Groundwater  used for human consumption causes histological and molecular changes in the thyroid that affect the morphogenesis process of xenopus laevis

Groundwater used for human consumption causes histological and molecular changes in the thyroid that affect the morphogenesis process of xenopus laevis | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Groundwater used for human consumption causes histological and molecular changes in the thyroid that affect the morphogenesis process of xenopus laevis Maria Fernanda Modarelli, Rodrigo Miguel Bilbao, Osvaldo Juan Ponzo This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3716033/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Groundwater is a reservoir of fresh water. In the goitrous areas the water used for drinking and cooking generally is high in nitrate content. The aim of the present study was to evaluate in that areas, chronic effects to expose of xenopus laevis larvae during a complete metamorphosis process to groundwater. For analyze the effects on thyroid function. The xenopus laevis larvae were divided into 3 work groups submerged, until the end of metamorphosis, in: drinking water as control (C), groundwater as group (G) and drinking water plus potassium perchlorate as positive control (PC). In (G) group metamorphosis process was completed in 37% and 0% in (PC) (p < 0.0001). In the 60 of Niewkoop and Faber, stage the wet weight and body length was in (G) smaller than (C) and (PC) (p < 0.0001). But was not difference in 62 for (G), (PC) was bigger than (C). In 60 stage was no significant differences on the length of the hind limbs. In 62 was bigger in (G) and (PC) than in (C) (p < 0.001). Thyroid hyperplasia was grade 2 in (G) and 3 (PC) (p < 0.001) in 60 stage and 3 in (G) and (PC) in the 62 (p < 0.0001). Follicular hypertrophy and colloid depletion in (G) and (PC) (p < 0.0001). In groundwater was found high nitrates concentrations. These findings suggest that nitrate contaminate groundwater impairs thyroid function of xenopus laevis larvae. These observation could contribute to the current discussion about the acceptable daily intake of nitrate. nitrates groundwater morphogenesis thyroid NIS carrier xenopus laevis Figures Figure 1 Figure 2 Introduction Due to the increasing use of nitrate-containing fertilizers, the nitrate content of drinking water in areas with intensive agriculture has increased surpassing the concentration of 50 mg/l NO3. It is higher than the level permitted in drinking water. The discovery of nitrate and nitrites in drinking water is an indicator of contamination. Nitrate ingestion not only causes methemoglobinemia, but also other negative and undesirable effects such as cancer formation, differences in vitamin A metabolism, and disorders of growth and development (Buchholz and Shi; 2018, Weyer et al.; 2001 ) Nitrate (NO3 –) and nitrites are found naturally in soil, water, plants and foods. At the moment these natural resources’ nitrate and nitrites contents are increasing due to the use of excessive fertilisers and industrial development (Jordão et al; 2002 ). Although nitrate is not as powerful a goitrogenic substance as thiocynate and perchlorate, which mainly inhibits the accumulation of iodide in the thyroid gland (Guiles et al; 2000), it may cause malfunction of the thyroid. Its potential detrimental effects on the thyroid axis implicate numerous physiological processes, including growth regulation, general metabolism, and metamorphosis in metamorphic animals (Babalola and Van Wyk; 2021). If the high nitrate and and others EDs production will not be stopped, they will continue to accumulate in the environment, drinking water, and food, thus becoming global environmental pollutants, potentially threating the human health and wildlife (Stubleski et al; 2021, Jian et al; 2018 ). During the metamorphosis of xenopus laevis, morphological changes are accompanied by thyroid histological changes (Denver et al; 2002, Brown and Cai; 2007) regulated by thyrotropin (TSH) levels (Carr et al; 2003, Thambirajah et al; 2019, Yue et al.; 2023 ); acting on expression of NIS (Dai et al; 1996 , Goleman et al; 2002, Carr et al; 2008 ) through the regulation of synthesis of transcription factors such as PAX8, TTF-1 and TTF-2 (Saito et al; 1997 , Ohno et al; 1999). Amphibian metamorphosis represents an adequate biological model to evaluate the action of thyroid disruptors (Grim et al, 2009, Mughal et al; 2018 ) and their consequences in the morphogenesis process in amphibians. Allowing to evaluate the risk that exposure to these substances to humans during the embryonic and fetal periods (Tata et al;1998, Yao et al; 2017 , Li et al; 2019). The novelty of this work consists of the use of groundwater samples from the city of Buenos Aires (Argentina), consumed by a population with a high prevalence of hypothyroidism and goiter, in a study of chronic in vivo exposure in Xenopus laevis larvae. during the metamorphosis process. The objective was to analyze the histological and biomolecular changes of the thyroid gland and their impact on the morphogenesis process of the larvae, caused by groundwater. Materials and methods Animals and experimental model In vivo chronic exposure model in xenopus laevis larvae that complied with the ARRIVE guidelines for laboratory animal experiments in accordance with the Animals (Scientific Procedures) Act of the National Research Council Guide for the Care and use of laboratory animals eighth edition. Approved by the Institutional Committee for the Care and Use of Laboratory Animals, of the Faculty of Medicine of the UBA, ( [email protected] . EXP-UBA 0003598/2013). Xenopus laevis larvae from the Endocrinology Laboratory of the Department of Physiology of the Faculty of Medicine of the University of Buenos Aires, from a single nest, were used. They were maintained, with a light-dark cycle: 12h-12h (Timer Master Clear), 22 ± 2ºC (Top House 3000 Fg portable air conditioner 3000 W Cold/Heat Tac-12chpa/k with timer) and water pH: 7 .2 to 8 (Metrhom 691 digital pH meter). They were fed ad libitum, with balanced food (Sera micron) and the rations were increased along with the growth of the tadpoles according to Table 1 of the feeding regime for Xenopus laevis tadpoles of the AMA guidelines. Rearing density did not exceed approximately four tadpoles/L culture water (static exposure system) or 10 tadpoles/L culture water. Under these conditions, tadpoles developed from stages 45NF/46NF to stage 51N, according to Nieuwkoop-Faber criteria (Nieuwkoop et al; 1956) in twelve days. We transferred the randomly selected newly hatched larvae to plastic containers filled with 500 ml of water per larva, so that each container had 13 larvae in group (C), divided into 2 containers with 5 larvae and 1 with 3. In Group (G) and (PC) had 18, divided into 3 containers of 6 larvae each. We maintained water levels at 800 ml during larval rearing to avoid evaporation effects and changed them every 48 hours to prevent deterioration (OECD; 2018a ). These experiment repeated for four times in the same condition, for reach number of animals necessary to consider the experiment valid. The water used to carry out the experiment was groundwater from wells 30 to 60 m deep in the southern suburb of the city of Buenos Aires (Argentina) considered suitable for human consumption. Were made microbiological, physicochemical were carried out at the Argentine Technological and Industrial Institute (INTI- www.inti.gob.ar ). For development of the experiment, the larvae were divided into 3 groups that define the following treatments: Control group (C): larvae submerged in drinking water, eliminating chlorine through a filter. Groundwater group (G): larvae submerged in groundwater and larvae submerged in drinking water with the addition of 0.007 mg/L potassium perchlorate (KCLO4) (thyroid disrupting inhibitor NIS) as Positive Control (PC) (Bertrand et al; 2004). All groups were observed during a period of complete biological metamorphosis (premetamorphosis, prometamorphosis and climax). Considering the metamorphosis process complete, when larvae reached 66NF stage. To evaluate morphological parameters were sampled on days 6, 14, 21 and 28. Tadpoles moved individually from the accumulation tank using a small net or strainer and transferred to a transparent measuring chamber (100 mm Petri dish) containing the water corresponding to each group. The animals were carefully handled during this transfer to minimize handling stress and avoid injuries. And I don't anesthetize them. The developmental stages of animals are determined using a binocular dissecting microscope. In each group, animals at stage 58NF, 60NF and 62NF were sacrificed, to analyze histological and biomolecular changes. The larvae were sacrificed by immersion in a solution of MS222 (200 mg/L) (Strykowski, JL.; 2015). The euthanized tadpoles are rinsed in water by body weight determination. Hindlimb length, snout to vent length, and developmental stage (using a binocular dissection microscope) are determined for each tadpole. After the sacrifice was extracted, part of the lower jaw tissue where the thyroid is anatomically located was decapitated and dissected for subsequent histological and biomolecular analysis. Tissues destined for histological processing were preserved in Bouin's solution and tissues for biomolecular determinations were frozen at -70ºC in lysis buffer (0.0625 M Tris, 3% SDS and 2% 2-mercaptoethanol plus cocktails of inhibitors). Protease Thermo Scientific: 78430). Morphological analysis Were analyzed: total time of metamorphosis, developmental stage according to the Nieuwkoop and Faber criteria, hind limb length, body length (snout-vent length), wet weight and mortality [21 OECD; 2009, AMA; 2009]. Histological analysis In thyroid glands of larvae, the histological changes evaluated were: degree of follicular hypertrophy, hyperplasia of the gland and colloidal according to Guide on Amphibian Thyroid Histology [22 OECD; 2007b, US, EPA; 2008b]. After sacrifice, tissues were fixed in Bouin's solution, then subjected to a progressive dehydration process in increasing concentrations of alcohol and finally in xylene, and subsequently submerged in paraffin. For later stained with hematoxylin-eosin and were analyzed by optical microscopy. To determine the degrees of follicular hyperplasia, a severity classification is incorporated with a range of four that include: 0, 1, 2 and 3 [23 OECD; 2009, AMA; 2009]. In this severity classification, the degrees of severity have been based on the percentage of affected cells and/or the percentage of affected tissue, where: Grade 0 (not significant): This grade is used for processes in which less than 20% of the tissue is affected. Grade 1 (level): This grade is used for processes in which between 30 and 50% of the tissue is affected. Grade 2 (moderate): This grade is used for processes in which between 60 and 80% of the tissue is involved. Grade 3 (severe) This grade is used for processes in which more than 80% of the tissue is involved. Western blot After thyroid extraction, tissue sample was homogenized and were cold processed with lysis buffer (1.51 g Tris, 6 g SDS, 2-beta mercaptoethanol 5 to 20 ml, pH 6.8) and protease inhibitor cocktail (ABCAM: ab271306-3m). Protein concentrations were measured used Bradford method. The supernatant was heated to 70°C and then centrifuged at 1600 rpm. Proteins were then separated by SDS/PAGE (BioRad Mini Protean 3 Cell tank kit) and transferred to a PVDF membrane (Amersham, UK). Once proteins were transferred, the membrane was washed with 1X TBS (20 mM TrisHCl, 150mM NaCl pH 7.8) and blocking was performed with a 0.2% solution of TBS-Tween-20 (TBS-T) and skimmed milk at 5% v/v for 1 hour at room temperature under shaking. The membrane was incubated with a rabbit anti-NIS-COOH-terminal polyclonal primary antibody (Millipore Corp. USA, CAT #ABC1453) at dilutions of 1:500 overnight at 4°C. After successive washes with TBS-T, membrane was blocked again for 30 to 40 minutes, three more washes with TBS-T were performed. It was incubated for 1h at room temperature under shaking, with peroxidase-conjugated mouse anti-rabbit polyclonal secondary antibody (Abcam ab205718ab205718), at a dilution of 1:1500. Finally, it was incubated for 5 minutes with chemiluminescence detection reagent (ECL) (Biorad, cat #170–5060 USA) and exposed to an X-ray plate (Kodak and GE). After the radiographic development technique, protein bands were quantified using Scion Image beta version 4.0.2 image analysis software. Statistical Analyzes GraphPad Instat (2019) software was used, with a significance of p < 0.05. To determine morphological differences between groups, an analytical comparison was made using ANOVA of multiple intergroup comparisons. Fisher post-test for 60NF stage and Kruskal-Wallis test with Tukey-Kramer post-test for 62NF stage was performed. For histological differences between groups, an ANOVA was performed with Fisher post-test for stage 60NF and Kruskal-Wallis test with Tukey-Kramer post-test for stage 62NF. Finally, for expression level of NIS transporter, an ANOVA was performed with Dunnett's posttest for stage 60 NF, with Kruskal-Wallis and Dunn test for stage 62 NF. Results Morphology larvae analysis Number of larvae that were able to complete the metamorphosis process was significantly different between the groups (C, G and PC). 