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Yersinia ruckeri sp. nov., the redmouth (RM) bacterium

Cultures of the redmouth (RM) bacterium, one of the etiological agents of redmouth disease in rainbow trout ( Salmo gairdneri ) and certain other fishes, were characterized by means of their biochemical reactions, by deoxyribonucleic acid (DNA) hybridization, and by determination of guanine-plus-cytosine (G+C) ratios in DNA. The DNA relatedness studies confirmed the fact that the RM bacteria are members of the family Enterobacteriaceae and that they comprise a single species that is not closely related to any other species of Enterobacteriaceae. They are about 30% related to species of both Serratia and Yersinia. A comparison of the biochemical reactions of RM bacteria and serratiae indicated that there are many differences between these organisms and that biochemically the RM bacteria are most closely related to yersiniae. The G+C ratios of RM bacteria were approximated to be between 47.5 and 48.5%. These values are similar to those of yersiniae but markedly different from those of serratiae. On the basis of their biochemical reactions and their G+C ratios, the RM bacteria are considered to be a new species of Yersinia , for which the name Yersinia ruckeri is proposed. Strain 2396-61 (= ATCC 29473) is designated the type strain of the species.

International Journal of Systematic Bacteriology

Methanolobus taylorii sp nov, a new methylotropic, estuarine methanogen

Previously published phylogenetic studies of 16S rRNA showed that methylotrophic, slightly halophilic, methanogenic strain GS-16 T (T = type strain) represents a new species of bacterium. We propose the name Methanolobus taylorii for this species; strain GS-16 is the type strain.

International Journal of Systematic Bacteriology

Desulfuromonas thiophila sp. nov., a new obligately sulfur-reducing bacterium from anoxic freshwater sediment

A mesophilic, acetate-oxidizing, sulfur-reducing bacterium, strain NZ27(T), was isolated from anoxic mud from a freshwater sulfur spring. The cells were ovoid, motile, and gram negative. In addition to acetate, the strain oxidized pyruvate, succinate, and fumarate. Sulfur flower could be replaced by polysulfide as an electron acceptor. Ferric nitrilotriacetic acid was reduced in the presence of pyruvate; however, this reduction did not sustain growth. These phenotypic characteristics suggested that strain NZ27(T) is affiliated with the genus Desulfuromonas. A phylogenetic analysis based on the results of comparative 16S ribosomal DNA sequencing confirmed that strain NZ27(T) belongs to the Desulfuromonas cluster in the recently proposed family 'Geobacteraceae' in the delta subgroup of the Proteobacteria. In addition, the results of DNA-DNA hybridization studies confirmed that strain NZ27(T) represents a novel species. Desulfuromonas thiophila, a name tentatively used in previous publications, is the name proposed for strain NZ27(T) in this paper.

International Journal of Systematic Bacteriology

Sulfurospirillum barnesii sp. nov. and Sulfurospirillum arsenophilum sp. nov., new members of the Sulfurospirillum clade of the ε-Proteobacteria

Two strains of dissimilatory arsenate-reducing vibrio-shaped bacteria are assigned to the genus Sulfurospirillum. These two new species, Sulfurospirillum barnesii strain SES-3(T) and Sulfurospirillum arsenophilum strain MIT-13(T), in addition to Sulfurospirillum sp. SM-5, two strains of Sulfurospirillum deleyianum, and Sulfurospirillum arcachonense, form a distinct clade within the ?? subclass of the Proteobacteria based on 16S rRNA analysis.

International Journal of Systematic Bacteriology

Comparative sensitivities of diagnostic procedures used to detect bacterial kidney disease in salmonid fishes

Kidney and spleen homogenates from each of 60 coho salmon ( Oncorhynchus kisutch ) and steelhead trout ( Salmo gairdneri ) were examined for detection of Renibacterium salmoninarum . The proportions of positives differed widely with the detection procedures used: in coho salmon, 5% were positive by the Gram-stain procedure, 10% by the direct fluorescent antibody test, 48% by bacteriological isolation, 65% by staphylococcal coagglutination, and 73% by counterimmunoelectrophoresis; in steelhead trout, 3% were positive by Gram-stain, 8.3% by fluorescent antibody, 17% by bacteriological isolation, and 67% by counterimmunoelectrophoresis. Renibacterium salmoninarum was not detected in either coho salmon or steelhead trout by immunodiffusion analysis.

