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E. B. Shotts

Publications and source records attributed to E. B. Shotts.

16 recordsLinked to original sources

Enteric redmouth bacterium: Comparison of isolates from different geographic areas

Characteristics of 18 strains of Yersinia ruckeri the causative agent of enteric redmouth disease, from a variety of hosts and a wide geographic range were compared. Except for variations in colonial morphology and in citrate, sorbitol, and trehalose reactions, the strains were morphologically and biochemically homogeneous. Sixteen strains reacted with antiserum against the first serotype reported, and one strain reacted with antiserum against a more recently reported serotype. One strain represented a third serotype. Evidence presented indicated that extension of the geographic range may not be entirely a result of the transport of carrier fish.

Journal of Fish Diseases↗

Identification of Flavobacterinium and Flexibacterin species by species-specific polymerase chain reaction primers to the 16S ribosomal RNA gene

Species-specific polymerase chain reaction (PCR) primers have been developed for the identification of the causative agents of warmwater and marine finrot in fish: Flavobacterium columnare ( Flexibacter columnaris ) and Flexibacter maritimus. Differences in gene sequence in the bacterial small-subunit (16S) ribosomal RNA (rRNA) were used to design the species-specific PCR primers. The previously reported species-specific PCR primers Psy1 and Psy2 for the identification of Flavobacterium psychrophilum ( Flexibacter psychrophila ), the causative agent of coldwater finrot, were also used to develop a speciation scheme for all three bacterial finrots. These three primer sets were successful in discriminating among yellow-pigmented bacteria as well as in speciating the three major pathogenic flexibacteria to fish. The primer sets were designed to produce uniquely sized subproducts of 16S rRNA for each species: Flavobacterium psychrophilum (1,100 base pairs, bp), F. columnare (800 bp), and Flexibacter maritimus (400 bp). These primers were shown to correctly speciate field isolates in double-blind experiments ( P = 0.01).

Journal of Aquatic Animal Health↗

Determination of phylogenetic relationships of Flavobacterinium psychrophilum (Flexibacterin psychrophilus), Flavobacterinium columnare (Flexibacterin columnaris), and Flexibacterin maritimus by sequence analysis of 16S ribosomal RNA genes amplified by polymerase chain reaction

The nearly complete small subunit 16S ribosomal RNA (rRNA) gene sequence amplified by polymerase chain reaction was determined for Flavobacterium psychrophilum (formerly Flexibacter psychrophilus ) by using automated nucleotide sequencing. The sequence was found to be 1,465 base pairs (bp) in length, a size consistent with previously determined sequences for 14 other bacterial species from various taxa, including the yellow-pigmented bacteria. Sequence signatures confirmed that this organism was a member of the bacterial division Bacteroides–Flavobacterium. Parsimonious and additive phylogenetic trees were constructed with homology, and pairwise evolutionary distances were used to estimate phylogenetic relationships. Data show that F. psychrophilum, F. columnare , and Flexibacter maritimus are closely related, have a common descent, and represent a distinct group within the division Bacteroides–Flavobacterium. This group also included other organisms from the genera Flavobacterium and Cytophaga. Further, Flavobacterium aquatile , the type species for the genus Flavobacterium , was also determined to be a member of this “ Flavobacterium–Cytophaga–Flexibacter subcomplex.” This supports previous assertions that the type strain of Flavobacterium should be changed to a more representative species such as Flavobacterium breve .

Journal of Aquatic Animal Health↗

Windrow composting as an effective method to dispose of large numbers of fish

During July 1998, at the USGS Leetown Science Center, Kearneysville, West Virginia, an epizootic occurred in 16-month-old Atlantic salmon (Salmo salar). The cause ofmortality was diagnosed as furunculosis, a serious disease in salmonid fishes caused by the bacterium Aeromonas salmonicida. The fish were being maintained as part ofongoing research and were held in uncovered 30-m-Iong concrete raceways, each supplied with about 757 L per minute of 12 ?C pathogen-free spring water. The means by which the fish became infected could not be determined, and there was no recent history offurunculosis in the hatchery system.

Biological Information and Technology Notes↗

Occurrence of Loma cf salmonae brook, brown and rainbow trout from Buford Trout Hatchery, Georgia, USA

During a 6 mo study of moribund trout from Buford hatchery, Buford, Georgia, USA, a Loma cf. salmonae microsporidian parasite was studied in the gills of brook trout Salvelinus fontinalis , brown trout Salmo trutta , and rainbow trout Oncorhynchus mykiss . This parasite was morphologically similar to L. salmonae and L. fontinalis but differed in spore size. Scanning and transmission electron microscopy demonstrated that xenomas were embedded in gill filaments. Transmission electron micrographs prepared from fresh tissue showed mature spores with 12 to 15 turns of their polar tube. Spore diameters for the Georgia strain from formalin-fixed gill tissues measured 3.5 (SD ±0.1) by 1.8 (SD ±0.1) µm. Electron micrographs of formalin-fixed, deparaffinized tissues of rainbow trout from Pennsylvania and West Virginia show spores with a diameter of 3.5 (±0.2) by 1.7 (±0.1) µm and 3.4 (±0.2) by 1.8 (±0.1) µm, respectively. Transmission electron micrographs of spores from Pennsylvania and West Virginia show that mature spores from both states had 13 to 15 turns of their polar tubes. Measurements from transmission electron micrographs prepared from alcohol-fixed tissues from Virginia fish contained spores with a diameter of 3.0 (±0.3) by 1.1 (±0.3) µm and 12 to 15 turns of their polar tubes. These measurements are consistent with L. salmonae and therefore suggest that the parasite is present on the east coast of the United States. During the height of the Georgia epizootic, the percentage of fish with observed xenomas reached 62.2% (N = 87), and the highest number of xenomas counted per 10 gill filaments was 133 (N = 87). The microsporidian epizootic occurred either during the autumn months or when intake river water quality reached combined iron-manganese concentrations as high as 1.01 (mean 0.44, SD ±0.42) mg -1 .

