Research
A Section 508–conformant HTML version of this article is available at http://dx.doi.org/10.1289/EHP351.
Poultry Consumption and Arsenic Exposure in the U.S. Population Anne E. Nigra,1,2,3 Keeve E. Nachman,3,4,5 David C. Love,3,4 Maria Grau-Perez,1,3 and Ana Navas-Acien1,2,3 1Department
of Environmental Health Sciences, Columbia University Mailman School of Public Health, New York, New York, USA; of Epidemiology, 3Department of Environmental Health Sciences, 4Center for a Livable Future, and 5Department of Health Policy and Management, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA
2Department
Background: Arsenicals (roxarsone and nitarsone) used in poultry production likely increase inorganic arsenic (iAs), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), and roxarsone or nitarsone concentrations in poultry meat. However, the association between poultry intake and exposure to these arsenic species, as reflected in elevated urinary arsenic concentrations, is unknown. Objectives: Our aim was to evaluate the association between 24-hr dietary recall of poultry consumption and arsenic exposure in the U.S. population. We hypothesized first, that poultry intake would be associated with higher urine arsenic concentrations and second, that the association between turkey intake and increased urine arsenic concentrations would be modified by season, reflecting seasonal use of nitarsone. Methods: We evaluated 3,329 participants ≥ 6 years old from the 2003–2010 National Health and Nutrition Examination Survey (NHANES) with urine arsenic available and undetectable urine arsenobetaine levels. Geometric mean ratios (GMR) of urine total arsenic and DMA were compared across increasing levels of poultry intake. Results: After adjustment, participants in the highest quartile of poultry consumption had urine total arsenic 1.12 (95% CI: 1.04, 1.22) and DMA 1.13 (95% CI: 1.06, 1.20) times higher than nonconsumers. During the fall/winter, participants in the highest quartile of turkey intake had urine total arsenic and DMA 1.17 (95% CI: 0.99, 1.39; p-trend = 0.02) and 1.13 (95% CI: 0.99, 1.30; p-trend = 0.03) times higher, respectively, than nonconsumers. Consumption of turkey during the past 24 hr was not associated with total arsenic or DMA during the spring/summer. Conclusions: Poultry intake was associated with increased urine total arsenic and DMA in NHANES 2003–2010, reflecting arsenic exposure. Seasonally stratified analyses by poultry type provide strong suggestive evidence that the historical use of arsenic-based poultry drugs contributed to arsenic exposure in the U.S. population. Citation: Nigra AE, Nachman KE, Love DC, Grau-Perez M, Navas-Acien A. 2017. Poultry consumption and arsenic exposure in the U.S. population. Environ Health Perspect 125:370–377; http://dx.doi.org/10.1289/EHP351
Introduction In populations with low arsenic levels in drinking water, exposure to inorganic arsenic (iAs) occurs mainly through diet, particularly through the consumption of rice and other grains, as well as some juices and wine [Davis et al. 2012; deCastro et al. 2014; U.S. Food and Drug Administration (FDA) 2014a; Navas-Acien et al. 2011]. iAs is a toxic and carcinogenic metalloid that occurs naturally in water, air, and soil and enters the food supply through geological releases, contaminated water, and anthropogenic sources such as pesticide residue, nonferrous metal smelting, and waste incineration [Agency for Toxic Substances and Disease Registry (ATSDR) 2007]. Increasing evidence suggests that mono- and dimethylated metabolites of inorganic arsenic [monomethylarsonate (MMA) and dimethylarsinate (DMA), respectively] may cause oxidative stress and cytotoxicity and may be carcinogenic depending on the valence state [Hughes 2002; International Agency for Research on Cancer (IARC) 2012a]. Little is known, however, about the potential independent contribution of poultry intake to arsenic exposure in human populations. Arsenic-based drugs (roxarsone, used in chickens, and nitarsone, used in turkeys) were
370
