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Title: ENHANCED GROSS VISUALIZATION OF CHICKEN PEYER’S PATCH: NOVEL STAINING TECHNIQUE APPLIED TO FRESH TISSUE SPECIMENS

Author
item Vaughn, Lara
item Holt, Peter
item Moore, Randle
item Gast, Richard

Submitted to: Avian Diseases
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/13/2006
Publication Date: 6/26/2006
Citation: Vaughn, L.E., Holt, P.S., Moore, R.W., Gast, R.K. 2006. Enhanced gross visualization of chicken peyer’s patch: Novel staining technique applied to fresh tissue specimens. Avaian Diseases. (50):298-302

Interpretive Summary: The lower intestinal tract contains tissues associated with the immune system. Within the portion of the intestinal tract known as the ileum, there are organized areas of lymphoid tissue, called Peyer's patches (Pp). The Peyer’s patches may serve as an important site for monitoring inflammatory and immunological responses of the host against intestinal (enteric) pathogens. Chicken Pp are often difficult to observe with the unaided eye and a simple technique to enhance visualization of the Pp is lacking. Therefore, we designed a new (novel) staining method that is quick, easy and accurate to aid in identification and recovery of the chicken Pp from fresh tissue specimens. Lower intestinal tracts were removed from white leghorn hens and commercial broilers. The region involving the ileum, ceca and colon was cut-out (excised) as one intact/whole segment. The segment was flushed with deionized-water to remove intestinal contents and then a dilute Eosin-Y stain solution was infused. After 1 minute, the Eosin-Y was gently extruded. Modified-Crystal Violet was then injected into the GI segment, where upon the lymphoid tissue area became obvious at the outer, exterior (serosal) surface of the intestine. The Pp in the lower, ending region of the ileum (distal ileum) appeared as a pale whitish-pink circular/oval area with surrounding gut tissue stained light purple. The exact Pp site could be easily observed at both the exterior (serosal) and inner (mucosal) surface of the intestine with the unaided eye. The Pp areas were cut out, then processed and prepared appropriately to allow viewing of the tissue in greater detail with a microscope. Examination under the microscope revealed large clusters of lymphocytes indicative of Pp. The new Eosin-Y + m-Crystal Violet staining technique promotes rapid identification and accurate recovery of chicken Pp lymphoid tissue from fresh tissue specimens.

Technical Abstract: The lower intestinal tract contains tissues associated with the immune system. Within the portion of the intestinal tract known as the ileum, there are organized areas of lymphoid tissue, called Peyer's patches (Pp). The Peyer’s patches may serve as an important site for monitoring inflammatory and immunological responses of the host against intestinal (enteric) pathogens. Chicken Pp are often difficult to observe with the unaided eye and a simple technique to enhance visualization of the Pp is lacking. Therefore, we designed a new (novel) staining method that is quick, easy and accurate to aid in identification and recovery of the chicken Pp from fresh tissue specimens. Lower intestinal tracts were removed from white leghorn hens and commercial broilers. The region involving the ileum, ceca and colon was cut-out (excised) as one intact/whole segment. The segment was flushed with deionized-water to remove intestinal contents and then a dilute Eosin-Y stain solution was infused. After 1 minute, the Eosin-Y was gently extruded. Modified-Crystal Violet was then injected into the GI segment, where upon the lymphoid tissue area became obvious at the outer, exterior (serosal) surface of the intestine. The Pp in the lower, ending region of the ileum (distal ileum) appeared as a pale whitish-pink circular/oval area with surrounding gut tissue stained light purple. The exact Pp site could be easily observed at both the exterior (serosal) and inner (mucosal) surface of the intestine with the unaided eye. The Pp areas were cut out, then processed and prepared appropriately to allow viewing of the tissue in greater detail with a microscope. Examination under the microscope revealed large clusters of lymphocytes indicative of Pp. The new Eosin-Y + m-Crystal Violet staining technique promotes rapid identification and accurate recovery of chicken Pp lymphoid tissue from fresh tissue specimens.