soad hassan - Academia.edu (original) (raw)
Papers by soad hassan
The current study aimed to serodiagnosis of monieziasis among sheep and goats at Beni Suif govern... more The current study aimed to serodiagnosis of monieziasis among sheep and goats at Beni Suif governorate by using ELISA. Affinity chromatography was adopted to purify Moniezia (M ) expansa adult worms extract, which showed more diagnostic activity than eggs extract in the diagnosis of monieziasis by ELISA. Adult, egg extracts and the affinity partially purified antigen were characterized by SDS polyacrylamide gel electrophoresis (SDS- PAGE). M. expansa adult worms extract showed 15 bands with molecular weight ranged from 315 -12 KDa, M. expansa eggs antigen showed 11 bands with molecular weight ranged from 315 -11.5 KDa, while affinity purified fraction showed simple electrophoretic profile only 3 bands, with molecular weight 164, 88 and 67 KDa. The serodiagnostic value of the M. expansa affinity partially purified fraction was checked against antibodies collected from sheep and goats at Beni Suif governorate. Percentage of monieziasis in sheep and goats was found as 69.7%and 74.4% re...
Tropical Biomedicine, 2020
The present study was conducted to detect the therapeutic effect of Moringa oleifera and Thymus v... more The present study was conducted to detect the therapeutic effect of Moringa oleifera and Thymus vulgaris oils on hepatic coccidiosis in experimentally infected rabbits. Also, immunomodulatory effect of the two oils was detected. Twenty-four Newzealand rabbits were used in this study and divided into 4 groups; healthy rabbits, experimentally infected rabbits with Eimeria stiedae oocysts, and two infected treated groups (one with moringa (200 mg/kg) and the other with thyme (500 mg/kg) oils). The results showed highly significant reduction in oocysts shedding (P<0.001 and P<0.05) in the two infected and treated rabbits than the infected non-treated rabbits in almost all days post infection (PI). Thyme oil was more potent and stopped oocysts shedding earlier at the day 34 PI compared to moringa oil at the day 41 PI. Microscopically, there was a damage in the oocysts shed by treated rabbits. Macroscopically, the livers of thyme oil treated rabbits showed more enhancement with protection percentage 75% than those treated with moringa oil in which protection percentage was 55%. The highest titer of antibodies was detected in moringa oil treated rabbits. It was concluded that both moringa and thyme oils had an anti-coccidial effect with thyme oil superiority. So, thyme oil could be useful as an alternative product for the control of rabbit coccidiosis.
Research Journal of Parasitology, 2016
A total number of 1379 female buffaloes reared at Lower Egypt was gynecologically examined by ult... more A total number of 1379 female buffaloes reared at Lower Egypt was gynecologically examined by ultrasonography and blood and fecal samples were collected during a period of three years (2004-2006). Out of these animals 896 (64.97%) were suffering from reproductive disorders. The recorded reproductive disorders were ovarian inactivity, endometritis, delayed puberty, mastitis, repeat breeding, retained placenta and abortions, vaginal/uterine prolapses and cystic ovaries. Eimeriosis was detected by coprological examination in 62.87% of the total examined samples. The incidence was high in animals having genital disorders (81.70%) as compared with healthy animals (27.95%). High infection rate was recorded in animals suffering from inactive ovaries (87.90%), abortion (78.57%), repeat breeding (72.41%) and endometritis (66.91%). Low incidence was reported in animals suffering from mastitis (48.72%), retained placenta (51.85%) and delayed puberty (57.89%). Eimeria sp oocyst antigen was partially purified by ionexchange chromatography in which DEADE cellulose was utilized. Two isolated fractions and crude antigen were characterized by SDS polyacrylamide gel electrophoresis which showed 14 bands in crude extract. While the isolated fractions revealed simple electrophoretic profile, fraction I resolved into 8 bands of molecular weight ranged from 15.1 to 180 KD and fraction II resolved into 6 bands of molecular weight ranged from 21.2 to 87.7 KD. Employing naturally infected animal sera in immunoblot assay, six immunogenic bands were detected in crude extract, four of them were relative in FI of molecular weight ranged from 25.1 to 124.6 KD, while the immunogenicity of F II reside in only two bands of 45.9 and 28 KD. Fraction I was more sensitive than fraction II by ELISA. Hence, fraction I was utilized for diagnosis of Eimeriosis with 100% sensitivity. It could be concluded that a tight association is found between the occurrence of reproductive disorders and instantaneous infections with Eimeriosis. Fraction I of Eimeria sp.oocyst partially purified antigen could be of helpful diagnostic value.
