[Trial on the biochemical identification of aerobic pathogenic bacteria].
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Gene-specific DNA probes can be used to identify enterotoxigenic Escherichia coli and Campylobacter jejunii by DNA hybridization. In addition, the DNA probes can be exploited to provide potentially important information concerning the epidemiology of enteric infections.
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A pathogen was isolated from natural dead pests of Ceracris kiangsu in Geleshan farm of Chongqin. Its pathogenecity was confirmed by the law of KOCK. It was identified as Pseudomonas pseudoalcaligenes according to its physiological and biochemical properties as well as the G + C content of DNA (63.73mol%). The results of preliminary bioassay show that the pathogen can infect the grasshoppers and Ceracris kiangsu, and also can infect other pests of grassland in a certain extent.
The authors examined the possibility of detecting M. tuberculosis cells in various types of diagnostic material (sputum, blood, bone marrow, bronchoalveolar lavage fluid) from tuberculosis patients using polymerase chain reaction (PCR). The developed PCR-based test systems helped detect M. tuberculosis in 48 (90.6%) out of 53 tuberculosis patients, in contrast to much slower microbiological methods which permitted detection of Mycobacteria in only 21 (39.6%) patients. High specificity and virtually no false-positive results of PCR were demonstrated in testing diagnostic material from patients with chronic nonspecific pulmonary diseases and from children with lympholeukemia and anemia.
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Mycobacteriosis has become a major concern for the commercial mariculture of the European sea bass Dicentrachus labrax in Israel. The disease remains asymptomatic for a long time, is virtually impossible to eradicate with antibiotics, stunts the growth of the fish and renders the fish unmarketable. The pathogen was identified as Mycobacterium marinum by direct sequencing and analysis of approximately 600 bp of the pathogen ribosomal encoding DNA (rDNA). The polymerase chain reaction technique was evaluated as a diagnostic tool for detecting the infection in D. labrax and found to be highly specific and sensitive.
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For the identification of the plant pathogenic bacterium Erwinia amylovora, the immuno gold staining (IGS) and immuno gold silver staining (IGSS) techniques are tested. The IGS and IGSS methods are at least as sensitive an indirect immunofluorescence and require less primary antiserum. Moreover they have the advantage that the preparations can be conserved permanently and unchanged. The preparation of the IGS can be observed with transmitted light or--with considerable better result--using epipolarization microscopy. The IGSS method deserves special attention because of its high contrast in normal brigth field microscopy with transmitted light.
Identification of the emerging pathogen Vibrio vulnificus biotype 3 has become a challenge for clinical laboratories in the last few years. In this study, the abilities of five commercial systems to identify this new species have been evaluated for the first time, using a unique collection of strains. Fifty-one well-documented wild strains of V. vulnificus biotype 3 were processed using API 20 NE, GNI+ Vitek 1 cards, ID-GNB Vitek 2 cards, Neg Combo 20 Microscan panels, and NMIC/ID-5 BD Phoenix panels. The numbers of strains identified as V. vulnificus by ID-GNB, NMIC/ID-5, and GNI+ were 50 (98.0%), 46 (90.2%), and 7 (13.7%), respectively. Neg Combo 20 Microscan panels and API 20 NE were unable to identify any of the strains of this emerging pathogen to the species level and mostly misidentifies them as other species of the Vibrionaceae family. Data on the phenotypic pattern of V. vulnificus biotype 3 when processed in all five systems as presented here could help clinical laboratories in identifying this new pathogen.
