Corynebacterium conjunctivae: A New Corynebacterium Species Isolated from the Ocular Surface of Healthy Horses.

Twenty-two unidentified Gram-positive, rod-shaped organisms were recovered from the conjunctival surface of apparently healthy horses and subjected to a polyphasic taxonomic analysis. Based on cellular morphology and biochemical criteria, the isolates were tentatively assigned to the genus Corynebacterium, although they did not match any recognized species. Comparative 16S rRNA gene sequencing studies demonstrated that all of the isolates were phylogenetically members of the genus Corynebacterium. The isolates shared 99.4 to 100% 16S rRNA gene sequence similarity among the strains and 96.5% similarity with Corynebacterium tapiri 2385/12T, which was the closest phylogenetically related species. The DNA G+C content was 58.4 mol%. The major fatty acids were C15:0, C16:0, C17:1 ω8c and C18:1 ω9c, while the predominant mycolic acids consisted of C30:0, C32:0 and C34:0. The isolates were distinguished from related Corynebacterium species by a number of phenotypic properties. On the basis of phenotypic and phylogenetic evidence, it is proposed that the unknown isolates from horses be classified in the genus Corynebacterium as Corynebacterium conjunctivae sp. nov. The type strain of C. conjunctivae is ICM19-01138T (DSM 109759T = CCUG 73728T).

A new genus and species of turtle blood fluke (Digenea: Schistosomatoidea) from the Mekong snail-eating turtle, Malayemys subtrijuga (Schlegel & Müller) (Testudines: Geoemydidae) in Vietnam, with a reassessment of related Asiatic turtle blood flukes and molecular phylogeny.

Platt sinuosus Roberts & Bullard n. g., n. sp. (type-species) infects the kidney and mesenteric blood vessels of Mekong snail-eating turtles, Malayemys subtrijuga (Schlegel & Müller), in the Mekong River Basin. Species of Platt Roberts & Bullard n. g. are unique by the combination of having a papillate ventral sucker, vasa efferentia that are dorsal to the gonads, a massive cirrus-sac that is directed anteriad or laterad, and a vitellarium that surrounds the intestinal caeca. The new species resembles Platt ocadiae (Takeuti, 1942) Roberts & Bullard n. comb. but differs from it by having an external seminal vesicle that overlaps with or is immediately posterior to the level of the ventral sucker. Seven species previously of Hapalorhynchus Stunkard, 1922 are reassigned herein to Platt: P. odhnerensis (Mehra, 1933) Roberts & Bullard n. comb.; P. yoshidai (Ozaki, 1939) Roberts & Bullard n. comb.; P. ocadiae; P. oschmarini (Belous, 1963) Roberts & Bullard n. comb.; P. sutlejensis (Mehrotra, 1973) Roberts & Bullard n. comb.; P. synderi (Platt & Sharma, 2012) Roberts & Bullard n. comb.; and P. tkachi (Platt & Sharma, 2012) Roberts & Bullard n. comb. A dichotomous key to Platt spp. is provided. Hapalorhynchus sheilae (Mehrotra, 1973) Bourgat, 1990 and Hapalorhynchus mica (Oshmarin, 1971) Bourgat, 1990 are considered as species inquirendae, and Hapalorhynchus indicus (Thapar, 1933) Price, 1934 and Hapalorhynchus macrotesticularis (Rohde, Lee, & Lim, 1968) Brooks & Sullivan, 1981 are considered as species incertae sedis. Phylogenetic analysis of the large subunit rDNA (28S) showed P. sinuosus and P. snyderi to be sister taxa distinct from a monophyletic Hapalorhynchus and Coeuritrema platti Roberts & Bullard, 2016.

Genetic studies to re-affiliate Edwardsiella tarda fish isolates to Edwardsiella piscicida and Edwardsiella anguillarum species.

Until 2012, the genus Edwardsiella was composed by three species Edwardsiella tarda, Edwardsiella hoshinae and Edwardsiella ictaluri. In 2013, Edwardsiella piscicida, compiling fish pathogenic strains previously identified as E. tarda was described, and more recently a new species isolated from diseased eel was reported, namely Edwardsiella anguillarum. The incorporation of these species into the genus makes necessary a revision of the taxonomic position of the isolates previously identified as E. tarda. Using AFLP technique, MLSA studies and in silico DNA-DNA hybridization, 46 of 49 E. tarda isolates were re-assigned as E. piscicida and 2 as E. anguillarum, whereas it was confirmed previous classification of the Edwardsiella types and reference strains used. The study of the taxonomic resolution of the genes 16S rRNA, adk, atpD, dnaJ, glnA, hsp60, tuf as well as the possible combinations among housekeeping genes, showed that the gene dnaJ was the more resolutive. In conclusion, the use of molecular techniques is necessary to accurately identify Edwardsiella isolates, especially when differentiating new species from E. tarda.

