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Aims The aim was to investigate the intestinal community of Enterococcus faecalis from healthy confined non-industrialized chicken. Methods and Results A total of 206 E. faecalis isolates were collected from cloacal swabs. The prevalence of E. faecalis from two confined flocks was 83 and 96%, while only 44 and 13% in two fancy breeder flocks. A total of 204 isolates were characterized by pulsed-field-gel-electrophoresis (PFGE) where 40 strains were selected for multi-locus sequence typing (MLST). In all, 19 PFGE patterns and 14 STs were obtained. Four STs were identified in each of the two confined flocks, with one shared ST, while seven STs were identified in the two fancy breeder flocks. Only six of the identified STs had previously been registered in chicken. Conclusion The majority of clonal lineages of E. faecalis associated with chicken reared in confinement and under backyard conditions were unrelated. Significance and Impact of the Study This study has provided new information on opportunistic E. faecalis in confined non-industrial chicken. Knowledge of population diversity and prevalence compared to conventional production is important to accesses, if certain clonal lineages are more likely to be associated with other intestinal E. faecalis lineages in chicken, and it is an important tool for gaining control of clones involved in disease and antibiotic resistance. [ABSTRACT FROM AUTHOR]

Aims The aim of this study was to investigate whether continuous contamination of light pasteurized egg products with Salmonella enterica serovar Tennessee ( S. Tennessee) at a large European producer of industrial egg products was caused by persistent contamination of the production facility and to characterize the persistent strains. Methods and Results Seventy-three S. Tennessee isolates collected from products over a 3-year period with intermittent contamination, and 15 control strains were compared by pulsed field gel electrophoresis ( PFGE) using two enzymes. Forty-five case isolates distributed throughout the full period were shown to belong to one profile type. Isolates representing different PFGE profiles were all assigned to ST 319 by multilocus sequence typing ( MLST). The case isolates did not show a higher ability to form biofilm on a plastic surface than noncase isolates. Characteristically, members of the persistent clone were weak producers of H2S in laboratory medium. S. Tennessee isolated from the case was able to grow better in pasteurized egg product compared with other serovars investigated. Conclusions It was concluded that the contamination was caused by a persistent strain in the production facility and that this strain apparently had adapted to grow in the relevant egg product. Significance and Impact of the Study S. Tennessee has previously been associated with persistence in hatching facilities. This is the first report of persistent contamination of an egg production facility with this serovar. [ABSTRACT FROM AUTHOR]

To characterize isolates of Salmonella Typhimurium DT41 obtained from infected flocks of broiler breeders by multiple-locus variable-number tandem-repeats analysis (MLVA) and compare results with a diverse strain collection from Germany and United Kingdom and isolates from Danish patients. A total of 102 isolates of Salm. Typhimurium phage type DT41 were MLVA typed. MLVA typing showed 4, 12, 25, 9 and 8 different alleles at the five MLVA loci 9, 5, 6, 10 and 3, respectively. A dendrogram based on MLVA types was constructed, and one large group, nine minor groups and 29 more unrelated MLVA types were obtained. The major group included 20 of the 30 human isolates. Isolates obtained from broiler breeders demonstrated major diversity, indicating the existence of several independent introductions of DT41 at farm level. When comparison was made to isolates included from Germany and England, DT41 seems to be ubiquitous in the wild fauna which might represent a risk factor for poultry. Transmission from Danish broilers to humans was not demonstrated, neither was the transmission from rearing farms to broiler breeder farms. Sources of infection at broiler breeder farm level remained unidentified. Major diversity was demonstrated for DT41 MLVA types. A persisting problem with infection of broiler breeder flocks with DT41 was not reflected in broiler flocks originating from these flocks. [ABSTRACT FROM AUTHOR]

Aims: To develop a RT-PCR method for detection of the multilocus sequence type 82 of Enterococcus faecalis associated with amyloid arthropathy (AA) in layers. Methods and Results: Bacteria were selected from lesions including AA in layers. The primers were designed based on the phosphate ATP binding cassette transporter ( pstS) and xanthine phosphoribosyltransferase ( xpt) genes and first tested against three isolates with known base pairs at the specific sites. Subsequently, 12 isolates were selected from our collection by one researcher, and RT-PCR was performed blinded. The sequence type (ST) was then confirmed by multilocus sequence analysis. Two single-nucleotide polymorphisms in the pstS and xpt genes allowed an unambiguous identification of ST82. As an alternative to DNA extraction, a boiling method for release of DNA from cells was used. Conclusions: The real-time PCR targeting ST82 enables rapid screening of Ent. faecalis cultured from suspect cases with results available after a few hours, much faster than multilocus sequence typing and pulse field gel electrophoresis. Significance and Impact of the Study: The new method allows a rapid screening of isolates with results available after only few hours. This RT-PCR method could be a useful tool for molecular epidemiological studies on the spread of arthropathic and amyloidogenic Ent. faecalis within and between birds more efficiently. [ABSTRACT FROM AUTHOR]

