Comprehensive insight into the alterations in the gut microbiome and the intestinal barrier as a consequence of iron deficiency anaemia.

BACKGROUND: Iron deficiency is the top leading cause of anaemia, whose treatment has been shown to deteriorate gut health. However, a comprehensive analysis of the intestinal barrier and the gut microbiome during IDA have not been performed to date. This study aims to delve further into the analysis of these two aspects, which will mean a step forward minimising the negative impact of iron supplements on intestinal health. METHODS: IDA was experimentally induced in an animal model. Shotgun sequencing was used to analyse the gut microbiome in the colonic region, while the intestinal barrier was studied through histological analyses, mRNA sequencing (RNA-Seq), qPCR and immunofluorescence. Determinations of lipopolysaccharide (LPS) and bacteria-specific immunoglobulins were performed to assess microbial translocation. RESULTS: Microbial metabolism in the colon shifted towards an increased production of certain amino acids, short chain fatty acids and nucleotides, with Clostridium species being enriched during IDA. Structural alterations of the colonic epithelium were shown by histological analysis. RNA-Seq revealed a downregulation of extracellular matrix-associated genes and proteins and an overall underdeveloped epithelium. Increased levels of serum LPS and an increased immune response against dysbiotic bacteria support an impairment in the integrity of the gut barrier during IDA. CONCLUSIONS: IDA negatively impacts the gut microbiome and the intestinal barrier, triggering an increased microbial translocation. This study emphasizes the deterioration of gut health during IDA and the fact that it should be addressed when treating the disease.

Fermented Goat's Milk Contributes to the Recovery of Iron Deficiency Anemia via Modulation of the Gut Microbiome.

Iron deficiency anemia (IDA) is a global public health concern affecting 1.6 billion people worldwide. The administration of iron supplements during the treatment of IDA adversely affects the intestinal barrier function and the composition and functionality of the intestinal microbiome, both of which are already altered during IDA. For this reason, it is of great interest to develop nutritional strategies aimed at alleviating these gut alterations associated with IDA and its treatment. In this sense, fermented goat's milk (FGM) was studied due to its nutritional quality. Our findings showed that in anemic animals the consumption of a FGM-based diet, compared to a standard diet, had positive modulatory effects on the intestinal microbiome. FGM-based diet restored intestinal dysbiosis, the intestinal barrier functionality, and bacterial translocation, contributing to a more efficient recovery of IDA. Therefore, FGM is a useful nutritional tool to ease intestinal alterations occurring during IDA and during its treatment.

Changes in Adiposity and Body Composition during Anemia Recovery with Goat or Cow Fermented Milks.

To date, no studies are available about adipose tissue modifications during anemia recovery; therefore, the aim of this study is to provide detailed information about adipose tissue homeostasis during anemia recovery with fermented milks. Forty male Wistar rats were placed on a pre-experimental period of 40 days, divided in two groups (normal-Fe diet and Fe-deficient diet). Then rats were fed fermented goat or cow milk-based diets with normal-Fe content during 30 days. Ghrelin and adiponectin decreased in both groups of animals fed fermented goat milk, whereas leptin and NEFA increased. UCP-1 decreased in anemic rats fed either fermented milk, and irisin greatly increased in both groups of animals fed fermented goat milk. Fermented goat milk reduces adiposity, inducing leptin elevation and ghrelin reduction. Conversely, plasma adiponectin concentrations decreased in animals fed fermented goat milk, showing an inverse correlation with NEFA, an important marker of lipid mobilization, indicating increased lipolysis. Irisin up-regulation in animals fed fermented goat milk contributes to a favorable metabolic profile and the browning of adipose tissue during anemia recovery.

