ABSTRACT
Piscirickettsia salmonis is the most important pathogen in salmonid mariculture in Chile. Since it was reported numerous piscirickettsiosis outbreaks have occurred differing in virulence and mortality. Genetic variability of P. salmonis isolates has been suggested as one factor to explain this. However until now isolates obtained from outbreaks have not been analyzed. Knowledge of genetic variability of P. salmonis is very limited and also a useful screening method for genetic variations in isolates without sequencing is not available. Here we report an electrophoretic analysis of internal transcribed spacer region (ITS) of eleven P. salmonis isolates obtained from different salmon species and places in southern Chile. When PCR products were submitted to polyacrylamide gel electrophoresis (PAGE) a characteristic electrophoretic pattern was observed, distinguishable from ITS of other bacteria, including fish pathogens. Even though this pattern is conserved in all isolates, a difference in ITS electrophoretic mobility was observed, determining clearly two groups: ITS with higher or with lower electrophoretic mobility, including LF-89 and EM-90 isolates, respectively. A higher ITS sequence homology inside each group was shown by heteroduplex mobility assay (HMA). Our results show that genetic variability between Chilean P. salmonis isolates allows the differentiation of two groups with similar behavior observed previously when six P. salmonis isolates from three geographic origins were analyzed by 16S, 23S and ITS sequencing. PAGE analysis of ITS and HMA could be a basis to develop an assay for screening genetic variability between P. salmonis isolates.
Subject(s)
DNA, Ribosomal Spacer/analysis , Gammaproteobacteria/genetics , Gammaproteobacteria/isolation & purification , Salmonidae/microbiology , Animals , Chile , Electrophoresis, Polyacrylamide Gel , Gammaproteobacteria/classification , Genotype , Nucleic Acid Heteroduplexes/analysis , Nucleic Acid Hybridization , Oncorhynchus kisutch/microbiology , Oncorhynchus mykiss/microbiology , Polymerase Chain Reaction , Polymorphism, Genetic , Salmo salar/microbiologyABSTRACT
The IgG antibody response specific to Helicobacter pylori was evaluated through ELISA in a group of 92 gastric patients colonized by this bacteria. 74 had gastritis and 19 gastroduodenal ulcer. Three control groups were studied in a similary way: normal adult volunteers (n=17), adults with E coli or S typhi bacteremia (n=30) and normal infants (n = 30). IgG antibody response to H pylori was demonstrated in 98% of colonized patients and 0% of infants. Asymptomatic individuals and those with bacteremia had high rates of antibody response (76 and 90% respectively), although this rate and also the titers of antibody response were significantly lower than that of colonized patients (p < 0.05). ELISA reactive sera from colonized patients and asymptomatic individuals evidenced a similar antibody pattern when tested by blotting. This profile was absent in non reactive sera, including those with high antibody titers to C jejuni. The presence of specific IgG antibodies to H pylory in the majority of colonized gastric patients and asymptomatic adults suggest that this infection is very common in our population
