Aeromonas shuberti as a cause of multi-organ necrosis in internal organs of Nile tilapia, Oreochromis niloticus.

A disease with white spots in internal organs of Nile tilapia occurred in Zhanjiang, southern China. Multiple, white nodules, 0.8-2.2 mm in diameter, were scattered throughout the liver, spleen and kidney of diseased fish. Signs of nodules reproduced after artificial infection with the isolated strain. Isolated bacteria were Gram-negative, facultative anaerobic, motile, short rod-shaped, with a length of 1.2-2.2 µm. Morphological and biochemical tests, as well as phylogenetic analysis, all strongly indicated that the isolate from tilapia is identical to Aeromonas schubertii (A. schubertii) which temporary named LF1708 strain. Antibiotic sensitivity assays showed the LF1708 is sensitive to 24 of 27 tested antibiotics. Pathogenicity test revealed that the isolate at the dose of 3.75 × 106 CFU/g killed 100% of experimental tilapia within 2 days and the dose of 1 × 107 CFU/g killed 100% of experimental zebrafish within 1 day. Histopathology of diseased tilapia infected with A. schubertii showed numerous necrotic lesions widely distributed in spleen, liver and kidney, and infiltration with a large number of bacteria. To our knowledge, this was the first report that associated A. schubertii with mortality in tilapia.

Transparent Tiger barb Puntius tetrazona, a fish model for in vivo analysis of nocardial infection.

Nocardiosis afflicts multiple species of cultured fish, resulting in substantial economic losses to the aquaculture industry, however, lack of detailed knowledge on disease pathogenesis has hampered the development of effective prevention and control strategies. In this study, we injected a green fluorescent protein (GFP)-labeled Nocardia seriolae strain into a transparent mutant strain of Tiger barb (Puntius tetrazona) to monitor tissue pathogen accumulation and tissue damage in vivo, and to clarify the relationship between pathogenic processes and overt symptoms. GFP-labeled bacteria were phagocytized by leukocytes and could proliferate within these cells, which in turn led to leukocyte aggregation, leukocyte death, and granuloma formation. In addition, intracellular bacteria could permanently colonize various tissues via leukocyte circulation, causing multi-organ infection as revealed by changes of tissue transparency. Histology revealed granulomatous lesions in organs such as muscle, kidney, and spleen that was corresponded to the tissue opacities in vivo. Confocal microscopy confirmed massive accumulations of GFP-labeled bacteria within these granulomas, which often contained a necrotic core. Tiger barb transparency allows for real-time observation of in vivo pathological changes within the same animal, and the pathogenic process can be evaluated based on the shape and size of body opacities. Thus, transparent Tiger barb is a promising model to study the pathogenesis of nocardiosis.

Identification of a transparent mutant tiger barb Puntius tetrazona and its use for in vivo observation of a Pleistophora sp. (Microsporidia) infection.

A transparent mutant tiger barb Puntius tetrazona was identified and characterized by its transparent body, which allows clear visualization of internal organs. Hybridization of this mutant with the albino variant produces a transparent and albinoid double phenotype, and the transparency of this mutant is controlled by a recessive allele. Light microscopic and ultrastructural examinations show that in contrast to normal individuals, transparent mutants lack iridophores, and light penetrates unimpeded through the body. Pleistophora sp. infection was observed in vivo, allowing live observation of parasite dissemination and the consequent pathological alterations in the fish body as well as the simultaneous acquisition of data on the dynamics and spatial pattern of pathogenic invasion. It is superior to common fish models, as dynamic experimental data can be obtained from individual fish.