Kentucky State University

Poster Title

Evaluation of DNA Damage in Blood Lymphocytes of Farm Workers During the Growing Season

Institution

Kentucky State University

Abstract

Chronic low level exposure to pesticides has been shown to cause many health conditions such as induction of oxidative stress, cytogenetic damage, and increased susceptibility to cancers in humans. DNA damage can be determined and quantified by Single Cell Gel Electrophoresis, also called Comet Assay. The objective of this study was to determine DNA damage in lymphocytes of farm workers who are continuously exposed to low levels of pesticides. Blood was collected once a month for six months, form June 2004 to November 2004 from farmers (n = 11) and unexposed controls (n = 8). Lymphocytes were separated on histopaque by centrifugation. An appropriate aliquot of washed lymphocytes were mixed with low melting agarose and coated on microscope slides. Cells were lysed, followed with lysis of DNA by treatment with NaOH (pH > 13.2). The fragmented DNA was electrophoresed at 40o C, at 300 milliamps, for 20 minutes, at pH > 13.2. The slides were stained with the fluorescent dye SYBR green and using LOATS software, DNA damage was determined. The initial data indicated no significant difference in tail lengths of comets from farm workers and control group.

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Evaluation of DNA Damage in Blood Lymphocytes of Farm Workers During the Growing Season

Chronic low level exposure to pesticides has been shown to cause many health conditions such as induction of oxidative stress, cytogenetic damage, and increased susceptibility to cancers in humans. DNA damage can be determined and quantified by Single Cell Gel Electrophoresis, also called Comet Assay. The objective of this study was to determine DNA damage in lymphocytes of farm workers who are continuously exposed to low levels of pesticides. Blood was collected once a month for six months, form June 2004 to November 2004 from farmers (n = 11) and unexposed controls (n = 8). Lymphocytes were separated on histopaque by centrifugation. An appropriate aliquot of washed lymphocytes were mixed with low melting agarose and coated on microscope slides. Cells were lysed, followed with lysis of DNA by treatment with NaOH (pH > 13.2). The fragmented DNA was electrophoresed at 40o C, at 300 milliamps, for 20 minutes, at pH > 13.2. The slides were stained with the fluorescent dye SYBR green and using LOATS software, DNA damage was determined. The initial data indicated no significant difference in tail lengths of comets from farm workers and control group.