Lack of cytogenetic effects in mice or mutations in Salmonella receiving sodium fluoride.

We have examined the possible effect of fluoride intake on chromosome damage. There was no evidence of increased frequency of chromosomal aberration in bone marrow or testis cells of mice with either 50 ppm fluoride intake over several generations or 100 ppm intake for 6 weeks compared to animals drinking distilled water. Fluoride was not found to be mutagenic in a widely used bacterial mutagenesis assay over a range of 0.1 to as high as 2000 microgram fluoride per plate.     

Cytochemical response of kidney, liver and nervous system of fluoride ions in drinking water.

Morphological and cytochemical studies on the squirrel monkey have been made after maintaining the sujects on pure distilled water and fluoridated distilled water for 18 months with the objective of determining the effect of fluoride on the activity of some hydrolytic and oxidative enzymes in the kidney, liver and nervous system. Daily water intake by individual animals was measured over the final 10 months of the animal's exposure to 0,1 and 5 ppm fluoride. Water consumption was considerably higher in the animals on higher fluoride intake. Whereas the nervous system remained totally unaffected by this experimental procedure, the liver showed a slightly enhanced acivity of Krebs citric acid cycle enzymes. The kidneys, however, showed significant cytochemical changes, especially in the animals on 5 ppm fluoride intake in their drinking water. In these animals, the glomeruli showed an increase in the activity of acid phosphatase and the enzymes belonging to the citric acid cycle and the pentose shunt, whereas lactate dehydrogenase, a resentative of the anaerobic glyoclytic pathway, remained unchanged or only slightly changed. These observations suggest that fluoride in concentrations as low as 5 ppm interferes to some extent with the intracellular metabolism of the excretory system.     

Influence of dietary lead on fluoride bioavailability in the rat

A factorial experiment was conducted with weanling rats fed a purified diet to determine the influence of dietary lead (0 or 100 ppm) as either lead acetate or lead carbonate on fluoride bioavailability (2 or 10 ppm as sodium fluoride). During the 6-wk study, both forms of lead depressed weight gain, regardless of the fluoride level, despite the fact that food intake was similar for all treatment groups. Both forms of lead produced a small, but significant, reduction in femur and second molar fluoride. This effect, however, could only be demonstrated in rats fed diets containing 10 ppm fluoride, indicating a significant interaction between lead and fluoride for these indices of fluoride bioavailability. This interactive effect between fluoride and lead was also demonstrated for apparent fluoride absorption. Both forms of dietary lead significantly increased the lead concentration of plasma, femur, liver, and kidney, and both forms of lead significantly increased the urinary excretion of delta-aminolevulinic acid. The level of dietary fluoride failed to influence these measurements. We therefore conclude that, although small amounts of dietary lead reduce fluoride bioavailability, small amounts of dietary fluoride do not appear to significantly influence the utilization of dietary lead.     

Lipid peroxidation and antioxidant systems in the blood of young rats subjected to chronic fluoride toxicity

Wistar albino rats were exposed to 30 or 100 ppm fluoride in drinking water during their fetal, weanling and post-weaning stages of life up to puberty. Extent of lipid peroxidation and response of the antioxidant systems in red blood cells and plasma to prolonged fluoride exposure were assessed in these rats in comparison to the control rats fed with permissible level (0.5 ppm) of fluoride. Rats treated with 100 ppm fluoride showed enhanced lipid peroxidation as evidenced by elevated malondialdehyde (MDA) levels in red blood cells but, 30 ppm fluoride did not cause any appreciable change in RBC MDA level. 30 ppm fluoride-intake resulted in increased levels of total and reduced glutathione in red blood cells and ascorbic acid in plasma while 100 ppm fluoride resulted in decreases in these levels. The activity of RBC glutathione peroxidase was elevated in both the fluoride-treated groups, more pronounced increase was seen with 100 ppm. Reduced to total glutathione ratio in RBC and uric acid levels in plasma decreased in both the groups. RBC superoxide dismutase activity decreased significantly on high-fluoride treatment. These results suggest that long-term high-fluoride intake at the early developing stages of life enhances oxidative stress in the blood, thereby disturbing the antioxidant defense of rats. Increased oxidative stress could be one of the mediating factors in the pathogenesis of toxic manifestations of fluoride.     

