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Enteric septicemia of catfish (ESC), caused by the gram‐negative enteric bacteria Edwardsiella ictaluri, is a significant threat to catfish aquaculture in the southeastern United States. Antibiotic intervention can reduce mortality; however, antibiotic use results in an imbalance, or dysbiosis, of the gut microbiota, which may increase susceptibility of otherwise healthy fish to enteric infections. Herein, recovery of the intestinal microbiota and survivability of channel catfish in response to ESC challenge was evaluated following a 10‐day course of florfenicol and subsequent probiotic or prebiotic supplementation. Following completion of florfenicol therapy, fish were transitioned to a basal diet or diets supplemented with a probiotic or prebiotic for the remainder of the study. Digesta was collected on Days 0, 4, 8 and 12, beginning on the first day after cessation of antibiotic treatment, and gut microbiota was characterized by Illumina sequencing of the 16S rRNA gene (V4 region). Remaining fish were challenged with E. ictaluri and monitored for 32 days post‐challenge. Florfenicol administration resulted in dysbiosis characterized by inflated microbial diversity, which began to recover in terms of diversity and composition 4 days after cessation of florfenicol administration. Fish fed the probiotic diet had higher survival in response to ESC challenge than the prebiotic (p =.019) and negative control (p =.029) groups. [ABSTRACT FROM AUTHOR]

Objective: Proliferative gill disease (PGD) in Channel Catfish Ictalurus punctatus and hybrid catfish (Channel Catfish × Blue Catfish I. furcatus) is attributed to the myxozoan Henneguya ictaluri. Despite evidence of decreased H. ictaluri transmission and impaired parasite development in hybrid catfish, PGD still occurs in hybrid production systems. Previous metagenomic assessments of clinical PGD cases revealed numerous myxozoans within affected gill tissues in addition to H. ictaluri. The objective of this study was to investigate the development and pathologic contributions of H. ictaluri and other myxozoans in naturally and experimentally induced PGD. Methods: Henneguya species‐specific in situ hybridization (ISH) assays were developed using RNAscope technology. Natural infections were sourced from diagnostic case submissions in 2019. Experimental challenges involved Channel Catfish and hybrid catfish exposed to pond water from an active PGD outbreak, and the fish were sampled at 1, 7, 10, 12, 14, 16, 18, and 20 weeks postchallenge. Result: Nine unique ISH probes were designed, targeting a diagnostic variable region of the 18S ribosomal RNA gene of select myxozoan taxa identified in clinical PGD cases. Partial validation from pure H. ictaluri, H. adiposa, H. postexilis, and H. exilis infections illustrated species‐specific labeling and no cross‐reactivity between different myxozoan species or the catfish hosts. After experimental challenge, mature plasmodia of H. ictaluri and H. postexilis formed in Channel Catfish but were not observed in hybrids, suggesting impaired or delayed sporogenesis in the hybridized host. These investigations also confirmed the presence of mixed infections in clinical PGD cases. Conclusion: Although H. ictaluri appears to be the primary cause of PGD, presporogonic stages of other myxozoans were also present, which may contribute to disease pathology and exacerbate respiratory compromise by further altering normal gill morphology. This work provides molecular confirmation and more resolute developmental timelines of H. ictaluri and H. postexilis in Channel Catfish and supports previous research indicating impaired or precluded H. ictaluri sporogony in hybrid catfish. Impact statementThis work implicates Henneguya ictaluri as the primary cause of proliferative gill disease, a devastating parasitic disease of U.S. catfish aquaculture, but demonstrates other myxozoan parasites can contribute to disease severity. The study better defines the developmental timeline of H. ictaluri and H. postexilis in Channel Catfish and supports research indicating impaired maturation of H. ictaluri in hybrid catfish. [ABSTRACT FROM AUTHOR]

Bolbophorus damnificus is a digenetic trematode causing significant economic losses within the United States commercial catfish industry. The indirect life cycle is complex, requiring piscivorous birds, aquatic snails, and fish to complete. With federal protections on many piscivorous birds and no FDA‐approved therapeutic treatment for trematode infections in fish, management within the catfish industry is primarily limited to controlling the snail host. Planorbella trivolvis and Biomphalaria havanensis are snails commonly found in commercial catfish ponds and known to transmit B. damnificus. This study evaluated a low‐cost snail trap prototype for monitoring populations of these snails in efforts to better inform treatment regimes. Snail traps were deployed into ponds on a commercial catfish farm and checked weekly throughout the production season. All snails were identified and counted to evaluate trap efficacy. Traps captured significantly more snails in foodfish ponds than in fingerling ponds; however, this trend was not present in monthly box samples. B. havanensis was rarely collected and was only found consistently within one pond, while P. trivolvis was routinely collected from all ponds, although abundance varied widely among ponds. Snail traps could be a useful, low‐cost method to monitor snail populations within ponds and better inform control treatments. [ABSTRACT FROM AUTHOR]

