Cross-protection induced by cross-reactive antigen against Fasciola gigantica and Trichinella spiralis infections

Trichinella spiralis/Fasciola gigantica cross-reactive fraction was purified from T. spiralis larval extract by affinity column chromatography in which CNBr-Sepharose 4B was coupled with F. gigantica antibodies. The fraction consisted of six polypeptides of 191KDa, 178KDa, 149KDa, 106KDa. 101 KDa and 32KDa as revealed by SDS-PAGE. Analysis of the free amino acids of the fraction revealed 17 amino acids with high proportions of tyrosine and glutamic. Immunization of rabbits subcutaneously with the cross-reactive fraction in Freund's adjuvant, followed by challenge with F. gigantica metacercariae resulted in reduction in worm burdens reached to 47.8%. While, immunization of rats with the same fraction in Freund's adjuvant, followed by infection with T. spiralis larvae resulted in reduction in worm count reached to 74%. IgG antibody response in rabbits increased due to immunization to reach its maximum value at the time of infection and then decreased gradually up to the end of the experiments; but remained higher than the level in non vaccinated control animals. In rat sera, IgG level increased due to vaccination but the level recorded its optimum value one week post infection and then decreased. Thus, the cross-reactive antigen proved cross-protection with the protection inducing capability against both diseases

Trichinella spiralis: affinity purified antigen based diagnosis and immunoprophylaxis

The current research introduces a trial to develop vaccine candidate against trichenosis. A method of affinity chromatography was adopted to purify a Trichinella spiralis larval extract. The isolated fraction resolved into six bands of 148 KDa, 133 KDa, 118.5 KDa, 101 KDa 98.5 KDa and 79.5 KDa as observed by SDS-PAGE. The diagnostic value of this fraction was checked against antibodies regularly collected from rats experimentally infected with trichinosis compared with that of crude larval extract by ELISA. The crude extract detected the antibodies as early as one week post infection and the maximum level was recorded four weeks post infection. The advantage of the isolated fraction over the crude extract in trichinosis diagnosis was clearly observed at high serum dilution reached to 1:4000. The protective value of the isolated fraction was also investigated. Rats immunized subcutaneously with affinity purified larval extract with Freund's adjuvant showed reduction in worm burden reached to 82%. IgG antibody response in immunized rats was higher than that of control infected animals as measured by ELISA. This response might be partially responsible for the observed protection

Identification and diagnostec evaluation of cross-reactive antigen between hydatid cyst fluid of echinococcusgranulosus and trichinella spiralis larval extract

A cross reactive fraction was isolated from hydatid cyst fluid antigen of E. granulosus using CNBr Sepharose 4B affinity chromatography in which anti-T. spiralis antibodies were coupled with the column. Biochemical characterization of the isolated fraction included the use of SDS-PAGE, isoelectric focusing and amino acid analysis. The fraction showed 5 polypeptides of 165, 95.5, 63.5, 30.6 and 24 KDa. The isoelectric points of these polypeptides were 7.8, 7.2, 6.7, 6.2 and 5.7. The fraction exhibited 17 amino acids and was rich in tyrosine [20.81] and glutamic [15.28] micro gram/100 mg. The fraction proved higher potency in the diagnosis of experimental trichinellosis in rats than echinococcosis in dogs by ELISA. All experimentally infected animals reacted positively, recording 100% diagnostic sensitivity. Collectively, the present study proved that the hydatid cyst fluid cross-reactive fraction could be used in the diagnosis of trichinellosis at different intervals of infection and as early as 1 week post infection

Protective effect of eimeria stiedae copro antigen against hepatic coccidiosis in rabbits

Immunization of rabbits against hepatic coccidiosis was tried. The animals were immunized twice with Eimeria stiedae coproantigen in freund's adjuvant with two week intervals. The rabbits were challenged orally with sporulated E.steidae oocysts one week post last injection. The protection was assessed by number of oocysts output, number of focal liver lesions, clinical sings and antibody response. The immunization resulted in 70% protection from infection and decline in oocysts count. High level of IgG response in immunized rabbits than control infected ones was occurred and being responsible for the recorded protection. The electrophoretic make up of the coproantigen and oocyst antigen showed different patterns of separation. Common as well as specific bands to each antigen were identified. Using the rabbit sera after 3 weeks post vaccination in immunoblot assay, immunogenic components were detected of molecular weight 155, 103, 74, 66, 44, 22KD with coproantigen and 155, 115, 57, 26 KD with oocyst antigen. While, the rabbit sera after 2 and 4 weeks post challenge reacted with oocyts antigen, in immunoblot assay, revealing immunogenic bands of molecular weight 155, 115, 57, 26,22 KD and 155, 115, 57, 25 KD respectively. Bands of 22 KD and 155 KD are partially responsible for eliciting host protective immune response where they were recognized by immunized sera in coproantigen and by immunized infected sera in oocyst antigen

Control of hepatic coccidiosis in rabbits using calendula micrantha officinalis and peganum harmala extracts

Control of hepatic coccidiosis in rabbits using two plant extracts [Calendula micrantha officinalis and Peganum harmala] was investigated. The LD[50] values of the two plant extracts against Eimeria stiedae oocyst viability in vitro were 31ppm for C.m.officinalis extract and I73pprn for P.harmala extract. The lethal effect of C.m.officinalis extract, which proved its potency in controlling E.stiedae oocysts as judged by LD[50], was probed by experimental infection of rabbits with C.m.officinalis treated oocyts. Occyst count and score lesions were considered as strong indications of plant extract efficacy in controlling hepatic coccidiosis. The curative effect of C.m.officinalis extract on hepatic coccidiosis was investigated by treating rabbits with a dose of 30ppm/ kg body weight. The present study indicated that, this plant extract could be effective as a new biological control agent of hepatic coccidial oocysts

