The Experts below are selected from a list of 1134 Experts worldwide ranked by ideXlab platform
Hongwei Wang - One of the best experts on this subject based on the ideXlab platform.
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Effects of Diclazuril on the expression of enolase in second-generation merozoites of Eimeria tenella.
Poultry science, 2020Co-Authors: Bianhua Zhou, Hai-yan Ding, Jing-yun Yang, Jun Chai, Hong-wei Guo, Hongwei WangAbstract:Abstract Eimeria tenella is an obligate intracellular parasite of the chicken cecum; it brings huge economic loss to the chicken industry. Enolase is a multifunctional glycolytic enzyme involved in many processes of parasites, such as infection and migration. In this study, the effect of Diclazuril on the expression of enolase in second-generation merozoites of E. tenella (EtENO) was reported. The prokaryotic expression plasmid pET-28a-EtENO was constructed and transformed into Escherichia coli BL21 (DE3). Then, it was subjected to expression under the induction of isopropyl-β-D-1-thiogalactopyranoside. The expressed products were identified and purified. The purified EtENO protein was used for antibody preparation. The EtENO mRNA and protein expression levels were analyzed via real-time PCR and Western blotting. Localization of EtENO on the merozoites was examined by immunofluorescence technique. The mRNA and protein expression levels of EtENO were decreased by 36.3 and 40.36%, respectively, by Diclazuril treatment. EtENO distributed in the surface, cytoplasm, and nucleus of the infected/control group. With Diclazuril treatment, it was significantly reduced in the surface and cytoplasm and even disappeared in the nucleus of the infected/Diclazuril group. These observations suggested that EtENO may play an important role in mechanism of Diclazuril anticoccidial action and be a potential drug target for the intervention with E. tenella infection.
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Eukaryotic elongation factor 2 is involved in the anticoccidial action of Diclazuril in the second-generation merozoites of Eimeria tenella.
Veterinary parasitology, 2019Co-Authors: Bianhua Zhou, Hai-yan Ding, Jing-yun Yang, Hong-wei Guo, Liu-shu Jia, Hongwei WangAbstract:Eimeria tenella, an obligate intracellular parasite, can actively invade the cecal epithelial cells of chickens and cause severe enteric disease. Eukaryotic elongation factor 2 (eEF2) plays a major role in protein synthesis and cell survival. This study aims to explore the exact mechanisms underlying Diclazuril inhibition in second-generation merozoites of E. tenella. The eEF2 cDNA of the second-generation merozoites of E. tenella (EtEF2) was cloned by reverse transcriptase polymerase chain reaction and rapid amplification of cDNA ends. Diclazuril-induced expression profiles of EtEF2 were also analyzed. The cloned full-length cDNA (2893 bp) of the EtEF2 nucleotide sequence encompassed a 2499 bp open reading frame (ORF) that encoded a polypeptide of 832 residues with an estimated molecular mass of 93.12 kDa and a theoretical isoelectric point of 5.99. The EtEF2 nucleotide sequence was submitted to the GenBank database with the accession number KF188423. The EtEF2 protein sequence shared 99 % homology with the eEF2 sequence of Toxoplasma gondii (GenBank XP_002367778.1). The GTPase activity domain and ADP-ribosylation domain were conserved signature sequences of the eEF2 gene family. The changes in the transcriptional and translational levels of EtEF2 were detected through quantitative real-time PCR and Western blot analyses. The mRNA expression level of EtEF2 was 2.706 fold increases and the protein level of EtEF2 was increased 67.31 % under Diclazuril treatment. In addition, the localization of EtEF2 was investigated through immunofluorescence assay. Experimental results demonstrated that EtEF2 was distributed primarily in the cytoplasm of second-generation merozoites, and its fluorescence intensity was enhanced after Diclazuril treatment. These findings indicated that EtEF2 may have an important role in understanding the signaling mechanism underlying the anticoccidial action of Diclazuril and could be a promising target for novel drug exploration.