Influence of dietary fluoride restriction on regulation of plasma nd soft tissue fluoride contents.

The adjustments in total fluoride concentration in plasma, bones, liver, and muscle were examined when rats were given a diet of very low fluoride content following a dietary regimen of elevated fluoride intake. The animals received a diet containing 34 ppm of fluoride and water with 50 ppm added fluoride in the 28-day initial period and in the depletion period they were given a diet containing only 0.21 ppm of fluoride and distilled water. The findings indicated a 12-fold increase in the fluoride content of the humeri after 28 days of high-flurodie intake with a greater increment by the epiphyses than by the diaphyses. During 21 days of the depletion period the skeletal fluoride was reduced by only 7.7% indicating a marked retention of fluoride during processes of bone remodeling and growth. The plasma, muscle, and liver total fluoride contents were significantly increased at the end of the period of high-fluoride intake, but these concentrations were found to be restored to base-line levels in 3-7 days of the depletion period. By comparison of the distribution of total fluoride with injected radiofluoride between tissue and plasma waters, it was concluded that muscle and liver contain bound fluoride that does not exchange completely with ionic fluoride.     

Effects of fluoride ingestion on white-tailed deer (Odocoileus virginianus)

The effects of the addition of 25 or 50 ppm fluoride (F), as sodium fluoride (NaF), to the rations of 5-mo-old male white-tailed deer were similar to those observed in domestic cattle fed similar amounts of fluoride. The ingestion of 50 ppm F for 2 yr resulted in the accumulation of over 7,000 ppm F in bone ash. Accumulation of fluoride in antlers was extensive and occurred more rapidly than in skeletal tissue. Fluoride ingestion resulted in lesions on the developing incisors that were similar, but not identical to those seen in other species. Increased molar wear in the deer fed 50 ppm F was minimal, and no gross pathology of the mandible was observed. Only mild hyperostosis of the long bones was evident.     

Absence of mutagenic and antimutagenic activities of fluoride in Ames salmonella assays

The mutagenicity of fluoride (as sodium fluoride, NaF) was investigated with Ames Salmonella/microsome assays in strains of TA97a, TA98, TA100, TA102 and TA1535. The concentrations of NaF tested ranged from 0.44 to 4421 micrograms/plate (0.1 to 1000 ppm F), both with and without microsome activation. In addition, the suggested antimutagenic effect of fluoride was evaluated with known mutagens at various concentrations of NaF (0.44-442.2 micrograms/plate, 0.1-100 ppm F). The data showed that NaF, in amounts from 0.44 to 442.2 micrograms/plate (0.1-100 ppm F), failed to significantly increase the number of the revertants over the number observed in the solvent (distilled deionized water) controls. Increases of NaF to, and beyond, 1100 micrograms/plate (250 ppm F) resulted in a toxic effect and a reduction of the revertants to various degrees among the strains. NaF in the presence of known mutagens did not significantly decrease the number of the revertants. The results of this study indicate that NaF does not have mutagenic or antimutagenic effects in the strains tested with Ames Salmonella assays.     

Temporal kinetics of fluoride accumulation: from fetal to adult deer

In June 2011, a volcano deposited about 100 million tons of tephra over parts of Chile and over 36 million ha of Argentina. Initially, fluoride was considered irrelevant; however, recently wild deer exhibited strong fluorosis, with fluoride level increasing 38-fold among severely affected deer. Whereas mothers averaged 2,151 ppm, their late-term fetuses had only 19.8 ppm, indicating a barrier to fluoride transport in utero. Levels among four age classes increased significantly, at a rate of about 1,000 ppm/year. The temporal kinetics of accumulation suggests that sources of available fluoride are highly effective. Thus, compared to prior background levels (63 ppm in adults) and to fetuses starting at about 20 ppm, 1-year-old calves averaged 1,035 ppm (maximum 1,830 ppm), 2-year olds averaged 2,151 ppm (maximum 2,513 ppm), and older deer averaged 2,806 ppm (maximum 5,175 ppm). As osteofluorosis occurs in deer with >4,000 ppm, accumulation of 1,000 ppm/year would result in adults reaching levels causing osteopathology in 1–2 years. Importantly, impacts may be further exacerbated by regional iodine and selenium deficiencies. Iodine deficiency may increase incidences of dental fluorosis and severity of damages, while selenium deficiency impacts iodine metabolism. Fluorosis will affect population dynamics, morbidity, predation susceptibility, and other ecosystem components like scavenger and plant communities.     

