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doi: 10.15389/agrobiology.2025.4.722eng

UDC: 636.085

Acknowledgements:
Lab studies were carried out at the Center for Collective Use of the BST RAS (http://цкп-бст.рф).
Supported financially by the Russian Science Foundation, project No. 23-16-00165.

 

INFLUENCE OF VARIOUS FORMS OF CALCIUM IN THE DIET ON THE MICRO- AND MACROELEMENT PROFILES OF ORGANS AND TISSUES IN RATS

Е.А. Sizova1, 2 , T.N. Kholodilina1, 2, К.S. Nechitailo1, 2,
K.V. Ryazantseva1, D.E. Shoshin1, 2, E.V. Yausheva1, S.V. Lebedev1

1Federal Research Centre of Biological Systems and Agrotechnologies RAS, 29, ul. 9 Yanvarya, Orenburg, 460000 Russia, e-mail izova.L78@yandex.ru (✉ corresponding author), olodilina@rambler.ru, k.nechit@mail.ru, reger94@bk.ru, aniilshoshin@mail.ru, asilena56@mail.ru, sv74@list.ru;
2Orenburg State University, 13, prosp. Pobedy, Orenburg, 460018 Russia

ORCID:
Sizova Е.А. orcid.org/0000-0002-5125-5981
Shoshin D.E. orcid.org/0000-0003-3086-681X
Kholodilina T.N. orcid.org/0000-0002-3946-8247
Yausheva E.V. orcid.org/0000-0002-1589-2211
Nechitailo К.S. orcid.org/0000-0002-8755-414X
Lebedev S.V. orcid.org/0000-0001-9485-7010
Ryazantseva K.V. orcid.org/0000-0001-5134-0396

Final revision received December 11, 2024
Accepted April 10, 2025

The problems of the body’'s calcium supply have several etiologies. Firstly, calcium in the diet should have a high biological availability which requires the selection of forms of calcium-containing substances that meet this criterion. Secondly, feed producers tend to over-saturate diets with calcium or it is added to improve physical properties during the processing of certain types of waste from the grain processing industry, e.g., soybean meal, but such inclusions are often ignored. Despite the widespread scientific study of the issue, the search for the most accessible form of calcium for the animal body is still relevant. In this work, the elemental profile of target organs in Wistar rats with an excess of various forms of calcium was established for the first time. The metabolic relationships of macro- and microelements, including calcium-conjugated ones, have been revealed, depending on the organ and the form of the injected substances. The aim of the work was studing the concentration of macro-, micro- and toxic elements in the liver, kidneys, femur and blood serum of Wistar rats with the additional use of various sources of calcium in the diet: chloride, citrate and carbonate. The work was performed in the vivarium of the Federal Research Center for Biological Systems and Agrotechnologies RAS (Orenburg) in 2024 on male Wistar rats aged 6 months. The duration of the experimental reference period was 21 days. After the preparatory period (10 days), 20 rats were randomly divided into four groups (n = 5): a control group and three experimental groups. In addition to the basic diet, 45% of the daily calcium requirement was administered to the animals of the experimental groups in the form of CaCl2 for group I, Ca3(C6H5O7)2 for group II, and CaCO3 for group III. The amount of feed consumed, according to the daily requirement for this type of animal, was 30 g/individual. The starting material for the preparation of various forms of calcium was limestone flour (CaCO3) (Akkerman Cement LLC, Russia). For the synthesis of CaCl2, the method of transferring Ca2+ ions into solution by reaction with hydrochloric acid was used. The citrate form of Ca3(C6H5O7)2 was obtained by reacting a solution of citric acid in one water (concentration less than 10 %) with limestone flour. Biomaterial (blood, liver, kidneys, femur) for the study was obtained after decapitation of rats under inhalation anesthesia 21 days after the start of the experiment. The elemental analysis of the substrates was performed by mass spectrometry on a single quadrupole mass spectrometer with inductively coupled plasma Agilent 7900 ICP-MS (Agilent, USA). Thermo-oxidative degradation of the organic matrix was carried out in a microwave TOPEX+ sample preparation system (PREEKEM, China). The content of Cr, Fe, Zn, and As was analyzed in the helium regime using a collision cell. The concentration of certain elements (Ca, P, Mg, Fe) in blood serum was determined using an automatic biochemical analyzer CS-T240 (DIRUI Industrial Co., Ltd., China). Analyzing the elemental profile of rat organs and tissues when various forms of calcium were added to the diet, we found that they accumulated toxic and conditionally toxic elements (Pb, Ba, Al) to a greater extent with dietary Ca chloride and carbonate, and to a lesser extent (Bi) when using the citrate form in the diet. The body lost toxic and conditionally toxic elements upon the use of Ca3(C6H5O7)2 (five elements, V, Al, Ba, Sr, Sn), CaCl2 (four elements, Cd, Al, Ba, Sn), CaCO3 (one element, Sn). In turn, the forms of calcium-containing substances did not affect the concentration of calcium-dependent elements. The experiment did not establish a critical decrease in their number. A tendency to a decrease in the content of iron and nickel in the femur was revealed with the accumulation of selenium and barium in the liver, and nickel and chromium in the kidneys in all groups. Calcium accumulated only in the liver after administration of CaCl2. There was a tendency to decrease the content of copper, iron, zinc, magnesium, and manganese in the blood serum when using all forms of calcium. The results obtained made it possible to evaluate the metabolism of calcium, its synergists and antagonists under the influence of the studied factor, which in practice can ensure the optimization of dietary rationing and the prioritization of a particular form of calcium, especially at the risk of its overdose.

