Search PubMed⌕ Search

Biomedical subjects

R E Olsen

Publications and source records attributed to R E Olsen.

At least 19 recordsLinked to original sources

Histological changes in intestine of Atlantic salmon (Salmo salar L.) following in vitro exposure to pathogenic and probiotic bacterial strains.

Furunculosis and vibriosis are diseases that cause severe economic losses in the fish-farming industry. The foregut of the Atlantic salmon (Salmo salar L.) was exposed in vitro to two fish pathogens, Aeromonas salmonicida (causative agent of furunculosis) and Vibrio anguillarum (causative agent of vibriosis), and to one probiotic strain, Carnobacterium divergens, at 6 x 10(4) or 6 x 10(6) viable bacteria per milliliter. Histological changes following bacterial exposure were assessed by light and electron microscopy. Control samples (foregut exposed to Ringer's solution only) and samples exposed only to C. divergens had a similar appearance to intact intestinal mucosal epithelium, with no signs of damage. However, exposure of the foregut to the pathogenic bacteria resulted in damaged epithelial cells, cell debris in the lumen, and disorganization of the microvilli. Co-incubation of the foregut with a pathogen and C. divergens did not reverse the damaging effects caused by the pathogen, although these were alleviated when probiotic bacteria were used. Based on these results, we suggest that the probiotic bacterium, C. divergens, is able to prevent, to some extent, pathogen-induced damage in the Atlantic salmon foregut.

Aeromonas salmonicida↗

Translocation of viable Aeromonas salmonicida across the intestine of rainbow trout, Oncorhynchus mykiss (Walbaum).

The pathogenic bacterium Aeromonas salmonicida is the causative agent of the destructive disease furunculosis in salmonids. Horizontal transmission in salmonids has been suggested to occur via the skin, gills and/or intestine. Previous reports are contradictory regarding the role of the intestine as a route of infection. The present study therefore investigates the possibility of bacterial translocation across intestinal epithelia using Ussing chamber technology, in vitro. Intestinal segments were exposed for 90 min to fluorescein isothiocyanate-labelled pathogenic A. salmonicida. Sampling from the serosal side of the Ussing chambers showed that bacteria were able to translocate across the intestinal epithelium in both the proximal and distal regions. Plating and subsequent colony counting showed that the bacteria were viable after translocation. During the 90 min exposure to A. salmonicida, the intestinal segments maintained high viability as measured by electrical parameters. The distal region responded to bacterial exposure by increasing the electrical resistance, indicating an increased mucus secretion. This study thus demonstrates translocation of live A. salmonicida through the intestinal epithelium of rainbow trout, suggesting that the intestine is a possible route of infection in salmonids.

Aeromonas salmonicida↗

Rainbow trout can discriminate between feeds with different oil sources.

The purpose of present two-choice trials was to examine the capacity of groups of juvenile rainbow trout to differentiate between two isolipidic diets containing distinct oils and to detect an eventual preference. The choice was offered by means of two self-feeders per tank. One feeder distributed a standard diet with fish oil (FO), the other a diet containing vegetable oil, either rich in linolenic acid (linseed oil, LO), linoleic acid (sunflower oil, SO), or oleic acid (rapeseed oil, RO). Each 15-day preference test was preceded by a 15-day adaptation period during which both feeders distributed the same diet. The tests were followed by a 10- to 15-day validation period in order to confirm that feeder solicitations were steered by the characteristics of the diets. Preferences were expressed as relative changes in feed demands for a specific feeder. Averaged over all groups, the preference tests demonstrated the capacity of rainbow trout to discriminate between a diet with FO and a diet containing vegetable oil, and indicated a general preference for the diet with FO over the other diets irrespective of whether they received the diet with fish oil (Experiment 1) or with vegetable oil (Experiment 2) prior to the preference test. The tests also indicated a difference in the extent of relative avoidance of each of the three vegetable oil diets. Diet LO was the most avoided, as indicated by the 37-39% decrease in demands for the feeder with diet LO (P<0.05). Diet RO was the best accepted, causing a decrease in feed demands of only 15-17% (P>0.05). The avoidance of diet SO at the end of the preference test was 30% (P>0.05) after an initially higher avoidance of 43% (P<0.05). It is believed that the metabolic consequences of the excess of linolenic or linoleic acid negatively affected the feed acceptances of diets LO and SO. Further work is needed to elucidate a possible interference of differences in palatability. In all groups, the lower demands for the vegetable oil diets were compensated by increased demands for diet FO. Hence, changes in diet selection had no effect on total feed or energy intakes, measured as the sum of both selections.

Analysis of Variance↗

Epithelium-associated bacteria in the gastrointestinal tract of Arctic charr (Salvelinus alpinus L.). An electron microscopical study.

