Date of Award

2011

Document Type

Thesis

Abstract

Natural Resistance Associate Macrophages Protein one (NRAMP1) gene encodes a transmembrane protein that regulates divalent cation transport that hinders intracellular pathogen proliferation. The objective of this study was to determine how NRAMP1 expression and other factors may be associated with coccidia infections in goats. Clinically healthy Boer, Spanish, and Spanish-Boer cross goats (N=140) from the NCA&T Small ruminant unit were used. The FAMACHA© score, body condition score, body weight, and age were recorded. Fecal samples were evaluated for the levels of haemonchus eggs and coccidia oocytes using McMaster slides. Blood samples were evaluated for packed cell volume and white blood cell differential counts. Genomic DNA was extracted from blood on FTA cards according to the manufacturer's protocol (N=40). RNA was extracted using the ZR Whole-Blood Total RNA Kit (N=12). The RNA samples were reverse-transcribed and the cDNA was obtained using the Ambion-Retroscript kit. Specific primers for NRAMP1, IL-8, IL-10, Interferon gamma, and CD14 were used for RT PCR, to detect NRAMP1 in genomic DNA and to determine CD14 and cytokine gene expression from isolated RNA. Samples amplified using primers for GAPDH were used as loading controls. xiv The effect of breed, age, and infection status on selected health parameters, gene identification and expression were analyzed using SAS ANOVA. Coccidia fecal egg count was significantly related to age (p<0.05). Expression of NRAMP1, IL-10 and IL-8 was significantly increased with increased fecal coccidia egg counts (p<0.05). haemonchus fecal egg count was significantly related to FAMACHA score, PCV, and to increased percentages of neutrophils, eosinophils, and monocytes in blood. Expression of CD14 and IL-8 was significantly increased in goats with high haemonchus fecal egg counts (p<0.0114). A 443bp region of the NRAMP1 gene was identified in genomic DNA from 50% of goats tested. This variation may be associated with gene polymorphisms. Distinct gene expression patterns may be associated with the innate immune response to coccidia and nematodes in goats. Age appropriate management strategies will help control coccidiosis in goats.

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