Effect of nano folic acid and folic acid on mortality rate and homocysteine concentration in broiler chickens

Document Type : Research Paper

Authors

1 Department of Animal Science, Faculty of Animal Science, Sari University of Agricultural Sciences and Natural Resources, Sari, Iran

2 Department of Animal Science, Faculty of Animal Science and Fisheries, Sari Agricultural Sciences and Natural Resources University, Sari, Iran.

3 Mycology and Medical Bacteriology Research Center, Kerman University of Medical Sciences, Kerman, Iran.

Abstract

The aim of this study was to investigate and compare the effect of nano folic acid supplementation and folic acid in reducing serum homocysteine level, mortality rate and some blood biochemical parameters in broiler chickens. A total of 250 male broiler chickens of Ross 308 hybrid were implemented for 42 days as a completely randomized design in a 2x2 factorial format with five treatments and five replications and ten chickens in each replication. Treatments include: 1) control ration, 2) Folic acid-free diet with folic acid supplement (4 mg per liter of water) in non-nano form, 3) Folic acid-free diet with folic acid supplement (4 mg per liter of water) in nano form, 4) control diet with folic acid supplement (4 mg per liter of water) in non-nano form, 5) Control diet with folic acid supplement (4 mg per liter of water) were in nano form. After six weeks, serum samples were collected and analyzed for homocysteine concentration. The results showed that regular folic acid significantly decreased homocysteine concentration in serum and decreased mortality (P<0.05), but in nano folic acid treatment, this reduction was more than other treatments. The interaction effect of nano folic acid and regular folic acid supplements significantly reduced serum homocysteine levels compared to the control (P<0.05). But in the control treatment, it had the highest level of homocysteine concentration and the highest mortality. Also, nano folic acid and folic acid significantly decreased the activity of alanine aminotransferase and aspartate aminotransferase compared to the control treatment (P<0.05). But it had no significant effect on the activity of alkaline phosphatase. In addition, folic acid and nano folic acid significantly increased the concentration of thyroid hormones (T3 and T4) compared to the control treatment (P<0.05). Therefore, we conclude that nano folic acid is more effective than normal folic acid in reducing the concentration of homocysteine in blood circulation and may be a promising alternative to optimize homocysteine metabolism and reduce cardiovascular risks associated with mortality in broilers.

Keywords

Main Subjects


Extended Abstract

Introduction

The purposes of using vitamins in the poultry industry are to maintain normal cellular function, health and prevention of diseases in the body. Since group B vitamins, especially folic acid, are used on a small scale in poultry feed, they may be destroyed due to improper storage conditions or heat from cooking in feed factories. Therefore, in order to solve this problem in the diet of broiler chickens, this nutrient must be fortified and the bioactive substance of the food must be protected. Existing techniques for encapsulation of nutrients or active ingredients include spray-drying microencapsulation, extrusion encapsulation, fluidized bed coating, colocalization, molecular encapsulation using liposome encapsulation, and hydrogel encapsulation. Therefore, the aim of this study was to investigate and compare the effect of Nano folic acid supplementation and regular folic acid in reducing homocysteine level and mortality rate in broiler chickens.

 

Materials and methods

In this study, in total, 250 one-day-old broiler chickens of the Ross 308 commercial strain were distributed into five groups and five replications with 2 x 2 factorial method in the form of a completely random design, and ten broiler chickens in each replication. The groups are: 1) control diet (with folic acid), 2) diet without folic acid with folic acid in water (4 mg/L) in non-Nano form, 3) diet without folic acid with folic acid in water (4 mg/L) in the form of nanoliposomes, 4) the control diet with folic acid in water (4 mg/L) in non-nano form, 5) the control diet with folic acid in water (4 mg/L) in the form of nanoliposomes. Birds had free access to food and water and received 23 hours of light per day. The initial rearing temperature was set at 33°C and decreased by 2°C weekly. The experiment lasted 6 weeks. There were three diets consisting of starter, grower and finisher that were fed as mass, which was based on corn/soybean meal that met the nutrient requirements determined by the NRC (1994) of the commercial hybrid Ross 308 with and without folic acid supplementation prepared.

 

Results and discussion

The effect of the form of folic acid consumption was significant on the reduction of homocysteine concentration and mortality, but the consumption form of nano folic acid caused a significant reduction of homocysteine and mortality. Also, the treatment of nano folic acid in the amount of 4 mg/kg along with the control diet significantly reduced homocysteine and mortality compared to the control group (p<0.05). The control diet along with folic acid in water (4 mg/L) reduced homocysteine and mortality in non- nano form compared to the control group. Also, the control diet along with folic acid in water (4 mg/L) in nano form significantly reduced the percentage of homocysteine and mortality compared to the control group (p<0.05).