Morphological and molecular confirmation of Myxobolus cerebralis myxospores infecting wild­caught and cultured trout in North Carolina (SE USA).

We used microscopy and molecular biology to provide the first documentation of infections of Myxobolus cerebralis (Myxozoa: Myxobolidae), the etiological agent of whirling disease, in trout (Salmonidae) from North Carolina (USA) river basins. A total of 1085 rainbow trout Oncorhynchus mykiss, 696 brown trout Salmo trutta, and 319 brook trout Salvelinus fontinalis from 43 localities across 9 river basins were screened. Myxospores were observed microscopically in pepsin-trypsin digested heads of rainbow and brown trout from the Watauga River Basin. Those infections were confirmed using the prescribed nested polymerase chain reaction (PCR; 18S rDNA), which also detected infections in rainbow, brown, and brook trout from the French Broad River Basin and the Yadkin Pee-Dee River Basin. Myxospores were 9.0-10.0 µm (mean ± SD = 9.6 ± 0.4; N = 119) long, 8.0-10.0 µm (8.8 ± 0.6; 104) wide, and 6.0-7.5 µm (6.9 ± 0.5; 15) thick and had polar capsules 4.0-6.0 µm (5.0 ± 0.5; 104) long, 2.5-3.5 µm (3.1 ± 0.3; 104) wide, and with 5 or 6 polar filament coils. Myxospores from these hosts and rivers were morphologically indistinguishable and molecularly identical, indicating conspecificity, and the resulting 18S rDNA and ITS-1 sequences derived from these myxospores were 99.5-100% and 99.3-99.8% similar, respectively, to published GenBank sequences ascribed to M. cerebralis. This report comprises the first taxonomic circumscription and molecular confirmation of M. cerebralis in the southeastern USA south of Virginia.

Comparative Phenotypic and Genotypic Analysis of Edwardsiella Isolates from Different Hosts and Geographic Origins, with Emphasis on Isolates Formerly Classified as E. tarda, and Evaluation of Diagnostic Methods.

Edwardsiella spp. are responsible for significant losses in important wild and cultured fish species worldwide. Recent phylogenomic investigations have determined that bacteria historically classified as Edwardsiella tarda actually represent three genetically distinct yet phenotypically ambiguous taxa with various degrees of pathogenicity in different hosts. Previous recognition of these taxa was hampered by the lack of a distinguishing phenotypic character. Commercial test panel configurations are relatively constant over time, and as new species are defined, appropriate discriminatory tests may not be present in current test panel arrangements. While phenobiochemical tests fail to discriminate between these taxa, data presented here revealed discriminatory peaks for each Edwardsiella species using matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) methodology, suggesting that MALDI-TOF can offer rapid, reliable identification in line with current systematic classifications. Furthermore, a multiplex PCR assay was validated for rapid molecular differentiation of the Edwardsiella spp. affecting fish. Moreover, the limitations of relying on partial 16S rRNA for discrimination of Edwardsiella spp. and advantages of employing alternative single-copy genes gyrB and sodB for molecular identification and classification of Edwardsiella were demonstrated. Last, sodB sequencing confirmed that isolates previously defined as typical motile fish-pathogenic E. tarda are synonymous with Edwardsiella piscicida, while atypical nonmotile fish-pathogenic E. tarda isolates are equivalent to Edwardsiella anguillarum Fish-nonpathogenic E. tarda isolates are consistent with E. tarda as it is currently defined. These analyses help deconvolute the scientific literature regarding these organisms and provide baseline information to better facilitate proper taxonomic assignment and minimize erroneous identifications of Edwardsiella isolates in clinical and research settings.

Emendation and new species of Hapalorhynchus Stunkard, 1922 (Digenea: Schistosomatoidea) from musk turtles (Kinosternidae: Sternotherus) in Alabama and Florida rivers.

