RESUMO
In this study, we screened differentially expressed genes in a multidrug-resistant isolate strain of Clostridium perfringens by RNA sequencing. We also separated and identified differentially expressed proteins (DEPs) in the isolate strain by two-dimensional electrophoresis (2-DE) and mass spectrometry (MS). The RNA sequencing results showed that, compared with the control strain, 1128 genes were differentially expressed in the isolate strain, and these included 227 up-regulated genes and 901 down-regulated genes. Bioinformatics analysis identified the following genes and gene categories that are potentially involved in multidrug resistance (MDR) in the isolate strain: drug transport, drug response, hydrolase activity, transmembrane transporter, transferase activity, amidase transmembrane transporter, efflux transmembrane transporter, bacterial chemotaxis, ABC transporter, and others. The results of the 2-DE showed that 70 proteins were differentially expressed in the isolate strain, 45 of which were up-regulated and 25 down-regulated. Twenty-seven DEPs were identified by MS and these included the following protein categories: ribosome, antimicrobial peptide resistance, and ABC transporter, all of which may be involved in MDR in the isolate strain of C. perfringens. The results provide reference data for further investigations on the drug resistant molecular mechanisms of C. perfringens.
Assuntos
Animais , Proteínas de Bactérias/genética , Clostridium perfringens/genética , Análise de Sequência de RNA/métodos , Genes MDR , Farmacorresistência Bacteriana Múltipla/genética , Espectrometria de Massas/métodos , Proteínas de Bactérias/metabolismo , Eletroforese em Gel Bidimensional/métodos , Regulação Bacteriana da Expressão Gênica/genética , Genoma Bacteriano/genética , Clostridium perfringens/classificação , Clostridium perfringens/efeitos dos fármacos , Clostridium perfringens/metabolismo , DNA Complementar , Proteoma/genética , Transcriptoma/genética , Ontologia GenéticaRESUMO
This study reports on comparisons between polymerase chain reaction [PCR] and conventional diagnostic methods for typing Clostridium perfringens toxins collected from Central Saudi Arabia. Fecal samples from 150 animals showing signs of enterotoxaemia were collected from 24 April 2009 to 25 September 2009, from different farms located in Riyadh, Kingdom of Saudi Arabia. Twenty-seven toxigenic strains of Clostridium perfringens were recovered from 150 fecal and intestinal content samples were identified and typed by conventional methods. All the strains were analyzed by PCR using specific primers for alpha, beta, epsilon and iota toxin genes. The experimental work was carried out at the Center of Excellence in Biotechnology, King Saud University, Riyadh, Kingdom of Saudi Arabia. The results revealed alpha toxin gene Clostridium perfringens type A in 22 [81.5%] strains out of 27 toxigenic strains, however, only 20 [74.1%] of them were identified previously as type A by classical method. One strain [3.7%] was identified as type C and 3 strains [11.1%] were identified as D by PCR typing. Moreover, PCR results confirmed the traditional methods in typing one strain as type B [3.7%]. Also, PCR method can detect 2 other strains of type A directly from the feces and intestinal contents of the examined chicken, which provide negative results in bacteriological examination. Polymerase chain reaction technique can be used as an alternative diagnostic method for detection and typing of Clostridium perfringens
Assuntos
Animais , Clostridium perfringens/classificação , Tipagem Molecular , Reação em Cadeia da PolimeraseRESUMO
A gene encoding beta toxin was amplified by polymerase chain reaction from C. perfringens type C isolate and cloned in pUC 19 vector. The nucleotide sequence was identical with C. perfringens type B beta toxin gene sequence. The Southern hybridization using labelled beta toxin gene probe revealed the presence of positive signals only in beta producing C. perfingens.
Assuntos
Animais , Toxinas Bacterianas/genética , Southern Blotting , Clonagem Molecular , Clostridium perfringens/classificação , Primers do DNA/química , DNA Bacteriano/análise , Genes Bacterianos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Mapeamento por Restrição , Análise de Sequência de DNARESUMO
Con la finalidad de probar el rendimiento de dos caldos de cultivo, usados comunmente para anaerobios, caldo tiaglicolato y caldo schedle. Se evaluaron ambos caldos con una cepa de clostridium perfringens, estableciéndose para dicho fin, curvas de crecimiento a temperaturas óptimas de crecimiento (37ºC y 46ºC) para este microorganismo. En el presente trabajo, no se encontraron diferencias significativas en cuanto al rendimiento de ambos caldos. Por lo que se concluye que, tanto en caldo tiaglicolato, usado para el aislamiento de clostridium perfringens en alimentos, y caldo Schedler, se obtuvieron buenos rendimientos.