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effect of Diclazuril on the bursa of fabricius morphology and siga expression in chickens infected with eimeria tenella
Korean Journal of Parasitology, 2015Co-Authors: Bianhua Zhou, Fuwei Yuan, Erjie Tian, Hongwei WangAbstract:The effects of Diclazuril on the bursa of Fabricius (BF) structure and secretory IgA (SIgA) expression in chickens infected with Eimeria tenella were examined. The morphology of the BF was observed by hematoxylin and eosin staining, while ultrastructural changes were monitored by transmission electron microscopy. E. tenella infection caused the BF cell volumes to decrease, irregularly arranged, as well as, enlargement of the intercellular space. Diclazuril treatment alleviated the physical signs of damages associated with E. tenella infection. The SIgA expression in BF was analyzed by immunohistochemistry technique. The SIgA expression increased significantly by 350.4% (P<0.01) after E. tenella infection compared to the normal control group. With the treatment of Diclazuril, the SIgA was relatively fewer in the cortex, and the expression level was significantly decreased by 46.7% (P<0.01) compared with the infected and untreated group. In conclusion, E. tenella infection in chickens induced obvious harmful changes in BF morphological structure and stimulated the expression of SIgA in the BF. Diclazuril treatment effectively alleviated the morphological changes. This result demonstrates a method to develop an immunological strategy in coccidiosis control.
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effect of Diclazuril on intestinal morphology and siga expression in chicken infected with eimeria tenella
Parasitology Research, 2014Co-Authors: Erjie Tian, Bianhua Zhou, Jing Zhao, Xueying Wang, Wen Deng, Hongwei WangAbstract:Secretory immunoglobulin A (SIgA), as a vital actor involving in the mucosal immunity, plays a key role in defending a variety of pathogenic infections, such as bacteria, viruses and parasites. Eimeria tenella is an obligate intracellular apicomplexan parasite contacting with the digestive tract mucosa and specially parasitizes chicken caecum, causing a severe form of coccidiosis. Coccidiosis is currently mainly controlled using chemotherapeutic agents. Diclazuril, a classic coccidiostat, was used widely in the poultry industry. Because of the rising problem of drug resistance, it is therefore crucial to understand the pattern of the SIgA expression in the action of Diclazuril against E. tenella. In this study, the intestinal morphology in the caecum was analyzed by haematoxylin-eosin (HE) staining, and the SIgA expression was examined by immunohistochemical technique. At the same time, the duodenum, jejunum and ileum tissues have also been evaluated. HE staining results showed that E. tenella infection caused severe damage characterized by structural disorder, haemorrhage, inflammatory cell infiltration, serous and fibrinous exudation in chicken caecum and invisible damage in the duodenum, jejunum and ileum. With the treatment of Diclazuril, the damage in the caecum was alleviated obviously. Immunohistochemical analysis demonstrated that the SIgA level in the infected group was increased in the duodenum (p < 0.05), jejunum and ileum, respectively, but decreased (p < 0.01) in the caecum, compared with the control group. Interestingly, the SIgA level was decreased in the duodenum (p < 0.05), jejunum and ileum but increased (p < 0.05) in the caecum in the infected/Diclazuril group in comparison to the infected group. The results showed that Diclazuril effectively alleviated the damage in the caecum induced by E. tenella and provided a cure for coccidiosis by improving the immune function in chickens.
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A novel serine/threonine protein phosphatase type 5 from second-generation merozoite of Eimeria tenella is associated with Diclazuril-induced apoptosis
Parasitology research, 2013Co-Authors: Bianhua Zhou, Hongwei Wang, Zhen-sheng Zhao, Mei Liu, Wenchao Yan, Jing Zhao, Zhe Zhang, Feiqun XueAbstract:Screening the anticoccidial drug targets is very important for developing novel drugs and revealing the molecular basis of drug resistance in coccidia. Due to high effectivity and safety, Diclazuril was used widely in the poultry industry. To assess the roles of the serine/threonine protein phosphatase type 5 of second-generation merozoites in Eimeria tenella (EtPP5) in the anticoccidial activity of Diclazuril against chicken coccidiosis, EtPP5 was cloned using reverse transcriptase polymerase chain reaction and rapid amplification of cDNA ends. Ultrastructural changes in second-generation merozoites and mRNA expression level of EtPP5 were monitored by transmission electron microscopy (TEM) and quantitative real-time PCR, respectively. The results showed that the full length of the cloned EtPP5 cDNA (2,495 bp) encompassed a 1,647-bp open reading frame encoding a polypeptide of 548 residues with an estimated molecular mass of 60.82 kDa and a theoretical isoelectric point of 5.89. Molecular analysis of EtPP5 reveals the presence of a C-terminal phosphatase domain and an extended N-terminal tetratricopeptide repeat motif, a typical feature of protein phosphatases. The cDNA sequence has been submitted to the GenBank database with accession number JX987508. EtPP5 shared 89% homology with the published sequence of a PP5 ortholog of Toxoplasma gondii at the amino acid level (GenBank XP_002364442.1). TEM observed that Diclazuril induced ultrastructural changes in second-generation merozoites. Quantitative real-time PCR analysis showed that compared with the control group, the level of EtPP5 mRNA expression was significantly downregulated by 51.4% by Diclazuril treatment. The high similarity of EtPP5 to previously described PP5 of other organisms, as well as its downregulated expression and connection with apoptosis in the second-generation merozoites induced by Diclazuril, suggests that it could act an important role in understanding the signaling mechanism underlining the Diclazuril-induced merozoites apoptosis.
