An early in vitro sampling method for fish immunization programs
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Erythrocytic inclusion body syndrome (EIBS), a viral erythrocytic necrosis (VEN)-like disease, has been observed in several areas in the Northwest. This virus disease is clinically diagnosed by microscopic examination of blood smears for intracytoplasmic erythrocytic inclusion bodies. Fish biologists involved in EIBS diagnostic work have been using several types of hematological stains. It became apparent that standardization of the staining procedure was needed. Comparative tests were conducted on blood smears and kidney imprints with the following commonly used blood stains: (1) Leishman-Giesma, (2) Pinacyanol chloride, (3) Powell 's Giemsa, (4) Harleco's Giemsa, (5) Diff Quik differential stain, (6) Wright's. Pinacyanol chloride stain was found to be the most consistent. The following staining procedure is recommended.
High incidences of Ichthyophonus hoferi, a parasite primarily of marine and estuarine fishes, have recently been reported in rockfishes and Pacific herring from the eastern North Pacific (Kent et al 2001, Hershberger et al 2002). Taxonomic position of I. hoferi remains unresolved, but recent phylogenetic studies have grouped the organism with Dermocystidium, Psorospermium, the rosette agent of salmonids, and Rhinosporidium in the Mesomycetozoa, a novel clade of protists near the animal-fungal divergence (Herr et al 1999). Genetic differences among isolates from the east coast of North America indicate that more than one species of Ichthyophonus exist (Rand et al 2000), and different species have likely been assigned the same name based on morphological characteristics. Therefore, hereafter in this manuscript, the organism will be referred to as Ichthyophonus .
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N-sodium-N-chloro-rho-toluenesulfonamide (chloramine-T) effectively controls bacterial gill disease (BGD) in cultured fishes, BGD, a common disease of hatchery-reared salmonids, causes more fish losses than any other disease among these species, This study describes a liquid chromatographic (LC) method that is capable of direct, simultaneous analysis of chloramine-T and its primary degradation product, rho-toluenesulfonamide (rho-TSA), in water. The procedure involves reversed-phase (C-18) LC analysis with ion suppression, using 0.01 M phosphate buffer at pH 3. The mobile phase is phosphate buffer-acetonitrile (60 + 40) at 1 mL/min. Both chemicals can be detected with a UV spectrophotometer at 229 nm; the method is linear up to 40 mg, chloramine-T or rho-TSA/L. Mean recoveries were 96.4 +/- 6.1% for water samples fortified with 0.03 mg chloramine-T/L and 95.3 +/- 4.6% for water samples fortified with 0.005 mg rho-TSA/L. Limits of detection without sample enrichment for chloramine-T and rho-TSA are 0.01 mg/L and 0.001 mg/L, respectively.
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