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Chicken eggshell (ES) waste is a rich source of calcium carbonate (CaCO 3 ); however, the potential of ES as dietary calcium (Ca) in old laying hens has not been explored. This study compared the effects of feeding limestone, cockle shell, oyster shell, fine ES, and coarse ES as the sole Ca source on production performance, egg quality, blood biochemical constituents, and tibia characteristics in old laying hens. A total of 450 ISA-Brown laying hens at 73 wk of age with similar egg production rate (EPR) were randomly assigned to 5 treatment groups (90 hens/group, 9 hens/replicate) for 7 wk. Dietary treatment groups comprised a corn-soybean meal based diet containing different Ca sources: (i) limestone (LS; < 2 mm and 2–4 mm mixed in the ratio of 3:7) as control, (ii) cockle shell (CS; 1–4 mm), (iii) oyster shell (OS; 3–16 mm), (iv) ES fine particles (ESF; < 1 mm), and (v) ES coarse particles (ESC; 3–5 mm). Results indicated that dietary inclusion of coarse ES particles significantly increased average egg weight ( P  < 0.001) and daily egg mass ( P  < 0.05), and decreased feed conversion ratio ( P  < 0.001) as compared with the other treatments. However, no significant differences in EPR, feed intake, cracked egg proportion, and mortality were observed among the dietary treatments ( P  > 0.05). Notably, the use of ESF led to a lower proportion of cracked eggs than ESC ( P  < 0.05). ESC fed hens produced the heaviest eggs whereas CS fed hens produced the lightest ( P  < 0.001); the particle size of ES also affected the egg weight ( P  < 0.05). The eggs from OS and ESC fed hens showed a greater albumen height in comparison to eggs from CS group ( P  < 0.05); but no significant difference was observed among the LS, OS, ESF, and ESC groups (P > 0.05). The yolk color was darker in the eggs of group ESF as compared with other dietary groups ( P  < 0.01). However, no significant effects on Haugh units and shell properties were observed among the treatments ( P  > 0.05). The blood biochemistry results were not affected by the dietary Ca ( P  > 0.05) except for lower levels of high-density lipoprotein percentage (HDL %) in OS and ESC fed hens ( P  < 0.05). The tibia characteristics including weight, length, width, and breaking strength did not differ among the dietary groups ( P  > 0.05). However, the ESC and OS fed hens showed higher tibia bone mineral density (BMD) than the other groups ( P  < 0.001). In conclusion, coarse ES as a sole Ca source had beneficial effects on the production performance, egg quality, and tibia BMD in old laying hens.

Background In recent years, high phosphate intakes were discussed critically. In the small intestine, a part of the ingested phosphate and calcium precipitates to amorphous calcium phosphate (ACP), which in turn can precipitate other intestinal substances, thus leading to a beneficial modulation of the intestinal environment. Therefore, we analysed faecal samples obtained from a human intervention study regarding gut-related parameters. Methods Sixty-two healthy subjects (men, n  = 30; women, n  = 32) completed the double-blind, placebo-controlled and parallel designed study (mean age: 29 ± 7 years; mean BMI: 24 ± 3 kg/m 2 ). Supplements were monosodium phosphate and calcium carbonate. During the first 2 weeks, all groups consumed a placebo sherbet powder, and afterwards a sherbet powder for 8 weeks according to the intervention group: P1000/Ca0 (1000 mg/d phosphorus), P1000/Ca500 (1000 mg/d phosphorus and 500 mg/d calcium) and P1000/Ca1000 (1000 mg/d phosphorus and 1000 mg/d calcium). After the placebo period and after 8 weeks of intervention faecal collections took place. We determined in faeces: short-chain fatty acids (SCFA) and fat as well as the composition of the microbiome (subgroup) and cyto- and genotoxicity of faecal water (FW). By questionnaire evaluation we examined tolerability of the used phosphorus supplement. Results Faecal fat concentrations did not change significantly due to the interventions. Concentrations of faecal total SCFA and acetate were significantly higher after 8 weeks of P1000/Ca500 supplementation compared to the P1000/Ca0 supplementation. In men, faecal total SCFA and acetate concentrations were significantly higher after 8 weeks in the P1000/Ca1000 group compared to the P1000/Ca0 one. None of the interventions markedly affected cyto- and genotoxic activity of FW. Men of the P1000/Ca1000 intervention had a significantly different gut microbial community compared to the men of the P1000/Ca0 and P1000/Ca500 ones. The genus Clostridium XVIII was significantly more abundant in men of the P1000/Ca1000 intervention group compared to the other groups. Supplementations did not cause increased intestinal distress. Conclusions The used high phosphorus diet did not influence cyto- and genotoxicity of FW and the concentrations of faecal fat independent of calcium intake. Our study provides first hints for a potential phosphorus-induced modulation of the gut community and the faecal total SCFA content. Trial registration The trial is registered at ClinicalTrials.gov as NCT02095392 .