Bianhua Zhou - One of the best experts on this subject based on the ideXlab platform.
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Diclazuril-induced expression of CDK-related kinase 2 in the second-generation merozoites of Eimeria tenella
2021Co-Authors: Bianhua Zhou, Hai-yan Ding, Jing-yun Yang, Jun Chai, Hong-wei Guo, Erjie TianAbstract:Abstract Background: Diclazuril is a classic anticoccidial drug. The key molecules of Diclazuril in anticoccidial action allows target screening for the development of anticoccidial drugs. In the present study, a Diclazuril anticoccidiosis animal model was established, and the transcription and translation levels of the CDK-related kinase 2 of Eimeria tenella (EtCRK2) were detected through quantitative real-time PCR and Western blot analysis, respectively. The localisation of EtCRK2 in merozoites was examined with immunofluorescence techniques.Results: The mRNA and protein expression levels of EtCRK2 decreased in the infected/Diclazuril group compared with those in the infected/control group. In addition, immunofluorescence analysis showed that EtCRK2 was localised in the cytoplasm of the merozoites. The fluorescence intensity of EtCRK2 in the infected/Diclazuril group was significantly weaker than that in the infected/control group.Conclusions: The anticoccidial drug Diclazuril against E.tenella affects the expression pattern of EtCRK2 molecule, and EtCRK2 is a potential target for new drug development.
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Effects of Diclazuril on the expression of enolase in second-generation merozoites of Eimeria tenella.
Poultry science, 2020Co-Authors: Bianhua Zhou, Hai-yan Ding, Jing-yun Yang, Jun Chai, Hong-wei Guo, Hongwei WangAbstract:Abstract Eimeria tenella is an obligate intracellular parasite of the chicken cecum; it brings huge economic loss to the chicken industry. Enolase is a multifunctional glycolytic enzyme involved in many processes of parasites, such as infection and migration. In this study, the effect of Diclazuril on the expression of enolase in second-generation merozoites of E. tenella (EtENO) was reported. The prokaryotic expression plasmid pET-28a-EtENO was constructed and transformed into Escherichia coli BL21 (DE3). Then, it was subjected to expression under the induction of isopropyl-β-D-1-thiogalactopyranoside. The expressed products were identified and purified. The purified EtENO protein was used for antibody preparation. The EtENO mRNA and protein expression levels were analyzed via real-time PCR and Western blotting. Localization of EtENO on the merozoites was examined by immunofluorescence technique. The mRNA and protein expression levels of EtENO were decreased by 36.3 and 40.36%, respectively, by Diclazuril treatment. EtENO distributed in the surface, cytoplasm, and nucleus of the infected/control group. With Diclazuril treatment, it was significantly reduced in the surface and cytoplasm and even disappeared in the nucleus of the infected/Diclazuril group. These observations suggested that EtENO may play an important role in mechanism of Diclazuril anticoccidial action and be a potential drug target for the intervention with E. tenella infection.
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Eukaryotic elongation factor 2 is involved in the anticoccidial action of Diclazuril in the second-generation merozoites of Eimeria tenella.