deliberately used in U.S. poultry production for decades, potentially representing an unnecessary and easily controllable source of exposure to iAs, MMA, DMA, and roxarsone/nitarsone in the population (Liu et al. 2016; Nachman et al. 2016; Silbergeld and Nachman 2008). The FDA withdrew marketing approvals for roxarsone and for two other arsenic-based feed additives in 2013, and they withdrew approval for nitarsone, which is used to prevent histomoniasis in turkeys, in December of 2015 (FDA 2014b, 2015; Abraham et al. 2013). Historical use of nitarsone in turkey production and of roxarsone in chicken production may thus have been a chronic source of arsenic exposure for the U.S. population and may be ongoing in other parts of the world (Yao et al. 2013). In 2010, it was estimated that ~88% of broiler chickens available at market had been treated with roxarsone (Nachman et al. 2012). A similar estimate is not available for nitarsone, but turkey industry representatives have reported that nitarsone was used seasonally during hot-weather months in young turkeys that were consumed during the fall/winter (Aubrey 2013). Analyses of chicken meat have shown that the use of roxarsone during chicken production likely volume
contributes to elevated iAs, DMA, and other unknown arsenic species in chicken meat and that the concentration of iAs increases with cooking (Nachman et al. 2013). Analyses of turkey meat have also shown that the use of nitarsone during turkey production likely contributes to elevated iAs, MMA, and other unknown arsenic species in turkey meat (Nachman et al. 2016). In a recent feeding study, iAs, MMA, DMA, roxarsone, and an unidentified roxarsone metabolite were elevated in the meat of chickens fed a roxarsone-supplemented diet compared with the levels of those compounds in chickens fed a non-roxarsone diet (Liu et al. 2016). However, it is unknown if consumption of poultry exposed to arsenic-based drugs results in increased arsenic exposure and internal dose in the population, as reflected in urinary excretion. The National Health and Nutrition Examination Survey (NHANES) collects 24-hr dietary recall information the same day that a spot urine sample is collected for total and speciated arsenic analysis [U.S. National Center for Health Statistics (NCHS) 2014]. Previous studies in NHANES have evaluated poultry consumption as a potential confounder of dietary arsenic exposure (Davis Address correspondence to A.E. Nigra, Department of Environmental Health Sciences, Columbia Mailman School of Public Health, 722 W. 168th St., 11th Floor, Room 1105, New York, NY 10032 USA. E-mail: [email protected], or A. NavasAcien, Department of Environmental Health Sciences, Columbia Mailman School of Public Health, 722 W. 168th St., 11th Floor, Room 1105, New York, NY 10032 USA. Telephone: 212-342-4712. E-mail: [email protected] Supplemental Material is available online (http:// dx.doi.org/10.1289/EHP351). This research was supported by a directed research grant from the Johns Hopkins Center for a Livable Future. A.N.-A and A.E.N. are supported by National Institutes of Health/National Institute for Environmental Health Sciences grants R01ES021367 and R01ES025216. The authors declare that they have no actual or potential competing financial interests. Received: 12 April 2016; Revised: 24 August 2016; Accepted: 19 September 2016; Published: 13 October 2016. Note to readers with disabilities: EHP strives to ensure that all journal content is accessible to all readers. However, some figures and Supplemental Material published in EHP articles may not conform to 508 standards due to the complexity of the information being presented. If you need assistance accessing journal content, please contact [email protected]. Our staff will work with you to assess and meet your accessibility needs within 3 working days.
125 | number 3 | March 2017 • Environmental Health Perspectives
Poultry and arsenic exposure
et al. 2012) or in analyses without accounting for other dietary sources of arsenic such as seafood (deCastro et al. 2014). Our objective was to evaluate whether consumption of poultry in the past 24 hr was associated with increased arsenic exposure as measured in urine by total arsenic and DMA during a time period when arsenic-based poultry drugs were approved for use in the United States. First, we hypothesized that poultry intake would be associated with higher arsenic exposure as determined in urine. Second, we hypothesized that the association between turkey intake and elevated urine arsenic would be modified by season (strongest for turkey consumed during the fall/winter and null for turkey consumed during the spring/ summer), whereas the association for chicken intake would persist across seasons. To our knowledge, this is the first study to evaluate the independent association between recent poultry consumption and arsenic exposure as reflected in urine total arsenic and DMA concentrations.