Influencia del suplemento con inulina enriquecida con fructooligosacáridos sobre el contenido y la densidad mineral ósea tras el parto y la lactación en ratas / Influence of supplementation with oligofructose-enriched inulin on bone mineral content and density after delivery and lactation in rats

Introducción: la gestación y lactancia están relacionadas con pérdidas temporales en la densidad mineral ósea (DMO) materna. Una suplementación con calcio podría resultar beneficiosa para evitar la pérdida de masa ósea del esqueleto materno. Otros nutrientes como los prebióticos han sido identificados como responsables de un incremento en la absorción de minerales, pudiendo condicionar la mineralización ósea. Objetivo: estudiar el efecto de la suplementación de la dieta materna con el prebiótico inulina enriquecida con oligofructosa, durante la gestación y la lactancia sobre el contenido mineral óseo (CMO) y la DMO al final del periodo de lactancia. Métodos: las ratas gestantes fueron alimentadas con dieta estándar (grupo CC), dieta fortificada en calcio (grupo Ca) o enriquecida con el prebiótico inulina enriquecida con oligofructosa (grupo Pre) hasta el final del periodo de lactancia. Posteriormente se evaluó el CMO y DMO por absorciometría de rayos X (DEXA) y el pH del contenido cecal. Resultados: en términos generales, el grupo Pre presenta los mayores valores absolutos de CMO y DMO de entre los tres grupos, siendo en la tibia significativamente diferentes en los grupos CC y Pre frente al grupo Ca. El pH del contenido cecal del grupo Pre es signifi cativamente inferior al de los grupos CC y Ca. Conclusión: la suplementación con inulina enriquecida con oligofructosa, en condiciones nutricionales no deficientes en calcio, durante la gestación y la lactancia, ejerce una protección del esqueleto materno en las ratas y puede ser considerada como una estrategia nutricional para proteger la masa ósea materna en el periodo perinatal (AU)

Introduction: Pregnancy and lactation are related with temporary decreases in maternal bone mineral density (BMD). Calcium supplementation could be beneficial to prevent bone loss of maternal skeleton. Other nutrients, such as prebiotics have showed to produce an increase of the mineral absorption and therefore affecting bone mineralization. Objective: To study the effect of maternal diet supplementation with prebiotic oligofructose-enriched inulin during gestation and lactation on the maternal bone mineral content (BMC) and BMD at the end of lactation. Methods: Pregnant rats were fed with standard diet (CC group), calcium fortified diet (Ca group) or with prebiotic oligofructose-enriched inulin supplemented diet until the end of the lactation period. At weaning, bone mineral content (BMC) and BMD were determined by dual-energy X-ray absorptiometry and the pH of the cecal content was also determined. Results: In absolute terms, the highest BMD and BMC were found in the Pre group as compared with the other two groups being significant in the tibia when compared Pre group and CC group with Ca group. The pH of the cecal content in the Pre group was also significantly lower as compared with the other two groups. Conclusion: Prebiotic oligofructose-enriched inulin supplementation, in calcium no-deficient conditions, during gestation and lactation exerts a protection on maternal skeleton during pregnancy and lactation in the rats and could be considered as a plausible nutritional option for protecting maternal bone mass during these periods (AU)

Maternal Dietary Supplementation with Oligofructose-Enriched Inulin in Gestating/Lactating Rats Preserves Maternal Bone and Improves Bone Microarchitecture in Their Offspring.

Nutrition during pregnancy and lactation could exert a key role not only on maternal bone, but also could influence the skeletal development of the offspring. This study was performed in rats to assess the relationship between maternal dietary intake of prebiotic oligofructose-enriched inulin and its role in bone turnover during gestation and lactation, as well as its effect on offspring peak bone mass/architecture during early adulthood. Rat dams were fed either with standard rodent diet (CC group), calcium-fortified diet (Ca group), or prebiotic oligofructose-enriched inulin supplemented diet (Pre group), during the second half of gestation and lactation. Bone mineral density (BMD) and content (BMC), as well as micro-structure of dams and offspring at different stages were analysed. Dams in the Pre group had significantly higher trabecular thickness (Tb.Th), trabecular bone volume fraction (BV/TV) and smaller specific bone surface (BS/BV) of the tibia in comparison with CC dams. The Pre group offspring during early adulthood had an increase of the lumbar vertebra BMD when compared with offspring of CC and Ca groups. The Pre group offspring also showed significant increase versus CC in cancellous and cortical structural parameters of the lumbar vertebra 4 such as Tb.Th, cortical BMD and decreased BS/BV. The results indicate that oligofructose-enriched inulin supplementation can be considered as a plausible nutritional option for protecting against maternal bone loss during gestation and lactation preventing bone fragility and for optimizing peak bone mass and architecture of the offspring in order to increase bone strength.