Veterinary parasitology, 2019Co-Authors: Bianhua Zhou, Hai-yan Ding, Jing-yun Yang, Hong-wei Guo, Liu-shu Jia, Hongwei WangAbstract:Eimeria tenella, an obligate intracellular parasite, can actively invade the cecal epithelial cells of chickens and cause severe enteric disease. Eukaryotic elongation factor 2 (eEF2) plays a major role in protein synthesis and cell survival. This study aims to explore the exact mechanisms underlying Diclazuril inhibition in second-generation merozoites of E. tenella. The eEF2 cDNA of the second-generation merozoites of E. tenella (EtEF2) was cloned by reverse transcriptase polymerase chain reaction and rapid amplification of cDNA ends. Diclazuril-induced expression profiles of EtEF2 were also analyzed. The cloned full-length cDNA (2893 bp) of the EtEF2 nucleotide sequence encompassed a 2499 bp open reading frame (ORF) that encoded a polypeptide of 832 residues with an estimated molecular mass of 93.12 kDa and a theoretical isoelectric point of 5.99. The EtEF2 nucleotide sequence was submitted to the GenBank database with the accession number KF188423. The EtEF2 protein sequence shared 99 % homology with the eEF2 sequence of Toxoplasma gondii (GenBank XP_002367778.1). The GTPase activity domain and ADP-ribosylation domain were conserved signature sequences of the eEF2 gene family. The changes in the transcriptional and translational levels of EtEF2 were detected through quantitative real-time PCR and Western blot analyses. The mRNA expression level of EtEF2 was 2.706 fold increases and the protein level of EtEF2 was increased 67.31 % under Diclazuril treatment. In addition, the localization of EtEF2 was investigated through immunofluorescence assay. Experimental results demonstrated that EtEF2 was distributed primarily in the cytoplasm of second-generation merozoites, and its fluorescence intensity was enhanced after Diclazuril treatment. These findings indicated that EtEF2 may have an important role in understanding the signaling mechanism underlying the anticoccidial action of Diclazuril and could be a promising target for novel drug exploration.
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effect of Diclazuril on the bursa of fabricius morphology and siga expression in chickens infected with eimeria tenella
Korean Journal of Parasitology, 2015Co-Authors: Bianhua Zhou, Fuwei Yuan, Erjie Tian, Hongwei WangAbstract:The effects of Diclazuril on the bursa of Fabricius (BF) structure and secretory IgA (SIgA) expression in chickens infected with Eimeria tenella were examined. The morphology of the BF was observed by hematoxylin and eosin staining, while ultrastructural changes were monitored by transmission electron microscopy. E. tenella infection caused the BF cell volumes to decrease, irregularly arranged, as well as, enlargement of the intercellular space. Diclazuril treatment alleviated the physical signs of damages associated with E. tenella infection. The SIgA expression in BF was analyzed by immunohistochemistry technique. The SIgA expression increased significantly by 350.4% (P<0.01) after E. tenella infection compared to the normal control group. With the treatment of Diclazuril, the SIgA was relatively fewer in the cortex, and the expression level was significantly decreased by 46.7% (P<0.01) compared with the infected and untreated group. In conclusion, E. tenella infection in chickens induced obvious harmful changes in BF morphological structure and stimulated the expression of SIgA in the BF. Diclazuril treatment effectively alleviated the morphological changes. This result demonstrates a method to develop an immunological strategy in coccidiosis control.
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effect of Diclazuril on intestinal morphology and siga expression in chicken infected with eimeria tenella
Parasitology Research, 2014Co-Authors: Erjie Tian, Bianhua Zhou, Jing Zhao, Xueying Wang, Wen Deng, Hongwei WangAbstract:Secretory immunoglobulin A (SIgA), as a vital actor involving in the mucosal immunity, plays a key role in defending a variety of pathogenic infections, such as bacteria, viruses and parasites. Eimeria tenella is an obligate intracellular apicomplexan parasite contacting with the digestive tract mucosa and specially parasitizes chicken caecum, causing a severe form of coccidiosis. Coccidiosis is currently mainly controlled using chemotherapeutic agents. Diclazuril, a classic coccidiostat, was used widely in the poultry industry. Because of the rising problem of drug resistance, it is therefore crucial to understand the pattern of the SIgA expression in the action of Diclazuril against E. tenella. In this study, the intestinal morphology in the caecum was analyzed by haematoxylin-eosin (HE) staining, and the SIgA expression was examined by immunohistochemical technique. At the same time, the duodenum, jejunum and ileum tissues have also been evaluated. HE staining results showed that E. tenella infection caused severe damage characterized by structural disorder, haemorrhage, inflammatory cell infiltration, serous and fibrinous exudation in chicken caecum and invisible damage in the duodenum, jejunum and ileum. With the treatment of Diclazuril, the damage in the caecum was alleviated obviously. Immunohistochemical analysis demonstrated that the SIgA level in the infected group was increased in the duodenum (p < 0.05), jejunum and ileum, respectively, but decreased (p < 0.01) in the caecum, compared with the control group. Interestingly, the SIgA level was decreased in the duodenum (p < 0.05), jejunum and ileum but increased (p < 0.05) in the caecum in the infected/Diclazuril group in comparison to the infected group. The results showed that Diclazuril effectively alleviated the damage in the caecum induced by E. tenella and provided a cure for coccidiosis by improving the immune function in chickens.