Methods Study Population We analyzed data from the 2003–2010 cycles of the NHANES, conducted by the NCHS. NHANES is a multi-stage, n ationally representative sample of the noninstitutiona lized population (NCHS 2014). Our study used data from the demographic questionnaire, the 24-hr dietary recall, the clinical examination, and the laboratory examination. All NHANES protocols were approved by the NCHS institutional review board (IRB), and all participants gave written informed consent (NCHS 2011a). Our study was exempt from IRB approval because we used de-identified, publicly available data. To capture a time period when urine arsenic measures were available while roxarsone, nitarsone, and other arsenic-based drugs were still available in poultry production, we restricted our analysis to 2003–2010 NHANES cycles. Urine arsenic was measured in a one-third subsample of all participants ≥ 6 years of age. From 10,451 participants in the NHANES 2003–2010 urine arsenic subsamples, we excluded 387 missing total urine arsenic, arsenobetaine, or DMA; 229 who were pregnant; 880 who were missing BMI, cotinine, urinary creatinine, or education information; and 5,626 with detectable arsenobetaine because seafood arsenicals markedly contribute to total arsenic exposure and DMA and make it difficult to evaluate the contribution of other foods to iAs exposure (NavasAcien et al. 2011). The final sample included 3,329 participants who were ≥ 6 years old. The response rate across the entire survey period was 76.5% (NCHS 2013).
Urine Arsenic
24-Hr Poultry Intake Assessment
Spot urine samples were collected during examination, poured in 5-mL cryovial vessels, frozen at ≤ –20°C, and shipped within 1 week on dry ice to the National Center for Environmental Health (NCEH) at the Centers for Disease Control and Prevention for analysis (NCHS 2005). Total arsenic concentrations were determined via quadrupole inductively coupled plasma–mass spectrometry with dynamic reaction cell (ICP-DRC-MS). Speciated arsenic concentrations (arsenite, arsenate, MMA, DMA, and arsenobetaine) were determined via highperformance liquid chromatography (HPLC) coupled to ICP-DRC-MS (NCHS 2007, 2009a, 2011b, 2011c). We used total arsenic and DMA concentrations in urine, but not arsenite, arsenate, or MMA as the limits of detection (LOD); percents of analytic sample below the LOD for these species [arsenite (1.2 μg/L, 97.7%), arsenate (1.0 μg/L, 96.8%), and MMA (0.9 μg/L, 74.0%)] were high compared to those in other studies evaluating urinary arsenic levels, and in most samples, these species were largely undetectable (Cubadda et al. 2012; NCHS 2007, 2009a, 2011b, 2011c; Scheer et al. 2012). Neither nitarsone nor roxarsone was measured in urine. For arsenobetaine, the LOD was 0.4 μg/L, and 37% of the participants had undetectable concentrations. Detectable arsenobetaine in NHANES has shown to be an excellent biomarker of recent seafood intake (NavasAcien et al. 2011). Restricting our study population to participants with undetectable arsenobetaine likely removed the contribution of seafood arsenicals to both total arsenic and DMA, which could then be interpreted as biomarkers of arsenic exposure not derived from seafood (Liu et al. 2016; Navas-Acien et al. 2011). The LOD for total arsenic ranged from 0.60 to 0.74 μg/L across the entire survey period, with an inter-assay coefficient of variation ranging from 3.0% to 19.4% for lots with mean concentrations of 3.6 to 8.15 μg/L (NCEH 2006a, 2007, 2008, 2010). For DMA across the entire survey period, the LOD was 1.7 μg/L with an inter-assay coefficient of variation ranging from 4.6% to 6.6% for lots with mean concentrations of 4.12 to 6.85 μg/L (NCEH 2004, 2006b, 2008, 2010). The percents of participants in the analytic sample below the LOD for poultry consumers and nonconsumers were 1.2% and 4.0%, respectively, for urine total arsenic and 28.2% and 32.0%, respectively, for DMA. Values below the LOD for total arsenic and DMA were replaced by the LOD divided by the square root of 2. The LOD for arsenobetaine was
A Section 508–conformant HTML version of this article is available at http://dx.doi.org/10.1289/EHP351.