Defluorination of fluoroacetate in the rat

Rats given 5 ppm F as FAc (equivalent to 26 ppm of NaFac) in the drinking water for approximately four months deposited as much fluoride in the skeletal system as did rats receiving 5 ppm F as NaF in the water. Little evidence could be found for the presence of organically bound fluoride in bone after ingesting FAc, though an appreciable proportion of skeletal fluoride deposited when NaF was ingested was shown not to respond to the fluoride ion electrode. The daily urinary excretion of total fluoride after FAc was somewhat greater than after NaF; about two thirds of this fluoride responded to the electrode, whereas more than 90 percent of the total fluoride after NaF was ionic in nature. The data are interpreted as showing that the rat is capable of splitting the C-F bond in FAc and/or in its fluoride-containing metabolites, with subsequent skeletal storage and renal excretion of the released fluoride ion. The chronic administration of this low level of FAc caused an early but temporary retardation of growth. The Krebs cycle was interfered with, as evidenced by increased concentrations of citrate in the kidney and urine. At termination of the experiment, histological examination of the testes showed that the FAc had induced severe damage characterized by massive disorganization of the tubules, nearly total loss of functional cells, absence of sperm, and damage to the Sertoli cells.     

Health risk assessment of fluoride in water distribution network of Mashhad,Iran

High and low levels of fluoride in drinking waters have been considered as a major public health issue in recent years. This cross-sectional study was conducted over five consecutive years (from 2012 to 2016) in the water distribution network of Mashhad city, Iran with the objectives of determining levels of fluoride and to perform health risk assessment among residents in the study area. Water samples were taken from 30 stations and were analyzed using UV-visible spectrophotometer. The mean annual concentrations of fluoride in all stations during five years of consecutive study were lower than the respective maximum permissible limits (1.5 mg/L) in water as regulated by the WHO. The human health risk assessment was performed by calculating the chronic daily intake and hazard quotient (HQ) of fluoride through human oral intake for adults (men and women) and children for each year during a five-year study. Health risk analysis in this study presented that the non-carcinogenic risk associated with fluoride exposure through drinking water in Mashhad was mostly acceptable because the mean HQ values of fluoride were lower than 1.     

Appropriate uses of fluorides for children: guidelines from the Canadian Workshop on the Evaluation of Current Recommendations Concerning Fluorides.

OBJECTIVE: To prevent fluorosis caused by excessive fluoride ingestion by revising recommendations for fluoride intake by children. OPTIONS: Limiting fluoride ingestion from fluoridated water, fluoride supplements and fluoride dentifrices. OUTCOMES: Reduction in the prevalence of dental fluorosis and continued prevention of dental caries. EVIDENCE: Before the workshop, experts prepared comprehensive literature reviews of fluoride therapies, fluoride ingestion and the prevalence and causes of dental fluorosis. The papers, which were peer-reviewed, revised and circulated to the workshop participants, formed the basis of the workshop discussions. VALUES: Recommendations to limit fluoride intake were vigorously debated before being adopted as the consensus opinion of the workshop group. BENEFITS, HARMS AND COSTS: Decrease in the prevalence of dental fluorosis with continuing preventive effects of fluoride use. The only significant cost would be in preparing new, low-concentration fluoride products for distribution. RECOMMENDATIONS: Fluoride supplementation should be limited to children 3 years of age and older in areas where there is less than 0.3 ppm of fluoride in the water supply. Children in all areas should use only a "pea-sized" amount of fluoride dentifrice no more than twice daily under the supervision of an adult. VALIDATION: These recommendations are almost identical to changes to recommendations for the use of fluoride supplements recently proposed by a group of European countries. SPONSORS: The workshop was organized by Dr. D. Christopher Clark, of the University of British Columbia, and Drs. Hardy Limeback and Ralph C. Burgess, of the University of Toronto, and funded by Proctor and Gamble Inc., Toronto, the Medical Research Council of Canada and Health Canada (formerly the Department of National Health and Welfare). The recommendations were formally adopted by the Canadian Dental Association in April 1993.     