Folic acid supplemented goat milk has beneficial effects on hepatic physiology, haematological status and antioxidant defence during chronic Fe repletion.

The aim of the current study was to asses the effect of goat or cow milk-based diets, either normal or Fe-overloaded and folic acid supplement on some aspects of hepatic physiology, enzymatic antioxidant defence and lipid peroxidation in liver, brain and erythrocyte of control and anaemic rats after chronic Fe repletion. 160 male Wistar rats were placed on 40 d in two groups, a control group receiving normal-Fe diet and the Fe-deficient group receiving low Fe diet. Lately, the rats were fed with goat and cow milk-based diets during 30 d, with normal-Fe content or Fe-overload and either with normal folic or folic acid supplemented. Fe-overload increased plasma alanine transaminase and aspartate transaminase levels when cow milk was supplied. Dietary folate supplementation reduced plasma transaminases levels in animals fed goat milk with chronic Fe overload. A remarkable increase in the superoxide dismutase activity was observed in the animals fed cow milk. Dietary folate supplement lead to a decrease on the activity of this enzyme in all the tissues studied with both milk-based diets. A concomitant increment in catalase was also observed. The increase in lipid peroxidation products levels in rats fed cow milk with Fe-overload, suggest an imbalance in the functioning of the enzymatic antioxidant defence. In conclusion, dietary folate-supplemented goat milk reduces both plasma transaminases levels, suggesting a hepatoprotective effect and has beneficial effects in situation of Fe-overload, improving the antioxidant enzymes activities and reducing lipid peroxidation.

Goat milk supplemented with folic acid protects cell biomolecules from oxidative stress-mediated damage after anaemia recovery in comparison with cow milk.

BACKGROUND: Fe overload is a common consequence of the anaemia treatment, increasing the oxidative stress and promoting the accumulation of damaged biomolecules, with the subsequently impairment of cell functions. Oxidative stress and the role of folic acid preventing free radical damage have been extensively studied; nevertheless, no studies are available about the influence of folic acid-supplemented goat milk consumption on the oxidative stress-mediated damage. AIM: The objective of the present study was to assess the influence of folic acid supplementation of goat milk- or cow milk-based diets, after Fe-overload treatment to palliate anaemia, on oxidative stress-mediated biomolecular damage in the liver, brain, erythrocytes, duodenal mucosa and plasma. METHODS: Control and anaemic rats were fed goat milk- or cow milk-based diets, either with normal Fe or Fe overload (450 mg/kg), and normal folic acid (2 mg/kg) or folic acid supplemented (40 mg/kg) for 30 days. RESULTS: During chronic Fe repletion, background DNA damage was significantly lower in anaemic rats fed folic acid-supplemented goat milk-based diet, as revealed by tail DNA (%), and folic acid-supplemented goat milk also had a beneficial effect, reducing the extent of lipid peroxidation in liver, plasma, erythrocytes and especially in brain and duodenal mucosa. Furthermore, protein oxidative damage was lower in anaemic rat duodenal mucosa for all goat milk-based diets. CONCLUSIONS: Folic acid supplement in goat milk avoids the undesirable effects of Fe overload during anaemia recovery in all the tissues studied, especially in the liver and duodenal mucosa, which are the tissues with higher exposition to dietary Fe.

A new insight to bone turnover: role of ω-3 polyunsaturated fatty acids.

Background. Evidence has shown that long-chain polyunsaturated fatty acids (LCPUFA), especially the ω -3 fatty acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are beneficial for bone health and turnover. Objectives. This review summarizes findings from both in vivo and in vitro studies and the effects of LC PUFA on bone metabolism, as well as the relationship with the oxidative stress, the inflammatory process, and obesity. Results. Some studies in humans indicate that LCPUFA can increase bone formation, affect peak bone mass in adolescents, and reduce bone loss. However, the cellular mechanisms of action of the LCPUFA are complex and involve modulation of fatty acid metabolites such as prostaglandins, resolvins and protectins, several signaling pathways, cytokines, and growth factors, although in certain aspects there is still some controversy. LCPUFA affect receptor activator of nuclear factor κ ß (RANK), a receptor found on the osteoclast, causing bone resorption, which controls osteoclast formation. Conclusions. Since fatty acids are an endogenous source of reactive oxygen species, free radicals alter the process of bone turnover; however, although there are clinical evidences linking bone metabolism and dietary lipids, more clinical trials are necessary to prove whether ω -3 PUFA supplementation plays a major role in bone health.