Keywords: forms of calcium, trace elements, macronutrients, liver, kidneys, femur, blood serum.

 

REFERENCES

  1. Walk C.L. The influence of calcium on phytase efficacy in non-ruminant animals. Animal Production Science, 2016, 56(8): 1345-1349 CrossRef
  2. Ravindran V., Abdollahi M.R., Bootwalla S.M. Nutrient analysis, metabolizable energy, and digestible amino acids of soybean meals of different origins for broilers. Poultry Science, 2014, 93(10): 2567-2577 CrossRef
  3. David L.S., Anwar M.N., Abdollahi M.R., Bedford M.R., Ravindran V. Calcium nutrition of broilers: current perspectives and challenges. Animals, 2023, 13(10): 1590 CrossRef
  4. Khan S., Mosvi S.N., Vohra S., Poddar N.K. Implication of calcium supplementations in health and diseases with special focus on colorectal cancer. Critical Reviews in Clinical Laboratory Sciences, 2024, 61(6): 496-509 CrossRef
  5. Ma B.N., Vo C.B. Role, absorption, and transport of calcium in the human body: a review. The Journal of Agriculture and Develoment, 2022, 21(4): 70-80 CrossRef
  6. Gromova O.A., Torshin I.Yu., Tomilova I.K., Gilel’s A.V., Demidov V.I. RMZh, 2016, 15: 1009-1017 (in Russ.).
  7. Wang T., Gu Q., Zhao J., Mei J., Shao M., Pan Y., Zhang J., Wu H., Zhang Z., Liu F. Calcium alginate enhances wound healing by up-regulating the ratio of collagen types I/III in diabetic rats. International Journal of Clinical and Experimental Pathology, 2015, 8(6): 6636-6645.
  8. Clines D.J.A., Dai Y., Heng, D., Aslanova F.B. Calcium. In: Encyclopedia of human nutrition (fourth edition). B. Caballero (ed.). Elsevier eBooks, 2023: 105-113 CrossRef
  9. Mann E., Schmitz-Esser S., Zebeli Q., Wagner M., Ritzmann M., Metzler-Zebeli B.U. Mucosa-associated bacterial microbiome of the gastrointestinal tract of weaned pigs and dynamics linked to dietary calcium-phosphorus. PLoS ONE, 2014, 9(1): e86950 CrossRef
  10. Bovee-Oudenhoven I.M., Wissink M.L., Wouters J.T., Van der Meer R. Dietary calcium phosphate stimulates intestinal lactobacilli and decreases the severity of a salmonella infection in rats. The Journal of Nutrition, 1999, 129(3): 607-612 CrossRef
  11. Heinola T., Jukola E., Näkki P., Sukura A.Consequences of hazardous dietary calcium deficiency for fattening bulls. Acta Veterinaria Scandinavica,2006, 48(1): 25 CrossRef
  12. Abaturov A.E., Kryuchko T.A., Krivusha E.L., Tkachenko O.Ya. Zdorov’e rebenka, 2018, 13(7): 681-690 (in Russ.).
  13. Bao S.F., Windisch W., Kirchgessner M. Calcium bioavailability of different organic and inorganic dietary Ca sources (citrate, lactate, acetate, oyster‐shell, eggshell, b‐tri‐Ca phosphate). Journal of Animal Physiology and Animal Nutrition, 1997, 78: 154-160 CrossRef
  14. Wiria M.S.S., Tran H.M., Nguyen P.H.B., Valencia O., Dutta S., Pouteau E. Relative bioavailability and pharmacokinetic comparison of calcium glucoheptonate with calcium carbonate. Pharmacol. Res. Perspect., 2020, 8(2): e00589 CrossRef
  15. Recker R.R., Hinders S., Davies K.M., Heaney R.P., Stegman M.R., Lappe J.M., Kimmel D.B. Correcting calcium nutritional deficiency prevents spine fractures in elderly women. Journal of Bone and Mineral Research, 1996, 11(12): 1961-1966 CrossRef
  16. Khatri K., Kaur M., Dhir T., Kankaria A., Arora H. Role of calcium &/or vitamin D supplementation in preventing osteoporotic fracture in the elderly: A systematic review & meta-analysis. Indian Journal of Medical Research, 2023, 158(1): 5-16 CrossRef