AIMS: The primary aim was to use transmission electron microscopy (TEM) and scanning electron microscopy (SEM) to define the location of epithelium-associated bacteria in the digestive tract of the salmonid fish, Arctic charr (Salvelinus alpinus). METHODS AND RESULTS: TEM and SEM examination of the gastrointestinal tract demonstrated substantial numbers of ovoid and rod-shaped bacterial cells associated with the microvillous brush borders of enterocytes. Bacteria were found at the tips of microvilli as well as between adjacent microvilli. Endocytosis of bacteria by epithelial cells was observed in two regions (pyloric caeca and midgut). CONCLUSION: Electron microscope examination of the gut is an important tool for evaluating the microbial ecology of the fish digestive tract ecosystem. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of the current study clearly demonstrate that the intestine is involved in bacterial endocytosis.

Animals↗

Identification and characterization of Carnobacteria isolated from fish intestine.

Eleven bacterial strains were isolated from the gastrointestinal tract of four fish species, Atlantic salmon (Salmo salar L.), Arctic charr (Salvelinus alpinus L.), Atlantic cod (Gadus morhua L.) and wolffish (Anarhichas lupus L.). All the strains were Gram-positive rods, non-sporing, catalase and oxidase-negative, able to grow at pH 9.0 but not on acetate containing media (pH < or = 5.4), and were fermentative. They had a high content of oleic acid (18:1 n-9) in cellular lipid, and were found to belong to the genus Carnobacterium by phenotypic criteria. The eleven carnobacteria strains were further identified on the basis of 16S rDNA sequence analysis and AFLP(TM) fingerprinting.

Aeromonas↗

Lactic acid bacteria associated with the digestive tract of Atlantic salmon (Salmo salar L.).

The present study reports the effect of excessive handling stress and starvation on the lactic acid bacteria associated with the digestive tract of Atlantic salmon (Salmo salar L.). A relatively low population level (approximately 2 x 103 bacteria per gram wet tissue) of viable adherent heterotrophic bacteria was associated with the digestive tract (foregut, midgut and hindgut). Of the 752 bacterial isolates isolated from diet, water and the digestive tract, 201 isolates belonged to the carnobacteria. Of these isolates, one from the diet, one from the rearing water and 80 from the gastrointestinal tract, were further identified on the basis of 16S rDNA sequence analysis. All these isolates were identified as being Carnobacterium piscicola-like. Daily repeated stress and starvation of the fish over 11 d had no influence on the total culturable bacterial numbers or population level of C. piscicola associated with the digestive tract. C. piscicola-like isolates colonizing the various intestinal regions (foregut, midgut and hindgut) were also screened for their ability to produce growth inhibitory compounds active against the fish pathogen Aeromonas salmonicida. Of the 199 C. piscicola isolates tested, 139 inhibited growth of the pathogen.

Aeromonas↗

The effect of diet on aerobic bacterial flora associated with intestine of Arctic charr (Salvelinus alpinus L.).

In order to extend the knowledge on the possible effect of diet on the gastrointestinal microbial community of fish, Arctic charr (Salvelinus alpinus L.) were fed diets containing high (23.7%) and low (6.4%) levels of carbohydrate. The number of viable aerobic and facultative aerobic bacteria associated with the digestive tract were not influenced by dietary regimen. A wide range of bacterial species was isolated, and the predominant bacterial species of both rearing groups were identified as Staphylococcus. There were, however, some differences in bacterial composition between the rearing groups, as well as inter-individual variations. For example, atypical Aeromonas salmonicida were isolated from the small and large intestine of two fish fed low dietary carbohydrate, while Aer. caviae-like isolates were found in the small intestine of four fish fed high carbohydrate. Non-motile Aeromonas spp. were found in the rearing group fed high dietary carbohydrate, but at low frequencies. Dietary manipulation seemed to influence the species composition of carnobacteria, Gram-positive rods, oxidase and catalase-negative and fermentative metabolism. Carnobacterium piscicola-like bacteria were only found in the small intestine, while C. mobile-like and Carnobacterium spp. were isolated from the large intestine of fish fed high carbohydrate. On the contrary, C. divergens-like isolates were found associated with the small and large intestine of fish fed low dietary carbohydrate.

Animals↗

Cross-pin femoral fixation: a new technique for hamstring anterior cruciate ligament reconstruction of the knee.