Influence of several sources and amounts of iron on DNA, lipid and protein oxidative damage during anaemia recovery.

The study was designed to assess the effect of several Fe amounts and sources on haematological parameters, DNA, lipid and protein oxidative damage during the course of Fe-deficiency anaemia recovery. Peripheral DNA damage was assessed using an alkaline comet assay. The brain, liver, erythrocyte and duodenal mucosa lipid peroxidation and protein damage were assessed in control and anaemic rats after Fe repletion with three different sources (FeSO4, haem Fe, and FeSO4 + haem Fe) and amounts (45, 12, and 31 mg Fe/kg diet) of Fe: F diet, H diet or C diet, respectively. After supplying the diets, the haematological parameters studied were recovered; being remarkable is the haemoglobin increase. The DNA damage was lower in rats with the H diet, as revealed by the percentage of DNA in head, tail and Olive tail moment compared in rats with the F (P < 0.001) and C (P < 0.05) diets. Lipid peroxidation was similar in all the tissues, except in the duodenal mucosa which was lower with H and C diets (P < 0.001). The animals fed with C diet showed lower oxidative protein damage in the duodenal mucosa (P < 0.001) and was also lower in the liver and erythrocytes for H and C diets (P < 0.001). No differences were found in the brain under our experimental conditions. In conclusion, Fe supplementation with low doses of haem Fe or combined forms of non-haem and haem Fe (FeSO4 + haem) are efficient in restoring the impaired haematological parameters and prevent the evoked oxidative stress associated with Fe supplements.

Positive influence of a natural product as propolis on antioxidant status and lipid peroxidation in senescent rats.

Given the importance of oxidative stress associated to aging, it would be interesting to assess the effect of oral supplementation with antioxidant substances capable of diminishing oxidative aggression and free radicals generation associated to this condition. This study investigated the effects of AIN-93 M diet supplemented either with 2 % of propolis, or with 4 % of a natural product obtained from lyophilizate vegetables, selected by its antioxidant properties, in senescent healthy Wistar rats fed ad libitum over 3 months. Propolis supplementation leads to a lower level of glucose and cholesterol concentrations together with a reduction in protein oxidation. Plasma thiobarbituric acid-reactive substance levels were lower in the rats consuming the natural vegetable product and propolis possibly due to its antioxidant components, neutralizing the free radical produced, and thus preventing cellular damage. The results of the present study suggest a synergic effect of overall propolis compounds reducing the oxidative stress and glucose and cholesterol plasma levels associated with aging.

Bile composition, plasma lipids and oxidative hepatic damage induced by calcium supplementation; effects of goat or cow milk consumption.

Calcium-fortified foods, especially milk and dairy products are recommended to be consumed daily for groups in risk of nutritional deficiency, including children, young adults, menopausal women, pregnant women and the elderly, however Ca-supplementation promotes gallstone formation because Ca is a nucleating factor. The objective of the current study was to assess the influence of cow or goat milk-based diets, either normal or Ca-supplemented, on bile composition, biochemical parameters and hepatic antioxidant status. Weanling male rats were randomly divided into six groups, fed standard, goat or cow milk-based diets, either with normal Ca content (5.0 g/kg), or Ca-supplemented (10.0 g/kg), for 2 weeks. Bile cholesterol concentration and output was higher in rats fed goat milk in comparison with those fed with standard and cow-milk-based diet. Ca-supplementation increased lithogenic index with the standard and cow-milk based diets, this change was not observed with the goat milk diet. Activities of plasma transaminases were also lower in the animals fed Ca-supplemented goat milk, in comparison with the other diets assayed. In general, Ca-supplement in the diet led to an increase in the hepatic oxidative damage, with an increase in the activities of all the antioxidant enzymes studied in the standard and cow milk diet, but not with goat milk. The habitual consumption of goat milk has positive effects on the plasma lipid profile, biliary composition and hepatic antioxidant defence. In addition, under our experimental conditions, Ca-supplementation of this type of milk does not increase the lithogenic index, or hepatic oxidative damage.