The distribution of fluoride in some antarctic seals

Summary The aim of this investigation was to find out why some antarctic seals tolerate fluoride levels in their food (krill) that would be toxic to man. The fluoride content of different soft and hard tissues, urine and feces from 16 L. carcinophagus, 13 L. weddelli and 2 A. tropicalis (urine only) were determined by the fluoride sensitive electrode or by a photometric method (urine). Results and conclusions are following: All soft tissues have low concentrations of fluoride in the same range as soft tissues from non antarctic and non krill eating marine or terrestrial vertebrates. But within the soft tissues there are significant differences in the fluoride content (blood: 0.001 ppm F/dw — brain: 9.1 ppm F/dw). A special tolerance for fluoride of soft tissues can be excluded. Compared with man or weddell-seals the crabeater-seals store two times more fluoride in their skeleton. Hard structures that stop their mineralisation early in the development like teeth have only a relatively low fluoride content. For the same reason only neglectable amounts of fluoride could be detected in hair. Fluoride excretion by the kidneys is efficient and is — besides the storage of fluoride in the skeleton — the second mechanism to guarantee a good blood homoiostasis for fluoride. Until now it is impossible to decide how much fluoride from the food is resorbed by the digestive tract.     

Continuing impacts on red deer from a volcanic eruption in 2011

In June of 2011, the Puyehue–Cordon Caulle volcanic eruption deposited large amounts of ashes in Chile and Argentina. Although ashes were initially considered innoxious based on water leachates, we found clinical cases of fluoride intoxication in red deer (Cervus elaphus) and domestic herbivores in Argentina. The diagnosis was corroborated by high bone fluoride concentrations. The dynamics of temporal accumulation of fluoride suggested an average increase of 1,000 ppm per year of fluoride in the bone. However, a few deer had fluoride levels, suggesting an accumulation rate of about 3,700 ppm per year. Via recent sampling of deer, we now confirm that bone levels have reached up to 10,396 ppm of fluoride after about 28 months of exposure. Tephra across various sites averaged 548 ppm of fluoride, and due to dry conditions and eolic redeposition of ashes particularly east of the continental divide, clinical fluorosis is expected to continue to intensify. The described impact will reverberate through several aspects of the ecology of the deer, including effects on population dynamics, morbidity, predation susceptibility, as well as other components of the ecosystem, including other herbivores, scavengers, and plant communities.     

Effects of Fluoridated Milk on Root Dentin Remineralization

Background

The prevalence of root caries is increasing with greater life expectancy and number of retained teeth. Therefore, new preventive strategies should be developed to reduce the prevalence of root caries. The aim of this study was to investigate the effects of fluoridated milk on the remineralization of root dentin and to compare these effects to those of sodium fluoride (NaF) application without milk.

Methods

Thirty extracted human molars were divided into 6 groups, and the root cementum was removed from each tooth. The dentin surface was demineralized and then incubated with one of the following six solutions: Sodium chloride NaCl, artificial saliva, milk, milk+2.5 ppm fluoride, milk+10 ppm fluoride and artificial saliva+10 ppm fluoride. Serial sections were cut through the lesions and investigated with polarized light microscopy and quantitative morphometry, scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS). The data were statistically evaluated using a one-way ANOVA for multiple comparisons.