Hapalorhynchus Stunkard, 1922 is emended based on morphological study of existing museum specimens (type and voucher specimens) and newly-collected specimens infecting musk turtles (Testudines: Kinosternidae: Sternotherus spp.) from rivers in Alabama and Florida (USA). Hapalorhynchus conecuhensis n. sp. is described from an innominate musk turtle, Sternotherus cf. minor, (type host) from Blue Spring (31°5'27.64″N, 86°30'53.21″W; Pensacola Bay Basin, Alabama) and the loggerhead musk turtle, Sternotherus minor (Agassiz, 1857) from the Wacissa River (30°20'24.73″N, 83°59'27.56″W; Apalachee Bay Basin, Florida). It differs from congeners by lacking a body constriction at level of the ventral sucker, paired anterior caeca, and a transverse ovary as well as by having a small ventral sucker, proportionally short posterior caeca, nearly equally-sized anterior and posterior testes, a small cirrus sac, and a uterus extending dorsal to the ovary and the anterior testis. Specimens of Hapalorhynchus reelfooti Byrd, 1939 infected loggerhead musk turtles, stripe-necked musk turtles (Sternotherus peltifer Smith and Glass, 1947), Eastern musk turtles (Sternotherus odoratus [Latreille in Sonnini and Latreille, 1801]), and S. cf. minor. Those of Hapalorhynchus cf. stunkardi infected S. minor and S. odoratus. Sternothorus minor, S. peltifer, and S. cf. minor plus S. minor and S. odoratus are new host records for H. reelfooti and H. cf. stunkardi, respectively. This is the first report of an infected musk turtle from the Coosa and Tallapoosa Rivers (Mobile-Tensaw River Basin), Pensacola Bay Basin, or Apalachee Bay Basin. Sequence analysis of the large subunit rDNA (28S) showed a strongly-supported clade for Hapalorhynchus.

Acipensericola glacialis n. sp. (Digenea: Aporocotylidae) from heart of lake sturgeon Acipenser fulvescens Rafinesque (Acipenseriformes: Acipenseridae) in the Great Lakes Basin, Lake Winnebago System, USA.

Acipensericola glacialis n. sp. infects the heart of lake sturgeon, Acipenser fulvescens (Rafinesque), in the Lake Winnebago System and differs from its only congener, Acipensericola petersoni Bullard, Snyder, Jensen & Overstreet, 2008, by having a dendritic intestine, deeply-lobed testes, a post-ovarian oötype, and a common genital pore that is medial to the dextral caecum. Acipensericola petersoni has a non-dendritic intestine, testes that are not deeply lobed, an oötype that is at level of the ovary (ventral to the ovary), and a common genital pore that is dorsal to the dextral caecum. Comparison of the large (28S) and small (18S) sub-unit ribosomal DNA and internal transcribed spacer 2 (ITS2) regions between specimens of A. glacialis n. sp. and A. petersoni revealed 13 (of 1,621 nt; 99.2% similarity in the 28S), 8 (of 1,841 nt; 99.9% similarity in the 18S), and 11 (of 442 nt; 97.5% similarity in the ITS2) nucleotide differences. Collectively, these results comprise an unexpectedly high degree of morphological and molecular similarity given the geographical (Mississippi River Basin vs Great Lakes Basin) and phylogenetic (Polyodontidae vs Acipenseridae) separation of these hosts but seemingly did not reject a previous hypothesis concerning lake sturgeon dispersal from the Mississippi Refugium following the Wisconsin glaciation ~18,000 years ago. The new species is the first nominal blood fluke described from a sturgeon.

Exotic "Gill Lice" Species (Copepoda: Lernaeopodidae: Salmincola SPP.) Infect Rainbow Trout (Oncorhynchus mykiss) and Brook Trout (Salvelinus fontinalis) in the Southeastern United States.