Poultry Consumption and Arsenic Exposure in the U.S. Population Anne E. Nigra,1,2,3 Keeve E. Nachman,3,4,5 David C. Love,3,4 Maria Grau-Perez,1,3 and Ana Navas-Acien1,2,3 1Department
of Environmental Health Sciences, Columbia University Mailman School of Public Health, New York, New York, USA; of Epidemiology, 3Department of Environmental Health Sciences, 4Center for a Livable Future, and 5Department of Health Policy and Management, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA
2Department
Background: Arsenicals (roxarsone and nitarsone) used in poultry production likely increase inorganic arsenic (iAs), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), and roxarsone or nitarsone concentrations in poultry meat. However, the association between poultry intake and exposure to these arsenic species, as reflected in elevated urinary arsenic concentrations, is unknown. Objectives: Our aim was to evaluate the association between 24-hr dietary recall of poultry consumption and arsenic exposure in the U.S. population. We hypothesized first, that poultry intake would be associated with higher urine arsenic concentrations and second, that the association between turkey intake and increased urine arsenic concentrations would be modified by season, reflecting seasonal use of nitarsone. Methods: We evaluated 3,329 participants ≥ 6 years old from the 2003–2010 National Health and Nutrition Examination Survey (NHANES) with urine arsenic available and undetectable urine arsenobetaine levels. Geometric mean ratios (GMR) of urine total arsenic and DMA were compared across increasing levels of poultry intake. Results: After adjustment, participants in the highest quartile of poultry consumption had urine total arsenic 1.12 (95% CI: 1.04, 1.22) and DMA 1.13 (95% CI: 1.06, 1.20) times higher than nonconsumers. During the fall/winter, participants in the highest quartile of turkey intake had urine total arsenic and DMA 1.17 (95% CI: 0.99, 1.39; p-trend = 0.02) and 1.13 (95% CI: 0.99, 1.30; p-trend = 0.03) times higher, respectively, than nonconsumers. Consumption of turkey during the past 24 hr was not associated with total arsenic or DMA during the spring/summer. Conclusions: Poultry intake was associated with increased urine total arsenic and DMA in NHANES 2003–2010, reflecting arsenic exposure. Seasonally stratified analyses by poultry type provide strong suggestive evidence that the historical use of arsenic-based poultry drugs contributed to arsenic exposure in the U.S. population. Citation: Nigra AE, Nachman KE, Love DC, Grau-Perez M, Navas-Acien A. 2017. Poultry consumption and arsenic exposure in the U.S. population. Environ Health Perspect 125:370–377; http://dx.doi.org/10.1289/EHP351
Introduction In populations with low arsenic levels in drinking water, exposure to inorganic arsenic (iAs) occurs mainly through diet, particularly through the consumption of rice and other grains, as well as some juices and wine [Davis et al. 2012; deCastro et al. 2014; U.S. Food and Drug Administration (FDA) 2014a; Navas-Acien et al. 2011]. iAs is a toxic and carcinogenic metalloid that occurs naturally in water, air, and soil and enters the food supply through geological releases, contaminated water, and anthropogenic sources such as pesticide residue, nonferrous metal smelting, and waste incineration [Agency for Toxic Substances and Disease Registry (ATSDR) 2007]. Increasing evidence suggests that mono- and dimethylated metabolites of inorganic arsenic [monomethylarsonate (MMA) and dimethylarsinate (DMA), respectively] may cause oxidative stress and cytotoxicity and may be carcinogenic depending on the valence state [Hughes 2002; International Agency for Research on Cancer (IARC) 2012a]. Little is known, however, about the potential independent contribution of poultry intake to arsenic exposure in human populations. Arsenic-based drugs (roxarsone, used in chickens, and nitarsone, used in turkeys) were
370