100% of the larvae submerged in drinking water (C), and 37.7% of those exposed to groundwater (G) completed their metamorphosis, while none of the larvae in the positive control group (PC) completed it (p < 0.0001). A delay in growth of the larvae was observed, from 60NF to 62NF stage. The appearance of the extremities (hind and forelegs) was delayed in the larvae of (G) and (PC) group, with respect to those of group (C) (p < 0.0001). The prometamorphosis time in (C) was 21 ± 6.19 days, in (G) 32 ± 2.25 days and 35 ± 1.33 days in (PC). A delay in prometamorphosis of 10 ± 2.68 days was observed in (G) and 14 ± 1.96 in (PC) days, compared to (C) group, being the difference highly significant (p < 0.001). These changes occurred especially in transition from 54NF to 60NF stage of prometamorphosis (p < 0.001), and there were no differences in transition from 60NF to 66NF stage in group (G). In climax duration were no significant differences between groups. However, in (PC) group, larvae remained in 62NF stage, without completing metamorphosis process. Wet Weight and snout to tail length of larvae In 60NF stage, it was observed that larvae of group (G), which managed to complete prometamorphosis, reached climax stage with a significantly lower Wet weight when compared to larvae of control group (p < 0.0001). However, larvae of group (PC) had a greater wet weight than those corresponding to group (G), but it was less than the weight of larvae (C) (p < 0.05). Snout to tail length was different between groups, being lower in (G) than in (C) and (PC) (p < 0.001) ( Table 1 ). In 62NF stage, larvae of group (G) reduced their wet weight vs (C) and (PC) (p < 0.001), but in this last group her body weight increased vs (C) and (G) (p < 0.0001). Snout to tail length was greater in (PC) than (C) and (G) (p < 0.0001) at 62NF stage (picture 1). In (G) was not significantly differences than in (C) ( Table 1 ). In 66NF stage, body weight was different, being lower in (G) vs (C) (p < 0.05). Snout to tail length at 66NF stage did not differ between groups. In (PC) group, the larvae did not reach 66NF, being arrested at 62NF stage, giving rise to giant tadpoles ( Table 1 ). Hind limb length During 60NF, differences considered extremely significant were observed in hind limb length between group (C) and (PC), with (C)<(PC) and (G)<(PC) (p < 0.0001). No very significant differences were observed during this stage between (C) and (G) ( Table 1 ). In stage 62NF, differences considered extremely significant were observed between group (C), (G) and (PC). At this stage the length of the hind legs was shorter in (G) when compared to (C) and (PC) (p < 0.001) and was greater in (PC) when compared to (C) and (PC) (p < 0.0001) ( Table 1 ). Table 1. Averages of measurements of wet weight, snout to tail length, hind limb leght of the larvae in stages 60NF, 62NF and 66NF. A significant difference can be observed between the groups at 60NF stage in wet weight was (G)<(PC)<(C), in 62NF (G)<(C)<(PC) and in 66NF (G)<(C). Snout to tail length in 60NF stage was (G)<(PC)=(C), in 62NF (G)<(C)<(PC) and in 66NF was not differ in weight between (C) and (G), in (PC) none reached 66NF stage. In 60NF stage hind limb lenght was not significant differences, in 62NF stage hind limb lenght was (G)<(C)<(PC). In 66NF was not differ in hind limb lenght between (C) and (G). **p<0.001, ***p<0.0001. Additional Observations Morphological alterations in adult frogs Morphological alterations observed in adult frogs, 2 years after completing metamorphosis, affected 85.7% of adult females (p < 0.0001), being objectiveable at eye and mouth level in 28.6%, column 14.3%, lower limbs 42.8%. The remaining 14.3% of the group of female frogs, although they did not present objectifiable macroscopic malformations, presented obesity. Male frogs were not affected by these macroscopic malformations. Histology changes Thyroid glandular hyperplasia Grade of follicular hyperplasia in stage 60NF was grade 0 in (C), 2 in (G) and grade 3 in (PC) (p < 0.001). In stage 62NF it was grade 0 in group (C) and 3 in groups (G) and (PC) (p (C)>(G) (p (C)>(G) (p < 0.001) (Fig. 1 ). Colloid area In 60NF stage there were no significant changes when analyzing this parameter. In (PC) calculate the colloidal area was not possible, because shape of the follicles does not allow obtaining a radius value. For this reason, they are not included in analysis of this variable. However, decrease was easily observable in 10X photographs, with colloid depletion being visible in most of the follicles (picture 1). Observing significant changes in glandular size in (PC), being greater in increasing order in (PC)>(G)>(C) (p (G)>(PC) (p < 0.02). In this stage, the glandular size is significantly greater in (PC) than in the other groups (p < 0.001). But the differences between (G) and (C) were not significant (Fig. 1 ). Histologic examination IN 60NF stage histologic examination in (G) group showed retention of lobular architecture and follicular hyperplasia. We observed that more than 76,2% of the follicles were empty. The follicular cells were columnar with pale cytoplasm and enlarged nuclei. Colloid was virtually absent. When colloid was present it was hardly ever in every follicular lumen. A scalloped pattern is observed in the colloid with a high degree of vacuolization (Fig. 1 ). In group (PC) the same changes were observed, but follicular hyperplasia was more intense than (G) and follicular hypertrophy was more severe than (G). For the same reason more than 91,3% of the follicles were empty. These changes were significantly different from those of (C) and (G) group. NIS expression When analyzing 60NF stage, higher level of expression of NIS were observed in (G) and (PC) groups vs (C) (p < 0.05) (Fig. 2 ). In 62NF stage, level expression of NIS carrier decreased in (G) and (PC) vs. (C), this difference being significant (p < 0.001). It is important to point out that, in some larvae of group (G), the NIS protein stopped expressing itself (Fig. 2 ). Biochemical determinations in groundwater Our working group took groundwater samples from the study area (southern suburb of the city of Buenos Aires - Argentina) from home wells at depths between 30 and 60 meters. Physicochemical, bacteriological and endocrine disruptor analyzes were performed. Due to its physicochemical characteristics, water under study was hard sulfated water with excess chlorides in 30-m wells, and semi-hard water with a high concentration of chlorides in 60-m wells. Both waters have high conductivity and a variable concentration of nitrates, which by themselves indicate that it is contaminated water. Nitrates were detected in concentrations between 45 mg/l to 83 mg/l. Other disruptors cannot be detected with detection limits used (Table 2 ). Table 2 Biochemical determinations in groundwater. Characteristic Drinking water Groundwater 30 m Groundwater 60 m Colour Colorless Colorless Greenish Smell without without without Sediment Very scarce Abundant Scarce pH 7.6 7.3 7.5 Residual active chlorine 0.00 ppm 0.00 ppm 0.00 ppm Conductivity 395 µg/cm 982 µg/cm 729 µg/cm Total dissolved solids 284 mg/L 707 mg/L 525 mg/L Total alcalinity (CO3Ca) 175 mg/L 409 mg/L 312 mg/L Total hardness (CO3Ca) 112 mg/L 336 mg/L 114 mg/L Chlorides (CL-) 29 mg/L 87 mg/L 36 mg/L Sulfates (SO4=) 20 mg/L 25 mg/L 20 mg/L Amonia (NH4+) No detectable No detectable No detectable Nitrites (NO2-) No detectable 0.01 mg/L 0.01 mg/L Nitrates(NO3-) < 5 mg/L 83 mg/L 24 mg/L Chrome (Cr + 6) No detectable No detectable No detectable Discussion Most of population of study area (southern suburb of the city of Buenos Aires - Argentina) obtains water for domestic use and irrigation from groundwater (Pampeano and Puelche aquifers), the most important source of fresh water in the region. Our working group detected nitrates in concentrations higher than those safe for human health in the water from these aquifers (Langer et al; 2006, Modarelli et al; 2022 ). A retrospective observational study carried out by our working group on 879 patients in this area, showed a prevalence for subclinical hypothyroidism of 57.8% and for goiter of 38.9%, in patients who consume water from underground wells; affecting women in their fertile stage to a greater extent (Modarelli et al; 2019) which could be due to chronic exposure to nitrates detected in groundwater. This situation has also been observed in other places on the planet by various authors, in relation to exposure to nitrates as well as other EDs (Colborn et al; 1993 , Gatseva et al; 2008, Huang et al; 2017b ). In people exposed to nitrates or perchlorates, iodine uptake by the NIS cotransporter is impaired (Tietge et al; 2005 , Zambrano-Fernández et al; 2022) which interferes with thyroid hormone synthesis and causes hypothyroxinemia (Braverman et al; 2005, Garcia-Torres et al; 2022, Serrano-Nacimento et al; 2022) As a consequence, in individuals, the release of pituitary TSH increases, this hormone being the main regulator of thyroid functions (Tonacchera et al.; 2004 , Dohan et al.; 2007). It has been known for years that prometamorphosis and climax in amphibians are regulated by T4/T3 levels, and that these in turn regulate TSH levels in larvae (Goos et al.; 1968 ). Authors such as Hemme (Hemme et al.;1972) and later Ospitz et al. (Opitz et al.; 2006), have described that the exposure of tadpoles compounds such as thiocyanates and perchlorates increases the expression of TSH mRNA, as well as the β receptor of this hormone. This elevation in TSH levels, stimulate the transcription of the NIS protein, which reaches its maximum expression level in xenopus larvae, between the 58NF to 60NF metamorphosis stages. In present study, xenopus laevis larvae chronically exposed to environmental groundwater samples containing nitrates in variable concentrations between 45mg/L − 83mg/L and was observed an increase in expression levels of the NIS cotransporter during 60NF stage at the larvae. This increase in protein expression could represent an adaptive mechanism for increased level of thyroid iodine absorption to and maintain the production of T4/T3 to reach metamorphic climax (De Groef et al; 2006). This could explain why duration of the climax to remain unchanged. Similar features of temporal, rapid, and persistent induction of NIS mRNA by TSH treatment have been reported in different rat thyroid follicle cell lines (Trapasso et al.; 1999 , Fozzatti et al.; 2007 ). However, at the 62NF stage a reduction in the expression level of the NIS cotransporter was observed, this would cause an asynchronous morphological development of the tadpoles and a reduction in the number of those that managed to complete metamorphosis. Consequently, those who reached the final stage of 66NF climax were smaller than those in the control group. The reduction in thyroid hormone levels, caused by the blockade of the NIS transporter exerted by nitrates, causes an increase in TSH levels. Studies in mammals have shown that its effects are mediated by the modulation exerted by thyroid hormones on specific gene expression during the process of embryonic development (Yamazzaki et al.; 2003, Montesinos et al.; 2016 ). Monitoring effected for two years, of larvae after completing metamorphosis, showed that only the females developed malformations that were involved eyes, mouth, lower limbs, and spine, which could be related to deficiency in T4 and T3 production that occurred during prometamorphosis, in larvae during their development. This reduction in thyroid hormone levels during metamorphosis could be generating changes in function of various proteins, through changes in DNA sequences that code for these proteins, affecting genes such as Xnkx-2.1., a homeobox gene expressed during early forebrain development, lung, and thyroid in xenopus laevis (Holleman et al.; 2000). Other studies indicated that the exposure to high levels of nitrate during pregnancy is a risk factor for spontaneous abortion, fetal death, prematurity, intrauterine growth restriction, low birth weight, congenital malformations, and neonatal death (Ward et al; 2018 ). Eskiocak, S et al (Eskiocak et al.; 2005 ) observed that in rats exposed to a concentration of 50 mg/L of nitrates, T4 levels decreased significantly in this group, but an increase was recorded with exposure to 100 mg/L. and in this group it was accompanied by an elevation of TSH. Our group observed changes of hyperplasia and hypertrophy in larval thyroid with an increase between 60NF and 62NF stages, when larvae were exposed to groundwater contaminated with nitrates at levels between 45 mg/L − 83 mg/L, above levels considered safe for human consumption (WHO; 2016 ). In the experiment by Eskiocak, S et al, the glands of rats would suffered a reduction in levels of iodine uptake, due to the competitive blockade exerted by nitrates, producing