Salmincola californiensis infected 25 of 31 (prevalence 0.8; intensity 2-35 [mean 6.6 ± standard deviation 7.7; n = 25]) rainbow trout, Oncorhynchus mykiss, from a private trout farm connected to the Watauga River, North Carolina. Salmincola edwardsii infected all of 9 (1.0; 2-43 [9.3 ± 13.0; 9]) brook trout, Salvelinus fontinalis, from Big Norton Prong, a tributary of the Little Tennessee River, North Carolina. Both lernaeopodids are well-known salmonid pathogens, but neither is native to, nor has been previously taxonomically confirmed from, the southeastern United States. Herein, we (1) use light and scanning electron microscopy to identify and provide supplemental morphological observations of these lernaeopodids, (2) furnish complementary molecular sequence data from the 28S rDNA (28S), and (3) document the pathological effects of gill infections. We identified and differentiated these lernaeopodids by the second antenna (exopod tip with large [S. californiensis] vs. slender [S. edwardsii] spines; endopod terminal segment with subequal ventral processes shorter than [S. californiensis] vs. longer than or equal to [S. edwardsii] dorsal hook), maxilliped palp (length typically ≤1/3 [S. californiensis] vs. 1/3-1/2 [S. edwardsii] subchela length exclusive of claw), and bulla (sub-circular and concave on manubrium's side [S. californiensis] vs. non-stellate [S. edwardsii]). Analysis of the 28S rDNA sequences confirmed our taxonomic assignments as demonstrated by 100% sequence similarity among the sympatric, morphologically-conspecific isolates. Histopathology revealed focal gill epithelial hyperplasia, obstruction of interlamellar water channels, lamellar fusion, and crypting of gill filaments. High intensity infections by either lernaeopodid are surveillance-worthy because they are potentially pathogenic to trout in the southeastern United States.

Two new species of Elopicola (Digenea: Aporocotylidae) from Hawaiian ladyfish, Elops hawaiensis (Eastern Sea) and Atlantic tarpon, Megalops atlanticus (Gulf of Mexico) with a comment on monophyly of elopomorph blood flukes.

Elopicola bristowi sp. n. infects the blood vascular system of Hawaiian ladyfish, Elops hawaiensis, in the Eastern Sea. It differs from the only nominal congener Elopicola nolancribbi by the combination of having rows of ventrolateral tegumental spines, a proportionally long oesophagus, anterior caeca, vasa efferentia coalescing ventral to the posterodextral margin of the testis, a post-testicular metraterm, a dextral common genital pore lateral to the oötype, and genitalia that are enantiomorphic relative to those of E. nolancribbi. Elopicola franksi sp. n. infects the heart and blood vascular system of Atlantic tarpon, Megalops atlanticus, in the Gulf of Mexico. It differs from its congeners by the combination of lacking ventrolateral tegumental spines and having an elongate body (6× longer than wide), a proportionally long oesophagus, a compact testis at level of the distal ends of the posterior caeca, and a post-testicular common genital pore at level of the oötype. Phylogenetic analyses based on the small subunit ribosomal DNA (18S), large subunit ribosomal DNA (28S), and internal transcribed spacer 2 (ITS2) genes revealed considerable genetic differences between these taxa. The 18S+28S tree showed a monophyletic Elopicola sister to all aporocotylids infecting fishes of Euteleosteomorpha. The ITS2 tree showed Paracardicoloides yamagutii as the sister taxon to Elopicola spp.

A new species of Spirorchis MacCallum, 1918 (Digenea: Schistosomatoidea) and Spirorchis cf. scripta from chicken turtle, Deirochelys reticularia (Emydidae), with an emendation and molecular phylogeny of Spirorchis.

Chicken turtles, Deirochelys reticularia (Latreille in Sonnini et Latreille) (Testudines: Emydidae) from Alabama, USA were infected by Spirorchis collinsi Roberts et Bullard sp. n. and Spirorchis cf. scripta. The new species is most easily differentiated from its congeners by the combination of having caeca that extend far beyond the genitalia, intercaecal genitalia positioned in the middle portion of the body, a testicular column that nearly abuts the caecal bifurcation, a cirrus sac positioned between the testes and ovary, a massive Mehlis' gland, an elongate, longitudinal metraterm that extends anteriad beyond the level of the ovary, a pre-ovarian genital pore, and a prominent, intercaecal Manter's organ. The specimens of S. cf. scripta differed from the holotype and published descriptions of Spirorchis scripta Stunkard, 1923 by several subtle morphological features, perhaps comprising intraspecific variation, but collectively warranted a detailed description herein. Based on examinations of the aforementioned specimens plus the holotype, paratypes and vouchers of morphologically-similar congeners, Spirorchis MacCallum, 1918 is emended to include the presence of oral sucker spines, a pharynx, lateral oesophageal diverticula ('plicate organ') and a median oesophageal diverticulum ('oeseophageal pouch'). Phylogenetic analysis of the nuclear large subunit rDNA (28S) recovered S. collinsi sister to Spirorchis picta Stunkard, 1923, > 99% similarity between S. cf. scripta and S. scripta, and a monophyletic Spirorchis MacCallum, 1918. No blood fluke infection has been reported previously from these drainages, Alabama, or this turtle species. This is the first new species of Spirorchis to be described from North America in 26 years.