Journal of Wildlife Diseases

General and specialized media routinely employed for primary isolation of bacterial pathogens of fishes

There are a number of significant diseases among cultured and free-ranging freshwater fishes that have a bacterial etiology; these represent a variety of gram-negative and gram-positive genera. Confirmatory diagnosis of these diseases involves primary isolation of the causative bacterium on bacteriologic media. Frequently used "general" bacteriologic media simply provide the essential nutrients for growth. For most of the major pathogens, however, there are differential and/or selective media that facilitate primary recovery. Some specialized media are available as "ready-to-use" from suppliers, while others must be prepared. Differential media employ various types of indicator systems, such as pH indicators, that allow diagnosticians to observe assimilation of selected substrates. An advantage to the use of differential media for primary isolation is that they hasten bacterial characterization by yielding the appropriate positive or negative result for a particular substrate, often leading to a presumptive identification. Selective media also incorporate agent(s) that inhibit the growth of contaminants typically encountered with samples from aquatic environments. Media that incorporate differential and/or selective components are ideally based on characters that are unique to the targeted bacterium, and their use can reduce the time associated with diagnosis and facilitate early intervention in affected fish populations. In this review, the concepts of general and differential/selective bacteriologic media and their use and development for fish pathogens are discussed. The media routinely employed for primary isolation of the significant bacterial pathogens of fishes are presented. ?? Wildlife Disease Association 2008.

Journal of Wildlife Diseases

An acute septicaemic disease of brown trout (Salmo trutta) and Atlantic salmon (Salmo salar) caused by a Pasteurella-like organism

Ulcerations of the skin associated with haemorrhagic petechiae of liver and kidneys, were the main signs of disease affecting salmon and brown trout in Norway. A death rate of 15–20% was estimated for the 5 month period of mid-March-August, although mortalities occurred throughout the year. Bacteriological examinations, involving 36 isolates, suggested the causual organism to be a Pasteurella although the DNA homology examination gave a G.C. ratio of 55.6 % which is high for Pasteurella as a group.

Journal of Fish Biology

Biography of Dr. John L. Fryer

There is probably no one in the fi sh health community who does not recognize the name of Professor John L. Fryer. Active in research for a period of more than 40 years, Dr Fryer has achieved internatio nal acclaim for his wor k on infectious diseases of Pacific salmon . The resea rch from his laboratory at Oregon State University (OSU) has spanned the disciplines of virology, parasitology, bacteriology, cell biology, immuno logy and fi sh physiology, result ing in more than 200 publications, two patents and recognition as one of the world's leading centres for researc h on infec tious diseases of salmon id fi sh.

Journal of Fish Diseases

Bench-top validation testing of selected immunological and molecular Renibacterium salmoninarum diagnostic assays by comparison with quantitative bacteriological culture

No gold standard assay exhibiting error-free classification of results has been identified for detection of Renibacterium salmoninarum , the causative agent of salmonid bacterial kidney disease. Validation of diagnostic assays for R. salmoninarum has been hindered by its unique characteristics and biology, and difficulties in locating suitable populations of reference test animals. Infection status of fish in test populations is often unknown, and it is commonly assumed that the assay yielding the most positive results has the highest diagnostic accuracy, without consideration of misclassification of results. In this research, quantification of R. salmoninarum in samples by bacteriological culture provided a standardized measure of viable bacteria to evaluate analytical performance characteristics (sensitivity, specificity and repeatability) of non-culture assays in three matrices (phosphate-buffered saline, ovarian fluid and kidney tissue). Non-culture assays included polyclonal enzyme-linked immunosorbent assay (ELISA), direct smear fluorescent antibody technique (FAT), membrane-filtration FAT, nested polymerase chain reaction (nested PCR) and three real-time quantitative PCR assays. Injection challenge of specific pathogen-free Chinook salmon, Oncorhynchus tshawytscha (Walbaum), with R. salmoninarum was used to estimate diagnostic sensitivity and specificity. Results did not identify a single assay demonstrating the highest analytical and diagnostic performance characteristics, but revealed strengths and weaknesses of each test.

Journal of Fish Diseases

An alternative bacteriological medium for the isolation of Aeromonas spp.

Two solid bacteriologic media were compared for cultivating Aeromonas spp. from piscine sources: the Rimler-Shotts (RS) medium and a starch-glutamate-ampicillin-penicillin-based medium (SGAP-10C) used for the recovery of Aeromonas spp. from water samples. The selective and differential capacities of the media were assessed March through October 1992 by recovery rate and phenotype of 99 isolates representing 15 genera of bacteria. Recovery frequency of Aeromonas spp. ( n = 62) was similar at 97% on RS and 95% on SGAP-10C. The SGAP-10C medium proved to be more specific than RS toward Aeromonas species (P ≤ 0.005). Use of SGAP-10C at 24 C for 48 hr offers a better choice for the laboratory recovery of Aeromonas spp. from clinical fish specimens.