a reduction in T4 levels. In the experiment developed by our group an increase in NIS transporter expression levels were observed in 60NF stage. As larvae have a high rate of metabolic demand during this stage of the metamorphosis, due to the critical changes that occur in their morphology (lung development, intestinal changes and tail resorption), they could develop, when exposed to a high concentration of nitrates, an increase in TSH levels. Giving rise to various events: hyperplasia, hypertrophy of the gland and stimulation of the synthesis of NIS transporter and therefore the absorption of iodine to maintain T4 production levels. If the inhibition of iodine uptake in the gland persists, the synthesis chain of the NIS transporter would end up being compromised at the intracellular level by inhibiting transcription factors such as TTF1 and/or PAX-8. This would cause a reduction in the synthesis of T4, the persistence of the stimulation exerted by TSH due to a decrease of negative feedback, which would explain the significant hyperplasia suffered in the follicular cells during 62NF stage, and the reduction in the expression levels of the NIS transporter during the 62NF stage. Conclusions Nitrates present in groundwater, in high concentrations, affect the functioning of the thyroid gland. Chronic exposure to these concentrations causes decreased and/or absent expression of NIS in exposed larvae. This generates histological alterations as a consequence of reduction in thyroxine levels, which translate into affectation of tadpole morphological development. This study shows the usefulness of the model to assess the safety of water considered suitable for human consumption. These observation could contribute to the current discussion about the acceptable daily intake of nitrate. Declarations Ethical Approval: Approved by the Institutional Committee for the Care and Use of Laboratory Animals, of the Faculty of Medicine of the UBA, ( [email protected] . EXP-UBA 0003598/2013). Acknowledgments: Dr. J. J. Lopez, Histology Department, School of Medicine of the University of Buenos Aires. Consent to Participate: the corresponding author confirms that the work described has not been published before (except in abstract form or as part of a published conference, review or thesis); that it is not being considered for publication elsewhere; that its publication has been approved by all co-authors; that its publication has been approved by the responsible authorities of the institution where the work is carried out. Consent to Publish: The author accepts responsibility for publishing this material on behalf of any and all co-authors. Funding: This work was supported by Endocrinology Laboratory, Institute of Physiology, Academic Unit II, School of Medicine, University of Buenos Aires. Buenos Aires. Argentina and with contribution of own funds. Competing interests : The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Author Contributions: All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Modarelli Maria Fernanda, Bilbao Rodrigo Miguel and Ponzo Osvaldo Juan. The first draft of the manuscript was written by Modarelli Maria Fernanda and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript. References Babalola OO, Van Wyk JH (Oct 2021) Exposure impacts of Imazapyr formulation on larval development and thyroid histology of Xenopus laevis. Environ Sci Pollut Res Int 28(37):50967–50974. 10.1007/s11356-021-14227-4 Epub 2021 May 11 Bertrand S, Brunet. FG, Escriva H, Parmentier G, Laudet V, Robinson-Rechavi M (Oct 2004) Evolutionary genomics of nuclear receptors: from twenty-five ancestral genes to derived endocrine systems. Mol Biol Evol 21(10):1923–1937. 10.1093/molbev/msh200 Epub 2004 Jun 30. PMID: 15229292 Braverman LE et al (Feb 2005) The effect of perchlorate, thiocyanate, and nitrate on thyroid function in workers exposed to perchlorate long-term. J Clin Endocrinol Metab 90(2):700–706. 10.1210/jc.2004-1821 Epub 2004 Nov 30. PMID: 15572417 Brown DD, Cai L (June 2007) Amphibian metamorphosis Review 306(1):20–33. https://doi.org/10.1016/j.ydbio.2007.03.021 Buchholz DR, Shi YB (Sep 2018) Dual function model revised by thyroid hormone receptor alpha knockout frogs. Gen Comp Endocrinol 265:214–218. 10.1016/j.ygcen.2018.04.020 Epub 2018 Apr 22. PMID: 29689262; PMCID: PMC6087486 Carr DL, Carr JA, Willis RE, Pressley TA (2008) Mar A perchlorate sensitive iodide transporter in frogs. Gen Comp Endocrinol. 1;156(1):9–14. 10.1016/j.ygcen.2008.01.005 . Epub 2008 Jan 12. PMID: 18275962; PMCID: PMC2279098 Carr DL, Laharrague F, Kahn B, Pressley TA, Carr JA (Apr 2003) Molecular characterization of a putative sodium/iodide symporter in the South African clawed frog, Xenopus laevis. Ann N Y Acad Sci 986:711–712. 10.1111/j.1749-6632 2003.tb07287. x. PMID: 12763923 Colborn T et al (1993) Developmental Effects of Endocrine Disrupting Chemicals in Wildlife and Humans. Rev Environ Health Perspect 101(5):378–384. 10.1289/ehp.93101378 Dai G, Levy O, Carrasco N (1996) Feb Cloning and characterization of the thyroid iodide transporter. Nature. 1;379(6564):458 – 60. 10.1038/379458a0 . PMID: 8559252 Denver RJ, Glennemeier KA, and., Boorse GC (Feb 2002) Endocrinology of complex life cycles: Amphibians. In: Pfaff DW, Arnold AP, Etgen AM, Fahrback SE, Rubin RT (eds) Hormones, brain and behavior. › biologia › public_html, pp 469–513. https://exa.unne.edu.ar De Groef B, Decallonne B, Van der Geyten S, Darras VM, Bouillon R (Jul 2006) Perchlorate versus other environmental sodium/iodide symporter inhibitors: potential thyroid-related health effects. Rev Eur J Endocrinol 155(1):17–25. 10.1530/eje.1.02190 Dohán O, Portulano C, Basquin C, Reyna-Neyra A, Amzel LM, Carrasco N (2007) The Na+/I- symporter (NIS) mediates electroneutral active transport of the environmental pollutant perchlorate. Proc Natl Acad Sci U S A 104(51):20250–20255. 10.1073/pnas.0707207104 Eskiocak S, Dundasr C, Basoglu T, (Jul (2005) The effects of taking chronic nitrate by drinking water on thyroid functions and morphology. Clin Exp Med 5(2):66–71. 10.1007/s10238-005-0068-1 Fozzatti L, Vélez ML, Lucero AM, Nicola JP, Mascanfroni ID, Macció DR et al (2007) Endogenous thyrocyte-produced nitric oxide inhibits iodide uptake and thyroid-specific gene expression in FRTL-5 thyroid cells. J Endocrinol 192(3):627–637. 10.1677/joe.1.06967 García Torres E et al (May 2022) Consumption of water contaminated by nitrate and its deleterious effects on the human thyroid gland: a review and update. Int J Environ Health Res 32(5):984–1001 Epub 2020 Aug 31. PMID: 32866080 Gatseva PD, Argirova MD (2008) High-nitrate levels in drinking water may be a risk factor for thyroid dysfunction in children and pregnant women living in rural Bulgarian areas. Int Hyg Environ Health 211(5):555–559. 10.1016/j.ijheh.2007.10.002 Epub 2007 Dec 27 Goleman WL, Carr JA, Anderson TA (Mar 2002) Environmentally relevant concentrations of ammonium perchlorate inhibit thyroid function and alter sex ratios in developing Xenopus laevis. Environ Toxicol Chem 21(3):590–597 PMID: 11878472 Goos HJ, Zwanenbeek HC, van Oordt PG (1968) Hypothalamic neurosecretion and metamorphosis in Xenopus laevis. II. The effect of thyroxine following treatment with propylthiouracil. Arch Anat Histol Embryol 51(1):267–274 PMID: 4905268 Guiles HJ (2000) Thyroid function. In: Bishop ML, DubenEngelkirk JL, Fody EP (eds) Clinical chemistry principles, procedures, correlations. Lippincott Williams and Wilkins, Philadelphia, pp 403–421 Grim C, Wolfe M, Braunbeck T et al (Jun 2009) Thyroid Histopathology Assessments for the Amphibian Metamorphosis Assay to Detect Thyroid-active Substances. Toxicol Pathol 37(4):415–424. 10.1177/0192623309335063 Epub 2009 Apr 22 Hemme L (1972) Die Differenzierungsgenese der TSH Zellen von Xenopus laevis unter Normalbedingungen und nach Thiouracilbehandlung. Z Zellforsch 125:353–377. https://doi.org/10.1007/BF00306632 Huang HB, Pan WH, Chang JW et al (2017b) ¿Does exposure to phthalates influence thyroid function and growth hormone homeostasis? The Taiwan Environmental Survey for Toxicants (TEST) 2013. Environ Res 153:63–72. https://doi.org/10.1016/j.envres.2016.11.014 Hollemann T, Pieler T, (Nov (2000) Xnkx-2.1: a homeobox gene expressed during early forebrain, lung and thyroid development in Xenopus laevis. Dev Genes Evol. 210(11):579 – 81. 10.1007/s004270000098 . PMID: 11180810 Jian JM, Chen D, Han FJ, Guo Y, Zeng L, Lu X, Wang F (2018) Sept A short review on human exposure to and tissue distribution of per- and polyfluoroalkyl substances (PFASs). Sci Total Environ. 636:1058–1069. doi: 10.1016/j.scitotenv.2018.04.380. Epub 2018 May 3. PMID: 29913568 Jordão CP, Pereira MG, Bellato CR, Pereira JL, Matos AT (2002) Oct Assessment of water systems for contaminants from domestic and industrial sewages. Environ Monit Assess. 2002;79(1):75–100. 10.1023/a:1020085813555 . PMID: 12381024 Langer P (2006) Increased thyroid volume and frequency of thyroid disorders signs in school children from nitrate polluted area. Chemosphere 62(4):559–564. 10.1016/j.chemosphere.2005.06.030 Epub 2005 Aug 10 Li S, Qiao K, Jiang Y, Wu Q, Coffin S, Gui W, Zhu G (Dec 2019) Disruptive effects of two organotin pesticides on the thyroid signaling pathway in Xenopus laevis during metamorphosis. Sci Total Environ 20:697:134140 Epub 2019 Aug 27. PMID: 31476497 Modarelli MF, Bilbao RM, Ponzo OJ, Water (2022) Contamination by Nitrates and its Thyroid Disruptive Action. Bioassay on Xenopus Laevis. GJMR B 22(2,10):34–87. https://medicalresearchjournal.org › Modarelli MF, Pozo OJ (2019) Relationship of subclinical hypothyroidism and goiter with the origin of water consumed by a population of the Buenos Aires suburbs. Med (Bs As) 79(1):11–19 PMID: 30694184 del Montesinos M M., et al (2016) Nitric oxide-repressed forkhead factor FoxE1 expression is involved in the inhibition of TSH-induced thyroid peroxidase levels. Mol Cell Endocrinol 420:105–115. 10.1016/j.mce.2015.11.020 Mughal BB, Demeneix BA, Fini JB (2018) Evaluating Thyroid Disrupting Chemicals in Vivo Using Xenopus laevis. Methods Mol Biol. 1801:183–192. 10.1007/978-1-4939-7902-8_15 . PMID: 29892825 Nieuwkoop PD, Faber J (1956) Normal table of Xenopus laevis (Daudin). A Systematical and Chronological Survey of the Development from the Fertilized Egg till the end of Metamorphosis. North Holland: Amsterdam 22(00):89129–89125. http://dx.doi.org/10.1016/S0168- OECD (2018a) Organisation for Economic Co-operation and Development. Revisedguidance document 150 on standardised test guidelines for evaluatingchemicals for endocrine disruption. OECD Series on Testing and Assessment. Paris France. 10.1787/9789264304741-en [accessed 15 September 2018] Ohno M, Zannini M, Levy O, Carrasco N, Di Lauro R (Mar 1999) The paired-domain transcription factor Pax8 binds to the upstream enhancer of the rat sodium/iodide symporter gene and participates in both thyroid-specific and cyclic-AMP-dependent transcription. Mol Cell Biol 19(3):2051–2060 PMID: 10022892; PMCID: PMC83998 Opitz R, Trubiroha A, Lorenz C et al (Jul 2006) Expression of sodium-iodide symporter mRNA in the thyroid gland of Xenopus laevis tadpoles: developmental expression, effects of antithyroidal compounds, and regulation by TSH Journal of Endocrinology. J Endocrinol 190(1):157–170. 10.1677/joe.1.06606 Saito T, Endo T, Nakazato M, Kogai T, Onaya T (1997) Thyroid-stimulating hormone-induced down-regulation of thyroid transcription factor 1 in rat thyroid FRTL-5 cells. Endocrinology. Feb;138(2):602-6. 10.1210/endo.138.2.4918 . PMID: 9002992 Serrano-Nascimento C, Nunes MT (2022) Oct 21 Perchlorate, nitrate, and thiocyanate: Environmental relevant NIS-inhibitors pollutants and their impact on thyroid function and human health. Front Endocrinol (Lausanne). 13:995503. 10.3389/fendo.2022.995503 . PMID: 36339434; PMCID: PMC9633673 Stubleski J, Salihovic S, Lind L, Lind PM, van Bavel B, Kärrman A (Oct 2016) Changes in serum levels of perfluoroalkyl substances during a 10-year follow-up period in a large population-based cohort. Environ Int 95:86–92 Epub 2016 Aug 16. PMID: 27542758 Tata JR, (Dec (1998) Amphibian metamorphosis as a model for studying the developmental actions of thyroid hormone. Cell Res 8(4):259–272. 10.1038/cr.1998.26 Thambirajah AA, Koide EM, Imbery JJ, Helbing CC (May 2019) Contaminant and Environmental Influences on Thyroid Hormone Action in Amphibian Metamorphosis Sec. Thyroid Endocrinol 14:1–29. https://doi.org/10.3389/fendo.2019.00276 Tietge JE et al (2005) Apr. Metamorphic inhibition of Xenopus laevis by sodium perchlorate: effects on development and thyroid histology. Environ Toxicol Chem. 24(4):926 – 33. 10.1897/04-105r.1 . PMID: 15839568 Tonacchera M et al (2004) Dec. Relative potencies and additivity of perchlorate, thiocyanate, nitrate, and iodide on the inhibition of radioactive iodide uptake by the human sodium iodide symporter. Thyroid. 14:1012–1019. 10.1089/thy.2004.14.1012 . PMID: 15650353 Trapasso F et al (1999) May Iodide symporter gene expression in normal and transformed rat thyroid cells. Eur J Endocrinol. 140(5):447 – 51. 