Øivind Bergh

Flexibacter ovolyticus sp. nov., a Pathogen of Eggs and Larvae of Atlantic Halibut, Hippoglossus hippoglossus L.

A psychrotrophic Flexibacter sp., Flexibacter ovolyticus sp. nov., was isolated from the adherent bacterial epiflora of Atlantic halibut (Hippoglossus hippoglossus L.) eggs and was shown to be an opportunistic pathogen for halibut eggs and larvae. The strains which we isolated had the enzymatic capacity to dissolve both the chorion and the zona radiata of the egg shells. A total of 35 isolates were characterized by using morphological and biochemical tests. These strains were rod shaped, gram negative, Kovacs oxidase positive, and pale yellow and exhibited gliding motility. They did not produce acid from any of the wide range of carbohydrates tested. Our isolates had the ability to degrade gelatin, tyrosine, DNA, and Tween 80. Starch, cellulose, and chitin were not degraded. The strains were catalase and nitrate reductase positive, did not produce H2S, and did not grow under anaerobic conditions. F. ovolyticus resembles Flexibacter maritimus, but differs from the latter species in several biochemical and physiological characteristics. DNAs from F. ovolyticus strains had guanine-plus-cytosine contents which ranged from 30.3 to 32.0 mol% (strains EKC001, EKD002T [T = type strain], and VKB004), and DNA-DNA hybridization studies revealed levels of relatedness between F. ovolyticus EKD002T and F. maritimus NCMB 2154T and NCMB 2153 of 42.7 and 30.0%, respectively. Compared with previously described Cytophaga and Flexibacter spp. with low guanine-plus-cytosine contents, F. ovolyticus constitutes a new species. Strain EKD002 (= NCIMB 13127) is the type strain of the new species.

Ontogeny of lymphoid organs and development of IgM-bearing cells in Atlantic halibut (Hippoglossus hippoglossus L.).

In teleost fish, the head kidney, thymus, and spleen are generally regarded as important immune organs. In this study, the ontogeny of these organs was studied in Atlantic halibut (Hippoglossus hippoglossus), larvae at various stages of development. We observed that the kidney was present at hatching, the thymus at 33days post hatch (dph), while the spleen was the last organ to be detected at 49dph. All three lymphoid organs were morphologically well developed during late metamorphic stages. Real time RT-PCR analysis showed that IgM mRNA expression could be observed at 66dph and later, which correlates well with in situ hybridisation data showing that a few IgM positive cells could be detected in the anterior kidney and spleen from 66dph. Our data also showed that the highest levels of IgM mRNA could be detected in halibut spleen. Immunostaining using a monoclonal antibody against halibut IgM detected IgM positive cells at 94dph in both the head kidney and the spleen, which is much later than the IgM mRNA. Numerous cells expressing both IgM mRNA and protein could be detected in the spleen and anterior kidney and also to some extent in thymus specimens from adult halibut.

Addition of bacteria bioencapsulated in Artemia metanauplii to a rearing system for halibut larvae

Theaim of this study was to determine whether it is possible to influence thespecies composition of the gut microflora in 70-days old halibut larvae(Hippoglossus hippoglossus L.) by addition of bacteriabioencapsulated in Artemia franciscana in two shortpulses.Two Vibrio strains, PB 1-11 and PB 6-1 were used. Twotreatments received each strain alone, a third treatment received the twobacterial strains in mixture, whereas in a control treatment no bacteria wereadded. Five fish tanks were included in each treatment. The bacteria werebioencapsulated in Artemia franciscana metanauplii duringashort-term incubation in bacterial suspensions of the specific strains, andwereadded in two pulses, on day 1 and day 10 of the experiment. Addition ofbacteriadid not increase the total number of colony-forming units (CFU) in the larvalgut. The total CFU in the water was lower in tanks added bioencapsulatedbacteria than in the control treatment (p < 0.05). Animmunocolony blot assay was used to measure the numbers of the specificbacteriaadded in samples of water, A. franciscana, and fishlarvae.At the end of the experiment on day 13, the strain PB 1-11 reached on average10% of the total CFU when added alone and 25% when added in mixture with PB6-1.Strain PB 6-1 reached at the end of the experiment 26.8% of the total CFU whenadded alone and 24.2% when added in mixture with PB 1-11, whereas thepercentages of PB 1-11 and PB 6-1 in the control fish were 12.7% and 10.8% ofthe total CFU, respectively. However, the differences between the experimentaltreatments compared with the control group were not significant(p > 0.05). The addition of bioencapsulated bacteria intwo pulses within a ten days period was therefore not sufficient to influencethe species composition of the microflora of the halibut larvae.

Mortality of scallop spat in cultivation, infested with tube dwelling bristle worms, Polydora sp.

Between 1995 and 1997, a traditional Norwegian oyster poll with an attachednursery was used for the production of scallop (Pecten maximus) spat.During summer 1997, four batches of 2 mm scallop spat were placed in thenursery. In June, hydroids (Obelia) caused problems with fouling andaccumulation of sediment in the nursery. Thereafter, the scallop spat wereheavily infested with tube dwelling polychaets (Polydora sp.). Thepolychaets were both introduced with poll water and reproduced inside thenursery. Infested spat suffered high mortalities. Although the spat could have been affected by the hydroids, as well as shortage of food, Polydorainfestations were considered the main cause of the mortalities. Dead andinfested spat were continuously sorted out and discarded, but the infestationproblems persisted. As a result, the scallop spat did not represent anycommercial value for the company, and in September, the nursery wasemptied. In total, approximately one million spat – representing onethird of Norways intensive scallop spat production in 1997, was lost.