  17. Kochanowski B.A. Effect of calcium citrate-malate on skeletal development in young, growing rats. Journal of Nutrition, 1990, 120(8): 876-881 CrossRef
  18. Karkach P.M., Kostiuk M.Y., Mashkin Y. Correction of calcium norms during the feeding day of laying-hens. Tehnologìâ Virobnictva ì Pererobki Produktìv Tvarinnictva, 2021, 1(164): 42-47 CrossRef
  19. Santana A.L.A., de Oliveira Carvalho P.L., Cristofori E.C., da Silva Chambo P.C., Barbizan M., Nunes R.V., Gregory C.R., Genova J.L. Supplementation of pig diets in the growth and termination phases with different calcium sources. Tropical Animal Health and Production, 2018, 50(3): 477-484 CrossRef
  20. Yang A., Wang K., Peng X., Lv F., Wang Y., Cui Y., Wang Y., Qu D., Zhou J., Si H. Effects of different sources of calcium in the diet on growth performance, blood metabolic parameters, and intestinal bacterial community and function of weaned piglets. Frontiers in Nutrition, 2022, 9: 885497 CrossRef
  21. Rotshteyn S. Effektivnoe zhivotnovodstvo, 2021, 2(168): 64-67 (in Russ.).
  22. Rubinov A.A., Mokhova E.V. Sbornik XVII Mezhdunarodnoy nauchno-prakticheskoy studencheskoy konferentsii «Khimiya i zhizn’» [Proc. XVII Int. Student Conf. «Chemistry and life»].Novosibirsk, 2018: 130-133 (in Russ.).
  23. Mako A.A., Mosuro A.O., Adedeji B.S., Jemiseye F.O., Abokede T. Comparative use of oyster shell and limestone as sources of calcium in the diet of laying chickens. Nigerian Journal of Animal Production, 2017, 44(1): 275-281 CrossRef
  24. Shafey T.M., McDonald M.W., Dingle J.G. Effects of dietary calcium and available phosphorus concentration on digesta pH and on the availability of calcium, iron, magnesium and zinc from the intestinal contents of meat chickens. British Poultry Science, 1991, 32(1): 185-194 CrossRef
  25. Tancharoenrat P., Ravindran V. Influence of tallow and calcium concentrations on the performance and energy and nutrient utilisation in broiler starters. Poultry Science, 2014, 93(6): 1453-1462 CrossRef
  26. Selle P.H., Cowieson A.J., Ravindran V. Consequences of calcium interactions with phytate and phytase for poultry and pigs. Livestock Science, 2009, 124(1-3): 126-141 CrossRef
  27. Stangl G.I., Kirchgessner M. Effect of different degrees of moderate iron deficiency on the activities of tricarboxylic acid cycle enzymes, and the cytochrome oxidase, and the iron, copper, and zinc concentrations in rat tissues. Zeitschrift für Ernahrungswissenschaft, 1998, 37(3): 260-268 CrossRef
  28. Walter A., Rimbach G., Most E., Pallauf J. Effects of citric acid supplements to a maize-soya diet on the in vitro availability of minerals, trace elements, and heavy metals. Journal of Veterinary Medicine Series A, 1998, 45(9): 517-24 CrossRef
  29. Bronner F. Calcium absorption—a paradigm for mineral absorption. The Journal of Nutrition, 1998, 128(5): 917-920 CrossRef.
  30. Vitti D.M., Silva Fiho S.S., Louvandini J.C.D., Dias H., Bueno R.S., Kebreab E. Phosphorus and calcium utilization in ruminants using isotope dilution technique. In: Phosphorus and calcium utilization and requirements in farm animals. D.M.S.S. Vitti, E. Kebreab (eds.). CABI Publishing, Wallingford, UK, 2010: 45-67 CrossRef
  31. Mutucumarana R.K., Ravindran V., Ravindran G., Cowieson A.J. Influence of dietary calcium concentration on the digestion of nutrients along the intestinal tract of broiler chickens. The Journal of Poultry Science, 2014, 51(4): 392-401 CrossRef
