agar and Fish-Diseases

Topics: Aeromonas; Agar; Ampicillin; Animals; Anti-Bacterial Agents; Cichlids; Fish Diseases; Ghana; Lakes; Tetracyclines

Four different colony morphologies were produced by Flavobacterium columnare strains on Shieh agar plate cultures: rhizoid and flat (type 1), non-rhizoid and hard (type 2), round and soft (type 3), and irregularly shaped and soft (type 4). Colonies produced on AO agar differed from these to some extent. The colony types formed on Shieh agar were studied according to molecular characteristics [Amplified Fragment Length Polymorphism (AFLP), Automated Ribosomal Intergenic Spacer Analysis (ARISA), and whole cell protein SDS-PAGE profiles], virulence on rainbow trout fingerlings, and adhesion on polystyrene and fish gills. There were no molecular differences between colony types within one strain. Type 2 was the most adherent on polystyrene, but type 1 was the most virulent. Adhesion of F. columnare strains used in this study was not connected to virulence. From fish infected with colony type 1, three colony types (types 1, 2 and 4) were isolated. Contrary to previous studies, our results suggest that strong adhesion capacity may not be the main virulence factor of F. columnare. Colony morphology change might be caused by phase variation, and different colony types isolated from infected fish may indicate different roles of the colony morphologies in the infection process of columnaris disease.

Topics: Agar; Animals; Bacterial Adhesion; Bacterial Proteins; Culture Media; DNA, Ribosomal Spacer; Electrophoresis, Polyacrylamide Gel; Fish Diseases; Flavobacteriaceae Infections; Flavobacterium; Gills; Oncorhynchus mykiss; Polymorphism, Restriction Fragment Length; Polystyrenes; Virulence

Piscirickettsia salmonis, a serious bacterial pathogen of farmed marine fish, previously considered culturable only in eukaryotic cell-culture systems, was grown for the first time on agar and broth containing enhanced levels of cysteine, thus greatly increasing the potential for isolation, in vitro culture and study of this organism. Virulence towards Atlantic salmon following passage on agar media was retained in a controlled laboratory trial. Of the studied temperatures, optimal growth on agar was observed at 22 degrees C.

Topics: Agar; Animals; Culture Media; Disease Outbreaks; Fish Diseases; Fisheries; Piscirickettsia; Piscirickettsiaceae Infections; Salmo salar

Piscirickettsia salmonis is the etiologic agent of piscirickettsiosis, an economically significant disease of fish. Isolation of P. salmonis by culturing on fish cell lines has been the standard technique since the initial isolation of the organism. The ability to grow P. salmonis on artificial media would relieve facilities of the cost of maintaining cell lines, permit isolation at fish culture sites with fewer contamination problems, and allow easier transport of isolates to diagnostic facilities for confirmation assays. This report describes the successful culture of P. salmonis on enriched blood agar.

Topics: Agar; Animals; Cell Culture Techniques; Fish Diseases; Piscirickettsia; Piscirickettsiaceae Infections; Salmonidae

In the present study, Anacker and Ordal agar, marine agar (MA), and Flexibacter maritimus medium (FMM) were compared with the dilute versions of Mueller-Hinton agar (DMHA) medium recommended by the National Committee for Clinical Laboratory Standards (NCCLS) for their use in disk diffusion tests with Tenacibaculum maritimum strains and to calculate the MICs of five drugs by the Etest method. Preliminary growth tests performed with 32 strains of this pathogen on each medium revealed that all strains failed to grow on DMHA, while the remaining media supported good growth of all isolates. In the susceptibility tests, which were carried out with the other three media, all strains were resistant to oxolinic acid and were highly susceptible to amoxicillin and trimethoprim-sulfamethoxazole, showing a good correspondence with the Etest values, which ranged from 0.064 to 0.75 and 0.006 to 1.5 mug/ml, respectively. Enrofloxacin and oxytetracycline produced significantly smaller inhibition zones and MICs on MA than on the other media assayed. However, fast, clear, and well-defined zones of inhibition were displayed for all strains at 24 h of incubation only on FMM by both the disk diffusion assay and Etest. In addition, FMM prepared with commercial sea salts instead of seawater was also suitable for bacterial isolation as well as for susceptibility testing. On the basis of these results, the use of FMM to determine the in vitro susceptibility of T. maritimum and its inclusion in a future revision of the NCCLS M42 report are recommended.