Skin lesions on yellowfin tuna Thunnus albacares from Gulf of Mexico outer continental shelf: Morphological, molecular, and histological diagnosis of infection by a capsalid monogenoid.

We characterize lesion-associated capsaline infections on yellowfin tuna, Thunnus albacares, in the Gulf of Mexico by comparing our specimens with published descriptions and museum specimens ascribed to Capsala biparasiticum and its synonyms: vouchers of C. biparasiticum from parasitic copepods; the holotype of Capsala neothunni; and vouchers of Capsala abidjani. Those from parasitic copepods differed by having a small, rounded body, large anterior attachment organs, closely spaced dorsomarginal body sclerites, small testes, and a short and wide testicular field. No morphometric feature in the holotype of C. neothunni ranged outside of that reported for the newly-collected specimens, indicating conspecificity of our specimens. The specimens of C. abidjani differed by having a large anterior attachment organ, few and dendritic testes, and a short, wide testicular field. Large subunit ribosomal DNA (28S) sequences grouped our specimens and Capsala sp. as sister taxa and indicated a phylogenetic affinity of Nasicola klawei. The haptoral attachment site comprised a crater-like depression surrounded by a blackish-colored halo of extensively rugose skin, with abundant pockmarked-like, irregularly-shaped oblong or semi-circular epidermal pits surrounding these attachment sites. Histology confirmed extensive folding of epidermis and underlying stratum laxum, likely epidermal hyperplasia, foci of weak cell-to-cell adhesions among apical malpighian cells as well as that between stratum germinativum and stratum laxum, myriad goblet cells in epidermis, rodlet cells in apical layer of epidermis, and lymphocytic infiltrates and melanin in dermis. The present study comprises (i) the first published report of this parasite from yellowfin tuna captured in the Gulf of Mexico-NW Atlantic Ocean Basin, (ii) confirmation of its infection on the skin (rather than on a parasitic copepod), (iii) the first molecular data for this capsaline, and (iv) the first observations of histopathological changes associated with a capsalid infection on a wild-caught epipelagic fish.

Diversity of bacteria cultured from the blood of lesser electric rays caught in the northern gulf of Mexico.

The prevalence and taxonomic diversity of bacteria cultured from the blood of apparently healthy Lesser Electric Rays Narcine bancroftii captured from open beach habitat in the north-central Gulf of Mexico are reported herein. The blood of 9 out of 10 Lesser Electric Rays was positive for bacteria, and bacterial isolates (n = 83) were identified by 16S rRNA gene sequencing. The majority of the isolates belonged to the phylum Proteobacteria (91.5%). Vibrio spp. comprised 53% of all isolates and were recovered from all Lesser Electric Rays with culture-positive blood. Among them, V. harveyi (n = 14) and V. campbellii (n = 11) were most common, followed by a group of unidentified Vibrio sp. (n = 10) related to V. nigripulchritudo. Isolates representing other species of Proteobacteria included Pseudoalteromonas (n = 13), Shewanella (n = 5), Amphritea (n = 3), Nautella (n = 3), and Arenibacter (n = 1). Higher bacterial diversity was observed in blood cultured on marine agar relative to blood agar, but gram-positive bacteria were isolated from the latter only. The 16S rRNA gene sequences of bacterial isolates were compared phylogenetically to those from related type strains. Most isolates were identified to the level of species, but some clustered independently from reference strains, likely representing new species of Vibrio, Amphritea, Shewanella, and Tenacibaculum. The present study is the first record of any bacterium from this ray species and reveals a taxonomically and phylogenetically diverse microbiota associated with its blood. Moreover, these data document that the presence of bacteria in elasmobranch blood is not coincident with clinical signs of disease, thereby rejecting the paradigm of septicemia indicating a disease condition in aquatic vertebrates.

Survival of Fish-Pathogenic Strains of Aeromonas hydrophila under Starvation.