10.1530/eje.0.1400447 . PMID: 10229912 Ward MH, Jones RR, Brender JD, de Kok TM, Weyer PJ, Nolan BT et al (2018) Drinking water nitrate and human health: An updated review. Int J Environ Res Public Health 15(7):1557. 10.3390/ijerph15071557 Weyer PJ et al (2001) Municipal drinking water nitrate level and cancer risk in older women: the Iowa Women's Health Study. Epidemiology. May;12(3):327 – 38. 10.1097/00001648-200105000-00013 . PMID: 11338313 WHO (2016) Nitrate and nitrite in drinking water: background document for development of WHO guidelines for drinking water quality. World Health Organization, Geneva Yamazaki K et al (2003) Feb. Genes regulated by thyrotropin and iodide in cultured human thyroid follicles: analysis by cDNA microarray. Thyroid. 13(2):149 – 58. 10.1089/105072503321319459 . PMID: 12699589 Yao X et al (2017) Optimization of the T3-induced Xenopus metamorphosis assay for detecting thyroid hormone signaling disruption of chemicals. J Environ Sci 52:314–324. 10.1016/j.jes.2016.09.020 Epub 2016 Nov 23 Yue B, Ning S, Miao H, Fang C, Li J, Zhang L et al (2023) Human exposure to a mixture of endocrine disruptors and serum levels of thyroid hormones: A cross-sectional study. J Environ Sci (China) 125:641–649. 10.1016/j.jes.2022.01.017 Zambrano-Fernández S, Zamora-Camacho FJ, Aragón P (Jan 2022) Direct and indirect effects of chronic exposure to ammonium on anuran larvae survivorship, morphology, and swimming speed. Chemosphere 287(Pt 3132349. 10.1016/j.chemosphere.2021.132349 Epub 2021 Sep 24. PMID: 34826957 Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3716033","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":271245219,"identity":"44fb939d-6297-4631-9722-89d449f3309d","order_by":0,"name":"Maria Fernanda Modarelli","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA4UlEQVRIiWNgGAWjYDAC5gMMDDxAmo+dsfEBkObhI6iFLQGihY2ZsdkApIWNBC0MbBJgAUI6dNu4Ex+8qbknx8bM3Fb5NcdOho2B+eGjG3i0mB3j3Ww451ixMdBhbbdltyUDHcZmbJyDT8v93m3SPGwJiW0gLZLbmIFaeNik8Wo5xgvU8i+hHqSlWHJbPZFaeNsSEkAOY/y47TBRWjYbzu1LMATa0izNuO04DxszIb8c49344M23BHl+9vaHH39uq7bnZ29++BifFhTAzAMmiVUOAow/SFE9CkbBKBgFIwYAAEaRPzG9JZxKAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-7313-0127","institution":"Universidad de Buenos Aires Facultad de Medicina","correspondingAuthor":true,"prefix":"","firstName":"Maria","middleName":"Fernanda","lastName":"Modarelli","suffix":""},{"id":271245220,"identity":"f24892e7-f16a-4e94-b069-b990e19d776d","order_by":1,"name":"Rodrigo Miguel Bilbao","email":"","orcid":"","institution":"University of Buenos Aires Faculty of Medicine: Universidad de Buenos Aires Facultad de Medicina","correspondingAuthor":false,"prefix":"","firstName":"Rodrigo","middleName":"Miguel","lastName":"Bilbao","suffix":""},{"id":271245221,"identity":"3805c69a-d15d-4ac9-848f-d8faed65eb9d","order_by":2,"name":"Osvaldo Juan Ponzo","email":"","orcid":"","institution":"University of Buenos Aires Faculty of Medicine: Universidad de Buenos Aires Facultad de Medicina","correspondingAuthor":false,"prefix":"","firstName":"Osvaldo","middleName":"Juan","lastName":"Ponzo","suffix":""}],"badges":[],"createdAt":"2023-12-06 16:29:04","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3716033/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3716033/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":50816796,"identity":"87aac8f2-3e8a-400f-ace3-72fa8631a3cb","added_by":"auto","created_at":"2024-02-07 19:50:24","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":912555,"visible":true,"origin":"","legend":"\u003cp\u003eLight microscopy with 10X and 40X magnification of Xenopus laevis thyroid glands at 60NF and 62NF stage. In 60NF A (10X) and A1 (40X): Control (C), B (10X) and B1 (40X): groundwater (G). C (10X) and C1 (40X): positive control (PC). In 62NF D (10X) and D1 (40X): Control (C), E (10X) and E1 (40X): groundwater (G). F (10X) and F1 (40X): positive control (PC). Thyroid glands are larger in group (PC) at 60NF stage and 62NF. At 62NF stage in (G), glands are smaller than in the other two. Glands of (PC) group in both stages underwent a marked reduction of colloidal area, (G) shows a greater number of filled follicles at 60NF and 62NF stages. PC: presents a markedly lower number of filled follicles than (C) and (G).\u003c/p\u003e","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3716033/v1/61b3ee1f612e1cadd8444b33.jpeg"},{"id":50816794,"identity":"32a1ab13-64ee-4ba9-b92a-7a0e59e4911c","added_by":"auto","created_at":"2024-02-07 19:50:21","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":93952,"visible":true,"origin":"","legend":"\u003cp\u003eLeft: expression level of the NIS. An increase is observed during the 60NF stage in groundwater (G) and positive control (PC) vs control (C). Followed by a reduction during the 62NF stage in both groups, being more pronounced in groundwater. Right: Western blot shows an increase in the thickness and optical density of the bands in (G1) and (PC1) at the 60NF stage and a reduction at 62NF, including the total disappearance of the band in (G2) and (PC2). Beta-actin was used as loading control. *p\u0026lt;0.05; **p\u0026lt;0.001 vs Control.\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-3716033/v1/e10a8b65c9c80f2cf4c76efb.png"},{"id":57261245,"identity":"1f7d6fc3-13bd-4e39-92a0-31d9181505db","added_by":"auto","created_at":"2024-05-28 09:44:48","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1594736,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3716033/v1/ff91b2f9-78ad-40eb-ae6b-78cee9d7d009.pdf"}],"financialInterests":"","formattedTitle":"Groundwater used for human consumption causes histological and molecular changes in the thyroid that affect the morphogenesis process of xenopus laevis","fulltext":[{"header":"Introduction","content":"\u003cp\u003eDue to the increasing use of nitrate-containing fertilizers, the nitrate content of drinking water in areas with intensive agriculture has increased surpassing the concentration of 50 mg/l NO3. It is higher than the level permitted in drinking water. The discovery of nitrate and nitrites in drinking water is an indicator of contamination. Nitrate ingestion not only causes methemoglobinemia, but also other negative and undesirable effects such as cancer formation, differences in vitamin A metabolism, and disorders of growth and development (Buchholz and Shi; 2018, Weyer et al.; \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2001\u003c/span\u003e) Nitrate (NO3 \u0026ndash;) and nitrites are found naturally in soil, water, plants and foods. At the moment these natural resources\u0026rsquo; nitrate and nitrites contents are increasing due to the use of excessive fertilisers and industrial development (Jord\u0026atilde;o et al; \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). Although nitrate is not as powerful a goitrogenic substance as thiocynate and perchlorate, which mainly inhibits the accumulation of iodide in the thyroid gland (Guiles et al; 2000), it may cause malfunction of the thyroid. Its potential detrimental effects on the thyroid axis implicate numerous physiological processes, including growth regulation, general metabolism, and metamorphosis in metamorphic animals (Babalola and Van Wyk; 2021).\u003c/p\u003e \u003cp\u003eIf the high nitrate and and others EDs production will not be stopped, they will continue to accumulate in the environment, drinking water, and food, thus becoming global environmental pollutants, potentially threating the human health and wildlife (Stubleski et al; 2021, Jian et al; \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eDuring the metamorphosis of xenopus laevis, morphological changes are accompanied by thyroid histological changes (Denver et al; 2002, Brown and Cai; 2007) regulated by thyrotropin (TSH) levels (Carr et al; 2003, Thambirajah et al; 2019, Yue et al.; \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2023\u003c/span\u003e); acting on expression of NIS (Dai et al; \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e1996\u003c/span\u003e, Goleman et al; 2002, Carr et al; \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2008\u003c/span\u003e) through the regulation of synthesis of transcription factors such as PAX8, TTF-1 and TTF-2 (Saito et al; \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e1997\u003c/span\u003e, Ohno et al; 1999).\u003c/p\u003e \u003cp\u003eAmphibian metamorphosis represents an adequate biological model to evaluate the action of thyroid disruptors (Grim et al, 2009, Mughal et al; \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) and their consequences in the morphogenesis process in amphibians. Allowing to evaluate the risk that exposure to these substances to humans during the embryonic and fetal periods (Tata et al;1998, Yao et al; \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2017\u003c/span\u003e, Li et al; 2019).\u003c/p\u003e \u003cp\u003eThe novelty of this work consists of the use of groundwater samples from the city of Buenos Aires (Argentina), consumed by a population with a high prevalence of hypothyroidism and goiter, in a study of chronic in vivo exposure in Xenopus laevis larvae. during the metamorphosis process. The objective was to analyze the histological and biomolecular changes of the thyroid gland and their impact on the morphogenesis process of the larvae, caused by groundwater.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eAnimals and experimental model\u003c/h2\u003e \u003cp\u003e In vivo chronic exposure model in xenopus laevis larvae that complied with the ARRIVE guidelines for laboratory animal experiments in accordance with the Animals (Scientific Procedures) Act of the National Research Council Guide for the Care and use of laboratory animals eighth edition. Approved by the Institutional Committee for the Care and Use of Laboratory Animals, of the Faculty of Medicine of the UBA, ([email protected]. EXP-UBA 0003598/2013).\u003c/p\u003e \u003cp\u003eXenopus laevis larvae from the Endocrinology Laboratory of the Department of Physiology of the Faculty of Medicine of the University of Buenos Aires, from a single nest, were used. They were maintained, with a light-dark cycle: 12h-12h (Timer Master Clear), 22\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026ordm;C (Top House 3000 Fg portable air conditioner 3000 W Cold/Heat Tac-12chpa/k with timer) and water pH: 7 .2 to 8 (Metrhom 691 digital pH meter). They were fed ad libitum, with balanced food (Sera micron) and the rations were increased along with the growth of the tadpoles according to Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e of the feeding regime for Xenopus laevis tadpoles of the AMA guidelines. Rearing density did not exceed approximately four tadpoles/L culture water (static exposure system) or 10 tadpoles/L culture water. Under these conditions, tadpoles developed from stages 45NF/46NF to stage 51N, according to Nieuwkoop-Faber criteria (Nieuwkoop et al; 1956) in twelve days. We transferred the randomly selected newly hatched larvae to plastic containers filled with 500 ml of water per larva, so that each container had 13 larvae in group (C), divided into 2 containers with 5 larvae and 1 with 3. In Group (G) and (PC) had 18, divided into 3 containers of 6 larvae each. We maintained water levels at 800 ml during larval rearing to avoid evaporation effects and changed them every 48 hours to prevent deterioration (OECD; \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2018a\u003c/span\u003e). These experiment repeated for four times in the same condition, for reach number of animals necessary to consider the experiment valid.\u003c/p\u003e \u003cp\u003eThe water used to carry out the experiment was groundwater from wells 30 to 60 m deep in the southern suburb of the city of Buenos Aires (Argentina) considered suitable for human consumption. Were made microbiological, physicochemical were carried out at the Argentine Technological and Industrial Institute (INTI-\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e\u003ca href=\"http://www.inti.gob.ar\" target=\"_blank\"\u003ewww.inti.gob.ar\u003c/a\u003e\u003c/span\u003e\u003cspan address=\"http://www.inti.gob.ar\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eFor development of the experiment, the larvae were divided into 3 groups that define the following treatments: Control group (C): larvae submerged in drinking water, eliminating chlorine through a filter. Groundwater group (G): larvae submerged in groundwater and larvae submerged in drinking water with the addition of 0.007 mg/L potassium perchlorate (KCLO4) (thyroid disrupting inhibitor NIS) as Positive Control (PC) (Bertrand et al; 2004). All groups were observed during a period of complete biological metamorphosis (premetamorphosis, prometamorphosis and climax). Considering the metamorphosis process complete, when larvae reached 66NF stage. To evaluate morphological parameters were sampled on days 6, 14, 21 and 28. Tadpoles moved individually from the accumulation tank using a small net or strainer and transferred to a transparent measuring chamber (100 mm Petri dish) containing the water corresponding to each group. The animals were carefully handled during this transfer to minimize handling stress and avoid injuries. And I don't anesthetize them.