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J. D. Shields

Publications and source records attributed to J. D. Shields.

3 recordsLinked to original sources

Infection experiments with Aphanomyces invadans in four species of estuarine fish

Along the eastern seaboard of the US, Atlantic menhaden, Brevoortia tyrannus, develop characteristic ulcerative lesions, a condition termed ulcerative mycosis. These lesions are identical to those seen across Asia in fish affected by epizootic ulcerative syndrome, a condition caused by the fungus-like oomycete Aphanomyces Invadans. Young-of-the-year menhaden inhabiting estuarine environments are the primary species affected in the USA and little is known about the factors involved in the initiation of the lesions, or why menhaden are predominantly infected. Atlantic menhaden, hogchoker, Trinectus maculatus, striped killifish, Fundulus majalis, and mummichog, Fundulus heteroclitus, were inoculated with A. invadans (80 zoospores per fish) to explore species differences in infection and lesion development. All four species developed lesions. Killifish developed frank lesions similar to those observed in menhaden but the gross lesions occurred later, approximately 5-10 days after those on menhaden. Hogchoker and mummichog did not develop gross skin ulcers; rather, their lesions appeared as reddened areas under the epidermis. Mummichogs also showed evidence of significant healing with a well-developed granuloma and significant myocyte regeneration. These experiments show that species barriers as well as ecological barriers can explain some of the factors involved in the development of lesions in, and specificity of the water mould for, menhaden.

Journal of Fish Diseases

Infectivity and pathogenicity of the oomycete Aphanomyces invadans in Atlantic menhaden Brevoortia tyrannus

Atlantic menhaden Brevoortia tyrannus develop characteristic skin ulcers in response to infection by the oomycete Aphanomyces invadans . To investigate pathogenicity , we conducted a dose response study. Juvenile menhaden were inoculated subcutaneously with 0, 1, 5, 10, 100, and 500 secondary zoospores per fish and monitored for 37 d post-injection (p.i.). Survival rates declined with increasing zoospore dose, with significantly different survivorship curves for the different doses. Moribund and dead fish exhibited characteristic ulcerous lesions at the injection site starting at 13 d p.i. None of the sham-injected control fish (0 zoospore treatment) died. The LD 50 (lethal dose killing 50% of exposed menhaden ) for inoculated fish was estimated at 9.7 zoospores; however, some fish receiving an estimated single zoospore developed infections that resulted in death. Menhaden were also challenged by aqueous exposure and confirmed that A. invadans was highly pathogenic by this more environmentally realistic route. Fish that were acclimated to culture conditions for 30 d, and presumably free of skin damage, then aqueously exposed to 100 zoospores ml -1 , exhibited 14% lesion prevalence with 11% mortality. Net-handled fish that were similarly infected had a significantly higher lesion prevalence (64%) and mortality (64%). Control fish developed no lesions and did not die. Scanning electron microscopy of fish skin indicated that zoospores adhered to intact epidermis, germinated and penetrated the epithelium with a germ tube. Our results indicate that A. invadans is a primary pathogen of menhaden and is able to cause disease at very low zoospore concentrations.

Diseases of Aquatic Organisms

Induction of skin ulcers in Atlantic menhaden by injection and aqueous exposure to the zoospores of Aphanomyces invadans

The infectivity and role of Aphanomyces invadans in the etiology of skin ulcers in Atlantic menhaden Brevoortia tyrannus were investigated with two laboratory challenges. In the first experiment, Atlantic menhaden received subcutaneous injections with secondary zoospores from one of three cultures of Aphanomyces : WIC (an endemic isolate of A. invadans in Atlantic menhaden from the Wicomico River, Maryland), PA7 (an isolate of A. invadans from striped snakehead Channa striata (also known as chevron snakehead), infected with epizootic ulcerative syndrome from Thailand), and ATCC-62427 (an isolate from Atlantic menhaden from North Carolina). Fish were injected with 1.9 × 10 2 (WIC-low), 1.9 × 10 3 (WIC-high), 5.2 × 10 2 (PA7), or 6.0 × 10 2 (ATCC-62427) zoospores and held in static water at 23.5°C (6‰ salinity) for 21 d. Both low and high doses of WIC caused incipient, granulomatous lesions after 5 d. Fish injected with the high-dose WIC died within 7 d. All fish injected with the low-dose WIC were dead after 10 d. Fish injected with zoospores of PA7 developed lesions after 9 d. Fish injected with the ATCC-62427 isolate or those that received subcutaneous injections of sterile water (controls) did not develop lesions. In the second experiment, fish were bath-exposed with zoospores of the WIC isolate after various trauma-inducing treatments. These treatments consisted of handling fish with a net (net stress, exposed for 2 h to either 70 or 700 zoospores/mL), physically removing a few scales (trauma, exposed for 1 h to 700 zoospores/mL), or acclimating fish with less handling (acclimated, untraumatized, exposed for 5.5 h to 110 zoospores/mL). Unexposed fish served as controls. Mortality ranged from 94% to 100% for net-handled and traumatized fish, with the prevalence of ulcerous lesions ranging from 70% to 79% in net-handled fish. However, mortality was 24% for the “untraumatized” fish and the prevalence of lesions was 32%. Fish injected with or exposed to bath challenges of zoospores developed lesions that were grossly and histologically identical to those observed in naturally infected Atlantic menhaden from several estuaries and rivers along the mid-Atlantic coast of the USA. The deeply penetrating ulcers were characterized by dermatitis, myofibrillar degeneration, and deep, necrotizing granulomatous myositis. Experimentally induced lesions, however, exhibited invasiveness, often involving the kidney. Injected or bath-exposed fish developed incipient granulomas after 5 d, which progressed to overt lesions over 7–9 d. We have here demonstrated that ulcerative skin lesions can be experimentally induced in Atlantic menhaden after exposure to oomycete zoospores of an endemic strain of A. invadans .

Journal of Aquatic Animal Health