Journal of Clinical & Cellular Immunology, 2015
Fasciolosis due to Fasciola gigantica or F. hepatica causes significant production losses in anim... more Fasciolosis due to Fasciola gigantica or F. hepatica causes significant production losses in animals, as well as being a zoonotic disease of global importance. In an attempt to develop vaccine against fasciolosis in rabbits, an immunoaffinity fraction of F. gigantica excretory secretory products was isolated. The fraction possesses 87.67% of the initial antigenic activities with 2051.5 fold increase in specific activity compared to crude extract. It consists of two bands of molecular weight 27 kDa and 23.5 kDa as revealed by SDS-PAGE. The detailed structural analyses of the pure fraction showed O-glycan [Ser-Arg-Ser-Arg-Ser-GlucNAc] using mass spectrometry. Vaccination of rabbits twice with the fraction resulted in 85% reduction in worm burden. It is also resulted in high antibody IgG levels as proved by ELISA. The highest IgG response was observed in vaccinated rabbits at two weeks post infection and remained stable to the end of the experiment. A significant expression of IL-4 and INF-γ was observed in vaccinated rabbits starting one week until thirteen weeks post infection. The level of IL-4 was significantly higher than the level of INF-γ throughout the experiment as measured by ELISA. Collectively, the current results suggest promising immunoprophylactic potentials of the immunoaffinity fraction of Fasciola gigantica excretory secretory products against fasciolosis in rabbits through induction of both cellular and humoral responses.
Veterinary World
Background and Aim: Cryptosporidiosis is a leading cause of diarrheal disease worldwide and is an... more Background and Aim: Cryptosporidiosis is a leading cause of diarrheal disease worldwide and is an animal and public health burden. This study aimed to evaluate the protective potential of affinity-purified Cryptosporidium parvum oocyst antigen as a vaccine candidate according to fecal oocyst shedding, humoral and cellular immune responses, histopathological changes, and the number of parasite developmental stages in ileal and hepatic tissues. Materials and Methods: We isolated oocysts from naturally infected buffalo calves and identified them molecularly as C. parvum isolates (GenBank: ON730707 and ON730708) by targeting the Cryptosporidium oocyst wall protein gene. We propagated the C. parvum oocysts in mice. In addition, we prepared crude antigen from the isolated oocysts by purification using cyanogen bromide-activated Sepharose-4B affinity chromatography coupled with rabbit hyperimmune serum. Then, we divided 81 parasite-free mice into three groups: (1) non-vaccinated non-infect...
Egyptian Journal of Veterinary Sciences
C RYPTOSPORIDIOSIS is a significant disease that causes diarrhea in humans and animals with relat... more C RYPTOSPORIDIOSIS is a significant disease that causes diarrhea in humans and animals with relatively high morbidity and mortality. The present study aimed to adopt a purified and potent antigen for the accurate diagnosis of cryptosporidiosis. A total of 278 animal hosts (60 newborn calves and 218 buffaloes) were used in the current study. Sixty fecal samples were collected from newborn calves aged less than one month raised in the Beni-Suef and Qalyubia governorates. The samples were examined under a microscope after modified Ziehl-Neelsen staining, and Cryptosporidium oocysts were isolated from naturally infected calves. These oocysts were used in mice experimental infection. The oocyst antigen and coproantigen were prepared from the mice feces. The diagnostic efficacy of the two prepared antigens was evaluated using an Enzyme Linked Immunosorbent Assay (ELISA) with experimentally infected mice sera. The crude oocyst antigen proved to have higher diagnostic potential than coproantigen, so, it was chosen for purification using Cyanogen Bromide-activated Sepharose-4B affinity chromatography coupled with rabbit hyperimmune serum raised against oocyst antigen. The affinity purified fraction and its crude Cryptosporidium antigen were evaluated using the ELISA. The resulting purified fraction was 6733 fold increase in binding activity compared with its crude antigen. Characterization of the isolated fraction was performed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), western blot, and amino acid analysis. SDS-PAGE clarified that the fraction contained three polypeptides of 94.7, 65, and 50 kDa, which were identified as immune-reactive components using a western blot analysis. The isolated fraction exhibited 17 amino acids and was rich in tyrosine, alanine, and phenylalanine. The affinity purified Cryptosporidium oocyst antigen effectively detected Cryptosporidium antibodies in experimentally infected mice sera and naturally infected buffalo sera with a sensitivity of 94.4% and 95.24 %, and a specificity of 100% and 93.33%, respectively. The purified fraction succeeded in diagnosis of cryptosporidiosis in 182 random serum samples collected from buffaloes with an incidence of 57.14 %. In conclusion, the affinity purified fraction of the Cryptosporidium oocyst antigen might be a good diagnostic candidate for cryptosporidiosis diagnosis and seroepidemiological surveillance.
World applied sciences journal, 2010
Isolation of immunodiagnostic fraction from Strongylus vulgaris adult worms was described by CNBr... more Isolation of immunodiagnostic fraction from Strongylus vulgaris adult worms was described by CNBr-Sepharose 4B affinity column chromatography. The isolation process resulted in a fraction with 2161.5 fold increase in binding activity compared to its crude extract. Characterization of the isolated fraction by SDS polyacrylamide gel electrophoresis and isoelectric focusing showed that the fraction consists of only two bands of 39 and 31 KDa with isoelectric points of 6.8 and 6.7. Comparative evaluation of the immunogenic binding activities of the crude extract, unbound and bound fractions by ELISA proved the potency of the bound fraction over the other two antigens. Diagnosis of S. vulgaris infection in horses by ELISA in which the bound fraction was utilized, recorded high infection percentage (73.7) as compared with the parasitological examination (27.6). The isolated fraction in the current study could be utilized successfully in the diagnosis of S. vulgaris infection using ELISA, ...