\u003c/p\u003e \u003cp\u003eThe developmental stages of animals are determined using a binocular dissecting microscope. In each group, animals at stage 58NF, 60NF and 62NF were sacrificed, to analyze histological and biomolecular changes.\u003c/p\u003e \u003cp\u003eThe larvae were sacrificed by immersion in a solution of MS222 (200 mg/L) (Strykowski, JL.; 2015). The euthanized tadpoles are rinsed in water by body weight determination. Hindlimb length, snout to vent length, and developmental stage (using a binocular dissection microscope) are determined for each tadpole. After the sacrifice was extracted, part of the lower jaw tissue where the thyroid is anatomically located was decapitated and dissected for subsequent histological and biomolecular analysis. Tissues destined for histological processing were preserved in Bouin's solution and tissues for biomolecular determinations were frozen at -70\u0026ordm;C in lysis buffer (0.0625 M Tris, 3% SDS and 2% 2-mercaptoethanol plus cocktails of inhibitors). Protease Thermo Scientific: 78430).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eMorphological analysis\u003c/h2\u003e \u003cp\u003eWere analyzed: total time of metamorphosis, developmental stage according to the Nieuwkoop and Faber criteria, hind limb length, body length (snout-vent length), wet weight and mortality [21 OECD; 2009, AMA; 2009].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eHistological analysis\u003c/h2\u003e \u003cp\u003eIn thyroid glands of larvae, the histological changes evaluated were: degree of follicular hypertrophy, hyperplasia of the gland and colloidal according to Guide on Amphibian Thyroid Histology [22 OECD; 2007b, US, EPA; 2008b].\u003c/p\u003e \u003cp\u003eAfter sacrifice, tissues were fixed in Bouin's solution, then subjected to a progressive dehydration process in increasing concentrations of alcohol and finally in xylene, and subsequently submerged in paraffin. For later stained with hematoxylin-eosin and were analyzed by optical microscopy. To determine the degrees of follicular hyperplasia, a severity classification is incorporated with a range of four that include: 0, 1, 2 and 3 [23 OECD; 2009, AMA; 2009]. In this severity classification, the degrees of severity have been based on the percentage of affected cells and/or the percentage of affected tissue, where:\u003c/p\u003e \u003cp\u003eGrade 0 (not significant): This grade is used for processes in which less than 20% of the tissue is affected.\u003c/p\u003e \u003cp\u003eGrade 1 (level): This grade is used for processes in which between 30 and 50% of the tissue is affected.\u003c/p\u003e \u003cp\u003eGrade 2 (moderate): This grade is used for processes in which between 60 and 80% of the tissue is involved.\u003c/p\u003e \u003cp\u003eGrade 3 (severe) This grade is used for processes in which more than 80% of the tissue is involved.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eWestern blot\u003c/h2\u003e \u003cp\u003eAfter thyroid extraction, tissue sample was homogenized and were cold processed with lysis buffer (1.51 g Tris, 6 g SDS, 2-beta mercaptoethanol 5 to 20 ml, pH 6.8) and protease inhibitor cocktail (ABCAM: ab271306-3m). Protein concentrations were measured used Bradford method. The supernatant was heated to 70\u0026deg;C and then centrifuged at 1600 rpm. Proteins were then separated by SDS/PAGE (BioRad Mini Protean 3 Cell tank kit) and transferred to a PVDF membrane (Amersham, UK). Once proteins were transferred, the membrane was washed with 1X TBS (20 mM TrisHCl, 150mM NaCl pH 7.8) and blocking was performed with a 0.2% solution of TBS-Tween-20 (TBS-T) and skimmed milk at 5% v/v for 1 hour at room temperature under shaking. The membrane was incubated with a rabbit anti-NIS-COOH-terminal polyclonal primary antibody (Millipore Corp. USA, CAT #ABC1453) at dilutions of 1:500 overnight at 4\u0026deg;C. After successive washes with TBS-T, membrane was blocked again for 30 to 40 minutes, three more washes with TBS-T were performed. It was incubated for 1h at room temperature under shaking, with peroxidase-conjugated mouse anti-rabbit polyclonal secondary antibody (Abcam ab205718ab205718), at a dilution of 1:1500. Finally, it was incubated for 5 minutes with chemiluminescence detection reagent (ECL) (Biorad, cat #170\u0026ndash;5060 USA) and exposed to an X-ray plate (Kodak and GE). After the radiographic development technique, protein bands were quantified using Scion Image beta version 4.0.2 image analysis software.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analyzes\u003c/h2\u003e \u003cp\u003eGraphPad Instat (2019) software was used, with a significance of p\u0026thinsp;\u0026lt;\u0026thinsp;0.05. To determine morphological differences between groups, an analytical comparison was made using ANOVA of multiple intergroup comparisons. Fisher post-test for 60NF stage and Kruskal-Wallis test with Tukey-Kramer post-test for 62NF stage was performed. For histological differences between groups, an ANOVA was performed with Fisher post-test for stage 60NF and Kruskal-Wallis test with Tukey-Kramer post-test for stage 62NF. Finally, for expression level of NIS transporter, an ANOVA was performed with Dunnett's posttest for stage 60 NF, with Kruskal-Wallis and Dunn test for stage 62 NF.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n \u003ch2\u003eMorphology larvae analysis\u003c/h2\u003e\n \u003cp\u003eNumber of larvae that were able to complete the metamorphosis process was significantly different between the groups (C, G and PC). 100% of the larvae submerged in drinking water (C), and 37.7% of those exposed to groundwater (G) completed their metamorphosis, while none of the larvae in the positive control group (PC) completed it (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001).\u003c/p\u003e\n \u003cp\u003eA delay in growth of the larvae was observed, from 60NF to 62NF stage. The appearance of the extremities (hind and forelegs) was delayed in the larvae of (G) and (PC) group, with respect to those of group (C) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001). The prometamorphosis time in (C) was 21\u0026thinsp;\u0026plusmn;\u0026thinsp;6.19 days, in (G) 32\u0026thinsp;\u0026plusmn;\u0026thinsp;2.25 days and 35\u0026thinsp;\u0026plusmn;\u0026thinsp;1.33 days in (PC).\u003c/p\u003e\n \u003cp\u003eA delay in prometamorphosis of 10\u0026thinsp;\u0026plusmn;\u0026thinsp;2.68 days was observed in (G) and 14\u0026thinsp;\u0026plusmn;\u0026thinsp;1.96 in (PC) days, compared to (C) group, being the difference highly significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). These changes occurred especially in transition from 54NF to 60NF stage of prometamorphosis (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), and there were no differences in transition from 60NF to 66NF stage in group (G). In climax duration were no significant differences between groups. However, in (PC) group, larvae remained in 62NF stage, without completing metamorphosis process.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\n \u003ch2\u003eWet Weight and snout to tail length of larvae\u003c/h2\u003e\n \u003cp\u003e\u003cem\u003eIn 60NF stage, it was observed that larvae of group (G), which managed to complete prometamorphosis, reached climax stage with a significantly lower Wet weight when compared to larvae of control group (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001). However, larvae of group (PC) had a greater wet weight than those corresponding to group (G), but it was less than the weight of larvae (C) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Snout to tail length was different between groups, being lower in (G) than in (C) and (PC) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (\u003c/em\u003eTable \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cem\u003e).\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eIn 62NF stage, larvae of group (G) reduced their wet weight vs (C) and (PC) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), but in this last group her body weight increased vs (C) and (G) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001). Snout to tail length was greater in (PC) than (C) and (G) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) at 62NF stage (picture 1). In (G) was not significantly differences than in (C) (\u003c/em\u003eTable \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cem\u003e).\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eIn 66NF stage, body weight was different, being lower in (G) vs (C) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Snout to tail length at 66NF stage did not differ between groups. In (PC) group, the larvae did not reach 66NF, being arrested at 62NF stage, giving rise to giant tadpoles (\u003c/em\u003eTable \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cem\u003e).\u003c/em\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003ch2\u003eHind limb length\u003c/h2\u003e\n \u003cp\u003e\u003cem\u003eDuring 60NF, differences considered extremely significant were observed in hind limb length between group (C) and (PC), with (C)\u0026lt;(PC) and (G)\u0026lt;(PC) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001). No very significant differences were observed during this stage between (C) and (G) (\u003c/em\u003eTable \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cem\u003e).\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eIn stage 62NF, differences considered extremely significant were observed between group (C), (G) and (PC). At this stage the length of the hind legs was shorter in (G) when compared to (C) and (PC) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and was greater in (PC) when compared to (C) and (PC) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) (\u003c/em\u003eTable \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cem\u003e).\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eTable 1.\u003c/em\u003e\u003c/strong\u003e\u003cem\u003e\u0026nbsp;Averages of measurements of wet weight, snout to tail length, hind limb leght of the larvae in stages 60NF, 62NF and 66NF. A significant difference can be observed between the groups at 60NF stage in wet weight was (G)\u0026lt;(PC)\u0026lt;(C), in 62NF (G)\u0026lt;(C)\u0026lt;(PC) and in 66NF (G)\u0026lt;(C). Snout to tail length in 60NF stage was (G)\u0026lt;(PC)=(C), in 62NF (G)\u0026lt;(C)\u0026lt;(PC) and in 66NF was not differ in weight between (C) and (G), in (PC) none reached 66NF stage. In 60NF stage hind limb lenght was not significant differences, in 62NF stage hind limb lenght was (G)\u0026lt;(C)\u0026lt;(PC). In 66NF was not differ in hind limb lenght between (C) and (G). **p\u0026lt;0.001, ***p\u0026lt;0.0001.\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003e\u003cem\u003e\u003cimg 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\" alt=\"image\"\u003e\u003c/em\u003e\u003c/em\u003e\u003cbr\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003eAdditional Observations\u003c/h2\u003e\n \u003cdiv id=\"Sec13\" class=\"Section3\"\u003e\n \u003ch2\u003eMorphological alterations in adult frogs\u003c/h2\u003e\n \u003cp\u003eMorphological alterations observed in adult frogs, 2 years after completing metamorphosis, affected 85.7% of adult females (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001), being objectiveable at eye and mouth level in 28.6%, column 14.3%, lower limbs 42.8%. The remaining 14.3% of the group of female frogs, although they did not present objectifiable macroscopic malformations, presented obesity. Male frogs were not affected by these macroscopic malformations.