Molecular characterization of "Candidatus Parilichlamydia carangidicola," a novel Chlamydia-Like epitheliocystis agent in yellowtail kingfish, Seriola lalandi (Valenciennes), and the proposal of a new family, "Candidatus Parilichlamydiaceae" fam. nov. (order Chlamydiales)
Stride M.C.,University of Tasmania |
Polkinghorne A.,Queensland University of Technology |
Miller T.L.,James Cook University |
Groff J.M.,University of California at Davis |
And 2 more authors.
Applied and Environmental Microbiology | Year: 2013
Three cohorts of farmed yellowtail kingfish (Seriola lalandi) from South Australia were examined for Chlamydia-like organisms associated with epitheliocystis. To characterize the bacteria, 38 gill samples were processed for histopathology, electron microscopy, and 16S rRNA amplification, sequencing, and phylogenetic analysis. Microscopically, the presence of membrane-enclosed cysts was observed within the gill lamellae. Also observed was hyperplasia of the epithelial cells with cytoplasmic vacuolization and fusion of the gill lamellae. Transmission electron microscopy revealed morphological features of the reticulate and intermediate bodies typical of members of the order Chlamydiales. A novel 1,393-bp 16S chlamydial rRNA sequence was amplified from gill DNA extracted from fish in all cohorts over a 3-year period that corresponded to the 16S rRNA sequence amplified directly from laser-dissected cysts. This sequence was only 87% similar to the reported "Candidatus Piscichlamydia salmonis" (AY462244) from Atlantic salmon and Arctic charr. Phylogenetic analysis of this sequence against 35 Chlamydia and Chlamydialike bacteria revealed that this novel bacterium belongs to an undescribed family lineage in the order Chlamydiales. Based on these observations, we propose this bacterium of yellowtail kingfish be known as "Candidatus Parilichlamydia carangidicola" and that the new family be known as "Candidatus Parilichlamydiaceae". © 2013, American Society for Microbiology.
Evenhuis J.P.,U.S. Department of Agriculture |
LaPatra S.E.,Clear Springs Foods Inc. |
Marancik D.,U.S. Department of Agriculture
Aquaculture | Year: 2014
Flavobacterium columnare is the etiologic agent of columnaris disease, a pervasive disease of fresh water finfish. During the past 4. years, losses that ranged from 5 to 50% in rainbow trout (Oncorhynchus mykiss) fry being reared at a constant 14.5. °C (mean weight, 0.2. g; ~. 400°days post-fertilization), have been occurring in Hagerman Valley, Idaho, USA. A total of 70 different F. columnare isolates were obtained from diseased fish and the water they were reared in. All of the isolates were confirmed to be genomovar I by 16S rRNA restriction fragment length polymorphism. Sequencing of the 16S rRNA, 16S-23S rDNA spacer region and the gyrase B subunit genes from these 70 strains revealed no sequence differences among these isolates. Whole-cell protein profiling by SDS-PAGE also indicated low variation between isolates. Virulence was assessed for a representative isolate and demonstrated a high degree of pathogenicity against rainbow trout fry at 15. °C. These results suggest the emergence of a highly successful F. columnare strain that can affect very early life stages of fish being reared at a constant 14.5. °C at a commercial rainbow trout farm in Idaho. © 2013.
Welch T.J.,U.S. Department of Agriculture |
LaPatra S.,Clear Springs Foods Inc.
Fish and Shellfish Immunology | Year: 2016
Enteric redmouth disease (ERM), caused by Yersinia ruckeri, has been controlled successfully using immersion-applied bacterin vaccines for several decades. While the host response to vaccination and the mechanism of protection of this vaccine have been elucidated, the bacterial components eliciting protection have remained unclear. Here we show that highly purified serotype O1 Y. ruckeri lipopolysaccharide (LPS) is sufficient to induce a protective response to experimental challenge in rainbow trout (Oncorhynchus mykiss). Dose response experiments demonstrated that Y. ruckeri LPS at doses of 1 ng/fish and above resulted in essentially complete protection and doses as low as 0.01 ng/fish (1.38 ng/kg) resulted in significant protection, thus demonstrating the extremely high potency of this immunogen. Analysis of the Y. ruckeri genome identified a cluster of putative O-antigen biosynthetic genes specific to serotype O1 strains. This cluster primarily consisted of genes encoding proteins predicted to function in the biosynthesis of legionamic acid, a nonulosonic acid known to be part of the O-polysaccharide repeat of O1 Y. ruckeri. Mutation of the nab2 gene, a nonulosonic acid biosynthesis gene (nab gene), resulted in production of severely truncated forms of LPS. Vaccination with bacterin vaccines derived from the nab2 mutant and its wild type parent strain demonstrated that LPS is a required component of the whole-cell bacterin vaccine and suggests that LPS is the only cellular component contributing to the protective response elicited by this vaccine. We speculate that the exceptionally high potency of Y. ruckeri LPS accounts for the unusual success of this vaccine when delivered by immersion. © 2016.