2 Abstract: Chromatographic analysis of Eimeria sp. crude antigen was done by Sephacryl S-200 col... more 2 Abstract: Chromatographic analysis of Eimeria sp. crude antigen was done by Sephacryl S-200 column. Four fractions were isolated by this approach. Comparative evaluation of the immunogenic binding activities of isolated fractions by ELISA proved the potency of fraction number one (F1) over the other three ones. Characterization of the isolated fraction (F1) by SDS polyacrylamide gel electrophoresis showed that the fraction consists of 3 bands of 225, 198, 114.5 KDa. Only one band of molecular weight 225 KDa was identified as immunogenic band by immunoblot assay in which cattle naturally infected sera were utilized. Diagnosis of Eimeria infection in cattle by ELISA in which F1 was utilized, recorded high infection percentage of 87.9% compared with coprological examination (24.2%). In conclusion: Fraction number one in the current research proved high diagnostic potential. It could be successfully utilized in the diagnosis of eimeriosis among cattle.
A developed ELISA was established to diagnose Fasciola gigantica infection in lactating buffaloes... more A developed ELISA was established to diagnose Fasciola gigantica infection in lactating buffaloes by detecting antibodies in both serum and milk samples. An immunogenic fraction was obtained from crude somatic antigen after fractionation on CNBr-Sepharose 4 B affinity column chromatography and used in detection of fasciolosis in lactating buffaloes. The prevalence of infection in 61 lactating buffaloes was established using serum and milk ELISAs compared with detection of Fasciola eggs in fecal samples. Serum ELISA gave the highest diagnostic value (68.9%) followed by milk ELISA (65.6%), while parasitological examination gave 36.1%. By SDS-PAGE, the isolated fraction resolved into five bands of molecular weights 97, 84, 65, 21 and 17 KDa. Immunoreactive bands of the isolated fraction were detected by immunoblot using naturally infected buffalo sera and positive defatted milk. Two bands of molecular weights 97 and 84 KDa were detected by positive sera while other two bands of molecul...
Toxoplasma gondii infection in humans is routinely assessed by serological means. Here, the autho... more Toxoplasma gondii infection in humans is routinely assessed by serological means. Here, the authors attempted to isolate fraction (LAb) from Toxoplasma gondii crude antigen (LA) prepared from Toxoplasma gondii strain isolated from infected horse tissues. The diagnostic activity of the isolated fraction was compared with RH strain and commercial kit antigens for detection of human toxoplasmosis. A total of 90 human serum samples was investigated by ELISA using the three compared antigens. The results showed that isolated horse fraction displayed highest diagnostic efficiency (80%) compared with RH strain (63%) and commercial kit (70%) antigens. By SDS- PAGE, LA was resolved into 11 bands of molecular weights ranged from 23- 207KDa. Five of them were detected in LAb with molecular weights ranged from (57-93KDa). Also, RH strain showed 11 bands of molecular weights (27-207 KDa). Immune-reactive bands of LA and LAb were identified by immunoblot assay using infected human sera. Two bands...
Toxoplasma gondii is a zoonotic protozoan coccidian parasite of both medical and veterinary impor... more Toxoplasma gondii is a zoonotic protozoan coccidian parasite of both medical and veterinary importance worldwide. Sero-prevalence of T. gondii infection in camels and cattle using purified antigen isolated from the crude extract of camel T. gondii tachyzoites of local strain is the purpose of the current study. Affinity purification process resulted in isolation of a single specific fraction identified by hyper immune serum of rabbit coupled with CNBr-Sepharose 4B. By SDS-PAGE, the fraction was resolved into three bands of 84, 78 and 65 KDa compared with 12 bands associated with crude extract of 114, 92, 84, 78, 65, 52, 43, 33, 26, 22, 18 and 15 KDa. The isolated fraction showed more diagnostic potency compared with crude extract by enzyme linked immune sorbent assay (ELISA) using two fold serially diluted rabbit hyper immune serum. Serological screening of apparently healthy 60 camels and 94 cattle with the isolated fraction using ELISA revealed higher prevalence of toxoplasmosis i...
Journal of Parasitic Diseases, 2019
The current study was designed to evaluate the in vivo fasciolicidal activity of Moringa (M.) ole... more The current study was designed to evaluate the in vivo fasciolicidal activity of Moringa (M.) oleifera leaf aqueous extract oral administration as well as its antibacterial activity against Clostridium (C.) novyi in sheep naturally co-infected with fascioliasis and C. novyi. Sheep naturally infected with fascioliasis were divided into 3 groups, heavily infected treated group, lightly infected treated group and mixed infection control (non-treated) group. Treatment groups were orally administered M. oleifera leaves aqueous extract at a dose of 150 mg/kg every 48 h for 21 days. Animal body weights, fecal egg count, serum levels of anti-Fasciola IgG, cytokines (IL-2, IL-17, IL-10), and bacterial count of C. novyi were evaluated. The results showed that treatment with M. oleifera improved the body weight gain and decreased fecal egg count in lightly and heavily infected groups compared to the nontreated group with 100% reduction in egg count in lightly infected sheep. Furthermore, the treatment with M. oleifera significantly reduced the serum levels of IgG, IL-2, and IL-17. Interestingly, elevated levels of IL-10 were recorded in both heavily and lightly infected sheep. The treatment with Moringa extract significantly decreased the fecal bacterial count of C. novyi in both heavily and lightly infected groups. In conclusion, this study highlights the potential beneficial effects of M. oleifera leaf against Fasciola (F.) gigantica and C. novyi.
