Flavobacterium psychrophilum causes bacterial cold-water disease (BCWD) in farmed rainbow trout ( Oncorhynchus mykiss ), with the multilocus sequence typing (MLST) clonal complex (CC) CC-ST10 accounting for the majority of outbreaks globally. The development of alternative strategies to antibiotic treatment of BCWD using bacteriophage-based control of F. psychrophilum , or virulence factors as targets for therapy, requires knowledge of the phage-sensitivity of outbreak strains and of universal traits contributing to their pathogenicity. To examine the association between virulence and both genetic (MLST sequence type (ST) and PCR-serotype) and phenotypic characteristics (adherence, antibiotic resistance, colony spreading motility, hemolytic and proteolytic activity), the median lethal dose (LD 50 ) of 26 geographically disparate F. psychrophilum isolates was determined in rainbow trout. Furthermore, the in vitro sensitivity of the isolates against five bacteriophages was determined by the efficiency of plating (EOP). The tested F. psychrophilum isolates were mainly represented by CC-ST10 genotypes (22 out of 26) and showed up to 3-log differences in LD 50 (8.9 × 10 3 to 3.1 × 10 6 CFU). No association between MLST ST and virulence was found because of a high variation in LD 50 within STs. All identified serotypes (0, 1, and 2) were pathogenic, but ten most virulent isolates belonged to serotype 1 or 2. Isolates of high (LD 50 < 10 5 CFU), moderate (LD 50 = 10 5 –10 6 CFU), and weak (LD 50 > 10 6 CFU) virulence were similar in phenotypic characteristics in vitro . However, the only non-virulent CC-ST10 isolate was deficient in spreading motility and proteolytic activity, indicating that the characteristics are required for pathogenicity in F. psychrophilum . Univariate correlation studies found only non-significant associations between LD 50 and the measured phenotypic characteristics, and the multivariable analysis did neither reveal any significant predictors of virulence. The majority of isolates (16 out of 26) were sensitive to at least four bacteriophages, with up to a 6-log variation in the EOP. Most CC-ST10 isolates (16 out of 22) were sensitive to the examined phages, including 5 out of the 7 most virulent isolates represented by prevalent and antibiotic-resistant STs. Our findings suggest that control of BCWD using lytic phages or interventions targeting shared characteristics of pathogenic F. psychrophilum strains should be further explored.

Nineteen Vibrio anguillarum-specific temperate bacteriophages isolated across Europe and Chile from aquaculture and environmental sites were genome sequenced and analyzed for host range, morphology and life cycle characteristics. The phages were classified as Siphoviridae with genome sizes between 46,006 and 54,201 bp. All 19 phages showed high genetic similarity, and 13 phages were genetically identical. Apart from sporadically distributed single nucleotide polymorphisms (SNPs), genetic diversifications were located in three variable regions (VR1, VR2 and VR3) in six of the phage genomes. Identification of specific genes, such as N6-adenine methyltransferase and lambda like repressor, as well as the presence of a tRNAArg, suggested a both mutualistic and parasitic interaction between phages and hosts. During short term phage exposure experiments, 28% of a V. anguillarum host population was lysogenized by the temperate phages and a genomic analysis of a collection of 31 virulent V. anguillarum showed that the isolated phages were present as prophages in >50% of the strains covering large geographical distances. Further, phage sequences were widely distributed among CRISPR-Cas arrays of publicly available sequenced Vibrios. The observed distribution of these specific temperate Vibriophages across large geographical scales may be explained by efficient dispersal of phages and bacteria in the marine environment combined with a mutualistic interaction between temperate phages and their hosts which selects for co-existence rather than arms race dynamics.

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Sequential biopsies (4-72 h) of early allergic and irritant patch test reactions have been examined immunohistologically for reactivity with 19 monoclonal antibodies against antigens on lymphoid cells in order to investigate the nature/origin of the infiltrating lymphoid cells and assess their state of activation/proliferation. The composition of the infiltrates was similar in allergic and irritant reactions and consisted of T-lymphocytes of helper/inducer types in association with T-cell accessory cells, i.e., Langerhans cells and HLA-DR-positive macrophages. No differences in expression of T-cell or macrophage associated antigens were seen in early as opposed to late biopsies. In contrast, the proportion of cells positive for markers associated with activation (interleukin-2 receptor) or proliferation (transferrin receptor, the Ki-67 nuclear antigen) of lymphoid cells was found to increase with time in both types of reaction. These data substantiate the view that T-cell immune reactions are implicated in both allergic and toxic patch test lesions, and indicate that the lymphocytes in the infiltrates are activated and proliferate.