Aims: The aims of the present investigation were to characterize and identify serovars of Riemerella anatipestifer and Riemerella-like isolates genetically and to test the specificity of PCR tests reported for the identification of R. anatipestifer. Methods and Results: A total of 50 isolates from poultry tentatively classified with Riemerella anatipestifer were characterized genetically by partial sequencing of rpoB and by nearly full sequencing of the 16S rRNA gene for selected isolates. The results obtained were compared with the data from 13 reference strains by phylogenetic analysis. A total of 41 isolates were identified as R. anatipestifer, three as Wautersiella falsenii like, a single isolate as Pelistega europaea, while five isolates were classified as new, unnamed taxa. None of the reported PCR tests for identification of R. anatipestifer were found specific. Conclusions: Characterization of R. anatipestifer and related bacteria by traditional methods is often inconclusive because of inconsistent reactions and phenotypic diversity. For the same reason, gene sequencing and phylogenetic analysis are essential to allow proper classification and identification as demonstrated in the present study. Significance and impact of the Study: The present investigations demonstrated that isolates of R. anatipestifer are often misidentified, and that new serovars should not be accepted unless they have been properly characterized by relevant genetic methods such as gene sequencing. In addition, we showed that the published PCR tests are not specific for this species. Finally, two new taxa were outlined, the final taxonomic positions of which remain to be identified. [ABSTRACT FROM AUTHOR]

Aims: The aim of the present investigation was to identify and characterize Pasteurella-like isolates obtained from clinically affected psittacine birds. Methods and Results: A total of 37 isolates from psittacine birds tentatively classified with the family Pasteurellaceae were characterized phenotypically. The genetic relationship was investigated by sequencing of partial rpoB and 16S rRNA genes for selected isolates. The results obtained were compared with the data from 16 reference strains. Nine isolates were identified as Gallibacterium spp., 16 as Volucribacter spp. or Volucribacter-like, while 11 isolates were classified as taxon 44 of Bisgaard. A single isolate was identified as Pasteurella multocida. Conclusions: Characterization of Pasteurellaceae by traditional methods is often inconclusive because of inconsistent reactions and phenotypic diversity. For the same reason, genotyping is essential to allow proper classification as demonstrated in the present study. Significance and Impact of the Study: Limited information exists on the isolation and significance of Pasteurellaceae associated with clinically affected psittacine birds showing signs of digestive and/or respiratory disorders. The present investigations demonstrated that these organisms are widely distributed among clinically affected birds, but isolation of these taxa cannot be unambiguously correlated with the symptoms observed. [ABSTRACT FROM AUTHOR]

Aim: To investigate if taxon 42 of Bisgaard isolated from pigs represents genuine [ Pasteurella] caballi, which was previously only isolated from horses. Methods and Results: A total of 15 field isolates from horses and pigs from five different countries representing three continents were subjected to extended phenotypical characterization. Although minor differences were observed between taxon 42 and [ P.] caballi, these differences did not allow phenotypic separation. Ribotyping based on HindIII digestion showed five profiles based on nine band positions. One [ P.] caballi strain and two taxon 42 strains shared the same profile. Ribotyping using HpaII gave a higher diversity with nine profiles based on ten band positions. While no profiles were shared between the taxon 42 and [ P.] caballi strains, pattern analysis showed that two of the taxon 42 isolates were most similar (91% similarity) with a [ P.] caballi isolate. The 16S rRNA gene sequencing of one strain of taxon 42 and one strain of [ P.] caballi was performed and compared with the published sequence for the type strain of [ P.] caballi. The three strains showed nearly identical sequences with at least 99.8% similarity. DNA re-associations measured by the micro-well method were 79 and 77%, respectively between the type strain of [ P.] caballi and two strains of taxon 42 representing distinct ribotypes and confirmed that taxon 42 belongs to [ P.] caballi. Conclusion: The present investigation documents that [ P.] caballi can be isolated from clinical respiratory specimens from pigs and the recognized association with respiratory infections in horses and horse bite infection in humans. Strains classified as taxon 42 are [ P.] caballi isolated from pigs and for both pigs and horses, lesions mainly include the respiratory tract. Significance and Impact of the Study: The results will improve the diagnostics and progress studies of virulence and epidemiology of [ P.] caballi. [ABSTRACT FROM AUTHOR]