LaFrentz B.R.,U.S. Department of Agriculture |
LaPatra S.E.,Clear Springs Foods Inc. |
Shoemaker C.A.,U.S. Department of Agriculture |
Klesius P.H.,U.S. Department of Agriculture
Diseases of Aquatic Organisms | Year: 2012
Flavobacterium columnare is a Gram-negative bacterium that causes columnaris disease and has significant economic impacts on aquaculture production worldwide. Molecular analyses have demonstrated that there is genetic diversity among F. columnare isolates. A review of the published literature that used restriction fragment length polymorphism analysis of the 16S rRNA gene revealed that all isolates typed from salmonids were Genomovar I. Our objective was to develop a laboratory challenge model for F. columnare in rainbow trout Oncorhynchus mykiss (Walbaum) and use the model to determine the virulence of Genomovar I and II isolates. Six F. columnare isolates were obtained from rainbow trout experiencing losses due to columnaris disease and were determined to be Genomovar I. Three of these were chosen for a preliminary assessment of virulence, and isolate 051-10-S5 was chosen for additional experiments to determine the reproducibility of the waterborne challenge model. In 2 independent experiments, cumulative percent mortalities (CPM) were 49 ± 10% and 50 ± 19%. Challenge of rainbow trout with Genomovar I and II isolates demonstrated a difference in the CPM, with the Genomovar II isolates inducing significantly higher CPM. This reproducible waterborne challenge model for columnaris disease in rainbow trout will be useful to investigate host- pathogen interactions, vaccine development, and other potential control strategies. This research also provides a basis for further defining the molecular diversity and virulence associated with F. columnare genomovars in rainbow trout and other salmonid species. © Inter-Research 2012.
Salinas I.,University of New Mexico |
LaPatra S.E.,Clear Springs Foods Inc. |
Erhardt E.B.,University of New Mexico
Developmental and Comparative Immunology | Year: 2015
Determining the earliest age at which farmed fish can be successfully vaccinated is a very important question for fish farmers. Nasal vaccines are novel mucosal vaccines that prevent aquatic infectious diseases of finfish. The present study investigates the ontogeny of the olfactory organ of rainbow trout by histology and aims to establish the earliest age for vaccination against infectious hematopoietic necrosis (IHN) and enteric red mouth (ERM) disease using the nasal route. Rainbow trout (. Oncorhynchus mykiss) were vaccinated intranasally (I.N) at three different ages: 1050° days (DD) (group A); 450 DD (group B); and 360 DD (group C), or 70, 30 and 24 days post-hatch (dph), respectively. The mean weights of groups A, B and C were 4.69 g, 2.9 g and 2.37 g, respectively. Fish received either a live attenuated IHN virus vaccine, ERM formalin killed bacterin or saline (mock vaccinated). Fish were challenged to the corresponding live pathogen 28 days post-vaccination. IHN vaccine delivery at 360 DD resulted in 40% mortality likely due to residual virulence of the vaccine. No mortality was observed in the ERM nasal delivery groups. Following challenge, very high protection rates against IHN virus were recorded in all three age groups with survivals of 95%, 100% and 97.5% in groups A, B and C, respectively. Survival against ERM was 82.5%, 87.5% and 77.5% in groups A, B and C, respectively. Survival rates did not differ among ages for either vaccine. Our results indicate the feasibility and effectiveness of nasal vaccination as early as 360 DD and vaccination-related mortalities when a live attenuated viral vaccine was used in the youngest fish. © 2015 Elsevier Ltd.
