Abstract
To determine the effect of the inclusion method on the histomorphometric evaluation of the gastrointestinal mucosa of horses, jejunum samples were collected using flank laparotomy. Sixteen mixed breed healthy adult horses, including four males and 12 females, aged 4-14 years with an average body weight of 248.40 ± 2.28 kg, were used. Jejunal biopsies were collected and analyzed by light microscopy using two methods: group 1 comprised biopsies fixed using 10% neutral formalin and embedded in paraffin; biopsies in group 2 were fixed in 2.5% glutaraldehyde in 0.1 M phosphate buffer pH 7.2, followed by inclusion in glycol methacrylate (GMA)-based plastic resin. Intestinal villi height, crypt depth, glandular mucosa thickness, total mucosal thickness, and villus/crypt ratio were then evaluated. For all the variables studied, with exception of the villus/crypt ratio, a significant difference (p < 0.05) was found between samples in groups 1 and 2. Processing samples for embedding in plastic resin was quicker and easier to perform compared to that for paraffin embedding. In addition, the epithelial lining of tissues in group 2 showed better resolution for conducting cytological studies under a light microscope. The difference between the studied variables can be attributed to tissue retraction caused by conventional processing for inclusion in paraffin. Therefore, the method of inclusion in GMA described in the present study appears to be a more reliable choice for morphometric evaluation of the intestinal mucosa of horses.
Highlights
The use of histological processing to assess the intestinal mucosa has been described in horses, poultry, and in humans (Lima et al, 2020; Milne et al, 2010; Rocchigiani et al, 2021)
Jejunal biopsies were collected and analyzed by light microscopy using two methods: group 1 comprised biopsies fixed using 10% neutral formalin and embedded in paraffin; biopsies in group 2 were fixed in 2.5% glutaraldehyde in 0.1 M phosphate buffer pH 7.2, followed by inclusion in glycol methacrylate (GMA)-based plastic resin
The difference between the studied variables can be attributed to tissue retraction caused by conventional processing for inclusion in paraffin
Summary
The use of histological processing to assess the intestinal mucosa has been described in horses, poultry, and in humans (Lima et al, 2020; Milne et al, 2010; Rocchigiani et al, 2021). This technique is used to evaluate the responses of the intestinal mucosa to nutritional support, congenital disorders (Ruemmele et al, 2006), treatment of gastrointestinal disorders, and morphometric assessments. The intestinal mucosa comprises of the epithelial lining, the lamina propria, and the mucosal muscle, which generally consist of a thin circular inner layer and a longitudinal outer layer of smooth muscle (Bezerra et al, 2016; Junqueira & Carneiro, 1999)
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