High-resolution genotyping of Salmonella strains by AFLP-fingerprinting.

High resolution AFLP fingerprinting, in which subsets of genomic restriction fragments are amplified by means of PCR, was used for the identification of different. Salmonella serotypes to investigate whether this technique is applicable in epidemiological studies. Seventy-eight different Salmonella strains comprising 62 serotypes were genetically identified by AFLP. Primer combination M00 (MseI primer without additional 3' nucleotides) and E11 (EcoRI primer with two additional 3' nucleotides) resulted in reproducible profiles containing approximately 50 bands. All serotypes were characterized by a unique profile. In addition, AFLP fingerprinting enabled phage type identification. Different strains previously identified as identical, using typing methods with lower resolution, could be distinguished, showing that AFLP fingerprinting is well suited for bacterial epidemiology and identification.

Discrepancy between Penner serotyping and polymerase chain reaction fingerprinting of Campylobacter isolated from poultry and other animal sources.

Thirty-four Campylobacter jejuni or coli strains, isolated from various livestock and darkling beetles from two Dutch poultry farms during different broiler production cycles, were subjected to Penner serotyping and polymerase chain reaction (PCR) fingerprint analysis. Ten different Penner serotypes were determined in the isolates. Visual scoring of the PCR fingerprints resulted in 14 clearly different profiles. Some strains with identical Penner serotypes exhibited different PCR fingerprints and conversely strains with different serotypes produced identical PCR fingerprints. Discrepancies between Penner serotyping and PCR fingerprinting were most obvious between isolates from different animal sources. Indications for the occurrence of genomic rearrangements were found. The inconsistency between serotyping and fingerprinting of Campylobacter strains suggests that conventional typing methods should be used in combination with fingerprinting if the epidemiological factors that contribute to Campylobacter colonization of live chickens are to be assessed reliably.

Polymerase chain reaction identification of Salmonella serotypes.

Seventy-seven Salmonella isolates comprising 61 different serotypes were subjected to polymerase chain reaction (PCR) fingerprinting using two primersets. Primerset L1/G1, amplifying the spacer regions between the 16S and 23S rRNA genes, resulted in simple PCR fingerprints. However, in some cases PCR amplification of different Salmonella serotypes with primerset L1/G1 resulted in identical fingerprint profiles. Fingerprints obtained with the ERIC primerset, that matches the enterobacterial repetitive intergenic consensus sequence, were more complicated but were serotype-specific. Consequently, fingerprinting with the ERIC primerset is applicable for typing Salmonella up to the serotype level. Fingerprinting with the L1 and G1 primers requires an additional treatment of the amplification product for accurate typing of salmonellas. Phage typing is not possible with either primerset.

Prevention of colonization by Salmonella enteritidis PT4 in broiler chickens.

Field experiments in The Netherlands and in Scandinavian countries have shown that an undefined microflora originating from SPF adult poultry will reduce considerably the colonization of young chicks by Salmonella. A commercial product from this so-called Nurmi concept, Broilact, was studied for its effectiveness in preventing infection of broilers with Salmonella enteritidis PT4 (S.e.). Two trials were carried out, in which the birds were exposed to S.e. via 'seeder' birds placed among them. The trial period was 21 days and each week one third of the chicks was killed and their caecal contents examined for salmonellas. The results of the first trial can be summarized as follows. (1) After 2 weeks the number of 'seeder' birds carrying the Salmonella decreased sharply; (2) the proportion of infected chicks in the Broilact-treated group was lower than in the non-treated group; (3) Counts of S.e. in the non-treated group were higher than in the Broilact group. Results of the second trial were comparable, although no salmonellas could be isolated after the second week.