Journal of Parasitic Diseases, 2019
This model could help to understand the regulation of intestinal secretory and inflammatory respo... more This model could help to understand the regulation of intestinal secretory and inflammatory responses, and could be useful for the screening of potential anticryptosporidial candidate compounds. Keywords Cryptosporidium parvum Á Buffaloes Á Tissue explant Á Moringa Á Scanning electron microscopy Á IFN-c Á IL-12 Á IL-14
![Research paper thumbnail of {"__content__"=>"Comparative ovicidal activity of leaf extracts on eggs.", "i"=>[{"__content__"=>"Moringa oleifera"}, {"__content__"=>"Fasciola gigantica"}]}](https://attachments.academia-assets.com/112357518/thumbnails/1.jpg)
](Veterinary world, 2018
Fasciolosis is an important zoonotic disease affecting the productive performance of farm animals... more Fasciolosis is an important zoonotic disease affecting the productive performance of farm animals in Egypt. The aim of the present study was comparing the ovicidal effect of different extracts as an alcoholic (Methanolic and Ethanolic) and aqueous leaf extracts on non-embryonated and developed eggs. Tested concentrations of extracts ranged from 12.5 to 800 mg/ml. Nitroxynil was used as reference drug with a dose of 100 mg/ml. alcoholic and aqueous extracts showed a concentration-dependent ovicidal effect on non-embryonated and developed eggs. Based on LC values, water extract showed the highest ovicidal activity since it registered the lowest values of 2.6 mg/ml on non-embryonated eggs. Non-embryonated eggs were more susceptible to aqueous extract than developed eggs. On the other hand, the developed eggs were more susceptible to ethanolic extract than non-embryonated eggs even the lowest LC (12.38 mg/ml). leaf extracts especially aqueous extract could be a promising step in the fie...
Due to its role in human Trichinellosis, there are increasing global requirements for reliable di... more Due to its role in human Trichinellosis, there are increasing global requirements for reliable diagnostic method for Trichinella spiralis infection in pigs. Consequently, there is a need for selection of potent diagnostic antigen that could be successfully utilized in the diagnosis. Currently, two antigens (somatic and excretory-secretory antigens of T. spiralis larvae) were evaluated for the diagnosis of pig trichinellosis using ELISA. The ES antigen exhibited highest diagnostic potential (88.3%) as compared to somatic antigen (68.3%). The diagnostic potential of ES antigen is attributed to five immunogenic bands identified by infected pig sera in immunoblot. These bands are of molecular weight 127, 58, 35, 20.5 and 12 KDa. While in somatic antigen six immunogenic bands of molecular weight 230, 89, 58, 27, 15 and 6 KDa were identified. In conclusion, monitoring for T. spiralis antibodies in swine farms should be based on screening pig sera by means of ELISA in which larval ES antigen was utilized.
International Journal of Zoological Research, 2016
Research Journal of Parasitology, 2016
The Journal of Zoology Studies, 2014
Moniezia expansa could be considered as the most important cestode parasites infecting sheep caus... more Moniezia expansa could be considered as the most important cestode parasites infecting sheep causing monieziasis which constitutes a problem in sheep breeding. The aim of the present study is to investigate on the anatomical cellular and tissue structure and development of the interproglottidal gland and tegument correspondig to the mechanism of its secretion. The detailed structures of the tegument and microtriches could not be seen by light microscoy examination and also the interproglottidal glands appear as interupted non differential cellular chain in between worm proglottides or segments. The ultrastructure study done by transmission electron microscopy (TEM) revealed that the glands develop separately from the tegument and compromising an endocrine character. It was observed that the gland consists of subglobular follicles or vesicles, which are embedded in parenchyma and pushing the tegument at the posterior border of the proglottid, formulated crypt-like structures with cen...
Journal of the Egyptian Society of Parasitology, 2008
Activity-guided fractionation of the methanol extract of Hedera canariensis (var. Gloire de Marin... more Activity-guided fractionation of the methanol extract of Hedera canariensis (var. Gloire de Maringo) Wild leaves afforded five saponins 1-5. Chemical and spectral methods (MS, 1HNMR, 13CNMR) showed that they are glycosides of hederagenin and oleanolic acid. The results showed that 4,5 exhibited molluscicidal properties, compound 1 was inactive. Mortality rate of exposed snails increased by increasing plant extract concentration. Lymnaea cailliaudi was more sensitive to plant extract than Biomphalaria alexandrina. The histopathological study revealed distinct damage in the structure of the stomach and ovotestis of treated L. cailliaudi snails. Saponin content was determined in term of haemolytic index.
