation and represents an initial step toward the development of an anti-urinary schistosomiasis pathology vaccine.

Xanthine oxidase is a commercially important enzyme with wide area of medical applications to develop diagnostic kits. Xanthine oxidase was extracted, purified and characterized from sheep liver (SLXO). The purification procedure involved acetone precipitation and chromatography on DEAE-cellulose and Sephacryl S-300 columns. The sheep liver xanthine oxidase was homogeneously purified 31.8 folds with 3.5 U/mg specific activity and 24.1% recovery. SLXO native molecular weight was 150 kDa and on SDS-PAGE appeared as single major band of 75 kDa representing a homodimer protein. Isoelectric focusing of the purified SLXO resolved into two closely related isoforms with pI values of 5.6 and 5.8. The apparent Km for xanthine oxidase at optimum pH 7.6 was found to be 0.9 mM xanthine. FeCl2 and NiCl2 increased the activity of SLXO, while CuCl2 and ZnCl2 were found to be potent inhibitors of the purified enzyme. Allopurinol inhibits SLXO competitively with one binding site on the purified molecule and Ki value of 0.06 mM.

AbstractThe hyaluronidase enzyme has been used in many such fields of medicine as ophthalmology, orthopaedia, internal medicine, gynecology, surgery, oncology and dermatology. In this study, the hyaluronidase enzyme was purified and characterized for the first time from Egyptian bee venom homogeneously using DEAE-cellulose and Sephacryl S-300 columns. Bee venom hyaluronidase specific activity was 411.7 units/mg protein with 49.9% yield and 3.23-fold purification. The molecular weight of the purified bee venom hyaluronidase native form was 37 kDa. The purified enzyme was found homogeneous on native PAGE and SDS-PAGE, with two congruent subunits of 18.4 kDa and isoelectric point (pI) of 8.6–8.8. The enzyme was found to be stable over a wide range of temperature (20–60°C) and pH (4.5–6.5), and its optimum activity at 37°C, pH 5.4 and 0.15 M NaCl. Km for bee venom hyaluronidase was 0.029 mg/ml hyaluronic acid and its activity was elevated in presence of MgCl2 and ZnCl2 and lowered in presence of FeCl2. Heparin inhibited the hyaluronidase enzyme noncompetitively with a Ki value of 2.9 units heparin and one binding site on the enzyme molecule.

Background Faba bean attacked by soil-borne pathogens causing root rot disease. This disease has serious damage to both plant stand and produced yield. The present study aimed to evaluate effectiveness of the bioagents; Trichoderma harzianum and some plant resistance inducers as fungicide alternatives against root rot disease incidence at both pre- and post-emergence growth stages. Results Under open greenhouse conditions, the incidence of faba bean root rot in pre- and post-emergence growth phases was considerably reduced by using six inorganic salts and five antioxidants individually or combining with each other or with the bio-stimulator T. harzianum that exceeded the used fungicide, Rhizolex-T. Application of enervit agitated the highest significant defensive impact during pre-emergence stage versus root rot incidence (5.0%), followed by calcium sulfate and [cysteine + T. harzianum] (6.7%). At post-emergence stage, majority of the treatments completely suppressed (100.0%) root rot incidence, except vitamax plus and the fungicide (Rizolex-T) which expressed by 91.7 and 18.8%, respectively. Duplicate irrigations of 23 treatments after faba bean dressing improved the synthesis of different protein contents with the 2nd of which enhanced higher protein contents than the 1st one, except [T. harzianum + vitamin E + vitamin C + enervit + selenium + vitamax plus], [T. harzianum + vitamax plus] and cysteine. Disodium phosphate induced the highest catalase (CAT) activity (1820.8 and 1677.2 U/g FWt) after both irrigations. [T. harzianum + vitamax plus] and vitamin E induced the highest peroxidase (POD) activity 217.4 and 356.9 U/g FWt after 1st and 2nd irrigations, respectively. Disodium phosphate and [T. harzianum + vitamin E + vitamin C + enervit + selenium + vitamax plus] induced the highest chitinase (CHIA) activity 52.8 and 54.4 U/g FWt after 1st and 2nd irrigations, respectively. Application of disodium phosphate, calcium sulfate, potassium metabisulfite, sodium sulfate, cysteine, [cysteine + potash alum], enervit, vitamin E, [vitamin E + vitamin C + enervit + selenium + vitamax plus], [T. harzianum + enervit], [T. harzianum + selenium], [T. harzianum + vitamin E], [T. harzianum + vitamin E + vitamin C + enervit + selenium + vitamax plus] and vitamin C stimulated the formation of new protein bands on SDS-PAGE after the 2nd irrigation treatment. Conclusions Such treatments are considered good and environmentally safe alternatives against root diseases for getting rid of the negative effects of fungicides.