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003ch2\u003eHistology changes\u003c/h2\u003e\n \u003cdiv id=\"Sec15\" class=\"Section3\"\u003e\n \u003ch2\u003eThyroid glandular hyperplasia\u003c/h2\u003e\n \u003cp\u003eGrade of follicular hyperplasia in stage 60NF was grade 0 in (C), 2 in (G) and grade 3 in (PC) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e\n \u003cp\u003eIn stage 62NF it was grade 0 in group (C) and 3 in groups (G) and (PC) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n \u003ch2\u003eHypertrophy of follicular epithelium\u003c/h2\u003e\n \u003cp\u003eIn 60NF stage, hypertrophy of follicular epithelium was greater in decreasing order in (PC)\u0026gt;(C)\u0026gt;(G) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e\n \u003cp\u003eIn 62NF stage, hypertrophy of follicular epithelium was significantly different between groups: (PC)\u0026gt;(C)\u0026gt;(G) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n \u003ch2\u003eColloid area\u003c/h2\u003e\n \u003cp\u003eIn 60NF stage there were no significant changes when analyzing this parameter. In (PC) calculate the colloidal area was not possible, because shape of the follicles does not allow obtaining a radius value. For this reason, they are not included in analysis of this variable. However, decrease was easily observable in 10X photographs, with colloid depletion being visible in most of the follicles (picture 1). Observing significant changes in glandular size in (PC), being greater in increasing order in (PC)\u0026gt;(G)\u0026gt;(C) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). Differences that were observed between (G) and (C) are not significant (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eIn stage 62NF colloidal area was in decreasing order (C)\u0026gt;(G)\u0026gt;(PC) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.02). In this stage, the glandular size is significantly greater in (PC) than in the other groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). But the differences between (G) and (C) were not significant (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\n \u003ch2\u003eHistologic examination\u003c/h2\u003e\n \u003cp\u003eIN 60NF stage histologic examination in (G) group showed retention of lobular architecture and follicular hyperplasia. We observed that more than 76,2% of the follicles were empty. The follicular cells were columnar with pale cytoplasm and enlarged nuclei. Colloid was virtually absent. When colloid was present it was hardly ever in every follicular lumen. A scalloped pattern is observed in the colloid with a high degree of vacuolization (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eIn group (PC) the same changes were observed, but follicular hyperplasia was more intense than (G) and follicular hypertrophy was more severe than (G). For the same reason more than 91,3% of the follicles were empty. These changes were significantly different from those of (C) and (G) group.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e\n \u003ch2\u003eNIS expression\u003c/h2\u003e\n \u003cp\u003eWhen analyzing 60NF stage, higher level of expression of NIS were observed in (G) and (PC) groups vs (C) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eIn 62NF stage, level expression of NIS carrier decreased in (G) and (PC) vs. (C), this difference being significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). It is important to point out that, in some larvae of group (G), the NIS protein stopped expressing itself (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eBiochemical determinations in groundwater\u003c/p\u003e\n \u003cp\u003eOur working group took groundwater samples from the study area (southern suburb of the city of Buenos Aires - Argentina) from home wells at depths between 30 and 60 meters. Physicochemical, bacteriological and endocrine disruptor analyzes were performed. Due to its physicochemical characteristics, water under study was hard sulfated water with excess chlorides in 30-m wells, and semi-hard water with a high concentration of chlorides in 60-m wells. Both waters have high conductivity and a variable concentration of nitrates, which by themselves indicate that it is contaminated water. Nitrates were detected in concentrations between 45 mg/l to 83 mg/l. Other disruptors cannot be detected with detection limits used (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eBiochemical determinations in groundwater.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"4\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCharacteristic\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDrinking water\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eGroundwater\u003c/p\u003e\n \u003cp\u003e30 m\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eGroundwater\u003c/p\u003e\n \u003cp\u003e60 m\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eColour\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eColorless\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eColorless\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGreenish\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eSmell\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ewithout\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ewithout\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ewithout\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eSediment\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eVery scarce\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAbundant\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eScarce\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003epH\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eResidual active chlorine\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00 ppm\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00 ppm\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.00 ppm\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eConductivity\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e395 \u0026micro;g/cm\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e982 \u0026micro;g/cm\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e729 \u0026micro;g/cm\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal dissolved solids\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e284 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e707 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e525 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal alcalinity (CO3Ca)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e175 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e409 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e312 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal hardness (CO3Ca)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e112 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e336 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e114 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eChlorides (CL-)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e29 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e87 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e36 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eSulfates (SO4=)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eAmonia (NH4+)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo detectable\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo detectable\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo detectable\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNitrites (NO2-)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo detectable\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.01 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.01 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eNitrates(NO3-)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;5 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e83 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e24 mg/L\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eChrome (Cr\u0026thinsp;+\u0026thinsp;6)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo detectable\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo detectable\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo detectable\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eMost of population of study area (southern suburb of the city of Buenos Aires - Argentina) obtains water for domestic use and irrigation from groundwater (Pampeano and Puelche aquifers), the most important source of fresh water in the region. Our working group detected nitrates in concentrations higher than those safe for human health in the water from these aquifers (Langer et al; 2006, Modarelli et al; \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). A retrospective observational study carried out by our working group on 879 patients in this area, showed a prevalence for subclinical hypothyroidism of 57.8% and for goiter of 38.9%, in patients who consume water from underground wells; affecting women in their fertile stage to a greater extent (Modarelli et al; 2019) which could be due to chronic exposure to nitrates detected in groundwater. This situation has also been observed in other places on the planet by various authors, in relation to exposure to nitrates as well as other EDs (Colborn et al; \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1993\u003c/span\u003e, Gatseva et al; 2008, Huang et al; \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2017b\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn people exposed to nitrates or perchlorates, iodine uptake by the NIS cotransporter is impaired (Tietge et al; \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2005\u003c/span\u003e, Zambrano-Fern\u0026aacute;ndez et al; 2022) which interferes with thyroid hormone synthesis and causes hypothyroxinemia (Braverman et al; 2005, Garcia-Torres et al; 2022, Serrano-Nacimento et al; 2022) As a consequence, in individuals, the release of pituitary TSH increases, this hormone being the main regulator of thyroid functions (Tonacchera et al.; \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2004\u003c/span\u003e, Dohan et al.; 2007).\u003c/p\u003e \u003cp\u003eIt has been known for years that prometamorphosis and climax in amphibians are regulated by T4/T3 levels, and that these in turn regulate TSH levels in larvae (Goos et al.; \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e1968\u003c/span\u003e). Authors such as Hemme (Hemme et al.;1972) and later Ospitz et al. (Opitz et al.; 2006), have described that the exposure of tadpoles compounds such as thiocyanates and perchlorates increases the expression of TSH mRNA, as well as the β receptor of this hormone. This elevation in TSH levels, stimulate the transcription of the NIS protein, which reaches its maximum expression level in xenopus larvae, between the 58NF to 60NF metamorphosis stages. In present study, xenopus laevis larvae chronically exposed to environmental groundwater samples containing nitrates in variable concentrations between 45mg/L \u0026minus;\u0026thinsp;83mg/L and was observed an increase in expression levels of the NIS cotransporter during 60NF stage at the larvae. This increase in protein expression could represent an adaptive mechanism for increased level of thyroid iodine absorption to and maintain the production of T4/T3 to reach metamorphic climax (De Groef et al; 2006). This could explain why duration of the climax to remain unchanged. Similar features of temporal, rapid, and persistent induction of NIS mRNA by TSH treatment have been reported in different rat thyroid follicle cell lines (Trapasso et al.; \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e1999\u003c/span\u003e, Fozzatti et al.; \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). However, at the 62NF stage a reduction in the expression level of the NIS cotransporter was observed, this would cause an asynchronous morphological development of the tadpoles and a reduction in the number of those that managed to complete metamorphosis. Consequently, those who reached the final stage of 66NF climax were smaller than those in the control group. The reduction in thyroid hormone levels, caused by the blockade of the NIS transporter exerted by nitrates, causes an increase in TSH levels. Studies in mammals have shown that its effects are mediated by the modulation exerted by thyroid hormones on specific gene expression during the process of embryonic development (Yamazzaki et al.; 2003, Montesinos et al.; \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Monitoring effected for two years, of larvae after completing metamorphosis, showed that only the females developed malformations that were involved eyes, mouth, lower limbs, and spine, which could be related to deficiency in T4 and T3 production that occurred during prometamorphosis, in larvae during their development. This reduction in thyroid hormone levels during metamorphosis could be generating changes in function of various proteins, through changes in DNA sequences that code for these proteins, affecting genes such as Xnkx-2.1., a homeobox gene expressed during early forebrain development, lung, and thyroid in xenopus laevis (Holleman et al.; 2000). Other studies indicated that the exposure to high levels of nitrate during pregnancy is a risk factor for spontaneous abortion, fetal death, prematurity, intrauterine growth restriction, low birth weight, congenital malformations, and neonatal death (Ward et al; \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eEskiocak, S et al (Eskiocak et al.; \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) observed that in rats exposed to a concentration of 50 mg/L of nitrates, T4 levels decreased significantly in this group, but an increase was recorded with exposure to 100 mg/L. and in this group it was accompanied by an elevation of TSH. Our group observed changes of hyperplasia and hypertrophy in larval thyroid with an increase between 60NF and 62NF stages, when larvae were exposed to groundwater contaminated with nitrates at levels between 45 mg/L \u0026minus;\u0026thinsp;83 mg/L, above levels considered safe for human consumption (WHO; \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). In the experiment by Eskiocak, S et al, the glands of rats would suffered a reduction in levels of iodine uptake, due to the competitive blockade exerted by nitrates, producing a reduction in T4 levels. In the experiment developed by our group an increase in NIS transporter expression levels were observed in 60NF stage. As larvae have a high rate of metabolic demand during this stage of the metamorphosis, due to the critical changes that occur in their morphology (lung development, intestinal changes and tail resorption), they could develop, when exposed to a high concentration of nitrates, an increase in TSH levels. Giving rise to various events: hyperplasia, hypertrophy of the gland and stimulation of the synthesis of NIS transporter and therefore the absorption of iodine to maintain T4 production levels. If the inhibition of iodine uptake in the gland persists, the synthesis chain of the NIS transporter would end up being compromised at the intracellular level by inhibiting transcription factors such as TTF1 and/or PAX-8. This would cause a reduction in the synthesis of T4, the persistence of the stimulation exerted by TSH due to a decrease of negative feedback, which would explain the significant hyperplasia suffered in the follicular cells during 62NF stage, and the reduction in the expression levels of the NIS transporter during the 62NF stage.