Results

The depth of the lesion decreased with increasing fluoride concentration and was the smallest after incubation with artificial saliva+10 ppm fluoride. SEM analysis revealed a clearly demarcated superficial remineralized zone after incubation with milk+2.5 ppm fluoride, milk+10 ppm fluoride and artificial saliva+10 ppm fluoride. Ca content in this zone increased with increasing fluoride content and was highest after artificial saliva+10 ppm fluoride incubation. In the artificial saliva+10 ppm fluoride group, an additional crystalline layer was present on top of the lesion that contained elevated levels of F and Ca.

Conclusion

Incubation of root dentin with fluoridated milk showed a clear effect on root dentin remineralization, and incubation with NaF dissolved in artificial saliva demonstrated a stronger effect.     

Effect of Fluoridated Water on Plasma Insulin Levels and Glucose Homeostasis in Rats with Renal Deficiency

Glucose intolerance in fluorosis areas and when fluoride is administered for the treatment of osteoporosis has been reported. Controlled fluoridation of drinking water is regarded as a safe and effective measure to control dental caries. However, the effect on glucose homeostasis was not studied so far. The aim of this study was to evaluate the effect of the intake of fluoridated water supply on glucose metabolism in rats with normal and deficient renal function. Male Sprague–Dawley rats were divided into eight groups of four rats. Renal insufficiency was induced in four groups (NX) which received drinking water containing 0, 1, 5, and 15 ppm F (NaF) for 60 days. Four groups with simulated surgery acted as controls. There were no differences in plasma glucose concentration after a glucose tolerance test between controls and NX rats and among rats with different intakes of fluoride. However, plasma insulin level increased as a function of fluoride concentration in drinking water, both in controls and in NX rats. It is concluded that the consumption of fluoridated water from water supply did not affect plasma glucose levels even in cases of animals with renal disease. However, a resistance to insulin action was demonstrated     

Inhibition of Streptococcus mutans biofilm accumulation and development of dental caries in vivo by 7-epiclusianone and fluoride

7-Epiclusianone (7-epi), a novel naturally occurring compound isolated from Rheedia brasiliensis, effectively inhibits the synthesis of exopolymers and biofilm formation by Streptococcus mutans. In the present study, the ability of 7-epi, alone or in combination with fluoride (F), to disrupt biofilm development and pathogenicity of S. mutans in vivo was examined using a rodent model of dental caries. Treatment (twice-daily, 60s exposure) with 7-epi, alone or in combination with 125 ppm F, resulted in biofilms with less biomass and fewer insoluble glucans than did those treated with vehicle-control, and they also displayed significant cariostatic effects in vivo (p < 0.05). The combination 7-epi + 125 ppm F was as effective as 250 ppm F (positive-control) in reducing the development of both smooth- and sulcal-caries. No histopathological alterations were observed in the animals after the experimental period. The data show that 7-epiclusianone is a novel and effective antibiofilm/anticaries agent, which may enhance the cariostatic properties of fluoride.     

Effect of boron as an antidote on dry matter intake, nutrient utilization and fluorine balance in buffalo (Bubalus bubalis) exposed to high fluoride ration

It is well known that excessive accumulation of fluorides can exert toxic effects on various tissues and organs so as to severely damage the health and production of animals. The aim of this study was to determine beneficial effect of boron on nutrient utilization in buffalo calves exposed to high fluoride (F) ration. For this purpose, we used three groups of four male Murrah buffalo calves (body weight 98-100 kg, aged 6-8 month) each. Control animal was given only basal diet and concentrate mixture. However, treatment I animals were fed basal diet, concentrate mixture, and F [as NaF, 60 ppm of dry matter (DM)]. The treatment II animals were fed basal diet, concentrate mixture, F (as NaF, 60 ppm of DM), and B (as sodium tetraborate, 140 ppm of DM). After 90 days of experimental feeding, a metabolism trial of 7 days duration was conducted to study the treatment effect on nutrient utilization of proximate nutrients, absorption, excretion, and retention of N, Ca, P, Fe, Zn, Cu, and F. Dietary F significantly (p < 0.05) depressed the dry matter intake and increased the apparent digestibility, absorption, and retention of F. However, boron supplementation significantly (p < 0.05) decreased the apparent digestibility, absorption, and retention of F and improved the dry matter intake, fecal excretion, and percent of absorbed F excreted via urine. Apparent digestibility of proximate nutrients (viz. DM, crude protein, crude fiber, ether extract, and nitrogen free extract) was unaffected on either F or F+B treatment. However, absorption and excretion of N, Ca, P, Fe, Zn, and Cu were affected significantly (p < 0.05) on F or F+B treatment. These findings suggest that fluoride-containing diet for short duration has effect on nutrient utilization, and boron at 140-ppm dose level, in general, antagonized the absorption and retention of F and also improved the feed intake in buffalo calves.     