In the mid‐2010s, Edwardsiella tarda was reaffiliated into three discrete taxa (E. anguillarum, E. piscicida, and E. tarda), obscuring previous descriptions of E. tarda‐induced pathology in fish. To clarify ambiguity regarding the pathology of E. tarda, E. piscicida, and E. anguillarum infections in US farm‐raised catfish, channel catfish (Ictalurus punctatus), blue catfish (I. furcatus), and channel × blue catfish hybrids were challenged with comparable doses of each bacterium. The most severe pathology and mortality occurred in fish challenged with E. piscicida, supporting previous reports of increased pathogenicity in commercially important ictalurids, while E. anguillarum and E. tarda warrant only minimal concern. Acute pathologic lesions among bacterial species were predominantly necrotizing and characteristic of gram‐negative sepsis but became progressively granulomatous over time. After 100 days, survivors were exposed to the approximate median lethal doses of E. piscicida and E. ictaluri, revealing some cross‐protective effects among E. piscicida, E. anguillarum, and E. ictaluri. In contrast, no fish that survived E. tarda challenge demonstrated any protection against E. piscicida or E. ictaluri. This work supports reports of increased susceptibility of channel, blue, and hybrid catfish to E. piscicida, while highlighting potential cross‐protective affects among fish associated Edwardsiella spp. [ABSTRACT FROM AUTHOR]

Advances in hybridization practices in U.S. catfish aquaculture have led to increased production of channel (Ictalurus punctatus) × blue catfish (Ictalurus furcatus) hybrids to capitalize on their more favorable production characteristics. However, the effects of typical channel catfish pathogens on hybrids are not well understood, including the digenean Bolbophorus damnificus, which has caused significant losses in channel catfish production. Three experiments were conducted to assess the longevity and site specificity of 2 life stages of B. damnificus impacting catfish production. The first experiment investigated the cercarial longevity and infectivity of B. damnificus over time. Channel catfish were individually challenged with 100 cercariae/fish with cercariae aged in 12-hr time intervals over 5 days (n = 5 fish/time point), with metacercarial cysts excised and enumerated 14 days postchallenge. There was a decrease in cercaria viability and encysted metacercariae over the first 36 hr, with the 12-hr time point having both the greatest cercaria survival and the highest number of metacercariae in exposed fish. The second experiment investigated the longevity of metacercariae within both channel and hybrid catfish. Fish (n = 30) were exposed to 2 treatments (75 or 150 cercariae/fish), and 2 fish from each treatment were sampled every 3 mo for 13 mo. Live metacercariae, based on motility observed after excystment, were found in both species up to 13 mo postchallenge, indicating the metacercariae of B. damnificus can persist throughout an entire growing season in both channel and hybrid catfish. The third experiment investigated the site specificity of metacercariae within both channel and hybrid catfish. Fish (n = 60/species) were challenged with 300 cercariae/fish and 9 fish/species were sampled after 90 days. Metacercariae were excised and enumerated from the anterior midsection (head and body), posterior midsection (trunk/caudal peduncle), ventral (belly), and caudal fin (tail) sections of each fish. Overall, the trunk/caudal peduncle had a 2-fold increase in the number of metacercariae excised, and although not significantly higher, results indicate this region should be the focal point of pondside assessment for the presence of B. damnificus because of ease of detection of encysted metacercariae. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Bolbophorus damnificus (Digenea: Bolbophoridae) causes significant losses in US catfish aquaculture. Little is known regarding the prevalence of infected snail hosts during outbreaks. To assess prevalence of snails observed shedding Bolbophorus spp. cercariae, as well as snails that were infected but not observed shedding cercariae, Planorbella trivolvis (n = 8,159) were collected from 14 catfish production ponds with B. damnificus activity in Mississippi, USA. Individual snails were placed in 10 ml of filtered (20 μm) reservoir pond water and observed for 48 hr for cercariae release. Identification of B. damnificus or Bolbophorus sp. type II cercariae was confirmed by duplex PCR. Genomic DNA was isolated from ~100 non‐shedding snails from each pond and the PCR assay used to identify the presence of B. damnificus or Bolbophorus sp. type II infected snails. The prevalence of snails shedding B. damnificus ranged from 0.23 to 13.6%. Observation of cercariae shedding underestimated true B. damnificus and Bolbophorus sp. type II prevalence in 42.9 and 64.3% of sampled ponds, respectively. Prevalence of B. damnificus infected snails was typically <3%, indicating numbers of infected snails during outbreaks are low and mitigation efforts may be successful simply by reducing snail numbers and not require complete eradication. [ABSTRACT FROM AUTHOR]

