Diversity in clinical isolates of Ictalurid herpesvirus 1 (IcHV1) from U.S. farm-raised catfish and virulence assessment in channel and channel × blue catfish hybrids.

Ictalurid herpesvirus 1 (IcHV1) is the most significant viral agent in U.S. catfish aquaculture. Little is known regarding the genetic stability and antigenic variability of IcHV1. Herein, the genetic and antigenic diversity of IcHV1 field isolates was assessed by restriction fragment length polymorphism (RFLP) analysis and serum neutralization assays. RFLP analysis identified two distinct genotypes (IcHV1A and IcHV1B), both discrete from blue catfish alloherpesvirus (BCAHV). Neutralization assays with anti-IcHV1 monoclonal antibody Mab-95 indicate shared antigenic determinants for IcHV1A and IcHV1B that are absent from BCAHV, which Mab-95 did not neutralize. Virulence assessments with representative isolates demonstrate significant differences between isolates within RFLP groups and pooled RFLP group data suggest IcHV1B (pooled survival [mean ± SE]: 58.3% ± 2.6) may be more virulent than IcHV1A (survival: 68.6% ± 2.4). Rechallenges with representative IcHV1A and IcHV1B isolates indicate a cross-protective effect, with fish surviving initial exposure to IcHV1A or IcHV1B showing robust protection when subsequently re-exposed to IcHV1A or IcHV1B. This work demonstrated significant differences in virulence between case isolates, identifying two discrete IcHV1 lineages, distinct from BCAHV, with similar virulence in channel and channel × blue catfish hybrids and a cross-protective effect in catfish surviving exposure to either lineage.

Edwardsiella piscicida identified in the Southeastern USA by gyrB sequence, species-specific and repetitive sequence-mediated PCR.

A new Edwardsiella taxon was recently described from fishes of Europe and Asia. Phenotypically similar to E. tarda, extensive genetic and phenotypic characterization determined this new strain does not belong to any established Edwardsiella taxa, leading to the adoption of a new taxon, E. piscicida. Concurrent research in the USA also identified 2 genetically distinct taxa within the group of organisms traditionally classified as E. tarda. Comparisons of gyrB sequences between US isolates and E. piscicida from Europe and Asia identified several US isolates with >99.6% similarity to the gyrB sequence of the E. piscicida type strain (ET883) but <87% similarity to the E. tarda type strain (ATCC #15947). A discriminatory PCR was developed for the identification of E. tarda and 2 genetic variants of E. piscicida (E. piscicida and E. piscicida-like species). Using these PCR assays, a survey was conducted of 44 archived bacterial specimens from disease case submissions to the Aquatic Research and Diagnostic Laboratory (Stoneville, MS, USA) between 2007 and 2012. All 44 isolates, originally identified phenotypically and biochemically as E. tarda, were identified as E. piscicida by PCR. Repetitive sequence-mediated PCR (rep-PCR) analysis of these archived specimens suggests they are largely homogenous, similar to what has been observed for E. ictaluri. The gyrB sequence data, coupled with the E. piscicida specific-PCR and rep-PCR data, confirms that E. piscicida has been isolated from fish disease cases in the southeastern USA. Moreover, our survey data suggests E. piscicida may be more prevalent in catfish aquaculture than E. tarda.

MYXOZOAN COMMUNITY COMPOSITION AND DIVERSITY IN CLINICAL CASES OF PROLIFERATIVE GILL DISEASE IN MISSISSIPPI CATFISH AQUACULTURE.