Journal of Wildlife Diseases

Lack of surface-associated microorganisms in a mixed species community of freshwater Unionidae

To determine whether unionids contain surface-attached endosymbiotic bacteria, ciliates, or fungi, we used scanning electron microscopy to examine the epithelial surface of various organs within the digestive systems and mantle cavity of temperate river and lake unionids on a seasonal basis. We also cultured material removed from the lumen of these same organs and from the mantle cavity to detect cellobiose-, cellulose-, and chitin- degrading microbes. No true endosymbiotic fauna were observed attached to the surface of the digestive or mantle tissues of any species of unionid. Microbial growth on cellulose or chitin bacteriological media varied by season and habitat, but not by unionid species or source of the isolate. Lake unionids did not contain microbes capable of digesting cellulose or chitin, whereas unionids from the river site did in March and August, but not in December. Since these cultured cellulose- and chitin-degrading bacteria were never found attached to any unionid tissues, they appeared to be transient forms, not true endosymbionts. Microbes capable of digesting cellobiose were found in every unionid collected in all seasons and habitats, but again, no microbes were directly observed attached to unionid tissues. If unionids, like most other vertebrates, lack digestive enzymes required to initiate primary bond breakage, then the lack of cellulolytic and chitinolytic endosymbionts would affect the ability to utilize cellulose or chitin foods. Thus, in captivity dry feeds based on corn, soybeans, or nauplii should be pre-digested to ensure maximum absorption of nutrients by unionids. The lack of celluloytic or chitinolytic endosymbionts should not affect relocation success, though the seasonal role of transient microbes in unionid nutrition requires further investigation.

Journal of Shellfish Research

Biosafety of parenteral Brucella abortus RB51 vaccine in bison calves

Vaccination is considered among the primary management tools for reducing brucellosis prevalence in Greater Yellowstone Area (GYA) ungulates. Before their use, however, vaccine safety and efficacy must be demonstrated. Twenty-seven female bison (Bison bison) calves (approx 5 months old) were vaccinated with Brucella abortus Strain RB51 (1.5 × 1010 colony forming units [CFU], subcutaneously) as part of routine management. We assessed the persistence, pathology, shedding, and transmission associated with RB51 by serial necropsy, bacteriology, histopathology, and serology of 20 of these 27 vaccinated calves, and RB51 serology of 10 nonvaccinated, commingling adult females. With the exception of 1 calf, RB51 dot-blot titers at necropsy were <1:80. Strain RB51 was cultured from lymph nodes in 4 of 4 calves at 14 weeks postvaccination (PV), 4 of 4 calves at 18 weeks PV, 1 of 4 calves at 22 weeks PV, 3 of 4 at 26 weeks PV, and 0 of 4 calves at 30 weeks PV. No gross lesions were observed. Mild histologic changes occurred only in a few draining lymph nodes early in sampling. Adverse clinical effects were not observed in vaccinates. Swabs from nasopharynx, conjunctiva, rectum, and vagina were uniformly culture negative for RB51. Strain RB51 dot-blot assays of bison cows were negative at a 1:20 dilution at 26 weeks PV. Our results suggest that RB51 persists longer in bison calves than in domestic cattle and is systemically distributed within lymphatic tissues. However, bison apparently clear the RB51 vaccine strain without shedding, transmission, or significant adverse reactions.

Journal of Wildlife Management

Pathology of brucellosis in bison from Yellowstone National Park

Between February 1995 and June 1999, specimens from seven aborted bison ( Bison bison ) fetuses or stillborn calves and their placentas, two additional placentas, three dead neonates, one 2-wk-old calf, and 35 juvenile and adult female bison from Yellowstone National Park (USA) were submitted for bacteriologic and histopathologic examination. One adult animal with a retained placenta had recently aborted. Serum samples from the 35 juvenile and adult bison were tested for Brucella spp. antibodies. Twenty-six bison, including the cow with the retained placenta, were seropositive, one was suspect, and eight were seronegative. Brucella abortus biovar 1 was isolated from three aborted fetuses and associated placentas, an additional placenta, the 2-wk-old calf, and 11 of the seropositive female bison including the animal that had recently aborted. Brucella abortus biovar 2 was isolated from one additional seropositive adult female bison. Brucella abortus was recovered from numerous tissue sites from the aborted fetuses, placentas and 2-wk-old calf. In the juvenile and adult bison, the organism was more frequently isolated from supramammary (83%), retropharyngeal (67%), and iliac (58%) lymph nodes than from other tissues cultured. Cultures from the seronegative and suspect bison were negative for B. abortus . Lesions in the B. abortus -infected, aborted placentas and fetuses consisted of necropurulent placentitis and mild bronchointerstitial pneumonia. The infected 2-wk-old calf had bronchointerstitial pneumonia, focal splenic infarction, and purulent nephritis. The recently-aborting bison cow had purulent endometritis and necropurulent placentitis. Immunohistochemical staining of tissues from the culture-positive aborted fetuses, placentas, 2-wk-old calf, and recently-aborting cow disclosed large numbers of B. abortus in placental trophoblasts and exudate, and fetal and calf lung. A similar study with the same tissue collection and culture protocol was done using six seropositive cattle from a B. abortus -infected herd in July and August, 1997. Results of the bison and cattle studies were similar.

Wyoming