Ocean warming and acidification threaten the future growth of coral reefs. This is because the calcifying coral reef taxa that construct the calcium carbonate frameworks and cement the reef together are highly sensitive to ocean warming and acidification. However, the global-scale effects of ocean warming and acidification on rates of coral reef net carbonate production remain poorly constrained despite a wealth of studies assessing their effects on the calcification of individual organisms. Here, we present global estimates of projected future changes in coral reef net carbonate production under ocean warming and acidification. We apply a meta-analysis of responses of coral reef taxa calcification and bioerosion rates to predicted changes in coral cover driven by climate change to estimate the net carbonate production rates of 183 reefs worldwide by 2050 and 2100. We forecast mean global reef net carbonate production under representative concentration pathways (RCP) 2.6, 4.5, and 8.5 will decline by 76, 149, and 156%, respectively, by 2100. While 63% of reefs are projected to continue to accrete by 2100 under RCP2.6, 94% will be eroding by 2050 under RCP8.5, and no reefs will continue to accrete at rates matching projected sea level rise under RCP4.5 or 8.5 by 2100. Projected reduced coral cover due to bleaching events predominately drives these declines rather than the direct physiological impacts of ocean warming and acidification on calcification or bioerosion. Presently degraded reefs were also more sensitive in our analysis. These findings highlight the low likelihood that the world’s coral reefs will maintain their functional roles without near-term stabilization of atmospheric CO₂ emissions.

Calcium carbonate formation is the primary pathway by which carbon is returned from the ocean–atmosphere system to the solid Earth 1 , 2 . The removal of dissolved inorganic carbon from seawater by precipitation of carbonate minerals—the marine carbonate factory—plays a critical role in shaping marine biogeochemical cycling 1 , 2 . A paucity of empirical constraints has led to widely divergent views on how the marine carbonate factory has changed over time 3 – 5 . Here we use geochemical insights from stable strontium isotopes to provide a new perspective on the evolution of the marine carbonate factory and carbonate mineral saturation states. Although the production of carbonates in the surface ocean and in shallow seafloor settings have been widely considered the predominant carbonate sinks for most of the history of the Earth 6 , we propose that alternative processes—such as porewater production of authigenic carbonates—may have represented a major carbonate sink throughout the Precambrian. Our results also suggest that the rise of the skeletal carbonate factory decreased seawater carbonate saturation states. Geochemical insights from a dataset of carbonate stable strontium isotopes suggest that porewater production of authigenic carbonates may have been an overlooked carbonate sink for much of Earth’s history.

Depending on toothpaste formulation, part of the fluoride is insoluble and would not be totally absorbable in the gastrointestinal tract, thus changing dental fluorosis risk estimation. This hypothesis was tested with formulations with either all fluoride in a soluble form (NaF/SiO 2 -based toothpaste, 1,100 µg F/g as labeled, 1,129.7 ± 49.4 µg F/g soluble fluoride as analyzed) or with around 20% of insoluble fluoride (Na 2 FPO 3 /CaCO 3 -based toothpaste, 1,450 µg F/g as labeled, 1,122.4 ± 76.4 µg F/g soluble fluoride as analyzed). Toothpastes were evaluated either fresh or after accelerated aging, which increased insoluble fluoride to 40% in the Na 2 FPO 3 /CaCO 3 -based toothpaste. In a blind, crossover clinical trial conducted in five legs, 20 adult volunteers ingested 49.5 µg of total fluoride/kg body weight from each formulation or purified water (control). Whole saliva and urine were collected as bioavailability indicators, and pharmacokinetics parameters calculated showed significantly (p < 0.05) lower fluoride bioavailability for Na 2 FPO 3 /CaCO 3 toothpaste, which was reduced further after aging. A significant correlation between the amount of soluble fluoride ingested, but not total fluoride, and fluoride bioavailability was found (r = 0.57, p < 0.0001). The findings suggest that the estimated fluorosis risk as a result of ingestion of Na 2 FPO 3 /CaCO 3 -based toothpastes should be calculated based on the toothpaste’s soluble rather than total fluoride concentration.

Calcium supplementation is known to increase bone mineral density and decrease fractures, but the relative efficacy of different forms of calcium supplementation is not established. We compared the effects of calcium carbonate and calcium citrate on markers of bone resorption in older postmenopausal women in an open-labeled crossover study. Forty women were randomized to receive 1000 mg/day of either calcium citrate or calcium carbonate for 12 weeks, followed by a 2-week washout without calcium supplements and 12 weeks treatment with the alternate calcium supplement. All women received vitamin D (900 IU/day). Thirty-four women (25 Caucasian, nine Hispanic) completed the study. No significant differences in the decrease in parathyroid hormone (PTH) or bone specific alkaline phosphatase or the increase in urinary calcium/creatinine were detected between the two treatments. However, calcium citrate supplementation decreased the collagen cross-link resorption markers, urinary N-telopeptide (-30%), C-telopeptide (-31%), free deoxypyridinoline (19%) and serum N-telopeptide (-8%), compared to no significant change following calcium carbonate supplementation (+2%, +3%, +2% and +2%, respectively; P<0.05). Calcium citrate decreased markers of bone resorption significantly more than calcium carbonate in postmenopausal women, although no differences in their effects in calcium excretion or PTH were detected.