Edwardsiella piscicida has emerged as an important pathogen of channel × blue catfish hybrids cultured in the southeastern United States. With increasing popularity of hybrid catfish and growing concern over E. piscicida, the crossprotective potential of a live‐attenuated E.ictaluri vaccine was evaluated. An immunization bath of E. piscicida (3 × 107 CFU/ml) protected channel catfish against immersion challenge of wild‐type E. ictaluri (1 × 107 CFU/ml) across multiple temperatures and feeding regimens (relative percent survival [RPS]: 89–100%). Channel catfish immunized orally with a live‐attenuated E. ictaluri vaccine/feed admix (3 × 107 CFU/g of feed) were protected against intracoelomic injections of both high (HD) (7.4x105 CFU/fish; RPS = 85%) and low doses (LD) (1.5 × 105 CFU/fish; RPS = 88%) of E. piscicida. Because of limited feed consumption during vaccination, oral immunization of hybrids was followed by a 30‐min immersion bath with cultured vaccine (5 × 106 CFU/ml). Combinatory oral/bath vaccination was ineffective against HD E. piscicida but was protective against LD E. piscicida (48% RPS) and E. ictaluri (96% RPS). Finally, naïve catfish that survived LD E. piscicida injections were protected against E. ictaluri immersion challenge (2 × 107 CFU/ml; RPS: Hybrids 55%; Channels 75%). Results suggest a live‐attenuated E. ictaluri vaccine can also protect channel and hybrid catfish against E. piscicida. [ABSTRACT FROM AUTHOR]

Abstract: Bolbophorus damnificus (Digenea: Bolbophoridae) causes significant economic losses in catfish aquaculture in the southeastern USA. To determine mortality trends associated with B. damnificus exposure, three separate disease challenges were conducted. The first challenge exposed channel catfish, Ictalurus punctatus, fingerlings (7–13 cm) to approximately 285 B. damnificus cercariae/L, while the second exposed fish to approximately 215 cercaria/L. Fish were monitored for morbidity and mortality for 28 d. In both studies, mortality first occurred 8 d postchallenge (PC) and peaked at 9 and 10 d PC, respectively. Mortality had subsided by 14 d PC, with only sporadic mortality occurring >15 d PC. Dead and moribund fish presented with lesions consistent with B. damnificus infection. A third challenge was performed to record histological changes associated with parasite development from initial exposure up to the cessation of mortality. Catfish (2–5 cm) were exposed as described above to a range covering 0, 35, 70, 100, 150, 175, and 215 cercariae/L. Each day, two fish were sampled from each treatment. Mortality trends were consistent with previous challenges. The mortality window coincided with parasite encapsulation, neovascularization, and development of the host‐derived cyst, suggesting that morbidity and mortality are linked to host pathophysiological changes in response to metacercariae encapsulation. [ABSTRACT FROM AUTHOR]

Abstract The severity of gill damage (percentage of gill lamellae with lytic lesions) was determined in juveniles from 10 USDA 103 line channel catfish Ictalurus punctatus full-sib families, 10 channel catfish×blue catfish I. furcatus backcross hybrid (7/8 channel catfish, 1/8 blue catfish) full-sib families and a mixed-family group of blue catfish placed in a commercial catfish pond experiencing proliferative gill disease (PGD)-related fish mortalities. An initial challenge was conducted with all families, and a second challenge was conducted using the two most susceptible (most gill damage) channel catfish and backcross hybrid families and the two most resistant (least gill damage) channel catfish and backcross hybrid families. In the initial challenge, percentage gill damage was not different between channel catfish (12.3%) and backcross hybrids (11.6%), but was lower in blue catfish (0.2%). Mean percentage gill damage in the second challenge was not different among resistant backcross hybrid families (6.9%), resistant channel catfish families (7.6%) and blue catfish (4.8%), but was higher in susceptible backcross hybrid and channel catfish families (19.0% and 11.9% respectively). The correlation among family means for gill damage from challenge 1 and challenge 2 was r =0.87. Consistent differences between channel catfish and blue catfish and between resistant and susceptible families within genetic groups for gill damage after PGD challenge suggest that there is a genetic component for resistance to PGD and that improving PGD resistance through selection may be possible. [ABSTRACT FROM AUTHOR]