Severe nutritional iron-deficiency anaemia has a negative effect on some bone turnover biomarkers in rats.

BACKGROUND: The role of iron (Fe) in bone formation and disease have not received much attention, a fact that is interesting given the known biochemical role that this mineral has upon collagen maturation together with the high prevalence of Fe-deficiency anaemia worldwide. AIM: To investigate the changes in bone formation, resorption and mineral content in developing rats with induced nutritional Fe-deficiency anaemia. METHODS: Thirty male Wistar rats were divided into two groups, a control group receiving AIN-93G diet with normal-Fe content and an anaemic group receiving AIN-93G diet with low-Fe content for 40 days. Both diets were prepared with an adequate calcium (Ca) and phosphorus (P) content. The most representative serum bone turnover biomarkers and femur and sternum calcium and phosphorus content, together with sternum Fe content were determined in both experimental groups. RESULTS: In anaemic rats, bone matrix formation diminished as revealed by the lower amount of procollagen type I N-terminal propeptide. Bone resorption process increased in Fe deficiency as shown by the increase of serum parathyroid hormone, tartrate-resistant acid phosphatase and levels of degradation products from C-terminal telopeptides of type I collagen released to the serum. In addition, mineralization process was affected by Fe deficiency, because Ca and P content in femur decreased markedly. CONCLUSIONS: Fe-deficiency anaemia had a significant impact upon bone, affecting bone mineralization, decreasing the matrix formation and increasing bone resorption, therefore it is of great interest to assess bone status in situation of Fe-deficiency anaemia.

Se bioavailability and glutathione peroxidase activity in iron deficient rats.

Little information is available on the relationship of Se deposit in target organs and GPx activity in iron deficiency anemia. As red blood cells (RBCs) play a crucial role on Se metabolism and during Fe deficiency anemia a lower count of RBCs is featured, we aimed to investigate the influence of this pathology on Se bioavailability and the relationship with antioxidant status. 20 male Wistar rats were randomly divided into two groups, a control group receiving AIN-93G diet with normal Fe content (45mg/kg diet) and the Fe-deficient group receiving AIN-93G diet with low Fe content (5mg/kg diet) for 40 days. Both diets were prepared with an adequate Se content (0.180mg/kg diet). The digestive and metabolic utilization of Se, the distribution in target organ, the GPx activity and TBARS production were measured after receiving the diets. Se retention increased (P<0.001) in the anemic group, fact that contributes to keep the enzymatic antioxidant activity of GPx in normal levels and the tendency observed is that stored Se increased in the organs, especially in kidney (P<0.01), however, a lower Se deposit was found in sternum of anemic rats (P<0.001). The lower count of RBCs featured in this pathology (P<0.001) causes a decrease of Se concentration in sternum meanwhile the increase in kidney deposit is a consequence of the lower urinary losses (P<0.001).

Effect of synergic dietary calcium enrichment and induced ferropenic anemia on antioxidant enzymes activity in rats.

OBJECTIVE: The aim of this study was to examine the synergism of dietary calcium enrichment (added to goat's or cow's milk) and induced nutritional ferropenic anemia on oxidative status. METHODS: Control rats and rats with induced nutritional ferropenic anemia were fed for 14 d with diets containing normal (5000 mg/kg) or double (10 000 mg/kg) the recommended calcium content. Thiobarbituric acid-reactive substances in plasma were measured, as were the activities of the antioxidant enzyme catalase, copper/zinc superoxide dismutase, and glutathione peroxidase in erythrocyte cytosol. RESULTS: Dietary calcium enrichment did not affect oxidative stress as assessed by thiobarbituric acid-reactive substances; however, it significantly upregulated the activities of some antioxidant enzymes examined in the erythrocyte cytosol. In particular, adding calcium to standard or milk-based diets significantly increased glutathione peroxidase activity in control and anemic rats and copper/zinc superoxide dismutase activity in control rats. CONCLUSION: The increased activities of glutathione peroxidase and copper/zinc superoxide dismutase induced by dietary calcium enrichment suggest that calcium supplementation may protect against oxidative stress even in nutritionally induced ferropenic anemia.