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eNitrates present in groundwater, in high concentrations, affect the functioning of the thyroid gland. Chronic exposure to these concentrations causes decreased and/or absent expression of NIS in exposed larvae. This generates histological alterations as a consequence of reduction in thyroxine levels, which translate into affectation of tadpole morphological development.\u003c/p\u003e \u003cp\u003eThis study shows the usefulness of the model to assess the safety of water considered suitable for human consumption.\u003c/p\u003e \u003cp\u003eThese observation could contribute to the current discussion about the acceptable daily intake of nitrate.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthical Approval:\u003c/strong\u003e Approved by the Institutional Committee for the Care and Use of Laboratory Animals, of the Faculty of Medicine of the UBA, ([email protected]. EXP-UBA 0003598/2013).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments:\u003c/strong\u003e Dr. J. J. Lopez, Histology Department, School of Medicine of the University of Buenos Aires.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Participate:\u003c/strong\u003e the corresponding author confirms that the work described has not been published before (except in abstract form or as part of a published conference, review or thesis); that it is not being considered for publication elsewhere; that its publication has been approved by all co-authors; that its publication has been approved by the responsible authorities of the institution where the work is carried out.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Publish:\u003c/strong\u003e The author accepts responsibility for publishing this material on behalf of any and all co-authors.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e This work was supported by Endocrinology Laboratory, Institute of Physiology, Academic Unit II, School of Medicine, University of Buenos Aires. Buenos Aires. Argentina and with contribution of own funds.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions:\u003c/strong\u003e All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Modarelli Maria Fernanda, Bilbao Rodrigo Miguel and Ponzo Osvaldo Juan. The first draft of the manuscript was written by Modarelli Maria Fernanda and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBabalola OO, Van Wyk JH (Oct 2021) Exposure impacts of Imazapyr formulation on larval development and thyroid histology of Xenopus laevis. Environ Sci Pollut Res Int 28(37):50967\u0026ndash;50974. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s11356-021-14227-4\u003c/span\u003e\u003cspan address=\"10.1007/s11356-021-14227-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003eEpub 2021 May 11\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBertrand S, Brunet. FG, Escriva H, Parmentier G, Laudet V, Robinson-Rechavi M (Oct 2004) Evolutionary genomics of nuclear receptors: from twenty-five ancestral genes to derived endocrine systems. Mol Biol Evol 21(10):1923\u0026ndash;1937. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1093/molbev/msh200\u003c/span\u003e\u003cspan address=\"10.1093/molbev/msh200\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003eEpub 2004 Jun 30. PMID: 15229292\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBraverman LE et al (Feb 2005) The effect of perchlorate, thiocyanate, and nitrate on thyroid function in workers exposed to perchlorate long-term. J Clin Endocrinol Metab 90(2):700\u0026ndash;706. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1210/jc.2004-1821\u003c/span\u003e\u003cspan address=\"10.1210/jc.2004-1821\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003eEpub 2004 Nov 30. PMID: 15572417\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBrown DD, Cai L (June 2007) Amphibian metamorphosis Review 306(1):20\u0026ndash;33. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ydbio.2007.03.021\u003c/span\u003e\u003cspan address=\"10.1016/j.ydbio.2007.03.021\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBuchholz DR, Shi YB (Sep 2018) Dual function model revised by thyroid hormone receptor alpha knockout frogs. Gen Comp Endocrinol 265:214\u0026ndash;218. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ygcen.2018.04.020\u003c/span\u003e\u003cspan address=\"10.1016/j.ygcen.2018.04.020\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003eEpub 2018 Apr 22. PMID: 29689262; PMCID: PMC6087486\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCarr DL, Carr JA, Willis RE, Pressley TA (2008) Mar A perchlorate sensitive iodide transporter in frogs. Gen Comp Endocrinol. 1;156(1):9\u0026ndash;14. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ygcen.2008.01.005\u003c/span\u003e\u003cspan address=\"10.1016/j.ygcen.2008.01.005\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. Epub 2008 Jan 12. PMID: 18275962; PMCID: PMC2279098\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCarr DL, Laharrague F, Kahn B, Pressley TA, Carr JA (Apr 2003) Molecular characterization of a putative sodium/iodide symporter in the South African clawed frog, Xenopus laevis. Ann N Y Acad Sci 986:711\u0026ndash;712. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1749-6632\u003c/span\u003e\u003cspan address=\"10.1111/j.1749-6632\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e2003.tb07287. x. PMID: 12763923\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eColborn T et al (1993) Developmental Effects of Endocrine Disrupting Chemicals in Wildlife and Humans. Rev Environ Health Perspect 101(5):378\u0026ndash;384. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1289/ehp.93101378\u003c/span\u003e\u003cspan address=\"10.1289/ehp.93101378\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDai G, Levy O, Carrasco N (1996) Feb Cloning and characterization of the thyroid iodide transporter. Nature. 1;379(6564):458\u0026thinsp;\u0026ndash;\u0026thinsp;60. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/379458a0\u003c/span\u003e\u003cspan address=\"10.1038/379458a0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 8559252\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDenver RJ, Glennemeier KA, and., Boorse GC (Feb 2002) Endocrinology of complex life cycles: Amphibians. In: Pfaff DW, Arnold AP, Etgen AM, Fahrback SE, Rubin RT (eds) Hormones, brain and behavior. \u0026rsaquo; biologia \u0026rsaquo; public_html, pp 469\u0026ndash;513. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://exa.unne.edu.ar\u003c/span\u003e\u003cspan address=\"https://exa.unne.edu.ar\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDe Groef B, Decallonne B, Van der Geyten S, Darras VM, Bouillon R (Jul 2006) Perchlorate versus other environmental sodium/iodide symporter inhibitors: potential thyroid-related health effects. Rev Eur J Endocrinol 155(1):17\u0026ndash;25. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1530/eje.1.02190\u003c/span\u003e\u003cspan address=\"10.1530/eje.1.02190\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDoh\u0026aacute;n O, Portulano C, Basquin C, Reyna-Neyra A, Amzel LM, Carrasco N (2007) The Na+/I- symporter (NIS) mediates electroneutral active transport of the environmental pollutant perchlorate. Proc Natl Acad Sci U S A 104(51):20250\u0026ndash;20255. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1073/pnas.0707207104\u003c/span\u003e\u003cspan address=\"10.1073/pnas.0707207104\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEskiocak S, Dundasr C, Basoglu T, (Jul (2005) The effects of taking chronic nitrate by drinking water on thyroid functions and morphology. Clin Exp Med 5(2):66\u0026ndash;71. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s10238-005-0068-1\u003c/span\u003e\u003cspan address=\"10.1007/s10238-005-0068-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFozzatti L, V\u0026eacute;lez ML, Lucero AM, Nicola JP, Mascanfroni ID, Macci\u0026oacute; DR et al (2007) Endogenous thyrocyte-produced nitric oxide inhibits iodide uptake and thyroid-specific gene expression in FRTL-5 thyroid cells. J Endocrinol 192(3):627\u0026ndash;637. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1677/joe.1.06967\u003c/span\u003e\u003cspan address=\"10.1677/joe.1.06967\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGarc\u0026iacute;a Torres E et al (May 2022) Consumption of water contaminated by nitrate and its deleterious effects on the human thyroid gland: a review and update. Int J Environ Health Res 32(5):984\u0026ndash;1001 Epub 2020 Aug 31. PMID: 32866080\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGatseva PD, Argirova MD (2008) High-nitrate levels in drinking water may be a risk factor for thyroid dysfunction in children and pregnant women living in rural Bulgarian areas. Int Hyg Environ Health 211(5):555\u0026ndash;559. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.ijheh.2007.10.002\u003c/span\u003e\u003cspan address=\"10.1016/j.ijheh.2007.10.002\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003eEpub 2007 Dec 27\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGoleman WL, Carr JA, Anderson TA (Mar 2002) Environmentally relevant concentrations of ammonium perchlorate inhibit thyroid function and alter sex ratios in developing Xenopus laevis. Environ Toxicol Chem 21(3):590\u0026ndash;597 PMID: 11878472\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGoos HJ, Zwanenbeek HC, van Oordt PG (1968) Hypothalamic neurosecretion and metamorphosis in Xenopus laevis. II. The effect of thyroxine following treatment with propylthiouracil. Arch Anat Histol Embryol 51(1):267\u0026ndash;274 PMID: 4905268\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGuiles HJ (2000) Thyroid function. In: Bishop ML, DubenEngelkirk JL, Fody EP (eds) Clinical chemistry principles, procedures, correlations. Lippincott Williams and Wilkins, Philadelphia, pp 403\u0026ndash;421\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGrim C, Wolfe M, Braunbeck T et al (Jun 2009) Thyroid Histopathology Assessments for the Amphibian Metamorphosis Assay to Detect Thyroid-active Substances. Toxicol Pathol 37(4):415\u0026ndash;424. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1177/0192623309335063\u003c/span\u003e\u003cspan address=\"10.1177/0192623309335063\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003eEpub 2009 Apr 22\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHemme L (1972) Die Differenzierungsgenese der TSH Zellen von Xenopus laevis unter Normalbedingungen und nach Thiouracilbehandlung. Z Zellforsch 125:353\u0026ndash;377. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF00306632\u003c/span\u003e\u003cspan address=\"10.1007/BF00306632\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHuang HB, Pan WH, Chang JW et al (2017b) \u0026iquest;Does exposure to phthalates influence thyroid function and growth hormone homeostasis? The Taiwan Environmental Survey for Toxicants (TEST) 2013. Environ Res 153:63\u0026ndash;72. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.envres.2016.11.014\u003c/span\u003e\u003cspan address=\"10.1016/j.envres.2016.11.014\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHollemann T, Pieler T, (Nov (2000) Xnkx-2.1: a homeobox gene expressed during early forebrain, lung and thyroid development in Xenopus laevis. Dev Genes Evol. 210(11):579\u0026thinsp;\u0026ndash;\u0026thinsp;81. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s004270000098\u003c/span\u003e\u003cspan address=\"10.1007/s004270000098\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 11180810\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJian JM, Chen D, Han FJ, Guo Y, Zeng L, Lu X, Wang F (2018) Sept A short review on human exposure to and tissue distribution of per- and polyfluoroalkyl substances (PFASs). Sci Total Environ. 636:1058\u0026ndash;1069. doi: 10.1016/j.scitotenv.2018.04.380. Epub 2018 May 3. PMID: 29913568\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJord\u0026atilde;o CP, Pereira MG, Bellato CR, Pereira JL, Matos AT (2002) Oct Assessment of water systems for contaminants from domestic and industrial sewages. Environ Monit Assess. 