The current study aimed to serodiagnosis of monieziasis among sheep and goats at Beni Suif govern... more The current study aimed to serodiagnosis of monieziasis among sheep and goats at Beni Suif governorate by using ELISA. Affinity chromatography was adopted to purify Moniezia (M ) expansa adult worms extract, which showed more diagnostic activity than eggs extract in the diagnosis of monieziasis by ELISA. Adult, egg extracts and the affinity partially purified antigen were characterized by SDS polyacrylamide gel electrophoresis (SDS- PAGE). M. expansa adult worms extract showed 15 bands with molecular weight ranged from 315 -12 KDa, M. expansa eggs antigen showed 11 bands with molecular weight ranged from 315 -11.5 KDa, while affinity purified fraction showed simple electrophoretic profile only 3 bands, with molecular weight 164, 88 and 67 KDa. The serodiagnostic value of the M. expansa affinity partially purified fraction was checked against antibodies collected from sheep and goats at Beni Suif governorate. Percentage of monieziasis in sheep and goats was found as 69.7%and 74.4% re...
Tropical Biomedicine, 2020
The present study was conducted to detect the therapeutic effect of Moringa oleifera and Thymus v... more The present study was conducted to detect the therapeutic effect of Moringa oleifera and Thymus vulgaris oils on hepatic coccidiosis in experimentally infected rabbits. Also, immunomodulatory effect of the two oils was detected. Twenty-four Newzealand rabbits were used in this study and divided into 4 groups; healthy rabbits, experimentally infected rabbits with Eimeria stiedae oocysts, and two infected treated groups (one with moringa (200 mg/kg) and the other with thyme (500 mg/kg) oils). The results showed highly significant reduction in oocysts shedding (P<0.001 and P<0.05) in the two infected and treated rabbits than the infected non-treated rabbits in almost all days post infection (PI). Thyme oil was more potent and stopped oocysts shedding earlier at the day 34 PI compared to moringa oil at the day 41 PI. Microscopically, there was a damage in the oocysts shed by treated rabbits. Macroscopically, the livers of thyme oil treated rabbits showed more enhancement with protection percentage 75% than those treated with moringa oil in which protection percentage was 55%. The highest titer of antibodies was detected in moringa oil treated rabbits. It was concluded that both moringa and thyme oils had an anti-coccidial effect with thyme oil superiority. So, thyme oil could be useful as an alternative product for the control of rabbit coccidiosis.
Research Journal of Parasitology, 2016
A total number of 1379 female buffaloes reared at Lower Egypt was gynecologically examined by ult... more A total number of 1379 female buffaloes reared at Lower Egypt was gynecologically examined by ultrasonography and blood and fecal samples were collected during a period of three years (2004-2006). Out of these animals 896 (64.97%) were suffering from reproductive disorders. The recorded reproductive disorders were ovarian inactivity, endometritis, delayed puberty, mastitis, repeat breeding, retained placenta and abortions, vaginal/uterine prolapses and cystic ovaries. Eimeriosis was detected by coprological examination in 62.87% of the total examined samples. The incidence was high in animals having genital disorders (81.70%) as compared with healthy animals (27.95%). High infection rate was recorded in animals suffering from inactive ovaries (87.90%), abortion (78.57%), repeat breeding (72.41%) and endometritis (66.91%). Low incidence was reported in animals suffering from mastitis (48.72%), retained placenta (51.85%) and delayed puberty (57.89%). Eimeria sp oocyst antigen was partially purified by ionexchange chromatography in which DEADE cellulose was utilized. Two isolated fractions and crude antigen were characterized by SDS polyacrylamide gel electrophoresis which showed 14 bands in crude extract. While the isolated fractions revealed simple electrophoretic profile, fraction I resolved into 8 bands of molecular weight ranged from 15.1 to 180 KD and fraction II resolved into 6 bands of molecular weight ranged from 21.2 to 87.7 KD. Employing naturally infected animal sera in immunoblot assay, six immunogenic bands were detected in crude extract, four of them were relative in FI of molecular weight ranged from 25.1 to 124.6 KD, while the immunogenicity of F II reside in only two bands of 45.9 and 28 KD. Fraction I was more sensitive than fraction II by ELISA. Hence, fraction I was utilized for diagnosis of Eimeriosis with 100% sensitivity. It could be concluded that a tight association is found between the occurrence of reproductive disorders and instantaneous infections with Eimeriosis. Fraction I of Eimeria sp.oocyst partially purified antigen could be of helpful diagnostic value.