Concrete cracks must be repaired promptly in order to prevent structural damage and to prolong the structural life of the building (or other such construction). Biological self-healing concrete is a recent alternative technology involving the biochemical reaction of microbial induced calcium carbonate precipitation (MICP). This study determined the most appropriate technique to encapsulate spores of Bacillus sphaericus LMG 22257 with sodium alginate so as to protect the bacterial spores during the concrete mixing and hardening period. Three techniques (extrusion, spray drying and freeze drying) to encapsulate the bacterial spores with sodium alginate were evaluated. The freeze-drying process provided the highest bacterial spore survival rate (100%), while the extruded and spray-dried processes had a lower spore survival rate of 93.8% and 79.9%, respectively. To investigate the viability of microencapsulated spores after being mixed with mortar, the decomposed urea analysis was conducted. The results revealed that the freeze-dried spores also showed the highest level of urea decomposition (metabolic activity assay used as a surrogate marker of spore germination and vegetative cell viability). Thus, the self-healing performance of concrete mixed with freeze-dried spores was evaluated. The results showed that the crack healing ratio observed from the mortar specimens with freeze-dried microencapsulated spores were significantly higher than those without bacteria. This study revealed that freeze drying has a high potential as a microencapsulation technique for application to self-healing concrete technology.

Climate change is increasing the need to limit levels of anthropogenic CO 2 released into the atmosphere. One approach being investigated is to generate products based on microbially induced carbonate precipitation (MICP), which can trap CO 2 as CaCO 3 . We recently identified a novel MICP pathway in bacteria that is initiated by Ca 2+ toxicity in cells, causing extracellular CO 2 to be trapped as CO 3 2- by Escherichia coli, although the yield of precipitated CaCO 3 remained low (in the milligram range). In this work, we used the E. coli Keio gene knock-out library to identify 54 genes involved in MICP in E. coli, which could be broadly characterized into four groups: central metabolism, iron metabolism, cell architecture, and transport. The role of central metabolism appears to be crucial in maintaining alkaline conditions surrounding the cell that promote CaCO 3 precipitation. The role of iron metabolism was less clear, although the results suggest that growth rate influences the initiation of MICP. While the impact of repeating polymeric structures on cell surfaces promoting MICP is well established, our results suggest that other structural features may play a role, including fimbriae and flagella. Finally, the results confirmed that Ca 2+ transport is central to MICP under calcium stress. The results further suggest that the ZntB efflux pump may play a previously unidentified role in Ca 2+ transport in E. coli. By overexpressing some of these genes, our work suggests that there are several previously unidentified cellular mechanisms that could serve as a target for enhanced MICP in E. coli. By incorporating these processes into MICP pathways in E. coli, it may be possible to increase the volume of CO 2 fixed using this pathway and yield potentially new products that can replace CO 2 intensive products, such as precipitated calcium carbonates (PCCs) for industry.

Biodeposition of minerals is a widespread phenomenon in the biological world and is mediated by bacteria, fungi, protists, and plants. Calcium carbonate is one of those minerals that naturally precipitate as a by-product of microbial metabolic activities. Over recent years, microbially induced calcium carbonate precipitation (MICP) has been proposed as a potent solution to address many environmental and engineering issues. However, for being a viable alternative to conventional techniques as well as being financially and industrially competitive, various challenges need to be overcome. In this review, the detailed metabolic pathways, including ammonification of amino acids, dissimilatory reduction of nitrate, and urea degradation (ureolysis), along with the potent bacteria and the favorable conditions for precipitation of calcium carbonate, are explained. Moreover, this review highlights the potential environmental and engineering applications of MICP, including restoration of stones and concrete, improvement of soil properties, sand consolidation, bioremediation of contaminants, and carbon dioxide sequestration. The key research and development questions necessary for near future large-scale applications of this innovative technology are also discussed.

Crystallization by particle attachment (CPA) of amorphous precursors has been demonstrated in modern biomineralized skeletons across a broad phylogenetic range of animals. Precisely the same precursors, hydrated (ACC-H₂O) and anhydrous calcium carbonate (ACC), have been observed spectromicroscopically in echinoderms, mollusks, and cnidarians, phyla drawn from the 3 major clades of eumetazoans. Scanning electron microscopy (SEM) here also shows evidence of CPA in tunicate chordates. This is surprising, as species in these clades have no common ancestor that formed a mineralized skeleton and appear to have evolved carbonate biomineralization independently millions of years after their late Neoproterozoic divergence. Here we correlate the occurrence of CPA from ACC precursor particles with nanoparticulate fabric and then use the latter to investigate the antiquity of the former. SEM images of early biominerals from Ediacaran and Cambrian shelly fossils show that these early calcifiers used attachment of ACC particles to form their biominerals. The convergent evolution of biomineral CPA may have been dictated by the same thermodynamics and kinetics as we observe today.