Calcium-enriched goats' milk aids recovery of iron status better than calcium-enriched cows' milk, in rats with nutritional ferropenic anaemia.

Ca-Fe interactions are known, but no studies are available about the effects of Ca-enriched goat or cow milk on Fe status in nutritional ferropenic anaemia (NFA). To examine this matter, control and Fe-deficient rats were fed for 14 d with goat or cow milk diets containing either normal or high Ca content (5000 or 10,000 mg/kg diet), and different indices and parameters related to iron status were measured. The apparent digestibility coefficient (ADC) and the Fe retention/intake (R/I) ratio were higher in control and anaemic rats fed goat milk diet (G diet), despite high-Ca content. Ca enrichment decreased Fe stores in liver and sternum in anaemic rats fed cow milk diet (C diet), however G diet did not modify Fe content in the organs studied in control and anaemic rats. In anaemic rats, Ca-supplementation decreased haematocrit, but platelets and serum Fe were not affected, however, in control rats platelets increased except for Ca-enriched G diet, this fact reveals that Ca-Fe interaction is minimized with G diet. Serum ferritin was always higher in rats fed G vs. C diet, both in control and anaemic rats fed either normal or Ca-enriched diets. Ca-supplementation decreased ferritin levels in control and anaemic rats fed C diet and also, though to a lesser extent, in those given the G diet. This indicates that with this G diet there is a better recovery of body Fe stores in anaemic rats, despite Ca-supplementation. In this study it is noteworthy that despite high Ca content, a goat milk diet resulted in minimal Ca-Fe interactions and did not adversely affect Fe status in rats with NFA.

Effects of goats' or cows' milks on nutritive utilization of calcium and phosphorus in rats with intestinal resection.

We analysed the effects of goats' milk (GM) on the nutritive utilization of Ca and P in rats with resection of 50 % distal small intestine in comparison with cows' milk (CM) and a standard non-milk diet. The three test diets contained 200 g protein and 100 g fat/kg. The apparent digestibility coefficient (ADC) of Ca and P were considerably higher in the two groups of rats given the GM diet than those given the other two diets. Ca and P retention did not decrease by effect of intestinal resection with GM diet. In both groups of animals, serum Ca and P levels and ionic Ca were higher in the case of the GM diet than the other two diets, whereas the parathyroid hormone levels were lower. Ca content in femur, sternum and longissimus dorsi muscle was higher in rats given the GM diet. P content in femur and sternum was higher among the two groups of rats given a milk-based diet (GM or CM), especially with GM diet. The GM diet has beneficial effects on nutritive utilization of Ca and P in control rats and those with resection of the distal small intestine.

Effect of dietary inclusion of goat milk on the bioavailability of zinc and selenium in rats.

The effects of dietary inclusion of freeze-dried goat and cow milk on the utilization of zinc and selenium, and on the metabolic fate of zinc, were studied in transected (control) and resected rats (resection of 50% of the distal small intestine). Intestinal resection reduced the apparent digestibility coefficient and zinc retention in the cow milk diet, whereas these biological indices were similar in transected (control) and resected rats with standard (without milk) and goat milk diets. The apparent digestibility coefficient and retention of selenium were not affected by intestinal resection in the animals fed with the three diets studied. In transected (control) and resected rats, the apparent digestibility coefficient and retention of zinc and selenium were higher for the goat milk diet than for the other two diets. Zinc deposits in the organs, expressed as microg/g dry weight were in order of decreasing concentrations: femur, testes, sternum, liver, kidney, heart, spleen, longissimus dorsi muscle and brain. Deposits were greatest with the goat milk diet, followed by the standard diet and were lowest for the rats given the cow milk diet, both for transected (control) and resected animals. We conclude that consumption of the goat milk diet produces a greater bioavailability of zinc and selenium and a greater deposit of zinc in key organs, for both the transected (control) and the resected rats, with respect to the standard diet and the cow milk diet.