An abundance of morphologically variable Henneguya species complicates the understanding of disease relationships between ictalurid catfish and myxozoan (Phylum: Cnidaria) parasites on North American aquaculture operations. Henneguya ictaluri, the cause of proliferative gill disease (PGD) in channel and hybrid catfish, is arguably the most important parasite of commercial catfish aquaculture in the southeastern United States. While research indicates arrested development and limited sporogenesis of H. ictaluri in channel (Ictalurus punctatus) × blue (Ictalurus furcatus) hybrid catfish, incidents of PGD persist in hybrid production systems. This work investigated the influence of fish host on myxozoan community composition and diversity within naturally infected gill tissues from diagnostic case submissions to the Aquatic Research and Diagnostic Laboratory in Stoneville, Mississippi, from 2017 to 2019. Gills collected from farm-raised catfish with clinical PGD were subjected to metagenomic amplicon sequencing of the myxozoan 18S SSU rDNA gene diagnostic variable region 3 (DVR3). Myxozoan community composition significantly differed between channel and hybrid catfish PGD cases, with channel catfish having more diverse community structures. Channel catfish gills had a greater relative abundance of H. ictaluri in 2017 and 2019, while no differences were observed in 2018. Importantly, H. ictaluri was present in all channel and hybrid catfish PGD cases across all years; however, H. ictaluri was not the most abundant myxozoan in almost half the cases examined, suggesting other myxozoan species may also contribute to PGD pathology. The detection of numerous known and unclassified myxozoan sequences in addition to H. ictaluri provides evidence PGD may involve mixed species infections. Furthermore, the presence of numerous unclassified myxozoan sequences in gill samples from clinical PGD cases indicates the number of described species from U.S. farm-raised catfish vastly underestimates the true myxozoan diversity present within the varied pond microcosms associated with catfish aquaculture.

Chronic pathology and longevity of Drepanocephalus spathans infections in juvenile Channel Catfish.

Drepanocephalus spathans (Digenea: Echinostomatidae) is a common parasite of the double-crested cormorant Phalacrocorax auritus. The cercariae of D. spathans have been shown infective to juvenile Channel Catfish Ictalurus punctatus. The developing metacercariae concentrate in the cranial regions, often occluding blood vessels at the base of the branchial arch, occasionally resulting in death. The purpose of this study was to determine how long metacercariae of D. spathans persist in experimentally challenged Channel Catfish. Two separate infectivity trials were conducted. In both trials, metacercariae persisted at least 49 d postinfection, although prevalence and intensity of infection decreased over time. In the first trial, juvenile catfish (1-3 g) were exposed over three consecutive days to 100, 100, and 80 cercariae/fish/d, respectively. Fish were sampled 7 d after the final exposure, and metacercariae were observed in 83.3% (five of six) of challenged fish. At 21 d postexposure, metacercariae were present in only 50% of exposed fish (three of six). No metacercaria were observed in fish sampled at 35 d, however, metacercariae were present in one of six (16.7%) fish sampled 49 and 70 d postexposure, respectively. A second challenge consisted of a 24-h pooled exposure of 500 cercariae per fish. Again, metacercariae were present in most (six of seven; 85.7%) fish at 7 d postexposure. At 21 d postexposure, metacercariae were only evident in one of seven (14.3%) sampled fish. No metacercariae were present in any fish at 35 d postchallenge, yet one of seven (14.3%) fish was positive at 49 d postchallenge. The second study was terminated at 63 d postchallenge, as all fish sampled (n = 14) were negative for metacercariae. These data suggest that cercariae of D. spathans are infective to juvenile Channel Catfish, although the infection appears short lived as metacercariae rarely persisted longer than 2 months.

The pathology associated with visceral toxicosis of catfish.

Visceral toxicosis of catfish (VTC) syndrome was recognized in the late 1990 s and recently has been associated with exposure to Clostridium botulinum type E neurotoxin. Tentative diagnosis is based on clinical presentation and gross findings, and is confirmed by bioassay. In April 2009, channel catfish (Ictalurus punctatus) from 2 different farms presented with abnormal swimming behavior and mortalities. Nine fish were submitted to the Aquatic Research and Diagnostic Laboratory (Stoneville, Mississippi) for evaluation. Bacterial cultures from these fish were negative. Necropsy findings included intestinal intussusceptions, ascites, pale proximal intestines with engorged serosal blood vessels, splenic congestion, and a reticular pattern to the liver. Significant histopathologic findings were limited to cerebral, splenic, and hepatic congestion, splenic lymphoid depletion and perivascular edema, vascular dilation and edema of the gastrointestinal tract, and perivascular edema in the anterior and posterior kidneys. Intoxication from C. botulinum type E neurotoxin was suspected based on the clinical signs and lack of gross and microbiological evidence of an infectious disease process. The toxicosis was confirmed with a positive bioassay using serum collected from the submitted fish.