LaPatra S.,Clear Springs Foods Inc. |
Kao S.,University of New Mexico |
Erhardt E.B.,University of New Mexico |
Salinas I.,University of New Mexico
Vaccine | Year: 2015
Farmed fish are susceptible to different infectious disease agents including viruses and bacteria. Thus, multivalent vaccines or vaccination programs against two or more pathogens are valuable tools in aquaculture. Recently, nasal vaccines have been shown to be very effective in rainbow trout. The current study investigates, for the first time, the use of the nasal route in dual vaccination trials against two important aquatic diseases, infectious hematopoietic necrosis virus (IHN) and enteric red mouth (ERM) disease. Rainbow trout received live attenuated IHN virus (IHNV) vaccine and the ERM bacterin using four different vaccine delivery methods and were challenged with virulent IHNV or Yersinia ruckeri 7 (100. deg day) and 28 (400. deg day) days post-vaccination. The highest survival rates against IHNV at day 7 were obtained by nasal vaccination either when IHNV and ERM were delivered separately into each nare or when they were premixed and delivered to both nasal rosettes (group D). Protection at 28 days against IHNV was similar in all four vaccinated groups. Early protection against ERM was highest in fish that received each vaccine in separate nares (group B), whereas protection at 28 days was highest in the i.p. vaccinated group (group E), followed by the nasally vaccinated group (group B). Survival results were supported by histological observations of the left and right olfactory organ which showed strong immune responses one day (14. deg days) after vaccination in group B vaccinated fish. These data indicate that dual vaccination against two different pathogens via the nasal route is a very effective vaccination strategy for use in aquaculture, particularly when each nare is used separately during delivery. Further long-term studies should evaluate the contribution of adaptive immunity to the protection levels observed. © 2015 Elsevier Ltd.
Plant K.P.,University of Idaho |
LaPatra S.E.,Clear Springs Foods Inc.
Developmental and Comparative Immunology | Year: 2011
Disease prevention is essential to the continued development of aquaculture around the world. Vaccination is the most effective method of combating disease and currently there are a number of vaccines commercially available for use in fish. The majority of aquatic vaccines are delivered by injection, which is by far the most effective method when compared to oral or immersion deliveries. However it is labor intensive, costly and not feasible for large numbers of fish under 20. g. Attempts to develop novel oral and immersion delivery methods have resulted in varying degrees of success but may have great potential for the future. © 2011 Elsevier Ltd.
Purcell M.K.,U.S. Geological Survey |
LaPatra S.E.,Clear Springs Foods Inc. |
Woodson J.C.,U.S. Geological Survey |
Kurath G.,U.S. Geological Survey |
Winton J.R.,U.S. Geological Survey
Fish and Shellfish Immunology | Year: 2010
The main objective of this study was to assess correlates of innate resistance in rainbow trout full-sibling families that differ in susceptibility to Infectious hematopoietic necrosis virus (IHNV). As part of a commercial breeding program, full-sibling families were challenged with IHNV by waterborne exposure at the 1 g size to determine susceptibility to IHNV. Progeny from select families (N = 7 families) that varied in susceptibility (ranging from 32 to 90% cumulative percent mortality (CPM)) were challenged again at the 10 g size by intra-peritoneal injection and overall mortality, early viral replication and immune responses were evaluated. Mortality challenges included 20-40 fish per family while viral replication and immune response studies included 6 fish per family at each time point (24, 48 and 72 h post-infection (hpi)). CPM at the 1 g size was significantly correlated with CPM at the 10 g size, indicating that inherent resistance was a stable trait irrespective of size. In the larger fish, viral load was measured by quantitative reverse-transcriptase PCR in the anterior kidney and was a significant predictor of family disease outcome at 48 hpi. Type I interferon (IFN) transcript levels were significantly correlated with an individual's viral load at 48 and 72 hpi, while type II IFN gene expression was significantly correlated with an individual's viral load at 24 and 48 hpi. Mean family type I but not type II IFN gene expression was weakly associated with susceptibility at 72 hpi. There was no association between mean family susceptibility and the constitutive expression of a range of innate immune genes (e.g. type I and II IFN pathway genes, cytokine and viral recognition receptor genes). The majority of survivors from the challenge had detectable serum neutralizing antibody titers but no trend was observed among families. This result suggests that even the most resistant families experienced sufficient levels of viral replication to trigger specific immunity. In summary, disease outcome for each family was determined very early in the infection process and resistance was associated with lower early viral replication.
Clear Springs Foods Inc. | Date: 2013-04-10
Fish, namely, fresh and frozen, not live fed a special diet for nutritional value; Meat; Processed meat; Seafood, namely, fresh and frozen, not live fed a special diet for nutritional value.
Clear Springs Foods Inc. | Date: 2013-11-18
Fish, namely, Fresh and frozen fish, not live; Meat; Processed meat; Seafood, not live.