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A Audicana

Publications and source records attributed to A Audicana.

9 recordsLinked to original sources

Suitability of PCR fingerprinting, infrequent-restriction-site PCR, and pulsed-field gel electrophoresis, combined with computerized gel analysis, in library typing of Salmonella enterica serovar enteritidis.

Strains of Salmonella enterica (n = 212) of different serovars and phage types were used to establish a library typing computerized system for serovar Enteritidis on the basis of PCR fingerprinting, infrequent-restriction-site PCR (IRS-PCR), or pulsed-field gel electrophoresis (PFGE). The rate of PCR fingerprinting interassay and intercenter reproducibility was low and was only increased when DNA samples were extracted at the same time and amplified with the same reaction mixtures. Reproducibility of IRS-PCR technique reached 100%, but discrimination was low (D = 0.52). The PFGE procedure showed an intercenter reproducibility value of 93.3%. The high reproducibility of PFGE combined with the previously determined high discrimination directed its use for library typing. The use of PFGE with enzymes XbaI, BlnI, and SpeI for library typing of serovar Enteritidis was assessed with GelCompar 4.0 software. Three computer libraries of PFGE DNA profiles were constructed, and their ability to recognize new DNA profiles was analyzed. The results obtained pointed out that the combination of PFGE with computerized analysis could be suitable in long-term epidemiological comparison and surveillance of Salmonella serovar Enteritidis, specially if the prevalence of genetic events that could be responsible for changes in PFGE profiles in this serovar was low.

Animals↗

Phage typing combined with pulsed-field gel electrophoresis and random amplified polymorphic DNA increases discrimination in the epidemiological analysis of Salmonella enteritidis strains.

Phage typing (PT) combined with pulsed-field gel electrophoresis (PFGE) and a random amplified polymorphic DNA (RAPD) fingerprinting method was used to characterize Salmonella enteritidis strains. Twenty-four epidemiologically unrelated isolates, sampled from diverse ecological niches and fifteen isolates from four well-defined outbreaks of foodborne gastroenteritis, were studied. Seven phage types, with a predominance of PT 4 (63% of isolates), were observed when analysing the epidemiologically unrelated group. PT 4 was detected in all of the ecological niches studied, including food and fecally polluted river and beach water. The discriminatory power for phage typing, the average probability that the typing system will assign a different type to two unrelated strains randomly sampled in the microbial population, was 0.62. Ten PFGE pattern types were obtained with Xba I restriction endonuclease enzyme among the unrelated isolates; thirteen isolates belonged to PFGE pattern type 1 and the rest of the PFGE types were assigned to one or two isolates. The Dice coefficient clustered the similarities of the PFGE patterns between 80-100%. PFGE showed a discriminatory power of 0.72. Five clearly distinct RAPD patterns were observed with the OPS-19 oligonucleotide, but the discrimination obtained was low (0.46). The combination of the three typing methods increased the number of types to seventeen, giving high discrimination (0.92). Seven of the isolates recovered from various ecological niches belonged to the combination PT 4/PFGE 1/RAPD A and other combinations were unique or included only two strains. The four epidemiologically well-defined foodborne outbreaks were associated with the PT 4 phage type. In two of the outbreaks, other phage types (PT 7a and RDNC) were also observed in two isolates. Most of the isolates belonging to the foodborne outbreaks had an identical PFGE pattern (PFGE pattern type 1), but a difference in a restriction band was observed in an isolate belonging to an outbreak. Two RAPD patterns were observed in the outbreaks; RAPD pattern type A was detected in three of the four outbreaks. When the combined typing method was applied to the study, high concordance was observed and most of the outbreak strains belonged to the combination PT 4/PFGE 1/RAPD A. It is concluded that the combination of phage type with PFGE and RAPD provides a powerful discriminatory tool for the epidemiological analysis of unrelated and related strains of S. enteritidis.

Bacteriophage Typing↗

Typing of Salmonella enteritidis of different phage types of PCR fingerprinting.

Fifty-nine isolates of Salmonella spp. were typed by PCR fingerprinting using three single primers: ERIC2, M13 and OPS-19. First, their discrimination power in a group of nine different serotypes were studied and considerable differences in the band patterns were obtained. Further, a panel of 51 isolates of Salmonella enteritidis with eight different phage types were analysed with the three primers. The discriminating power increased by combining the patterns of the three primers, and in this case it was possible to distinguish between some phage types of Salm. enteritidis, but not all of them were differentiated.

Bacterial Typing Techniques↗

Modification of kanamycin-esculin-azide agar to improve selectivity in the enumeration of fecal streptococci from water samples.

Kanamycin-esculin-azide agar was modified by increasing the concentration of sodium azide to 0.4 g liter-1 and replacing kanamycin sulfate with 5 mg of oxolinic acid liter-1. The modification, named oxolinic acid-esculin-azide (OAA) agar, was compared with Slanetz-Bartley and KF agars by using drinking water and seawater samples. The OAA agar showed higher specificity, selectivity, and recovery efficiencies than those obtained by using the other media. In addition, no confirmation of typical colonies was needed when OAA agar was used, which significantly shortens the time of sample processing and increases the accuracy of the method.

Agar↗

Q fever: prevalence of antibodies to Coxiella burnetii in the Basque country.

The aim of this study was to ascertain the prevalence of Coxiella burnetii infection in the Basque population. To this end a stratified sampling was carried out taking as a criterion the size of the population in a given area of residence. Residents in sparsely populated areas were found to have a prevalence of 38.5%, significantly greater than in highly populated areas (odds ratio [OR] = 1.56; 95% confidence interval [CI]: 1.08-2.27 and OR = 1.58; 95% CI: 1.09-2.30). The prevalence was significantly higher in males (36.3%) than in females (29%), and was found to increase significantly with age. Current or previous participation in activities relating to agriculture and/or livestock farming or even having contact with cattle, goats or sheep were also found to be risk factors for the infection. A fuller study of the determinants of chronic Q fever should be initiated.

Adolescent↗

The role of hens' eggs in outbreaks of salmonellosis in north Spain.

Most outbreaks of salmonellosis occurring in Spain are transmitted by hens' eggs. This prompted a study on the incidence of Salmonella in eggs. Salmonella was found in 5 of 372 eggs associated with foodborne disease; one strain was isolated from the inside of the egg. In 998 eggs not associated with human salmonellosis, Salmonella was found on 6 occasions, including once inside the egg. These percentages are in general higher than those found in other Western countries. The possibility of eggs being contaminated before laying is discussed and an approach for controlling salmonellosis transmitted in this way is presented.

Animals↗

Semisolid media for isolation of Salmonella spp. from coastal waters.

The use of two semisolid media (semisolid Rappaport and semisolid Rappaport-Vassiliadis) for the isolation of Salmonella spp. from coastal waters was compared with the use of conventional media. Two hundred and fifty six samples were studied; Salmonella spp. were detected in 83. The semisolid media were the most sensitive, detecting 73 samples instead of the 53 detected by the conventional media (P less than 0.001). The rate of isolation of Salmonella spp. showed an increase of 56.6% when the semisolid media were added to the conventional media, it being possible to detect 54.2% of the total organisms 1 day earlier.

Culture Media↗