Journal of Clinical & Cellular Immunology, 2015
Fasciolosis due to Fasciola gigantica or F. hepatica causes significant production losses in anim... more Fasciolosis due to Fasciola gigantica or F. hepatica causes significant production losses in animals, as well as being a zoonotic disease of global importance. In an attempt to develop vaccine against fasciolosis in rabbits, an immunoaffinity fraction of F. gigantica excretory secretory products was isolated. The fraction possesses 87.67% of the initial antigenic activities with 2051.5 fold increase in specific activity compared to crude extract. It consists of two bands of molecular weight 27 kDa and 23.5 kDa as revealed by SDS-PAGE. The detailed structural analyses of the pure fraction showed O-glycan [Ser-Arg-Ser-Arg-Ser-GlucNAc] using mass spectrometry. Vaccination of rabbits twice with the fraction resulted in 85% reduction in worm burden. It is also resulted in high antibody IgG levels as proved by ELISA. The highest IgG response was observed in vaccinated rabbits at two weeks post infection and remained stable to the end of the experiment. A significant expression of IL-4 and INF-γ was observed in vaccinated rabbits starting one week until thirteen weeks post infection. The level of IL-4 was significantly higher than the level of INF-γ throughout the experiment as measured by ELISA. Collectively, the current results suggest promising immunoprophylactic potentials of the immunoaffinity fraction of Fasciola gigantica excretory secretory products against fasciolosis in rabbits through induction of both cellular and humoral responses.
Veterinary World
Background and Aim: Cryptosporidiosis is a leading cause of diarrheal disease worldwide and is an... more Background and Aim: Cryptosporidiosis is a leading cause of diarrheal disease worldwide and is an animal and public health burden. This study aimed to evaluate the protective potential of affinity-purified Cryptosporidium parvum oocyst antigen as a vaccine candidate according to fecal oocyst shedding, humoral and cellular immune responses, histopathological changes, and the number of parasite developmental stages in ileal and hepatic tissues. Materials and Methods: We isolated oocysts from naturally infected buffalo calves and identified them molecularly as C. parvum isolates (GenBank: ON730707 and ON730708) by targeting the Cryptosporidium oocyst wall protein gene. We propagated the C. parvum oocysts in mice. In addition, we prepared crude antigen from the isolated oocysts by purification using cyanogen bromide-activated Sepharose-4B affinity chromatography coupled with rabbit hyperimmune serum. Then, we divided 81 parasite-free mice into three groups: (1) non-vaccinated non-infect...
Egyptian Journal of Veterinary Sciences
C RYPTOSPORIDIOSIS is a significant disease that causes diarrhea in humans and animals with relat... more C RYPTOSPORIDIOSIS is a significant disease that causes diarrhea in humans and animals with relatively high morbidity and mortality. The present study aimed to adopt a purified and potent antigen for the accurate diagnosis of cryptosporidiosis. A total of 278 animal hosts (60 newborn calves and 218 buffaloes) were used in the current study. Sixty fecal samples were collected from newborn calves aged less than one month raised in the Beni-Suef and Qalyubia governorates. The samples were examined under a microscope after modified Ziehl-Neelsen staining, and Cryptosporidium oocysts were isolated from naturally infected calves. These oocysts were used in mice experimental infection. The oocyst antigen and coproantigen were prepared from the mice feces. The diagnostic efficacy of the two prepared antigens was evaluated using an Enzyme Linked Immunosorbent Assay (ELISA) with experimentally infected mice sera. The crude oocyst antigen proved to have higher diagnostic potential than coproantigen, so, it was chosen for purification using Cyanogen Bromide-activated Sepharose-4B affinity chromatography coupled with rabbit hyperimmune serum raised against oocyst antigen. The affinity purified fraction and its crude Cryptosporidium antigen were evaluated using the ELISA. The resulting purified fraction was 6733 fold increase in binding activity compared with its crude antigen. Characterization of the isolated fraction was performed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), western blot, and amino acid analysis. SDS-PAGE clarified that the fraction contained three polypeptides of 94.7, 65, and 50 kDa, which were identified as immune-reactive components using a western blot analysis. The isolated fraction exhibited 17 amino acids and was rich in tyrosine, alanine, and phenylalanine. The affinity purified Cryptosporidium oocyst antigen effectively detected Cryptosporidium antibodies in experimentally infected mice sera and naturally infected buffalo sera with a sensitivity of 94.4% and 95.24 %, and a specificity of 100% and 93.33%, respectively. The purified fraction succeeded in diagnosis of cryptosporidiosis in 182 random serum samples collected from buffaloes with an incidence of 57.14 %. In conclusion, the affinity purified fraction of the Cryptosporidium oocyst antigen might be a good diagnostic candidate for cryptosporidiosis diagnosis and seroepidemiological surveillance.
World applied sciences journal, 2010
Isolation of immunodiagnostic fraction from Strongylus vulgaris adult worms was described by CNBr... more Isolation of immunodiagnostic fraction from Strongylus vulgaris adult worms was described by CNBr-Sepharose 4B affinity column chromatography. The isolation process resulted in a fraction with 2161.5 fold increase in binding activity compared to its crude extract. Characterization of the isolated fraction by SDS polyacrylamide gel electrophoresis and isoelectric focusing showed that the fraction consists of only two bands of 39 and 31 KDa with isoelectric points of 6.8 and 6.7. Comparative evaluation of the immunogenic binding activities of the crude extract, unbound and bound fractions by ELISA proved the potency of the bound fraction over the other two antigens. Diagnosis of S. vulgaris infection in horses by ELISA in which the bound fraction was utilized, recorded high infection percentage (73.7) as compared with the parasitological examination (27.6). The isolated fraction in the current study could be utilized successfully in the diagnosis of S. vulgaris infection using ELISA, ...