deliberately used in U.S. poultry production for decades, potentially representing an unnecessary and easily controllable source of exposure to iAs, MMA, DMA, and roxarsone/nitarsone in the population (Liu et al. 2016; Nachman et al. 2016; Silbergeld and Nachman 2008). The FDA withdrew marketing approvals for roxarsone and for two other arsenic-based feed additives in 2013, and they withdrew approval for nitarsone, which is used to prevent histomoniasis in turkeys, in December of 2015 (FDA 2014b, 2015; Abraham et al. 2013). Historical use of nitarsone in turkey production and of roxarsone in chicken production may thus have been a chronic source of arsenic exposure for the U.S. population and may be ongoing in other parts of the world (Yao et al. 2013). In 2010, it was estimated that ~88% of broiler chickens available at market had been treated with roxarsone (Nachman et al. 2012). A similar estimate is not available for nitarsone, but turkey industry representatives have reported that nitarsone was used seasonally during hot-weather months in young turkeys that were consumed during the fall/winter (Aubrey 2013). Analyses of chicken meat have shown that the use of roxarsone during chicken production likely volume
contributes to elevated iAs, DMA, and other unknown arsenic species in chicken meat and that the concentration of iAs increases with cooking (Nachman et al. 2013). Analyses of turkey meat have also shown that the use of nitarsone during turkey production likely contributes to elevated iAs, MMA, and other unknown arsenic species in turkey meat (Nachman et al. 2016). In a recent feeding study, iAs, MMA, DMA, roxarsone, and an unidentified roxarsone metabolite were elevated in the meat of chickens fed a roxarsone-supplemented diet compared with the levels of those compounds in chickens fed a non-roxarsone diet (Liu et al. 2016). However, it is unknown if consumption of poultry exposed to arsenic-based drugs results in increased arsenic exposure and internal dose in the population, as reflected in urinary excretion. The National Health and Nutrition Examination Survey (NHANES) collects 24-hr dietary recall information the same day that a spot urine sample is collected for total and speciated arsenic analysis [U.S. National Center for Health Statistics (NCHS) 2014]. Previous studies in NHANES have evaluated poultry consumption as a potential confounder of dietary arsenic exposure (Davis Address correspondence to A.E. Nigra, Department of Environmental Health Sciences, Columbia Mailman School of Public Health, 722 W. 168th St., 11th Floor, Room 1105, New York, NY 10032 USA. E-mail: [email protected], or A. NavasAcien, Department of Environmental Health Sciences, Columbia Mailman School of Public Health, 722 W. 168th St., 11th Floor, Room 1105, New York, NY 10032 USA. Telephone: 212-342-4712. E-mail: [email protected] Supplemental Material is available online (http:// dx.doi.org/10.1289/EHP351). This research was supported by a directed research grant from the Johns Hopkins Center for a Livable Future. A.N.-A and A.E.N. are supported by National Institutes of Health/National Institute for Environmental Health Sciences grants R01ES021367 and R01ES025216. The authors declare that they have no actual or potential competing financial interests. Received: 12 April 2016; Revised: 24 August 2016; Accepted: 19 September 2016; Published: 13 October 2016. Note to readers with disabilities: EHP strives to ensure that all journal content is accessible to all readers. However, some figures and Supplemental Material published in EHP articles may not conform to 508 standards due to the complexity of the information being presented. If you need assistance accessing journal content, please contact [email protected]. Our staff will work with you to assess and meet your accessibility needs within 3 working days.
125 | number 3 | March 2017 • Environmental Health Perspectives
Poultry and arsenic exposure
et al. 2012) or in analyses without accounting for other dietary sources of arsenic such as seafood (deCastro et al. 2014). Our objective was to evaluate whether consumption of poultry in the past 24 hr was associated with increased arsenic exposure as measured in urine by total arsenic and DMA during a time period when arsenic-based poultry drugs were approved for use in the United States. First, we hypothesized that poultry intake would be associated with higher arsenic exposure as determined in urine. Second, we hypothesized that the association between turkey intake and elevated urine arsenic would be modified by season (strongest for turkey consumed during the fall/winter and null for turkey consumed during the spring/ summer), whereas the association for chicken intake would persist across seasons. To our knowledge, this is the first study to evaluate the independent association between recent poultry consumption and arsenic exposure as reflected in urine total arsenic and DMA concentrations.
