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Sugarcane farmers and cutters have been reported to be at high risk of acute kidney injury. This cross-sectional study aimed to assess acute kidney injury biomarkers, and cross-shift change among 150 sugarcane cutters and 98 sugarcane farmers in Thailand. Physical health examination, environmental measurements, and measured urinary neutrophil gelatinase-associated lipocalin (NGAL) were assessed. Cross-shift change of urine creatinine (uCr), albumin creatinine ratio (ACR), and NGAL between sugarcane farmers and cutters was compared. Factors influencing abnormal post-shift ACR and NGAL were analyzed by binary logistic regression adjusted with covariates. Sugarcane cutters were significantly more likely to take nonsteroidal anti-inflammatory drugs (NSAID) and herbs to relieve pain than sugarcane farmers. Males were more likely to be current smokers and drinkers although 62% of cutters and 56% of farmers were female. Sugarcane farmers and cutters had similar post-shift estimated glomerular filtration rate (eGFR) and ACR levels. Cross-shift measurements showed that both sugarcane cutters and sugarcane farmers had significantly higher post-shift geometric mean (GM) urinary NGAL, but that the cross-shift changes of NGAL were significantly higher for sugarcane cutters (182%) vs. sugarcane farmers (112%). Water intake less than recommendation of subjects was a significant predictor of a higher risk for an abnormal post-shift ACR after controlling for covariates, while being a sugarcane cutter was a predictor of a lower risk of an abnormal risk of a post-shift NGAL after controlling for covariates. Measurements of heat stress (WBGT plus workload) found that sugarcane cutters exceeded recommended exposures while sugarcane farmers were compliance to the TLV. These findings suggest that to protect sugarcane farmers and cutters, preventative measures are needed.