 

Conclusions

The present study shows that nano folic acid and regular folic acid are the most effective vitamin B9 for reducing high plasma homocysteine concentration and reducing losses in broiler chickens. Nano folic acid showed significantly better efficacy than conventional folic acid due to its smaller particle size and higher bioavailability.

 

Author Contributions

For research articles with several authors, a short paragraph specifying their individual contributions must be provided. The following statements should be used “Conceptualization, X.X. and Y.Y.; methodology, X.X.; software, X.X.; validation, X.X., Y.Y. and Z.Z.; formal analysis, X.X.; investigation, X.X.; resources, X.X.; data curation, X.X.; writing—original draft preparation, X.X.; writing—review and editing, X.X.; visualization, X.X.; supervision, X.X.; project administration, X.X.; funding acquisition, Y.Y. All authors have read and agreed to the published version of the manuscript.” Please turn to the CRediT taxonomy for the term explanation. Authorship must be limited to those who have contributed substantially to the work re-ported.

All authors contributed equally to the conceptualization of the article and writing of the original and subsequent drafts.

Data Availability Statement

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Acknowledgements

The Acknowledgments section should be a few sentences at the end, but it is important to recognize those people (organizations and individuals) who made considerable impact on the research, provided significant help to the author to formulate and complete the experiment, and improved the research at any stage (from providing access to equipment or field sites to editing the manuscript). However, this is an optional section.

In this section, you can acknowledge any support given which is not covered by the author contribution or funding sections. This may include administrative and technical support, or donations in kind (e.g., materials used for experiments).

The authors would like to thank all participants of the present study.

Ethical considerations

The study was approved by the Ethics Committee of the University of ABCD (Ethical code: IR.UT.RES.2024.500). The authors avoided data fabrication, falsification, plagiarism, and misconduct.

Conflict of interest

The author declares no conflict of interest.

 

Conflict of interest

The author declares no conflict of interest.