Topics: Agar; Animals; Anti-Bacterial Agents; Bacteriological Techniques; Culture Media; Fish Diseases; Fishes; Flavobacteriaceae; Flavobacteriaceae Infections; Microbial Sensitivity Tests; Seawater

The survival of skin penetrating cercariae depends on information on the direction to move toward deeper layers in the epidermis (the direction of further migration) and toward the surface (direction which must be avoided when migrating). We tested the hypothesis that parasites can use their photo-sensitivity for orientation away from the light-exposed skin surface towards darker locations. Cercariae of species invading humans ( Schistosoma mansoni), birds ( Trichobilharzia ocellata) and fish ( Diplostomum spathaceum) oriented towards light sources when free swimming in cuvettes. However, they shifted to a negative photo-orientation when migrating in agar substrates after penetration and transformation to schistosomula. This is a first hint that parasites may use photo-orientation when they navigate in host tissues.

Topics: Agar; Animals; Bird Diseases; Darkness; Ducks; Fish Diseases; Host-Parasite Interactions; Humans; Light; Movement; Schistosoma mansoni; Schistosomatidae; Trematoda; Trematode Infections; Water

Rainbow trout fry syndrome and cold-water disease are serious diseases in farmed salmonid fish. In the present study, three methods were compared, for the detection of the causative pathogen, Flavobacterium psychrophilum in water. The methods included traditional agar plate cultivation on tryptone yeast extract salts (TYES) agar, immunofluorescence antibody technique (IFAT) and nested PCR. The three methods were subsequently used for the detection of F. psychrophilum from fish farm environments. The nested PCR was the most sensitive method used for a detection of F. psychrophilum. As low as 3 CFU estimated by agar plate cultivation or 41 cells estimated by IFAT of F. psychrophilum per ml of non-sterile well water were needed for a detection using the nested PCR method. The obtained detection limits for the agar plate cultivation and the IFAT was 32 CFU/ml and 410 cells/ml, respectively. Using IFAT and nested PCR F. psychrophilum was detected most frequently in water samples from fish farms, but the pathogen was isolated from only a few samples using agar plate cultivation. In the present study IFAT and nested PCR proved to be rapid, specific and sensitive methods compared to traditional agar plate cultivation for the detection of F. psychrophilum from environmental samples. It is suggested that IFAT and nested PCR provide effective tools for the examination of F. psychrophilum in the environment.

Topics: Agar; Animals; Fish Diseases; Fish Products; Fishes; Flavobacterium; Fluorescent Antibody Technique; Fresh Water; Polymerase Chain Reaction; Sensitivity and Specificity; Water Microbiology

To investigate the relationships between water temperature, bacterial growth, virulence and antigen expression in Vibrio salmonicida, the causal agent of cold water vibriosis in Atlantic salmon (Salmo salar L.).. The significance of sea temperature was investigated using historical clinical and oceanographic data. An upper threshold for disease of approx. 10 degrees C was established. The effects of culture temperature and media type on bacterial growth were studied on solid and in liquid media. The highest rates of cell division were identified at 15 degrees C on solid media and 10 degrees C in liquid media. Outer membrane protein (OMP) expression and serological response in Atlantic salmon were studied using sodium dodecyl sulphate-polyacrylamide gel electrophoresis, Western blotting and enzyme-linked immunosorbent assay. A novel 76-kDa OMP produced in unshaken cultures at 10 degrees C was not found to stimulate a specific humoral response.. Diagnostic agar plate-based incubation of suspected V. salmonicida should be carried out at 15 degrees C. High yield broth cultures for vaccine production should be incubated at 10 degrees C or lower.. This study is, to the best of our knowledge, the first to identify different optimal temperatures in a bacterial species cultured on physically different types of media. The evidence presented suggests that V. salmonicida and possibly other bacteria destined for vaccine use in poikilothermic organisms should be cultured at temperatures consistent with that at which disease occurs.