Feiqun Xue - One of the best experts on this subject based on the ideXlab platform.
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Proteomic analysis of the effect of Diclazuril on second-generation merozoites of Eimeria tenella.
Parasitology research, 2013Co-Authors: Xiao-jiong Shen, Keyu Zhang, Xiaoyang Wang, Yin-chun Liu, Hua-jing Zhang, Chao Fan, Chengzhong Fei, Feiqun XueAbstract:Diclazuril has long been used as an effective benzeneacetonitrile anticoccidial for the control of Eimeria tenella that causes coccidiosis. However, the molecular mechanism underlying the anticoccidial effects of Diclazuril remains elusive. In this study, a proteomic analysis of the effect of Diclazuril on second-generation merozoites of E. tenella was performed. Using two-dimensional gel electrophoresis and real-time quantitative polymerase chain reaction (RT-PCR), 13 target proteins were found to be significantly affected by Diclazuril treatment, with 11 of these proteins being identified as annotated proteins from E. tenella or other Apicomplexa parasites. These proteins contribute to various functions, including metabolism, protein synthesis, and host cell invasion. Using RT-PCR, we identified the potential pattern of transcriptional regulation induced by Diclazuril, and we suggest some promising targets for the intervention of E. tenella infection.
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A novel serine/threonine protein phosphatase type 5 from second-generation merozoite of Eimeria tenella is associated with Diclazuril-induced apoptosis
Parasitology research, 2013Co-Authors: Bianhua Zhou, Hongwei Wang, Zhen-sheng Zhao, Mei Liu, Wenchao Yan, Jing Zhao, Zhe Zhang, Feiqun XueAbstract:Screening the anticoccidial drug targets is very important for developing novel drugs and revealing the molecular basis of drug resistance in coccidia. Due to high effectivity and safety, Diclazuril was used widely in the poultry industry. To assess the roles of the serine/threonine protein phosphatase type 5 of second-generation merozoites in Eimeria tenella (EtPP5) in the anticoccidial activity of Diclazuril against chicken coccidiosis, EtPP5 was cloned using reverse transcriptase polymerase chain reaction and rapid amplification of cDNA ends. Ultrastructural changes in second-generation merozoites and mRNA expression level of EtPP5 were monitored by transmission electron microscopy (TEM) and quantitative real-time PCR, respectively. The results showed that the full length of the cloned EtPP5 cDNA (2,495 bp) encompassed a 1,647-bp open reading frame encoding a polypeptide of 548 residues with an estimated molecular mass of 60.82 kDa and a theoretical isoelectric point of 5.89. Molecular analysis of EtPP5 reveals the presence of a C-terminal phosphatase domain and an extended N-terminal tetratricopeptide repeat motif, a typical feature of protein phosphatases. The cDNA sequence has been submitted to the GenBank database with accession number JX987508. EtPP5 shared 89% homology with the published sequence of a PP5 ortholog of Toxoplasma gondii at the amino acid level (GenBank XP_002364442.1). TEM observed that Diclazuril induced ultrastructural changes in second-generation merozoites. Quantitative real-time PCR analysis showed that compared with the control group, the level of EtPP5 mRNA expression was significantly downregulated by 51.4% by Diclazuril treatment. The high similarity of EtPP5 to previously described PP5 of other organisms, as well as its downregulated expression and connection with apoptosis in the second-generation merozoites induced by Diclazuril, suggests that it could act an important role in understanding the signaling mechanism underlining the Diclazuril-induced merozoites apoptosis.