Abstract Aims: Characterization and classification of members of Pasteurellaceae isolated from birds by extended phenotypic characterization and 16S rDNA gene sequence comparison. Methods and Results: A total of 95 avian isolates were subjected to extended phenotypic characterization. Thirteen bacterial strains selected from main phenotypic clusters and isolated from parrot, parakeet, budgerigar, partridge, pheasant, chicken, duck, hawk and gull were subsequently characterized by 16S rDNA gene sequencing. Eight of the sequenced strains were classified with six taxa of Bisgaard of which two (34 and 40) have not been published before, and the properties of four others (14, 22, 26 and 32) changed upon the characterization of these new isolates. Of the remaining strains, one was identified as a phenotypic variant in maltose and dextrin of Pasteurella gallinarum another as a trehalose positive variant of taxon 3 of Bisgaard. The remaining three strains sequenced were not closely related to existing taxa of Pasteurellaceae . However, they were found to belong to the Avian cluster with 92–97% 16S rDNA gene sequence similarity. Conclusion: The study allowed the classification of bacteria isolated from birds by the integrated use of extended phenotypic characterization and 16S rDNA gene sequence analysis. Only the application of 16S rDNA gene sequencing allows a correct identification of variant strains. Significance and Impact of the Study: The description of new taxa within the bacterial family Pasteurellaceae will subsequently allow additional isolates of these taxa to be identified and improve the diagnosis and epidemiological understanding of bacteria causing disease in birds. [ABSTRACT FROM AUTHOR]

Abstract Aims: To investigate contamination of surfaces on a poultry slaughter line from infected poultry and subsequent cross-contamination of non-infected poultry. Methods and Results: A broiler slaughterhouse was investigated for the presence of Salmonella on 17 defined points over two 1-week periods. Flocks supplied to slaughter and neck skin samples from processed chicken were likewise investigated. Salmonella was detected in 10 out of 18 flocks at ante-mortem inspection, while seven flocks tested positive in the finished products. Equipment at all but one control point at the slaughter line tested positive at least once during the study. The chicken receiving area was the most contaminated. By comparison of typing results from serotyping, plasmid profile typing and phage typing, direct evidence for cross-contamination with Salm. serotype Typhimurium, Salm. Serotype 4·12:b:– and Salm. serotype Virchow on the slaughter line was obtained for four of the flocks. The cleaning procedure in place did not remove all Salmonella from the contaminated areas. Conclusions: Evidence for contamination of equipment on a slaughter line and subsequent cross-contamination to non-infected chicken was provided by typing methods. Significance and Impact of the Study: This study has provided detailed information on cross-contamination on a slaughter line by the use of phage typing and plasmid profiling. The study stresses the importance of controlling Salmonella in the primary production, as contamination of the equipment on the slaughter line will act as a vehicle to contaminate finished products. Cleaning procedures on slaughter lines cannot be expected to control this problem with the current equipment. [ABSTRACT FROM AUTHOR]

Aims: To relate sensory shelf-life of iced whole and gutted squid to bacterial growth and chemical changes. Methods and Results: Cooked mantles from whole and gutted individuals were rejected after 10 and 12 days of storage, respectively, due to ammoniacal off-odours. Rate of production of both ammonia and trimethylamine was highest in the whole lot. Agmatine, which was only present in trace amounts in freshly-caught squid, increased rapidly in both lots. The main microflora at the time of sensory rejection of iced whole squid included Gram-negative, motile and non-fermentative rods, which were psychrophilic and had a requirement for NaCl. 16S rDNA sequence analyses identified the strains as belonging to the genus Pseudoalteromonas . Shewanella putrefaciens , Pseudoalteromonas sp. and Pseudomonas sp. dominated in spoiled gutted squid. Identification of strains from the stomach and digestive gland of recently-captured squid showed that the main flora consisted of Photobacterium phosphoreum. Conclusions: Spoilage of iced squid is likely to result from a combination of autolytic and bacterial changes. Agmatine seems to be an excellent freshness indicator. Photobacterium phosphoreum may contribute to spoilage through activity in the digestive gland, followed by diffusion of volatile compounds and amines to the mantle. Significance and Impact of the Study: Due to the psychrophilic nature of P. phosphoreum and Pseudoalteromonas sp., spread-plating and low temperature incubation are recommended for bacteriological evaluation of iced squid. [ABSTRACT FROM AUTHOR]

The article focuses on young age colorectal cancer and identification of hereditary non-polyposis colorectal cancer (HNPCC) cohorts. It mentions that prophylactic measures such as colonoscopy and subsequent polypectomy are available in hereditary colorectal cancer. The major steps toward the resolution of the consequences of HNPCC include localization and identification of mismatch repair genes and genes involved in the adenoma-carcinoma multistep sequence, molecular genetic diagnosis and analysis of microsatellite instability.