RESUMO
Coccidiostat and antibiotics are widely used in poultry industry, but their effects on cecum microbial community and metabolomics in chickens infected with coccidia have been rarely studied. In this study, we analyzed the changes of microbiota and metabolomic which associated with Eimeria tenella infection in 8 days of age chickens in the presence or absence of ethanamizuril, sulfachlorpyridazine or their combinations treatment for 3 consecutive days. 16S rRNA gene sequencing and LC-MS/MS analyses were used to profile the cecal microbiome and metabolome in each group of chickens on 7 days post-infection. The results showed that coccidial infection induced significant perturbations in the distribution of microbial taxonomy and the metabolism of physiological functional molecules in cecal contents. Ethanamizuril treatment seemed to transform microbiota into a steady state conducive to animal health, and sulfachlorpyridazine treatment alleviated the growth of potentially harmful bacteria such as Escherichia-Shigella. The change trends of metabolites such as n-carbamoylglutamic acid were consistent with the anticoccidial effect of ethanamizuril. The combinations of ethanamizuril and sulfachlorpyridazine at low-dose had little effect on gut microbiota, metabolism and anticoccidial effect. These data indicate that the cecal microbiota and metabolic status of chickens infected with E. tenella following ethanamizuril treatment could be used to monitor the response to drug efficacy. This study provides a new system approach to elucidate the microbiota, metabolic and therapeutic effects of the combination of coccidiostat and antibiotics in the context of avian coccidiosis.
Assuntos
Coccidiose , Coccidiostáticos , Eimeria tenella , Microbioma Gastrointestinal , Doenças das Aves Domésticas , Sulfacloropiridazina , Animais , Eimeria tenella/genética , Galinhas/microbiologia , Sulfacloropiridazina/uso terapêutico , Coccidiostáticos/farmacologia , Coccidiostáticos/uso terapêutico , RNA Ribossômico 16S/genética , Cromatografia Líquida , Doenças das Aves Domésticas/tratamento farmacológico , Doenças das Aves Domésticas/microbiologia , Espectrometria de Massas em Tandem , Ceco/microbiologia , Coccidiose/tratamento farmacológico , Coccidiose/veterinária , Microbioma Gastrointestinal/genética , Metabolômica , Antibacterianos/farmacologia , Antibacterianos/uso terapêuticoRESUMO
BACKGROUND: Nitazoxanide not only exhibits a broad spectrum of activities against various pathogens infecting animals and humans but also induces cellular autophagy. Currently, the pattern of action and subcellular targets of nitazoxanide-induced cellular autophagy are still unclear. METHODS: To identify potential targets of nitazoxanide in mammalian cells, we developed an af-finity chromatography system using tizoxanide, a deacetyl derivative of nitazoxanide, as a ligand. Affinity chromatography was performed using VERO cell extracts on tizoxanide-biotin, and the isolated binding proteins were identified by mass spectrometry. Candidate target proteins ob-tained using affinity chromatography were co-analysed with the drug affinity response target sta-bility method. Fluorescent probes obtained by coupling rhodamine B to nitazoxanide were used for intracellular localisation of the binding targets. Solvent-induced protein precipitation profiling and thermal proteome profiling were used to further validate the binding proteins. RESULTS: The joint analysis of the drug affinity response target stability method and affinity chro-matography resulted in the screening of six possible candidate target proteins. Fluorescent probes localised the nitazoxanide-binding protein around the nuclear membrane. Molecular docking re-vealed that the binding proteins mainly formed hydrogen bonds with the nitro group of nitazoxa-nide. Solvent-induced protein precipitation profiling and thermal proteome profiling further vali-dated SEC61A, PSMD12, and PRKAG1 as potential target proteins of nitazoxanide. CONCLUSION: The data supports the idea that nitazoxanide is a multifunctional compound with multiple targets.
RESUMO
Eight mono- or disaccharide analogues derived from BLM disaccharide, along with the corresponding carbohydate-dye conjugates have been designed and synthesized in this study, aiming at exploring the effect of a gulose residue on the cellular binding/uptake of BLM disaccharide and it possible uptake mechanism. Our evidence is presented indicating that, for the cellular binding/uptake of BLM disaccharide, a gulose residue is an essential subunit but unrelated to its chemical nature. Interestingly, d-gulose-dye conjugate is able to selectively target A549 cancer cells, but l-gulose-dye conjugate fails. Further uptake mechanism studies demonstrate d-gulose-dye derivatives similar to BLM disaccharide-dye ones behave in a temperature- and ATP-dependent manner, and are partly directed by the GLUT1 receptor. Moreover, d-gulose modifying gemcitabine 53a exhibits more potent antitumor activity compared to derivatives 53b-c in which gemcitabine is decorated with other monosaccharides. Taken together, the monosacharide d-gulose conjugate offers a new strategy for solving cytotoxic drugs via the increased tumor targeting in the therapy of lung cancer.