2 Abstract: Chromatographic analysis of Eimeria sp. crude antigen was done by Sephacryl S-200 col... more 2 Abstract: Chromatographic analysis of Eimeria sp. crude antigen was done by Sephacryl S-200 column. Four fractions were isolated by this approach. Comparative evaluation of the immunogenic binding activities of isolated fractions by ELISA proved the potency of fraction number one (F1) over the other three ones. Characterization of the isolated fraction (F1) by SDS polyacrylamide gel electrophoresis showed that the fraction consists of 3 bands of 225, 198, 114.5 KDa. Only one band of molecular weight 225 KDa was identified as immunogenic band by immunoblot assay in which cattle naturally infected sera were utilized. Diagnosis of Eimeria infection in cattle by ELISA in which F1 was utilized, recorded high infection percentage of 87.9% compared with coprological examination (24.2%). In conclusion: Fraction number one in the current research proved high diagnostic potential. It could be successfully utilized in the diagnosis of eimeriosis among cattle.
A developed ELISA was established to diagnose Fasciola gigantica infection in lactating buffaloes... more A developed ELISA was established to diagnose Fasciola gigantica infection in lactating buffaloes by detecting antibodies in both serum and milk samples. An immunogenic fraction was obtained from crude somatic antigen after fractionation on CNBr-Sepharose 4 B affinity column chromatography and used in detection of fasciolosis in lactating buffaloes. The prevalence of infection in 61 lactating buffaloes was established using serum and milk ELISAs compared with detection of Fasciola eggs in fecal samples. Serum ELISA gave the highest diagnostic value (68.9%) followed by milk ELISA (65.6%), while parasitological examination gave 36.1%. By SDS-PAGE, the isolated fraction resolved into five bands of molecular weights 97, 84, 65, 21 and 17 KDa. Immunoreactive bands of the isolated fraction were detected by immunoblot using naturally infected buffalo sera and positive defatted milk. Two bands of molecular weights 97 and 84 KDa were detected by positive sera while other two bands of molecul...
Toxoplasma gondii infection in humans is routinely assessed by serological means. Here, the autho... more Toxoplasma gondii infection in humans is routinely assessed by serological means. Here, the authors attempted to isolate fraction (LAb) from Toxoplasma gondii crude antigen (LA) prepared from Toxoplasma gondii strain isolated from infected horse tissues. The diagnostic activity of the isolated fraction was compared with RH strain and commercial kit antigens for detection of human toxoplasmosis. A total of 90 human serum samples was investigated by ELISA using the three compared antigens. The results showed that isolated horse fraction displayed highest diagnostic efficiency (80%) compared with RH strain (63%) and commercial kit (70%) antigens. By SDS- PAGE, LA was resolved into 11 bands of molecular weights ranged from 23- 207KDa. Five of them were detected in LAb with molecular weights ranged from (57-93KDa). Also, RH strain showed 11 bands of molecular weights (27-207 KDa). Immune-reactive bands of LA and LAb were identified by immunoblot assay using infected human sera. Two bands...
Toxoplasma gondii is a zoonotic protozoan coccidian parasite of both medical and veterinary impor... more Toxoplasma gondii is a zoonotic protozoan coccidian parasite of both medical and veterinary importance worldwide. Sero-prevalence of T. gondii infection in camels and cattle using purified antigen isolated from the crude extract of camel T. gondii tachyzoites of local strain is the purpose of the current study. Affinity purification process resulted in isolation of a single specific fraction identified by hyper immune serum of rabbit coupled with CNBr-Sepharose 4B. By SDS-PAGE, the fraction was resolved into three bands of 84, 78 and 65 KDa compared with 12 bands associated with crude extract of 114, 92, 84, 78, 65, 52, 43, 33, 26, 22, 18 and 15 KDa. The isolated fraction showed more diagnostic potency compared with crude extract by enzyme linked immune sorbent assay (ELISA) using two fold serially diluted rabbit hyper immune serum. Serological screening of apparently healthy 60 camels and 94 cattle with the isolated fraction using ELISA revealed higher prevalence of toxoplasmosis i...