Methods Study Population We analyzed data from the 2003–2010 cycles of the NHANES, conducted by the NCHS. NHANES is a multi-stage, n ationally representative sample of the noninstitutiona lized population (NCHS 2014). Our study used data from the demographic questionnaire, the 24-hr dietary recall, the clinical examination, and the laboratory examination. All NHANES protocols were approved by the NCHS institutional review board (IRB), and all participants gave written informed consent (NCHS 2011a). Our study was exempt from IRB approval because we used de-identified, publicly available data. To capture a time period when urine arsenic measures were available while roxarsone, nitarsone, and other arsenic-based drugs were still available in poultry production, we restricted our analysis to 2003–2010 NHANES cycles. Urine arsenic was measured in a one-third subsample of all participants ≥ 6 years of age. From 10,451 participants in the NHANES 2003–2010 urine arsenic subsamples, we excluded 387 missing total urine arsenic, arsenobetaine, or DMA; 229 who were pregnant; 880 who were missing BMI, cotinine, urinary creatinine, or education information; and 5,626 with detectable arsenobetaine because seafood arsenicals markedly contribute to total arsenic exposure and DMA and make it difficult to evaluate the contribution of other foods to iAs exposure (NavasAcien et al. 2011). The final sample included 3,329 participants who were ≥ 6 years old. The response rate across the entire survey period was 76.5% (NCHS 2013).
Urine Arsenic
24-Hr Poultry Intake Assessment
Spot urine samples were collected during examination, poured in 5-mL cryovial vessels, frozen at ≤ –20°C, and shipped within 1 week on dry ice to the National Center for Environmental Health (NCEH) at the Centers for Disease Control and Prevention for analysis (NCHS 2005). Total arsenic concentrations were determined via quadrupole inductively coupled plasma–mass spectrometry with dynamic reaction cell (ICP-DRC-MS). Speciated arsenic concentrations (arsenite, arsenate, MMA, DMA, and arsenobetaine) were determined via highperformance liquid chromatography (HPLC) coupled to ICP-DRC-MS (NCHS 2007, 2009a, 2011b, 2011c). We used total arsenic and DMA concentrations in urine, but not arsenite, arsenate, or MMA as the limits of detection (LOD); percents of analytic sample below the LOD for these species [arsenite (1.2 μg/L, 97.7%), arsenate (1.0 μg/L, 96.8%), and MMA (0.9 μg/L, 74.0%)] were high compared to those in other studies evaluating urinary arsenic levels, and in most samples, these species were largely undetectable (Cubadda et al. 2012; NCHS 2007, 2009a, 2011b, 2011c; Scheer et al. 2012). Neither nitarsone nor roxarsone was measured in urine. For arsenobetaine, the LOD was 0.4 μg/L, and 37% of the participants had undetectable concentrations. Detectable arsenobetaine in NHANES has shown to be an excellent biomarker of recent seafood intake (NavasAcien et al. 2011). Restricting our study population to participants with undetectable arsenobetaine likely removed the contribution of seafood arsenicals to both total arsenic and DMA, which could then be interpreted as biomarkers of arsenic exposure not derived from seafood (Liu et al. 2016; Navas-Acien et al. 2011). The LOD for total arsenic ranged from 0.60 to 0.74 μg/L across the entire survey period, with an inter-assay coefficient of variation ranging from 3.0% to 19.4% for lots with mean concentrations of 3.6 to 8.15 μg/L (NCEH 2006a, 2007, 2008, 2010). For DMA across the entire survey period, the LOD was 1.7 μg/L with an inter-assay coefficient of variation ranging from 4.6% to 6.6% for lots with mean concentrations of 4.12 to 6.85 μg/L (NCEH 2004, 2006b, 2008, 2010). The percents of participants in the analytic sample below the LOD for poultry consumers and nonconsumers were 1.2% and 4.0%, respectively, for urine total arsenic and 28.2% and 32.0%, respectively, for DMA. Values below the LOD for total arsenic and DMA were replaced by the LOD divided by the square root of 2. The LOD for arsenobetaine was