Topics: Agar; Animals; Antibodies, Bacterial; Bacterial Outer Membrane Proteins; Bacterial Vaccines; Blotting, Western; Cold Temperature; Enzyme-Linked Immunosorbent Assay; Fish Diseases; Salmon; Temperature; Vibrio; Vibrio Infections

Two hundred and eighty four presumptive but not confirmed Vibrio vulnificus isolates grown on cellobiose-polymixin B-colistin agar (CPC) at 40 degrees C, recovered from sea water samples from Valencia, Spain, during a microbiological survey for V. vulnificus, were phenotypically identified. Most of the isolates (91%) corresponded to Vibrio species. V. harveyi (24%) and V. splendidus(19%) were the most abundant species identified, followed by V. navarrensis (13%), V. alginolyticus (8%) and V. parahaemolyticus (5%). The ability to grow on CPC agar and ferment cellobiose of several V. vulnificus strains from different origins and serovars, including reference strains, was tested. Most serovar E isolates and 25% of non-serovar E isolates could not grow on CPC agar.

Topics: Agar; Animals; Bacterial Typing Techniques; Bacteriological Techniques; Cellobiose; Colistin; Culture Media; Eels; Fermentation; Fish Diseases; Mediterranean Sea; Polymyxin B; Seasons; Species Specificity; Vibrio; Water Microbiology

Among 58 aerobic actinomycetes isolated from different sources and geographical locations, none of 23 Nocardia asteroides isolates, at 18 N. farcinica isolates, 1 of 5 N. otitidiscaviarum isolates, and 1 of 4 Rhodococcus species isolates opacified Middlebrook 7H10 medium. Within the N. asteroides complex, this characteristic, together with growth at 45 degrees C and resistance to each of erythromycin, cefotaxime, and tobramycin, provides a simple means of distinguishing N. farcinica from N. asteroides.

Topics: Actinomycetales; Agar; Animals; Australia; Bacteriological Techniques; Cefotaxime; Culture Media; Drug Resistance, Microbial; Drug Resistance, Multiple; Erythromycin; Fish Diseases; Fishes; Humans; Nephelometry and Turbidimetry; Nocardia; Nocardia asteroides; Nocardia Infections; Species Specificity; Tobramycin

Coomassie Brilliant Blue Agar was used to quantify the frequency of the A-layer phenotype in different isolates of Aeromonas salmonicida. Hydrophilic, non-clumping isolates of A. salmonicida consisted predominantly of the A-layer minus phenotype. These bacteria were avirulent by intraperitoneal injection into susceptible brook trout (Salvelinus fontinalis) and could not be reisolated from infected fish. By contrast, hydrophobic, clumping isolates were predominantly of the A-layer positive phenotype, highly virulent in brook trout, and easily recovered from dead or moribund fish. A-layer positive and negative clones of A. salmonicida were derived by plating bacteria on Coomassie Blue Agar. The plating showed clearly that Coomassie Blue Agar could be used as a highly selective in vitro screening method to reclaim the virulence of certain isolates of A. salmonicida having a relatively low percentage of A-layer positive phenotypes.

Topics: Aeromonas; Agar; Animals; Bacterial Infections; Bacterial Proteins; Electrophoresis, Polyacrylamide Gel; Fish Diseases; Phenotype; Rosaniline Dyes; Trout; Virulence

Charcoal is an effective replacement for serum in media for the isolation and culture of Renibacterium salmoninarum, the causative agent of bacterial kidney disease in salmonid fish. The medium, KDM-C, contains 10 g of peptone, 0.5 g of yeast extract, 1 g of L-cysteine hydrochloride, 1 g of activated charcoal, and 15 g of agar per liter and is adjusted to pH 6.8 with NaOH before autoclaving. Eight strains of R. salmoninarum grew from dilute inocula as well on KDM-C as on a standard serum-containing medium (KDM-2). The medium was effective for both primary isolations from fish and repeated transfers and has potential value for antigen preparation and physiological studies.

Topics: Agar; Animals; Bacterial Infections; Charcoal; Culture Media; Fish Diseases; Gram-Positive Bacteria; Kidney Diseases; Salmonidae

Topics: Agar; Animals; Antigens; Culture Media; Fish Diseases; Fungi; Immune Sera; Immunodiffusion; Salmonidae