2002;79(1):75\u0026ndash;100. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1023/a:1020085813555\u003c/span\u003e\u003cspan address=\"10.1023/a:1020085813555\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 12381024\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLanger P (2006) Increased thyroid volume and frequency of thyroid disorders signs in school children from nitrate polluted area. Chemosphere 62(4):559\u0026ndash;564. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.chemosphere.2005.06.030\u003c/span\u003e\u003cspan address=\"10.1016/j.chemosphere.2005.06.030\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003eEpub 2005 Aug 10\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi S, Qiao K, Jiang Y, Wu Q, Coffin S, Gui W, Zhu G (Dec 2019) Disruptive effects of two organotin pesticides on the thyroid signaling pathway in Xenopus laevis during metamorphosis. Sci Total Environ 20:697:134140 Epub 2019 Aug 27. PMID: 31476497\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eModarelli MF, Bilbao RM, Ponzo OJ, Water (2022) Contamination by Nitrates and its Thyroid Disruptive Action. Bioassay on Xenopus Laevis. GJMR B 22(2,10):34\u0026ndash;87. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://medicalresearchjournal.org \u0026rsaquo;\u003c/span\u003e\u003cspan address=\"https://medicalresearchjournal.org \u0026rsaquo;\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eModarelli MF, Pozo OJ (2019) Relationship of subclinical hypothyroidism and goiter with the origin of water consumed by a population of the Buenos Aires suburbs. Med (Bs As) 79(1):11\u0026ndash;19 PMID: 30694184\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003edel Montesinos M M., et al (2016) Nitric oxide-repressed forkhead factor FoxE1 expression is involved in the inhibition of TSH-induced thyroid peroxidase levels. Mol Cell Endocrinol 420:105\u0026ndash;115. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.mce.2015.11.020\u003c/span\u003e\u003cspan address=\"10.1016/j.mce.2015.11.020\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMughal BB, Demeneix BA, Fini JB (2018) Evaluating Thyroid Disrupting Chemicals in Vivo Using Xenopus laevis. Methods Mol Biol. 1801:183\u0026ndash;192. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/978-1-4939-7902-8_15\u003c/span\u003e\u003cspan address=\"10.1007/978-1-4939-7902-8_15\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 29892825\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNieuwkoop PD, Faber J (1956) Normal table of Xenopus laevis (Daudin). A Systematical and Chronological Survey of the Development from the Fertilized Egg till the end of Metamorphosis. North Holland: Amsterdam 22(00):89129\u0026ndash;89125. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.1016/S0168-\u003c/span\u003e\u003cspan address=\"10.1016/S0168-\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOECD (2018a) Organisation for Economic Co-operation and Development. Revisedguidance document 150 on standardised test guidelines for evaluatingchemicals for endocrine disruption. OECD Series on Testing and Assessment. Paris France. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1787/9789264304741-en\u003c/span\u003e\u003cspan address=\"10.1787/9789264304741-en\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e[accessed 15 September 2018]\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOhno M, Zannini M, Levy O, Carrasco N, Di Lauro R (Mar 1999) The paired-domain transcription factor Pax8 binds to the upstream enhancer of the rat sodium/iodide symporter gene and participates in both thyroid-specific and cyclic-AMP-dependent transcription. Mol Cell Biol 19(3):2051\u0026ndash;2060 PMID: 10022892; PMCID: PMC83998\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOpitz R, Trubiroha A, Lorenz C et al (Jul 2006) Expression of sodium-iodide symporter mRNA in the thyroid gland of Xenopus laevis tadpoles: developmental expression, effects of antithyroidal compounds, and regulation by TSH Journal of Endocrinology. J Endocrinol 190(1):157\u0026ndash;170. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1677/joe.1.06606\u003c/span\u003e\u003cspan address=\"10.1677/joe.1.06606\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaito T, Endo T, Nakazato M, Kogai T, Onaya T (1997) Thyroid-stimulating hormone-induced down-regulation of thyroid transcription factor 1 in rat thyroid FRTL-5 cells. Endocrinology. Feb;138(2):602-6. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1210/endo.138.2.4918\u003c/span\u003e\u003cspan address=\"10.1210/endo.138.2.4918\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 9002992\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSerrano-Nascimento C, Nunes MT (2022) Oct 21 Perchlorate, nitrate, and thiocyanate: Environmental relevant NIS-inhibitors pollutants and their impact on thyroid function and human health. Front Endocrinol (Lausanne). 13:995503. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fendo.2022.995503\u003c/span\u003e\u003cspan address=\"10.3389/fendo.2022.995503\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 36339434; PMCID: PMC9633673\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eStubleski J, Salihovic S, Lind L, Lind PM, van Bavel B, K\u0026auml;rrman A (Oct 2016) Changes in serum levels of perfluoroalkyl substances during a 10-year follow-up period in a large population-based cohort. Environ Int 95:86\u0026ndash;92 Epub 2016 Aug 16. PMID: 27542758\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTata JR, (Dec (1998) Amphibian metamorphosis as a model for studying the developmental actions of thyroid hormone. Cell Res 8(4):259\u0026ndash;272. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/cr.1998.26\u003c/span\u003e\u003cspan address=\"10.1038/cr.1998.26\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eThambirajah AA, Koide EM, Imbery JJ, Helbing CC (May 2019) Contaminant and Environmental Influences on Thyroid Hormone Action in Amphibian Metamorphosis Sec. Thyroid Endocrinol 14:1\u0026ndash;29. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fendo.2019.00276\u003c/span\u003e\u003cspan address=\"10.3389/fendo.2019.00276\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTietge JE et al (2005) Apr. Metamorphic inhibition of Xenopus laevis by sodium perchlorate: effects on development and thyroid histology. Environ Toxicol Chem. 24(4):926\u0026thinsp;\u0026ndash;\u0026thinsp;33. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1897/04-105r.1\u003c/span\u003e\u003cspan address=\"10.1897/04-105r.1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 15839568\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTonacchera M et al (2004) Dec. Relative potencies and additivity of perchlorate, thiocyanate, nitrate, and iodide on the inhibition of radioactive iodide uptake by the human sodium iodide symporter. Thyroid. 14:1012\u0026ndash;1019. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1089/thy.2004.14.1012\u003c/span\u003e\u003cspan address=\"10.1089/thy.2004.14.1012\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 15650353\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTrapasso F et al (1999) May Iodide symporter gene expression in normal and transformed rat thyroid cells. Eur J Endocrinol. 140(5):447\u0026thinsp;\u0026ndash;\u0026thinsp;51. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1530/eje.0.1400447\u003c/span\u003e\u003cspan address=\"10.1530/eje.0.1400447\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 10229912\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWard MH, Jones RR, Brender JD, de Kok TM, Weyer PJ, Nolan BT et al (2018) Drinking water nitrate and human health: An updated review. Int J Environ Res Public Health 15(7):1557. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/ijerph15071557\u003c/span\u003e\u003cspan address=\"10.3390/ijerph15071557\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWeyer PJ et al (2001) Municipal drinking water nitrate level and cancer risk in older women: the Iowa Women's Health Study. Epidemiology. May;12(3):327\u0026thinsp;\u0026ndash;\u0026thinsp;38. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1097/00001648-200105000-00013\u003c/span\u003e\u003cspan address=\"10.1097/00001648-200105000-00013\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 11338313\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWHO (2016) Nitrate and nitrite in drinking water: background document for development of WHO guidelines for drinking water quality. World Health Organization, Geneva\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYamazaki K et al (2003) Feb. Genes regulated by thyrotropin and iodide in cultured human thyroid follicles: analysis by cDNA microarray. Thyroid. 13(2):149\u0026thinsp;\u0026ndash;\u0026thinsp;58. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1089/105072503321319459\u003c/span\u003e\u003cspan address=\"10.1089/105072503321319459\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 12699589\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYao X et al (2017) Optimization of the T3-induced Xenopus metamorphosis assay for detecting thyroid hormone signaling disruption of chemicals. J Environ Sci 52:314\u0026ndash;324. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.jes.2016.09.020\u003c/span\u003e\u003cspan address=\"10.1016/j.jes.2016.09.020\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003eEpub 2016 Nov 23\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYue B, Ning S, Miao H, Fang C, Li J, Zhang L et al (2023) Human exposure to a mixture of endocrine disruptors and serum levels of thyroid hormones: A cross-sectional study. J Environ Sci (China) 125:641\u0026ndash;649. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.jes.2022.01.017\u003c/span\u003e\u003cspan address=\"10.1016/j.jes.2022.01.017\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZambrano-Fern\u0026aacute;ndez S, Zamora-Camacho FJ, Arag\u0026oacute;n P (Jan 2022) Direct and indirect effects of chronic exposure to ammonium on anuran larvae survivorship, morphology, and swimming speed. Chemosphere 287(Pt 3132349. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.chemosphere.2021.132349\u003c/span\u003e\u003cspan address=\"10.1016/j.chemosphere.2021.132349\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003eEpub 2021 Sep 24. PMID: 34826957\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"nitrates, groundwater, morphogenesis, thyroid, NIS carrier, xenopus laevis","lastPublishedDoi":"10.21203/rs.3.rs-3716033/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3716033/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eGroundwater is a reservoir of fresh water. In the goitrous areas the water used for drinking and cooking generally is high in nitrate content. The aim of the present study was to evaluate in that areas, chronic effects to expose of xenopus laevis larvae during a complete metamorphosis process to groundwater. For analyze the effects on thyroid function. The xenopus laevis larvae were divided into 3 work groups submerged, until the end of metamorphosis, in: drinking water as control (C), groundwater as group (G) and drinking water plus potassium perchlorate as positive control (PC). In (G) group metamorphosis process was completed in 37% and 0% in (PC) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001). In the 60 of Niewkoop and Faber, stage the wet weight and body length was in (G) smaller than (C) and (PC) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001). But was not difference in 62 for (G), (PC) was bigger than (C). In 60 stage was no significant differences on the length of the hind limbs. In 62 was bigger in (G) and (PC) than in (C) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). Thyroid hyperplasia was grade 2 in (G) and 3 (PC) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) in 60 stage and 3 in (G) and (PC) in the 62 (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001). Follicular hypertrophy and colloid depletion in (G) and (PC) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001). In groundwater was found high nitrates concentrations. These findings suggest that nitrate contaminate groundwater impairs thyroid function of xenopus laevis larvae. These observation could contribute to the current discussion about the acceptable daily intake of nitrate.\u003c/p\u003e","manuscriptTitle":"Groundwater used for human consumption causes histological and molecular changes in the thyroid that affect the morphogenesis process of xenopus laevis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-02-07 19:50:14","doi":"10.21203/rs.3.rs-3716033/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"7d3e1fc7-0eb3-421f-820f-59a21e9faeab","owner":[],"postedDate":"February 7th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-05-28T09:36:42+00:00","versionOfRecord":[],"versionCreatedAt":"2024-02-07 19:50:14","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3716033","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3716033","identity":"rs-3716033","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}