Cross - reaction: a common trait among helminthes

The cross-reaction between three important zoonotic helminths, Fasciola gigantica, Trichinella spiralis and Echinococcus granulosus was proved by ELISA. Cross-binding activities in the prepared antisera were strongly directed towards protoscolices and hydatid fluid antigens of E. Granulosus rather than to F. gigantica and T. spiralis antigens. Two sets of polypeptides were identified in each antigen by immunoblot [species-specific and cross-reactive]. The cross-reactive components in F. gigantica antigen were 205 KD, 178 KD, 166 KD, 106 KD, 100 KD, 65 KD, 45 KD and 32 KD. While, cross-reactive molecules in T. spiralis antigen were 205 KD, 191 KD, 166 KD, 148 KD, 132 KD and 32 KD. In protoscolices antigen, six cross-reactive components were identified [205 KD, 191 KD, 149 KD, 106 KD, 45 KD and 32 KD]. Moreover, 205 KD, 190 KD, 177 KD, 149 KD, 103 KD and 33 KD were detected in hydatid fluid antigen by heterologous antisera. Three polypeptides of 205 KD, 149 KD and 32 KD showed broad immunogenicity with the developed antisera raising the prospect of being putative common immunoprophylactic

Comparative immunodiagnostic approach of toxocariasis in buffalo calves [Jorunal Article]

Five Toxocara vitulorum antigens were utilized to diagnose natural toxocariasis in buffalo calves by ELISA. Adult antigen was proved to be the most potent one. The second potent antigen was egg antigen, followed by excretory-secretory antigen of male worms, then female worms. The coproantigen was the least potent one. The electrophoretic make-up of the antigens, examined by SDS-PAGE, revealed different patterns of separation. Common as well as specific component[s] to each antigen were identified. Employing naturally infected buffalo calf sera in immunoblot assay, five immunogenic components were detected in the adult antigen of molecular weight 191 KD, 166 KD, 102 KD, 65 KD and 54 KD. The reactive polypeptides in egg antigen were 191 KD, 105 KD, 99 KD and 79 KD. In coproantigen, six bands were identified. These components were of a molecular weight 191 KD, 178 KD, 166 KD, 124 KD, 96 KD and 65 KD. Five components of molecular weight 191 KD, 166 KD, 102 KD, 96 KD and 65 KD were immunogenic in excretory/secretory antigen of male worms. Only four polypeptides of 191 KD, 166 KD, 102 KD and 66 KD were identified in excretory/secretory female antigen

Molecular identity of major cross-reactive adult antigens in Fasciola gigantica, Toxocara vitulorum and Monieza expansa

Cross reactivity between Fasciola gigantica, Toxocara vitulorum and Moniezia expansa, whole worm extracts was proved by ELISA. Intense cross-reaction was observed between F. gigantica and M. expansa rather than between each of them and T. vitulorum. As judged by immunoblot, the cross-reactive antigens in F. gigantica which recognized by T. vitulorum antisera was 109 kD, while this component in addition to another one of 52 kD were detected by M. Expansa sera in the same extract. Furthermore, T. vitulorum antigen which cross-reacted with F. gigantica, was 133 kD and with M. expansa was 143 kD. Antigens responsible for cross-reactivity in M. expansa were 130 kD and 210 kD to T. vitulorum and F. Gigantica, respectively

Structural characterization and immunolocalization of egg antigens cross react with Toxicara vitulorum, Fasciola gigantica and moniezia expansa mature flukes

A structural homology between eggs of Toxocara vitulorum, Fasciola gigantica and Moniezia expansa was proved by the use of SDS-PAGE. In immunoblot, nine, eleven and seven polypeptides were recognized in F. gigantica, M. expansa and T. vitulorum eggs, respectively, by their respective rabbit anti-adult antisera. Moreover, components of 240 KD and 206 KD were recognized in the three eggs by different anti-adult antisera. The anatomic localization of the cross-reactive epitopes in eggs was determined by indirect immunofluorescence microscopy. The cross- reactive epitopes were mainly associated with embryonated cells of F. gigantica, egg shell, larvae and vitelline membranes of T. vitulorum and egg shell and granular layer of M. expansa

Effect of bacillus thuringiensis on the immature stage of ornithodors savingnyii [argasidae]

The potential activity for the two varieties of Bacillus thuringiensis [kurstaki and israelesis] was evaluated against fourth instar nymph of Ornithodoros savignyli. Bacillus thuringiensis [Dipel-2x] was more effective than B. thuringiensis israelensis [Vectobac]. The effect of this bacterium on the haemolymph protein alteration of previously mentioned tick using SDS-PAGE electrophoresis, and the histopathological changes of the nymphs were also studied

Studies on the molluscicidal activity of calendula micrantha officinalis [compositae] on fascioliasis transmitting snails

The molluscicidal activity of ethanolic extracts of calendula M. officinalis [flowers and leaves] was used as plant molluscicides against Lymnaea cailliaudi. The results indicated that flowers extract possessed a molluscicidal activity more than leaves extract and the LC50 was 35 and 52.17 ppm, respectively. The mortality rate of exposed snails was increased by prolongation of the exposure time. The molluscicidal effect resulted in enhancing energy utilization and nutrient consumption, since glucose, lipids proteins and triglycerids were greatly reduced. The stomach and digestive gland of the treated L. cailliaudi snails were greatly altered