Abstract The survival of Aeromonas hydrophila under low-nutrient conditions was investigated in this study. The behavior of three strains isolated from Common Carp Cyprinus carpio (China) and Channel Catfish Ictalurus punctatus (USA) was compared when cells were starved at different temperatures (4, 15, 25, and 35°C) over a 4-week period. Temperature played a major role in cell survival, and cell viability decreased rapidly at 4°C. Conversely, cells stored at 15°C did not lose viability over time. Warmer temperatures (25°C and 35°C) decreased cell numbers by approximately one order of magnitude. Ultrastructural changes in cell morphology were observed in starved cells. Our data confirm that A. hydrophila can persist in the aquatic environment for extended periods, but survival is strongly influenced by temperature. Received December 13, 2013; accepted March 22, 2014.

Diversity and ancestry of flatworms infecting blood of nontetrapod craniates "fishes".

We herein review all published molecular studies (life history, taxonomy, and phylogeny) and summarize all GenBank sequences and primer sets for the "fish blood flukes". Further, by analysing new and all available sequence data for the partial D1-D2 domains of 28S from 83 blood fluke taxa, we explore the evolutionary expansion of flatworm parasitism in the blood of craniates. Based on this analysis, the blood flukes infecting marine bony fishes (Euteleostei) are monophyletic. The clade comprising the chondrichthyan blood fluke plus the marine euteleost blood flukes is the sister group to tetrapod blood flukes (spirorchiids and schistosomes). The innominate blood fluke cercariae from freshwater gastropods were monophyletic and sister to the clade comprising spirorchiids and schistosomes, but low nodal support indicated that they may represent a distinct blood fluke lineage with phylogenetic affinities also to fish blood flukes. Blood flukes that utilize gastropod intermediate hosts were monophyletic (unidentified gastropod cercariae+tetrapod blood flukes) and those utilizing bivalves and polychaetes were monophyletic (marine fish blood flukes). Low or no taxon sampling among blood flukes of basal fish lineages and primary division freshwater fish lineages are significant data gaps needing closure. We also note that no record of an infection exists in a hagfish (Myxiniformes), lamprey (Petromyzontiformes), or nontetrapod sarcopterygiian, i.e., coelacanth (Coelacanthimorpha) or lungfish (Dipnoi). The present phylogenetic analysis reiterated support for monophyly of Schistosomatidae and paraphyly of spirorchiids, with the blood flukes of freshwater turtles basal to those of marine turtles and schistosomes.

Evaluation of global gene expression during cold shock in the human pathogen Vibrio vulnificus.

Vibrio vulnificus can adapt to cold temperatures by changing the expression profiles of certain genes and their resulting proteins. In this study, the complete V. vulnificus transcriptome was analyzed under cold shock by looking at gene expression changes occurring during the shift from 35°C to 4°C. A DNA microarray-based global transcript profiling of V. vulnificus showed that 165 genes out of 4,488 altered their expression profiles by more than twofold. From 35°C to 10°C, an overall gene repression was observed while changes occurring below 10°C mainly resulted in upregulation. The highest induction observed occurred in two of the five categorized cold-shock genes, cspA and cspB, which showed a complementary expression pattern during cold shock suggesting a homologous role. Other genes showing a significant fold increase included ribosomal genes, protein folding regulators, and membrane genes. Repressions were observed in all orthologous groups. Genes with top fold changes in repression include those coding for catalytic enzymes responsible for non temperature-related stress regulation. These included antioxidants, sugar uptake, and amino acid scavengers. V. vulnificus maintained a high level of cspA and cspB transcripts during the entire experiment suggesting that these class I cold-shock genes are required beyond the initial phase of the acclimation period.

First report of Yersinia ruckeri biotype 2 in the USA.

A polyphasic characterization of atypical isolates of Yersinia ruckeri (causative agent of enteric redmouth disease in trout) obtained from hatchery-reared brown trout Salmo trutta in South Carolina was performed. The Y. ruckeri isolates were biochemically and genetically distinct from reference cultures, including the type strain, but were unequivocally ascribed to the species Y. ruckeri, based on API 20E, VITEK, fatty acid methyl ester profiles, and 16S rRNA gene sequencing analysis. These isolates were nonmotile and unable to hydrolyze Tween 20/80 and were therefore classified as Y. ruckeri biotype 2. Genetic fingerprint typing of the isolates via enterobacterial repetitive intergenic consensus (amplified by polymerase chain reaction) and fragment length polymorphism showed biotype 2 as a homogeneous group distinguishable from other Y. ruckeri isolates. This is the first report of Y. ruckeri biotype 2 in the USA.