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a novel serine threonine protein phosphatase type 5 from second generation merozoite of eimeria tenella is associated with Diclazuril induced apoptosis
Parasitology Research, 2013Co-Authors: Bianhua Zhou, Hongwei Wang, Zhen-sheng Zhao, Mei Liu, Wenchao Yan, Jing Zhao, Zhe Zhang, Feiqun XueAbstract:Screening the anticoccidial drug targets is very important for developing novel drugs and revealing the molecular basis of drug resistance in coccidia. Due to high effectivity and safety, Diclazuril was used widely in the poultry industry. To assess the roles of the serine/threonine protein phosphatase type 5 of second-generation merozoites in Eimeria tenella (EtPP5) in the anticoccidial activity of Diclazuril against chicken coccidiosis, EtPP5 was cloned using reverse transcriptase polymerase chain reaction and rapid amplification of cDNA ends. Ultrastructural changes in second-generation merozoites and mRNA expression level of EtPP5 were monitored by transmission electron microscopy (TEM) and quantitative real-time PCR, respectively. The results showed that the full length of the cloned EtPP5 cDNA (2,495 bp) encompassed a 1,647-bp open reading frame encoding a polypeptide of 548 residues with an estimated molecular mass of 60.82 kDa and a theoretical isoelectric point of 5.89. Molecular analysis of EtPP5 reveals the presence of a C-terminal phosphatase domain and an extended N-terminal tetratricopeptide repeat motif, a typical feature of protein phosphatases. The cDNA sequence has been submitted to the GenBank database with accession number JX987508. EtPP5 shared 89% homology with the published sequence of a PP5 ortholog of Toxoplasma gondii at the amino acid level (GenBank XP_002364442.1). TEM observed that Diclazuril induced ultrastructural changes in second-generation merozoites. Quantitative real-time PCR analysis showed that compared with the control group, the level of EtPP5 mRNA expression was significantly downregulated by 51.4% by Diclazuril treatment. The high similarity of EtPP5 to previously described PP5 of other organisms, as well as its downregulated expression and connection with apoptosis in the second-generation merozoites induced by Diclazuril, suggests that it could act an important role in understanding the signaling mechanism underlining the Diclazuril-induced merozoites apoptosis.
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Effect of the Diclazuril on Hsp90 in the second-generation merozoites of Eimeria tenella.
Veterinary parasitology, 2011Co-Authors: Xiao-jiong Shen, Chenzhong Fei, Congcong Wang, Qingxu Zhu, Wenli Zheng, Minqi Qiu, Feiqun XueAbstract:Eimeria tenella (E. tenella) is one of the most virulent pathogens of coccidiosis. In apicomplexan parasites, Hsp90 (Heat shock protein 90) is essential for the invasion and survival in host cells. In this study, the effect of Diclazuril, an effective benzeneacetonitrile anticoccidial agent, on the expression of Hsp90 in the second-generation merozoites of E. tenella was investigated. We inoculated 8 × 10(4) oocysts/chicken suspended in 1 ml of distilled water, and chickens were challenged with E. tenella oocysts and provided with normal feed as Control group; chickens challenged with E. tenella oocysts and provided with 1mg/kg Diclazuril in feed from 96 h to 120 h after inoculation as treatment group. Then the second-generation merozoites were obtained after 120 h from the infected caeca. Our results showed that the transcription level of mzHsp90 was reduced by 29.7% in the Diclazuril treatment group, accompanied by reduced level of mzHsp90 protein in second-generation merozoites prepared from infected chickens. We also found that the subcellular localization of mzHsp90 was more dispersed in these merozoites. Moreover, we demonstrated that the effects of Diclazuril on mzHsp90 expression were direct by in vitro experiments. Taken together, our data provide insights into the molecular mechanisms of Diclazuril in the chemotherapy of E. tenella, and suggest that mzHsp90 represents a promising target for the intervention with E. tenella infection.
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eimeria tenella effects of Diclazuril treatment on microneme genes expression in second generation merozoites and pathological changes of caeca in parasitized chickens
Experimental Parasitology, 2010Co-Authors: Bianhua Zhou, Hongwei Wang, Keyu Zhang, Lifang Zhang, Xiaoyang Wang, Feiqun XueAbstract:The effects of Diclazuril on mRNA expression levels of invasion-related microneme genes were examined in second-generation merozoites of Eimeria tenella (E. tenella) by quantitative real-time (QRT) PCR. Diclazruil treatment of infected chickens significantly decreased the number of second-generation merozoites by 65.13%, and resulted in downregulation of EtMIC genes: EtMIC1 by 65.63%, EtMIC2 by 64.12%, EtMIC3 by 56.82%, EtMIC4 by 73.48%, and EtMIC5 by 78.17%. SEM images of caecum tissue from uninfected chickens showed regular intestinal villus structure. In infected chickens, a distinct loss of the superficial epithelium, with a flattened mucosa and large-area necrosis and anabrosis, was evident. In diclazruil-treated chickens, a decrease in merozoite number and a visibly improved appearance of the caeca were noted. These improvements appeared to be mediated in part by downregulation of the expression of invasion-related EtMIC genes in response to Diclazuril.