A new method of femoral fixation for arthroscopic anterior cruciate ligament (ACL) reconstruction using semitendinosus and gracilis (ST/G) tendons is presented. Biomechanical evaluation of the method was performed with pullout tests using animal tissues simulating fixation of ST/G tendons passing around a 2.5-mm pin placed transversely through a femoral tunnel. Clinical assessment of 22 patients was performed according to IKDC, Tegner, Lysholm, isokinetic strength, and KT1000 standards. Fixation strengths ranged from 725 N to 1,600 N for yield and maximal loads to failure for 35-mm to 70-mm cross-pins. Clinical results at mean follow-up of 30 months showed one traumatic failure, IKDC 86% normal or nearly normal, Tegner 6 ave, Lysholm 93 ave, KT1000 86% < 3 mm side-to-side difference on maximal manual evaluation, and isokinetic strength testing of quadriceps 8% deficit with hamstrings 8% deficit. Cross-pin fixation of ST/G tendons may be one of the strongest modes of femoral graft attachment yet devised. Our clinical outcomes in this initial series compare well with previous reports of ACL reconstruction regardless of graft or technique.

Adolescent↗

The effect of dietary fatty acids on lactic acid bacteria associated with the epithelial mucosa and from faecalia of Arctic charr, Salvelinus alpinus (L.).

Arctic charr, Salvelinus alpinus (L.), held in fresh water, were fed four experimental diets containing different polyunsaturated fatty acids (PUFA). In addition, one group fed a diet containing only coconut oil as sole lipid source served as control. The population of aerobic heterotrophic bacteria associated with the epithelial mucosa and the faecalia was estimated using the dilution plate technique. Generally, the population level of adherent bacteria increased along the digestive tract (stomach, small intestine and large intestine). Adherent Gram-negative and Gram-positive bacteria seemed to be present at equal levels in all parts of the alimentary tract. Lactic acid bacteria dominated among the Gram-positive bacteria, and they were detected in all regions of fish fed the PUFA supplemented diets. The frequency of lactic acid bacteria was highest in the digestive tract of fish fed diets with added 7.0% linolenic acid (18:3 n-3) or 4% of a PUFA mix. A lower frequency of lactic acid bacteria was found in fish fed dietary linoleic acid (18:2 n-6), and they were absent or present in low numbers in fish fed the coconut oil diet. It is suggested that dietary fatty acids affect the attachment sites for the gastrointestinal microbiota, possibly by modifying the fatty acid composition of the intestine wall. Numerical taxonomy procedures showed that the lactic acid bacteria Carnobacterium spp. and a Carnobacterium piscicola-like strain were predominant, with smaller numbers of Lactobacillus plantarum, Streptococcus spp. and Leuconostoc mesenteroides present. Seven strains of Carnobacterium spp. were further identified on the basis of 16S rDNA sequence analysis, and all these strains were identified as Carnobacterium piscicola.

Animal Feed↗

Effect of prior infection with virulent Shigella flexneri 2a on the resistance of monkeys to subsequent infection with Shigella sonnei.

All virulent shigellae have large plasmids. Plasmid-associated genes encode the expression of membrane-associated proteins (MAP), some of which correlate with the ability to invade susceptible epithelial cells. These MAP are serologically related in all of the shigella serotypes and evoke an antibody response after infection. To determine whether the MAP have a significant role in protection, 24 monkeys were infected with virulent Shigella flexneri 2a. After recovery, one group (with controls) was rechallenged with S. flexneri 2a; another group (with controls) was fed Shigella sonnei. The animals that were rechallenged with S. flexneri 2a were protected, while those that were fed S. sonnei experienced the same incidence of disease as controls. No differences in serum immune response to MAP after primary infection with S. flexneri were detected in immunoblots using lysates of S. flexneri or S. sonnei or in ELISA using water extracts of these strains.

Animals↗

Colorimetric determination of disulfiram in tablets: collaborative study.

The colorimetric reaction of cuprous iodide and disulfiram has been collaboratively studied in 8 laboratories to determine the drug in tablet form at 2 dosage levels. The disulfiram was extracted from the tablet matrix with methylene chloride. After a basic wash with 1 M NaOH, an aliquot of the methylene chloride solution was reacted with solid cuprous iodide, and the absorbance of the resulting color was measured. Single assays on 2 synthetic preparations of known disulfiram content were performed with average recoveries of 98.4 and 99.2% for 93.3 and 105.3 mg, respectively. Duplicate determinations on 2 commercial tablet preparations declared at 250 and 500 mg gave mean and standard deviation values of 246.6 +/- 6.40 and 493.6 +/- 12.0 mg, respectively. The results agreed closely with those obtained by the author using the NF iodometric titration procedure. The method has been adopted official first action.

Chromatography, Thin Layer↗

Platelet cryopreservation: a simple routine.

Cryopreservation of platelets as a blood-bank routine procedure offers functional platelets on demand. A simple method of freezing, thawing, and administering platelets which is within reach of any hospital with routine blood-banking facilities is outlined. Sixteen frozen-thawed platelet transfusions are reported.

Blood Platelets↗