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A comparative histopathological, immunohistochemical and nested-PCR study for diagnosis of infectious pancreatic necrosis in the naturally infected cultured rainbow trout (Oncorhynchus mykiss)

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Abstract

To confirm the diagnosis of the infectious pancreatic necrosis, the pancreas, liver, kidney and spleen specimens from 140 rainbow trout (Oncorhynchus mykiss) of weight ranges from 15 to 250 g were processed for routine histopathological and immunohistochemical (IHC) studies together with a nested-PCR using primers that amplify a 164-bp product. Of the 140 fish samples, 37 (26.4%) had a final diagnosis of IPN on the basis of hematoxylin and eosin staining (H&E), while 39/45 (86.6%) were positive by IHC method. With the nested-PCR, 59/140 (42.1%) fish samples were positive. The IHC and nested-PCR showed higher prevalence than histopathology (P < 0.05). Based on the nested-PCR, as the fish weight increased, IPN positive results decreased (P < 0.05). However, IHC method detected IPNV constantly in these weight ranges. Only IPNV serotype Sp was identified by nucleotide sequencing and immunohistochemistry, and antiserum to IPNV serotype Ab and IHNV showed negative results in IHC. This is the first comparative diagnostic study of IPN at different weight ranges in cultured salmonids.

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Abbreviations

IPN:

Infectious pancreatic necrosis

IPNV:

Infectious pancreatic necrosis virus

H&E:

Hematoxylin and eosin

IHC:

Immunohistochemistry

RT-PCR:

Reverse transcription-polymerase chain reaction

TBST:

Tris-buffered saline with Tween

BSA:

Bovine serum albumin

AEC:

3-Amino-9-ethylcarbazole

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Acknowledgments

This study was funded by grants from the Shiraz University, School of Veterinary Medicine. We thank Dr M. Ansarilari and Dr A. Tamadon for their assistance in statistical analysis. Mr. L. Shirvani and Mr. G. Yousefi from the Department of Pathology, Mr. M.R. Sorbi from the Division of Immunology, Mr. M.S. Freidoni from the Aquatic Animal Health Unit, School of Veterinary Medicine, Shiraz University are also acknowledged for their technical assistance.

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Authors and Affiliations

  1. Department of Pathobiology, School of Veterinary Medicine, Shiraz University, P. O. Box: 71345-1731, Shiraz, Iran

    Ahmad Oryan, Nasrollah Ahmadi & Arsalan Hosseini

  2. Aquatic Animal Health Unit, Department of Clinical Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

    Mostafa Akhlaghi

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  1. Ahmad Oryan
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  2. Nasrollah Ahmadi
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  3. Mostafa Akhlaghi
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  4. Arsalan Hosseini
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Correspondence to Nasrollah Ahmadi.

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Oryan, A., Ahmadi, N., Akhlaghi, M. et al. A comparative histopathological, immunohistochemical and nested-PCR study for diagnosis of infectious pancreatic necrosis in the naturally infected cultured rainbow trout (Oncorhynchus mykiss). Aquacult Int 20, 725–734 (2012). https://doi.org/10.1007/s10499-012-9499-3

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  • DOI: https://doi.org/10.1007/s10499-012-9499-3

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