Shahriar Sedghi - One of the best experts on this subject based on the ideXlab platform.
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tu1261 short term relief of rectal pain with diazepam cream a novel new approach
Gastroenterology, 2014Co-Authors: Mudit Chowdhary, Minh Hang, Ali Keshavarzian, Shahriar SedghiAbstract:Toxoplasmsis is one of the most common foodborne gastrointestinal and congenital illnesses in United States. Two billion people globally are estimated to be infected with Toxoplasma with severe and unknown consequences. Safe and effective preventive or therapeuticmeasures are needed to impact the disease. Diclazuril, an herbicide related compound, specifically targets chlorophyll complex in the organisms which is not present in man and animals. We hypothesized that Diclazuril to be safe and effective against gastrointestinal Toxoplasmosis in our murine pregnancy model. Methods: Programmed pregnant mice were infected with escalating doses of organisms from Type II strain (most common strain from man in U.S) and treated with varied doses of Diclazuril, or sham. Dams were monitored daily for distress, pain, and abortion and samples collected at the end of experiment. Results: Infected dams developed anemia (Controls vs infected-dams p<0.05) and ascities with moderate to severe GI related complications in organisms' dose dependent manner. Diclazuril dose dependently protected dams from ascities and anemia. Infected dams showed splenomegaly, with massive infiltration of epithelioid cells and loss of germinal structure with significant increases in weight and length of splenic tissues and improved in dams treated with Diclazuril. Infected dams exhibited moderate (score 0 to 4 scale=3+0.02) to severe hepatitis (3.5+0.01) with influx of inflammatory and plasma cells, dysplastic and necrotized hepatocytes in organisms' dose escalatingmanner. Diclazuril treatment dose dependently protected dams from hepatitis, also in organisms' dose related manner (respectively infected-treated versus infected-shamtreated controls P<0.001, P<0.01 and P<0.05). Colonic tissues significantly shortened in length with infiltration of lymphocytes, and macrophages in mucosal structure which improved in Diclazuril treated dams. Diclazuril protected fetuses but had no effect on abdominal pain related behavior. Conclusions: Diclazuril treatment dose dependently protected dams from mild to moderate gastrointestinal complications and their fetuses against adverse effects of Toxoplasmosis. This is the first report describing of Diclazuril safety and efficacy in pregnancy and gastrointestinal Toxoplasmosis. Supported by NIH-NIDCR (HO) and KSTC (TT).
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Tu1261 Short Term Relief of Rectal Pain With Diazepam Cream: A Novel New Approach
Gastroenterology, 2014Co-Authors: Mudit Chowdhary, Minh Hang, Ali Keshavarzian, Shahriar SedghiAbstract:Toxoplasmsis is one of the most common foodborne gastrointestinal and congenital illnesses in United States. Two billion people globally are estimated to be infected with Toxoplasma with severe and unknown consequences. Safe and effective preventive or therapeuticmeasures are needed to impact the disease. Diclazuril, an herbicide related compound, specifically targets chlorophyll complex in the organisms which is not present in man and animals. We hypothesized that Diclazuril to be safe and effective against gastrointestinal Toxoplasmosis in our murine pregnancy model. Methods: Programmed pregnant mice were infected with escalating doses of organisms from Type II strain (most common strain from man in U.S) and treated with varied doses of Diclazuril, or sham. Dams were monitored daily for distress, pain, and abortion and samples collected at the end of experiment. Results: Infected dams developed anemia (Controls vs infected-dams p
L Mortier - One of the best experts on this subject based on the ideXlab platform.
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Liquid chromatographic tandem mass spectrometric determination of five coccidiostats in poultry eggs and feed.