Diseases of Aquatic Organisms↗

Plasmid-mediated romet resistance of Edwardsiella ictaluri

Romet®, a potentiated sulfa drug composed of five parts sulfadimethoxine and one part ormetoprim, is used to treat channel catfish Ictalurus punctatus infected with Edwardsiella ictaluri , the causal agent of enteric septicemia of catfish (ESC). Recently, several Romet-resistant isolates of E. ictaluri were isolated from channel catfish that had died from ESC in Virginia and Mississippi. This antimicrobial resistance was determined to be plasmid-encoded, as shown by agarose gel electrophoresis of plasmid DNA, and by the fact that the plasmid and associated antimicrobial resistance could be transferred to a recipient by single-step conjugation. Size of the R plasmid was approximately 55 kilobase pairs. Plasmids also conferred resistance to tetracycline, oxytetracycline, streptomycin, trimethoprim, and SXT (another potentiated sulfonamide, composed of trimethoprim and sulfamethoxazole). Each Romet-resistant E. ictaluri isolate was mated with a plasmid recipient, the Romet-sensitive Escherichia coli isolate 1932. Transconjugates were selected by plating on Mueller-Hinton agar that contained antimicrobials, and the mean transfer frequency was 1.483 × 10 −3 transconjugates per donor cell in the mating mixture. Each E. coli transconjugate was then mated with 20 Romet-sensitive isolates of E. ictaluri to assess the propensity of these E. ictaluri isolates to accept the R plasmid. The average transfer frequency when E. ictaluri isolates of channel catfish origin (N = 14) were used as recipients was 1.197 × 10 −2 transconjugates per donor cell in the mating mixture; mean transfer frequency when E. ictaluri isolates not originating from channel catfish were used was 1.019 × 10 -3 transconjugates per donor cell, which was significantly less (P = 0.0002).

Journal of Aquatic Animal Health↗

Comparison of plasmids isolated from Romet-30-resistant Edwardsiella ictaluri and tribrissen-resistant Escherichia coli

Edwardsiella ictaluri , the etiological agent of enteric septicemia of channel catfish (ESC) is the leading cause of bacterial disease in commercially raised channel catfish Ictalurus punctatus . The only drug approved by U.S. Food and Drug Administration for use against ESC is Romet-30. Recently, several isolates were obtained that had a naturally occurring resistance to Romet-30. On further characterization these isolates were shown to possess a 55-kilobase (kb) plasmid that encodes resistance to the drug. We compared Romet-30-resistant E. ictaluri and a Tribrissen-resistant strain of Escherichia coli (strain 1898) isolated from equine cystitis. Antimicrobial profiles, plasmid screening, restriction digest, and Southern blot analysis indicated that the two plasmids are very similar. The resistance afforded to the E. coli was encoded by a 55-kb plasmid. Each of the R plasmids conferred resistance to Romet-30, tetracycline, and Terramycin (oxytetracycline). The R plasmid from E. coli strain 1898 was transferred to Romet-30-sensitive isolates of E. ictaluri by single-step conjugation, rendering the transconjugates resistant to Romet30, tetracycline, and Terramycin. Each plasmid was cleaved into fragments by restriction enzymes, then electrophoresed in an agarose gel and transferred to nitrocellulose. A labeled probe prepared from the R plasmid of E. ictaluri isolate 5-90-156, one of the original Romet-30-resistant E. ictaluri isolates, was added. Restriction enzyme digestion resulted in an equal number of fragments of equal mobility for the two plasmids, and the probe hybridized with the same fragments of each plasmid. These results indicate that the R plasmids of E. ictaluri and E. coli are similar, if not identical.

Journal of Aquatic Animal Health↗

Post-spawning mortality of rainbow trout (Salmo gairdneri) associated with Lactobacillus

A Lactobacillus sp. was consistently isolated from dead and moribund postspawning brood stock rainbow trout. Pathology was characterized by massive chronic inflammation throughout the abdominal cavity and formation of pseudomembranes. The bacteria were most abundant in spent testes. Repeated handling was considered to be the major stressor leading to this infection and subsequent high mortality.

Journal of Wildlife Diseases↗

Bacterial diseases of fishes: Diagnostic procedures for gram-negative pathogens

A diagnostic scheme employing 10 media, 4 reagents, and 5 specific antiserums has been devised for identification of gram-negative pathogens of fishes. The scheme is not intended for detailed speciation of organisms, but for generic identification of organisms associated with infected fishes. Diagnosis can be accomplished in 24 h with selective media and antiserums, and in no more than 72 h with other media and reagents.

Journal of the Fisheries Research Board of Canada↗