The performance of two commercial chromogenic media for the isolation and presumptive identification of urinary tract pathogens, the CPS ID2 (bioMérieux, France) and the CHROMagar Orientation (BBL Becton Dickinson, USA), was evaluated and compared with that of cystine-lactose-electrolyte-deficient agar and tryptic soy agar with 5% sheep blood. The detection, determination of bacterial counts, and presumptive identification of bacteria causing urinary tract infections were evaluated in 3,000 urine specimens. The two chromogenic media showed excellent correlation with the standard media for the detection and the bacterial count of urinary pathogens. The Escherichia coli strains produced the expected colour on the CHROMagar Orientation and the CPS ID2 media in 99% and 90% of the cases, respectively. The Klebsiella-Enterobacter-Citrobacter and the Proteus-Morganella-Providencia groups were easily identified on both chromogenic media, but further biochemical tests were needed to differentiate them to a species level. Both media enabled the differentiation, with varying degrees of difficulty, of Pseudomonas spp. strains from members of the family Enterobacteriaceae. All isolates of Enterococcus spp. were correctly identified and were easily distinguished from the Streptococcus agalactiae isolates. Staphylococcus saprophyticus isolates were easy to identify only on the CHROMagar Orientation medium. No substantial difference was observed when comparing the results of the susceptibility tests, which were performed according to the standardized disk diffusion method as described by the National Committee for Clinical Laboratory Standards, for colonies recovered from the blood agar versus those recovered from the chromogenic media. In conclusion, the CPS ID2 and CHROMagar Orientation media enabled excellent detection, count determination, and presumptive identification of urinary pathogens, both in pure and mixed cultures, and reliable and accurate antimicrobial susceptibility testing directly from primary isolates. Moreover, these media allowed a remarkable reduction in the workload and a significant savings of time. On the basis of their performance, these media can replace the standard primary plating media used in the routine diagnosis of urinary tract infections.
The Quantum II, originally designed by Abbott Diagnostics for automated rapid identification of members of Enterobacteriaceae, was adapted for the identification of bacterial fish pathogens. The instrument operates as a spectrophotometer at a wavelength of 492.600 nm. A sample cartridge containing 20 inoculated biochemical chambers is inserted in the path of the analyzing beam. Reactions are converted into a 7-digit octal biocode, relayed via a sensor to the memory module, and compared to biocodes preprogrammed in the memory. An identification is then printed. Presently, the Quantum II is capable of identifying human strains of Aeromonas hydrophila and Edwardsiella tarda. This study was initiated to determine the feasibility of expanding the use of the Quantum II to include identification of bacterial fish pathogens. Ten to 50 isolates of Edwardsiella ictaluri, Serratia liquefaciens, Yersinia ruckeri, Aeromonas hydrophila, typical Aeromonas salmonicida, and atypical Aeromonas salmonicida were utilized to determine optimal incubation conditions, relative stability of the biochemicals, and ability to obtain consistent biocode numbers. After sorting the octal biocodes from the 169 isolates into groupings using a cluster analysis technique, it was shown by a Chi-square goodness of fit test that isolates of a given species were sorted into the same cluster group at a frequency of at least 99%. Results of this study illustrate the usefulness of the Quantum II BID system for the identification of bacterial fish pathogens not contained within the system's memory module.
Paracoccidioides brasiliensis, a thermodimorphic fungus, is the causative agent of the prevalent systemic mycosis in Latin America, paracoccidioidomycosis. We present here a survey of expressed genes in the yeast pathogenic phase of P. brasiliensis. We obtained 13,490 expressed sequence tags from both 5' and 3' ends. Clustering analysis yielded the partial sequences of 4,692 expressed genes that were functionally classified by similarity to known genes. We have identified several Candida albicans virulence and pathogenicity homologues in P. brasiliensis. Furthermore, we have analyzed the expression of some of these genes during the dimorphic yeast-mycelium-yeast transition by real-time quantitative reverse transcription-PCR. Clustering analysis of the mycelium-yeast transition revealed three groups: (i) RBT, hydrophobin, and isocitrate lyase; (ii) malate dehydrogenase, contigs Pb1067 and Pb1145, GPI, and alternative oxidase; and (iii) ubiquitin, delta-9-desaturase, HSP70, HSP82, and HSP104. The first two groups displayed high mRNA expression in the mycelial phase, whereas the third group showed higher mRNA expression in the yeast phase. Our results suggest the possible conservation of pathogenicity and virulence mechanisms among fungi, expand considerably gene identification in P. brasiliensis, and provide a broader basis for further progress in understanding its biological peculiarities.