REFERENCES
Aronson, D. C., Onkenhout, W., Raben, A. M. T. J., Oudenhoven, L. F. I. J., Brommer, E. J. P., & Van Bockel, J. H., (1994). Impaired homocysteine metabolism: a risk factor in young adults with atherosclerotic arterial occlusive disease of the leg. British Journal of Surgery, 81(8), 1114-1118.
Bagheri, S., H. Janmohammadi, R. Maleki, & A. Ostadrahimi., (2019). Laying hen performance, egg quality improved and yolk 5-methyltetrahydrofolate content increased by dietary supplementation of folic acid. Animal Nutrition. 5(2), 130-133.
Bhalerao, S., Hegde, M., Katyare, S., & Kadam, S., (2014). Promotion of omega-3 chicken meat production: an Indian perspective. World's Poultry Science Journal, 70(2), 365-374.
Blancquaert, D., Navarrete, O., Storozhenko, S., De Steur, H., Van Daele, J., Dong, W., Lei, C., Zhang, C., Stove, C., Gellynck, X. & Viaene, J., (2013). Biofortified rice to fight folate deficiency. Handbook of Food Fortification and Health: From Concepts to Public Health Applications Volume 1, 321-334.
Boushey, C.J., Beresford, S.A., Omenn, G.S. & Motulsky, A.G., (1995). A quantitative assessment of plasma homocysteine as a risk factor for vascular disease: probable benefits of increasing folic acid intakes. Jama, 274(13), 1049-1057.
Bowes, V.A., Julian, R.J., Leeson, S. & Stirtzinger, T., (1988). Research note: effect of feed restriction on feed efficiency and incidence of sudden death syndrome in broiler chickens. Poultry Science, 67(7), 1102-1104.
Brouwer, I.A., van Dusseldorp, M., Thomas, C.M., Duran, M., Hautvast, J.G., Eskes, T.K. & Steegers Theunissen, R.P., (1999). Low-dose folic acid supplementation decreases plasma homocysteine concentrations: a randomized trial. The American Journal of Clinical Nutrition, 69(1), 99-104.
Brigden, J. L., & Riddell, C., (1975). A survey of mortality in four broiler flocks in western Canada. The Canadian Veterinary Journal, 16(7), 194.
Classen, H.L., Riddell, C. & Robinson, F.E., (1991). Effects of increasing photoperiod length on performance and health of broiler chickens. British Poultry Science, 32(1), 21-29.
Dibaji, S.M., Seidavi, A., Asadpour, L., & DaSilva, F.M., (2014). Effect of a synbiotic on the intestinal microflora of chickens. Journal of Applied Poultry Research, 23(1), 1-6.
Emmert, J.L., Garrow, T.A. & Baker, D.H., (1996). Hepatic betaine-homocysteine methyltransferase activity in the chicken is influenced by dietary intake of sulfur amino acids, choline and betaine. The Journal of Nutrition, 126(8), 2050-2058.
Farooqi, I., & O Rahilly, S., (2004). Monogenic human obesity syndromes. Recent progress in Hormone Research, 59, 409-424.
Finkelstein, J.D. & Martin, J.J., (1986). Methionine metabolism in mammals. Adaptation to methionine excess. Journal of Biological Chemistry, 261(4), 1582-1587.
Gouda, A., Amer, S. A., Gabr, S., & Tolba, S. A., (2020). Effect of dietary supplemental ascorbic acid and folic acid on the growth performance, redox status, and immune status of broiler chickens under heat stress. Tropical Animal Health and Production, 52, 2987-2996.
Haïssaguerre, M., Shah, D. C., Jaïs, P., Shoda, M., Kautzner, J., Arentz, T., & Clémenty, J., (2002). Role of Purkinje conducting system in triggering of idiopathic ventricular fibrillation. The Lancet, 359(9307), 677-678.
Hebert, K., J. House, & W. Guenter., (2005). Effect of dietary folic acid supplementation on egg folate content and the performance and folate status of two strains of laying hens. Poultry Science, 84(10), 1533-1538.
Hogan, Albert G. and E. M. Parrot. Anemia in chicks due to vitamin deficiency. Journal of Biological Chemistry (Proceedings) 128:xlvi-xlvii. 19 39.
House, J., Braun, K., Ballance, D., O'connor, C., & Guenter, W., (2002). The enrichment of eggs with folic acid through supplementation of the laying hen diet. Poultry Science, 81(9), 1332-1337.
House, J.D., O'Connor, C.P. & Guenter, W., (2003). Plasma homocysteine and glycine are sensitive indices of folate status in a rodent model of folate depletion and repletion. Journal of Agricultural and Food Chemistry, 51(15), 4461-4467.
Husseiny, E., (1981). Effect of ambient temperature on mineral retention and balance of the broiler chicks. Poultry Science, 60(1), 1651.
Hulan, H.W., Proudfoot, F.G. & Mcrae, K.B., (1980). Effect of vitamins on the incidence of mortality and acute death syndrome (“flip-over”) in broiler chickens. Poultry Science, 59(4), 927-931.
Imaeda, N., (1999). Characterization of serum enzyme activities and electrolyte levels in broiler chickens after death from sudden death syndrome. Poultry Science, 78(1), 66-69.
Jing, M., Munyaka, P., Tactacan, G., Rodriguez-Lecompte, J., & House, J., (2014). Performance, serum biochemical responses, and gene expression of intestinal folate transporters of young and older laying hens in response to dietary folic acid supplementation and challenge with Escherichia coli lipopolysaccharide. Poultry Science, 93(1), 122-131.
Julian, R.J., (2000). Physiological, management and environmental triggers of the ascites syndrome: A review. Avian Pathology, 29(6), 519-527.
Kang, S. S., (1996). Treatment of hyperhomocystinemia: physiological basis. The Journal of Nutrition, 126(suppl_4), 1273S-1275S.
Kawada, M., Hirosawa, R., Yanai, T., Masegi, T. & Ueda, K., (1994). Cardiac lesions in broilers which died without clinical signs. Avian Pathology, 23(3), 503-511.
Kim, Y.I., (2007). Folate and colorectal cancer: An evidence‐based critical review. Molecular Nutrition & Food Research. 51(3), 267-292.