The effect of high fluoride intake on tissue trace elements and histology of testicular tubules in the rat

1. Male Wistar rats were exposed to fluoride (F) at concentrations of 100- and 200 ppm in their drinking water for 6- and 16 weeks.2. The high F intake caused several-fold increase in the F concentrations in the testes and bone as compared with control rats, both after the 6- and 16wk exposure; the bone F, but not testicular F, appeared to increase with dose and time.3. F exposure (100- and 200 ppm) decreased significantly the concentrations of zinc (Zn) in the testes, plasma, liver and kidneys particularly in the 16 wk groups; in the bone Zn tended to increase, however.4. The iron concentrations of the testes and plasma were not affected by F, whereas those of the liver, kidneys and bone appeared to increase under the influence of F.5. The concentrations of copper and manganese in the testes, liver and kidneys were not changed by F exposure.6. Fifty percent of the 100- and 200 ppm F rats after 16 weeks exhibited histopathologic changes in the germinal epithelium of the testes, which resembled those in Zn-deficient rats.7. The data suggest that a deprivation of testicular Zn due to a high F intake may be directly responsible for the injury of testicular tubules.     

Evaluation of Fluoride-Induced Oxidative Stress in Rat Brain: A Multigeneration Study

Multigenerational evaluation was made in rats on exposure to high fluoride (100 and 200 ppm) to assess neurotoxic potential of fluoride in discrete areas of the brain in terms of lipid peroxidation and the activity of antioxidant enzyme system. The rats were given fluoride through drinking water (100 and 200 ppm) and maintained subsequently for three generations. Fluoride treatment significantly increased the lipid peroxidation and decreased the activity of antioxidant enzymes viz, catalase, superoxide dismutase, glutathione peroxidase, glutathione S-transferase, and glutathione level in first-generation rats and these alterations were more pronounced in the subsequent second and third-generation rats in both the doses tested. Decreased feed and water consumption, litter size and organ (brain) somatic index, marginal drop in body growth rate and mortality were observed in all three generations. Decreased antioxidant enzyme activity and increased malondialdehyde levels found in the present study might be related to oxidative damage that occurs variably in discrete regions of the brain. Results of this study can be taken as an index of neurotoxicity in rats exposed to water fluoridation over several generations.     

Fluoride in tissues of Krill Euphausia superba Dana and Meganyctiphanes norvegica M. Sars in relation to the moult cycle

Summary The fluoride content of whole animals and different tissues of the euphausiid species Euphausia superba and Meganyctiphanes norvegica was analyzed by two different and improved methods of isolation and determination. In contrast to other authors our findings show that the internal organs (muscle, hepatopancreas and hemolymph) contain less than 6 ppm d.w. fluoride this being the same order of magnitude as for vertebrates. The high concentrations reported by other authors must be mainly due to contamination of the soft tissue during storage (post-mortem migration of fluoride from shell) and/or contamination caused by minute fractions of cuticle during dissection. Over 99% of the total fluoride content is located in the cuticle (i.e. integument) of the euphausiids (2600 ppm/d.w. in E. superba and 3300 ppm/d.w. in M. norvegica in pleon cuticle). Analysis of F^- levels in relation to the moulting cycle showed that the uptake in both euphausiids occurs at a comparable and fast rate during the same physiological phase shortly after moult, parallel to the general construction of the cuticle. The internal organs show homeostasis in respect to fluoride. Accordingly, no internal deposition takes place, and F^- is reaccumulated from the external medium at each moult.This work was supported by grants from the DFG No. Ad 24/9 and Bu 548/1