  32. Wilkinson S., Bradbury E., Thomson P., Bedford M., Cowieson A. Nutritional geometry of calcium and phosphorus nutrition in broiler chicks. The effect of different dietary calcium and phosphorus concentrations and ratios on nutrient digestibility. Animal, 2014, 8(7): 1080-1088 CrossRef
  33. López-Alarcón M., Hernández M., Sousa V., Moreno Á., Villapando S. Iron bioavailability and utilization in rats are lower from lime-treated corn flour than from wheat flour when they are fortified with different sources of iron. The Journal of Nutrition, 2003, 133(1): 154-159 CrossRef
  34. Matsuzaki H., Katsumata S.I., Uehara M., Suzuki K., Nakamura K. Effects of high calcium intake on bone metabolism in magnesium-deficient rats. Magnesium Research, 2005, 18(2): 97-102.
  35. Prather T.A., Miller D.D. Calcium carbonate depresses iron bioavailability in rats more than calcium sulfate or sodium carbonate. Journal of Nutrition, 1992, 122(2): 327-332 CrossRef
  36. Pabón M., Lönnerdal B. Effect of citrate on zinc bioavailability from milk fractions and infant formulas. Nutrition Research, 1993, 13(1): 103-111 CrossRef
  37. Takasugi S., Matsui T., Yano H. Effects of excess calcium as a different form on mineral metabolism in rats. Animal Science Journal, 2005, 76(5): 469-474 CrossRef
  38. Zhang B., Coon C. The relationship of calcium intake, source, size, solubility in vitro and in vivo, and gizzard limestone retention in laying hens. Poultry Science, 1997, 76(12): 1702-1706 CrossRef
  39. Bertinato J., Lavergne C., Plouffe L.J., El Niaj H.A. Small increases in dietary calcium above normal requirements exacerbate magnesium deficiency in rats fed a low magnesium diet. Magnesium Research, 2014, 27(1): 35-47 CrossRef
  40. Takasugi S., Matsui T., Omori H., Yano H. Excess calcium increases bone zinc concentration without affecting zinc absorption in rats. Biological Trace Element Research, 2007, 116(3): 311-20 CrossRef
  41. Anwar M.N., Ravindran V., Morel P.C.H., Ravindran G., Cowieson A.J. Effect of limestone particle size and calcium to non-phytate phosphorus ratio on true ileal calcium digestibility of limestone for broiler chickens. British Poultry Science, 2016, 57(5): 707-713 CrossRef
  42. Dabak J.D., Gazuwa S.Y., Ubom G.A. Nephroprotective potential of Ca and Mg against Cd and Pb in Rats. Asian J. Exp. Biol. Sci., 2012, 3(1): 214-221.
  43. Dabak J.D., Gazuwa S.Y., Ubom G.A. Comparative nephroprotective effects of concomitant administration of Ca, Mg and the combination of Ca and Mg against Cd and Pb Co-intoxicated rats. IOSR Journal of Environmental Science, Toxicology and Food Technology (IOSR-JESTFT), 2016, 10(12): 70-76.
  44. Al-Qahtani W.S., Virk P. Effect of calcium supplementation on lead-induced genotoxicity and bioaccumulation in suckling Wistar rats. Toxicological & Environmental Chemistry, 2015, 97(7): 968-988 CrossRef
  45. Haque M.M., Awal M.A., Mostofa M., Sikder M.M.H., Hossain M.A. Effects of calcium carbonate, potassium iodide and zinc sulphate in lead induced toxicities in rat model. Bangladesh Journal of Veterinary Medicine, 2008, 4(2): 123-127 CrossRef
  46. Brzóska M.M., Moniuszko-Jakoniuk J. The influence of calcium content in diet on cumulation and toxicity of cadmium in the organism. Archives of Toxicology, 1997, 72(2): 63-73 CrossRef

 

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