Kumar, K. A., Lalitha, A., Pavithra, D., Padmavathi, I. J., Ganeshan, M., Rao, K. R., Venu, L., Balakrishna, N., Shanker, N. H., & Reddy, S. U., (2013). Maternal dietary folate and/or vitamin B12 restrictions alter body composition (adiposity) and lipid metabolism in Wistar rat offspring. The Journal of nutritional biochemistry, 24(1), 25-31.
Korte, M., Sgoifo, A., Ruesink, W., Kwakernaak, C., Van Voorst, S., Scheele, C. W., & Blokhuis, H. J. (1999). High carbon dioxide tension (PCO2) and the incidence of cardiac arrhythmias in rapidly growing broiler chickens. Veterinary Record, 145(2), 40-43.
Kutlu, H. R., & Forbes, J. M., (1993). Changes in growth and blood parameters in heat-stressed broiler chicks in response to dietary ascorbic acid. Livestock Production Science, 36(4), 335-350.
Matte, J. J., Girard, C. L., & Brisson, G. J., (1992). The role of folic acid in the nutrition of gestating and lactating primiparous sows. Livestock Production Science, 32(2), 131-148.
Milne, D., Canfield, W., Mahalko, J., & Sandstead, H., (1984). Effect of oral folic acid supplements on zinc, copper, and iron absorption and excretion. The American Journal of Clinical Nutrition, 39(4), 535-539.
Olkowski, A.A. & Classen, H.L., (1995). Sudden death syndrome in broiler chickens: a review. Poultry and Avian Biology Reviews (United Kingdom).
Ononiwu, J.C., Thomson, R.G., Carlson, H.C. & Julian, R.J., (1979). Pathological studies of “sudden death syndrome” in broiler chickens. The Canadian Veterinary Journal, 20(3), 70.
Pancharuniti, N., Lewis, C.A., Sauberlich, H.E., Perkins, L.L., Go, R.C., Alvarez, J.O., Macaluso, M., Acton, R.T., Copeland, R.B., Cousins, A.L. & Gore, T.B., (1994). Plasma homocystine, folate, and vitamin B-12 concentrations and risk for early-onset coronary artery disease. The American Journal of Clinical Nutrition, 59(4), 940-948.
Perna, A.F., Ingrosso, D., Lombardi, C., Acanfora, F., Satta, E., Cesare, C.M., Violetti, E., Romano, M.M. and De Santo, N.G., (2003). Possible mechanisms of homocysteine toxicity. Kidney International, 63, S137-S140.
Rezaei, M., & Kazemi Fard, M., (2019). Effects of different levels of tomato powder with and without addition of enzymes on performance, blood parameters and antioxidant status of japanese quails. Research on Animal Production, 10(23), 11-21.
Riddell, C., (1991). Developmental, metabolic, and miscellaneous disorders. Diseases of  Poultry, 839-841.
Sahin, K., M. Onderci, N. Sahin, & M. Gursu., (2003). Dietary vitamin C and folic acid supplementation ameliorates the detrimental effects of heat stress in Japanese quail. The Journal of Nutrition, 133(6),1882-1886.
Sahin, K., Sahin, N., & Yaralioglu, S., (2002). Effects of vitamin C and vitamin E on lipid peroxidation, blood serum metabolites, and mineral concentrations of laying hens reared at high ambient temperature. Biological Trace Element Research, 85, 35-45.
SAS Institute., (2008). sas user,s guide statistics. SAS Institute Inc., cary, NC., USA.
Samuels, S.E., (2003). Diet, total plasma homocysteine concentrations and mortality rates in broiler chickens. Canadian Journal of Animal Science, 83(3), 601-604.
Scott, T.A., (2002). Evaluation of lighting programs, diet density, and short term use of mash as compared to crumbled starter to reduce incidence of sudden death syndrome in broiler chicks to 35 d of age. Canadian Journal of Animal Science, 82(3), 375-383.
Selhub, J., (1999). Homocysteine metabolism. Annual Review of Nutrition, 19(1), 217-246.
Shahraki, E., Kazemi fard, Rezai, M., Ansari, Z., Barani, M., (2024). Comparison of nano folic acid and folic acid on performance, carcass characteristics, blood parameters and microbial population of broiler chickens. Research on animal production. (Article in press, avaible on URL: https://rap.sanru.ac.ir/article-1-1403-en.html).
Siddiqui, M.F.M.F., Patil, M.S., Khan, K.M., Khan, L.A. & Mafsu, A.M., (2009). Sudden death syndrome–an overview. Veterinary World, 2(11), 444-447.
Tactacan, G., M. Jing, S. Thiessen, & J. Rodriguez-Lecompte., (2010). Characterization of folate-dependent enzymes and indices of folate status in laying hens supplemented with folic acid or 5-methyltetrahydrofolate. Poultry Science, 89(4), 68.
Toue, S., Kodama, R., Amao, M., Kawamata, Y., Kimura, T. and Sakai, R., (2006). Screening of toxicity biomarkers for methionine excess in rats. The Journal of  Nutrition, 136(6) 1716S-1721S.
Towbin, J.A., (2001). Molecular genetic basis of sudden cardiac death Cardiovascular Pathology 10, 283–295.
Volk, M., M. Herceg, B. Kralj, S. Meknic, & V. Tadic, 1974. Investigations of fetal syncope of fowl in broiler. 1. Incidence, clinical symptoms, pathomorphological finding and pathogenesis. Vet. Arhiv. 44:14–23.
Xie, M., Hou, S.S., Huang, W. & Fan, H.P., (2007). Effect of excess methionine and methionine hydroxy analogue on growth performance and plasma homocysteine of growing Pekin ducks. Poultry Science, 86(9), 1995-1999.
Yeo, L. K., Olusanya, T. O. B., Chaw, C. S., & Elkordy, A. A., (2018). Brief effect of a small hydrophobic drug (Cinnarizine) on the physicochemical characterisation of niosomes produced by thin-film hydration and microfluidic methods. Pharmaceutics, 10(4), 185. https://www.mdpi.com/1999-4923/10/4/185.
Zhang, Y., Jing, W., Zhang, N., Hao, J. & Xing, J., (2020). Effect of maternal folate deficiency on growth performance, slaughter performance, and serum parameters of broiler offspring. The Journal of Poultry Science, 57(4), 270-276.