Detection and quantification of Spironucleus barkhanus in experimentally infected Atlantic salmon Salmo salar

Guo, Fu Ci; Woo, P. T. K.

doi:10.3354/dao061175

Cited by 12 publications

(7 citation statements)

References 12 publications

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“…Three techniques -Wet Mount Examination (WME), Hematocrit Centrifuge Technique (HCT; Woo 1969) and Hemocytometer (HCM; Archer 1965) -were used for the detection and quantitation of parasites in the blood. WME was used for mucus detection; WME and HCM were used for tissues and organs (Guo & Woo 2004).…”

Section: Methodsmentioning

confidence: 99%

Experimental infections of Atlantic salmon Salmo salar with Spironucleus barkhanus

Guo

Woo

2004

Dis. Aquat. Org.

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Atlantic salmon Salmo salar L. (Salmonidae) were experimentally infected with Spironucleus barkhanus (Diplomonadida: Hexamitidae). Parasites were found in the blood 1 to 8 wk after infection, after which they disappeared from the blood and were found mainly in the internal organs (e.g. spleen and liver), eye socket or muscles. Mortality (38 out of 40 infected fish) occurred when fish had lesions in internal organs and/or on the body surface. Uninfected fish cohabiting with infected fish became infected after 4 wk, indicating direct transmission. There was no difference in susceptibility to spironucleosis between 3 different families of Atlantic salmon. All families developed the disease with a similar pattern of parasitaemia in the blood, similar clinical signs and gross pathology, and with very high mortality (29 out of 30). Clinical signs of systemic spironucleosis may include anemia, skin blisters, muscle ulcerations or unilateral exophthalmia. Gross pathologies include hemorrhaging of internal organs, splenomegaly or deformed (globulated) spleen, or granulomatous lesions in the spleen and liver.

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Section: Methodsmentioning

confidence: 99%

Experimental infections of Atlantic salmon Salmo salar with Spironucleus barkhanus

Guo

Woo

2004

Dis. Aquat. Org.

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“…For example, Watanabe et al (1998) reported successful prevention of vertical transmission of pathogenic viruses by selection of specific pathogen-free spawners, as indicated by results from an antibody-detection ELISA. However, antibody-detection ELISA has not been accepted internationally because the ELISA system for fish antibody detection has problems, such as low reproducibility partly due to high background optical density (OD) caused by non-specific reactions between fish antibodies and antigens (Olesen et al 1991, Höglund & Pilström 1995, Knopf et al 2000, Kibenge et al 2002, Guo & Woo 2004. We have experienced similar problems with ELISA systems used for detection of fish antibodies against pathogenic viruses, infectious hematopoietic necrosis virus, viral hemorrhagic septicemia virus, fish nodavirus, lymphocystis disease virus and koi herpesvirus, etc.…”

Section: Introductionmentioning

confidence: 95%

Non-specific adsorption of fish immunoglobulin M (IgM) to blocking reagents on ELISA plate wells

Kim

Nishizawa

Yoshimizu³

2007

Dis. Aquat. Org.

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Enzyme-linked immunosorbent assay (ELISA) is a popular technique for quantifiable detection of specific antibodies in warm-blooded animals, but it has not been accepted for detection of fish antibodies because of its low reproducibility, which is due in part to high background optical density (OD) measurements. In the present study, we report that the high background of a fish antibody- detection ELISA resulted from non-specific adsorption of fish immunoglobulin M (IgM) to blocking reagents on the ELISA plate wells. Four fish sera (from rainbow trout Oncorhynchus mykiss, masu salmon O. masou, Japanese flounder Paralichthys olivaceus and koi Cyprinus carpio) were poured into ELISA plate wells pre-blocked with several blocking reagents (skim milk, soybean milk, bovine serum albumin, fetal bovine serum, gelatin and Roche BlockingReagent®) and then washed out in order to measure the remaining fish IgM on the ELISA plate wells. Significant amounts of fish IgMs (OD absorbance at 492 nm: 0.3 to 1.1) remained on the ELISA plate wells with no antigenic protein except blocking reagents. The amount of remaining fish IgMs on the ELISA plate wells decreased significantly following treatment of fish sera with skim milk. However, the specific immuno-reactivity of fish IgM was not reduced by such treatment. Thus, we conclude that treatment of fish sera with skim milk is useful in reducing the high background OD often observed in fish IgM detection ELISA

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“…Several studies using ELISA systems for the detection of specific fish antibodies have been reported for aquatic animals (Jorgensen et al 1991;Yoshimizu et al 1992;Dixon et al 1994;Pilström 1994, 1995;LaPatra 1996;Nishida et al 1998;Watanabe et al 1998;Swain and Nayak 2003;Okuda et al 2006;Kim et al 2007Kim et al , 2008Kim et al , 2009Takami et al 2010). However, use of an ELISA for fish antibody detection can be problematic because of difficulties such as low reproducibility, which is partly due to high background optical density (OD) caused by nonspecific reactions between fish antibodies and antigens Pilström 1994, 1995;Knopf et al 2000;Kibenge et al 2002;Guo and Woo 2004). It was reported that one of the most important causes of those problems was the nonspecific adsorption of fish antibodies with ELISA blocking reagents, but nonspecific adsorption can be suppressed by the pretreatment of fish sera with a skim milk solution (Kim et al 2007).…”

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confidence: 99%

Cell Culture Medium Inhibits Antigen Binding Used in an ELISA for Detection of Antibodies against Nervous Necrosis Virus

Choi

Gye

et al. 2014

J Aqua Anim Hlth

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We investigated the optimum dilution of nervous necrosis virus (NNV) for use as antigens to detect antibodies by an enzyme-linked immunosorbent assay (ELISA) in the Sevenband Grouper Epinephelus septemfasciatus. The ELISA values for a standardized suspension of antigens diluted with L-15 medium containing 1% fetal bovine serum decreased gradually with the dilution of the antigens, whereas those for the antigens diluted with distilled water (DW) initially increased with the dilution of the antigens, peaked at a 320-fold dilution, and then decreased thereafter. Additional studies revealed that binding of NNV antigens to ELISA wells was inhibited by fetal bovine serum and other substances in the L-15 medium. Sera obtained from Sevenband Grouper vaccinated with live NNV vaccine and survivors from natural NNV-infection were subjected to antibody detection by ELISA. All of the sera were positive by ELISA when the standardized suspension was diluted 320-fold, whereas sera from five out of the six survivors and two out of the six vaccinated fish were negative or weakly positive by ELISA using NNV antigens diluted 10-fold. We therefore concluded that cultured NNV solutions prepared in cell culture media may need to be diluted with distilled water for use in ELISA. Received January 28, 2014; accepted April 16, 2014.

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Detection and quantification of Spironucleus barkhanus in experimentally infected Atlantic salmon Salmo salar

Cited by 12 publications

References 12 publications

Experimental infections of Atlantic salmon Salmo salar with Spironucleus barkhanus

Experimental infections of Atlantic salmon Salmo salar with Spironucleus barkhanus

Non-specific adsorption of fish immunoglobulin M (IgM) to blocking reagents on ELISA plate wells

Cell Culture Medium Inhibits Antigen Binding Used in an ELISA for Detection of Antibodies against Nervous Necrosis Virus

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