Journal of Parasitic Diseases, 2019
The current study was designed to evaluate the in vivo fasciolicidal activity of Moringa (M.) ole... more The current study was designed to evaluate the in vivo fasciolicidal activity of Moringa (M.) oleifera leaf aqueous extract oral administration as well as its antibacterial activity against Clostridium (C.) novyi in sheep naturally co-infected with fascioliasis and C. novyi. Sheep naturally infected with fascioliasis were divided into 3 groups, heavily infected treated group, lightly infected treated group and mixed infection control (non-treated) group. Treatment groups were orally administered M. oleifera leaves aqueous extract at a dose of 150 mg/kg every 48 h for 21 days. Animal body weights, fecal egg count, serum levels of anti-Fasciola IgG, cytokines (IL-2, IL-17, IL-10), and bacterial count of C. novyi were evaluated. The results showed that treatment with M. oleifera improved the body weight gain and decreased fecal egg count in lightly and heavily infected groups compared to the nontreated group with 100% reduction in egg count in lightly infected sheep. Furthermore, the treatment with M. oleifera significantly reduced the serum levels of IgG, IL-2, and IL-17. Interestingly, elevated levels of IL-10 were recorded in both heavily and lightly infected sheep. The treatment with Moringa extract significantly decreased the fecal bacterial count of C. novyi in both heavily and lightly infected groups. In conclusion, this study highlights the potential beneficial effects of M. oleifera leaf against Fasciola (F.) gigantica and C. novyi.
Journal of Parasitic Diseases, 2019
This model could help to understand the regulation of intestinal secretory and inflammatory respo... more This model could help to understand the regulation of intestinal secretory and inflammatory responses, and could be useful for the screening of potential anticryptosporidial candidate compounds. Keywords Cryptosporidium parvum Á Buffaloes Á Tissue explant Á Moringa Á Scanning electron microscopy Á IFN-c Á IL-12 Á IL-14
Veterinary world, 2018
Fasciolosis is an important zoonotic disease affecting the productive performance of farm animals... more Fasciolosis is an important zoonotic disease affecting the productive performance of farm animals in Egypt. The aim of the present study was comparing the ovicidal effect of different extracts as an alcoholic (Methanolic and Ethanolic) and aqueous leaf extracts on non-embryonated and developed eggs. Tested concentrations of extracts ranged from 12.5 to 800 mg/ml. Nitroxynil was used as reference drug with a dose of 100 mg/ml. alcoholic and aqueous extracts showed a concentration-dependent ovicidal effect on non-embryonated and developed eggs. Based on LC values, water extract showed the highest ovicidal activity since it registered the lowest values of 2.6 mg/ml on non-embryonated eggs. Non-embryonated eggs were more susceptible to aqueous extract than developed eggs. On the other hand, the developed eggs were more susceptible to ethanolic extract than non-embryonated eggs even the lowest LC (12.38 mg/ml). leaf extracts especially aqueous extract could be a promising step in the fie...
Due to its role in human Trichinellosis, there are increasing global requirements for reliable di... more Due to its role in human Trichinellosis, there are increasing global requirements for reliable diagnostic method for Trichinella spiralis infection in pigs. Consequently, there is a need for selection of potent diagnostic antigen that could be successfully utilized in the diagnosis. Currently, two antigens (somatic and excretory-secretory antigens of T. spiralis larvae) were evaluated for the diagnosis of pig trichinellosis using ELISA. The ES antigen exhibited highest diagnostic potential (88.3%) as compared to somatic antigen (68.3%). The diagnostic potential of ES antigen is attributed to five immunogenic bands identified by infected pig sera in immunoblot. These bands are of molecular weight 127, 58, 35, 20.5 and 12 KDa. While in somatic antigen six immunogenic bands of molecular weight 230, 89, 58, 27, 15 and 6 KDa were identified. In conclusion, monitoring for T. spiralis antibodies in swine farms should be based on screening pig sera by means of ELISA in which larval ES antigen was utilized.
International Journal of Zoological Research, 2016
Research Journal of Parasitology, 2016
The Journal of Zoology Studies, 2014
Moniezia expansa could be considered as the most important cestode parasites infecting sheep caus... more Moniezia expansa could be considered as the most important cestode parasites infecting sheep causing monieziasis which constitutes a problem in sheep breeding. The aim of the present study is to investigate on the anatomical cellular and tissue structure and development of the interproglottidal gland and tegument correspondig to the mechanism of its secretion. The detailed structures of the tegument and microtriches could not be seen by light microscoy examination and also the interproglottidal glands appear as interupted non differential cellular chain in between worm proglottides or segments. The ultrastructure study done by transmission electron microscopy (TEM) revealed that the glands develop separately from the tegument and compromising an endocrine character. It was observed that the gland consists of subglobular follicles or vesicles, which are embedded in parenchyma and pushing the tegument at the posterior border of the proglottid, formulated crypt-like structures with cen...
Journal of the Egyptian Society of Parasitology, 2008
Activity-guided fractionation of the methanol extract of Hedera canariensis (var. Gloire de Marin... more Activity-guided fractionation of the methanol extract of Hedera canariensis (var. Gloire de Maringo) Wild leaves afforded five saponins 1-5. Chemical and spectral methods (MS, 1HNMR, 13CNMR) showed that they are glycosides of hederagenin and oleanolic acid. The results showed that 4,5 exhibited molluscicidal properties, compound 1 was inactive. Mortality rate of exposed snails increased by increasing plant extract concentration. Lymnaea cailliaudi was more sensitive to plant extract than Biomphalaria alexandrina. The histopathological study revealed distinct damage in the structure of the stomach and ovotestis of treated L. cailliaudi snails. Saponin content was determined in term of haemolytic index.