Journal of Chromatography B, 2005Co-Authors: L Mortier, Els Daeseleire, Carlos Van PeteghemAbstract:Abstract A method is described which permits the quantitative detection of the chemical coccidiostats halofuginone, robenidine, Diclazuril, nicarbazin and dimetridazole and its main metabolite 2-hydroxydimetridazole in poultry eggs and feed. Sample preparations were kept very simple and are based upon extraction with an organic solvent. Sample extracts were injected into the liquid chromatography tandem mass spectrometry (LC–MS/MS) system on a C18 column and a gradient elution was performed. Dimetridazole-D3 and Diclazuril-bis, a structural analogue of Diclazuril, were used as internal standards. Detection was performed on a triple quadrupole mass spectrometer in the selected reaction monitoring mode after ionisation in the positive or negative electrospray ionisation mode. Argon was applied as collision gas for collision induced dissociation. Validation of the methods was performed based on Commission Decision 2002/657/EC [Official Journal of the European Communities L221 (2002) 8].
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Detection of Residues of the Coccidiostat Diclazuril in Poultry Tissues by Liquid Chromatography−Tandem Mass Spectrometry after Withdrawal of Medicated Feed
Journal of agricultural and food chemistry, 2005Co-Authors: L Mortier, Els Daeseleire, G. Huyghebaert, K. Grijspeerdt, Carlos Van PeteghemAbstract:A liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the quantitative determination of Diclazuril in poultry tissues and feed is presented. A simple clean up with an organic solvent was carried out. A reversed-phase C(18) column was used for the high-performance liquid chromatography (HPLC) to separate the analyte with a gradient of acetonitrile and water as mobile phase. The precursor ions produced by electrospray negative ionization were selected for collisional dissociation. Validation of the methods was performed based on Commission Decision 2002/657/EC (Off. J. Eur. Communities 2002, L221, 8-36). For the detection of Diclazuril in poultry meat, the decision limit was found to be 0.5 microg/kg. An animal experiment was set up in which 70 chickens were held for 6 weeks. From day 22 until day 32, they were fed feed containing 730 microg/kg Diclazuril. From day 33 until day 42, every day six chickens were slaughtered, and breast, thigh, and liver were analyzed. Average steady-state concentrations of 94, 135, and 722 microg/kg in breast, thigh, and liver were obtained, respectively. Nine days after withdrawal of the medicated feed, Diclazuril was still present in the different sample types.
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detection of residues of the coccidiostat Diclazuril in poultry tissues by liquid chromatography tandem mass spectrometry after withdrawal of medicated feed
Journal of Agricultural and Food Chemistry, 2005Co-Authors: L Mortier, Els Daeseleire, G. Huyghebaert, K. Grijspeerdt, Carlos Van PeteghemAbstract:A liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the quantitative determination of Diclazuril in poultry tissues and feed is presented. A simple clean up with an organic solvent was carried out. A reversed-phase C(18) column was used for the high-performance liquid chromatography (HPLC) to separate the analyte with a gradient of acetonitrile and water as mobile phase. The precursor ions produced by electrospray negative ionization were selected for collisional dissociation. Validation of the methods was performed based on Commission Decision 2002/657/EC (Off. J. Eur. Communities 2002, L221, 8-36). For the detection of Diclazuril in poultry meat, the decision limit was found to be 0.5 microg/kg. An animal experiment was set up in which 70 chickens were held for 6 weeks. From day 22 until day 32, they were fed feed containing 730 microg/kg Diclazuril. From day 33 until day 42, every day six chickens were slaughtered, and breast, thigh, and liver were analyzed. Average steady-state concentrations of 94, 135, and 722 microg/kg in breast, thigh, and liver were obtained, respectively. Nine days after withdrawal of the medicated feed, Diclazuril was still present in the different sample types.
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determination of the coccidiostat Diclazuril in poultry feed and meat by liquid chromatography tandem mass spectrometry
Analytica Chimica Acta, 2005Co-Authors: L Mortier, Els Daeseleire, Carlos Van PeteghemAbstract:Abstract A liquid chromatographic-tandem mass spectrometric method (LC–MS/MS) for the detection of the coccidiostat Diclazuril in poultry meat and feed was developed. After an appropriate extraction with an organic solvent, the samples were injected into the LC–MS/MS system on a C 18 column. A gradient with acetonitrile and water, each containing 0.1% formic acid, was applied. A structure analogue of Diclazuril was used as internal standard. The precursor ions produced by negative electrospray ionisation were selected for collisional dissociation with argon. For Diclazuril, two product ions were recorded with multiple reaction monitoring. Validation of the methods was performed based on commission decision 2002/657/EC [1] . The methods were applied to real-life samples: meat of a home-bred chicken and its feed were analysed.