Effect of adding different levels of synbiotic to milk on performance, digestibility, blood parameters and fecal score on Dalaq sucking lambs

Document Type : Research Paper

Authors

1 Dept. of Animal Nutrition, Faculty of animal Science, Gorgan University of agricultural Science,

2 Ph.D. Student, Dept. of Animal and Poultry Nutrition, Faculty of Animal Science, Gorgan University of Agricultural Sciences and Natural Resources

3 Gorgan University of Agricultural and Natural Resources

Abstract

Abstract
Introduction: Management of suckling lambs in the first trimester guarantees their health and future production. Animal specific physiological status during this period includes the ability to absorb large molecules, especially immunoglobulins, from the gut and to be prone to intestinal infections and diarrhea. Artificial changes in the lamb intestine flora may improve the intestinal environment. The use of antibiotics in the diet of animals kills beneficial bacteria and does not support the cellular immune system well, and overuse of antibiotics increases bacterial resistance to them. Therefore, replacing antibiotics with other compounds such as prebiotics, probiotics and synbiotics is considered as a suitable solution. These compounds have positive effects such as reducing intestinal infections and diarrhea by increasing the microbial balance of the intestine and increasing the absorption of nutrients including immunoglobulins. Synbiotics is a combination of probiotics and prebiotics that reduce the population of pathogenic microbes in the gastrointestinal tract by increasing their interaction. These compounds can also improve food conversion ratio. Synbiotics can have a positive effect on lamb health and growth rate.
Materials and Methods: The present study was conducted to investigate the effect of adding different levels of synbiotics to milk on performance, digestibility, blood parameters and fecal score on Dalaq sucking lambs. In this study, 24 suckling lambs with an average weight of 5 ±1.7 kg were selected and then all lambs were examined thoroughly to ensure their health and performance. Lambs were randomly used in four equal experimental groups (6 heads). Treatments include: 1) control treatment (without adding synbiotic to milk), 2) treatment receiving 2.5 g of synbiotic, 3) treatment receiving 5 g of synbiotic and 4) treatment receiving 7.5 g of synbiotic. The total duration of the experiment was 90 days, but feeding and recording of functional factors started from day 20 of the period .The lambs were kept in metabolic cages and the diets used in this experiment were prepared and adjusted based on NRC and were given to the lambs in the morning (8 o'clock) and in the evening (16 o'clock) as an appetite suppressant. Daily feed was provided to the lambs in a completely mixed form. Throughout the experiment, lambs had free access to clean drinking water. Daily feed was provided to the animals in a completely mixed form. In this study, a commercial synbiotic product (GmbH, BIOMIN) of Herzogenburg, Austria including Enterococcus faecium, fructooligosaccharide and seaweed extract through milk substitute was used. Milk was given to lambs twice a day (10% of body weight) by pacifiers. lambs were weighed one day per week to evaluate changes in weight. Dry matter intake was measured and fecal score was recorded daily. Feces samples were collected for digestibility in the final week for 5 days. lambs feces were evaluated daily. Feces scores were determined based on 1-firm and consistent, 2-soft and loose, 3-loose and watery, 4-watery with some blood and 5-watery with blood and mucus. Blood sampling was performed on day 83 so that all lambs were sampled intravenously before the morning meal and with dietary restriction for 12 to 14 hours and the blood was taken in two separate tubes, one containing heparin to obtain plasma and the other. Without heparin, the serum was poured to obtain it. The concentrations of blood Parameter were also determined by spectrophotometer.
Results and Discussion: The results showed that increasing the level of synbiotics in milk leads to an increase in dry matter intake, daily weight and end-of-period weight (P˂0.05) but the feed conversion ratio was not affected by the use of synbiotics (P> 0.05). The digestibility of crude protein, insoluble fiber in acidic detergent, insoluble fiber in neutral detergent, dry matter, organic matter and ether extract did not show a significant difference between experimental treatments (P> 0.05). Immunoglobulins in the blood of lambs increased with increasing synibiotic level in milk (P˂0.05) so that in 7.5 g of cinebbiotic treatment they had the highest and the control treatment had the lowest concentration of immunoglobulins (P˂0.05). Total protein, albumin, globulin and albumin to globulin ratios were not affected by experimental treatments (P<0.05) but cholesterol, glucose, triglyceride, ammonia nitrogen, high-density lipoprotein, and high-density lipoproteins. Lipoproteins with very low density were affected by experimental treatments (P˂0.05) so that increasing the level of synbiotic caused an increase in triglyceride, lipoproteins with low density and very low lipoproteins (P˂0.05). Further consumption of synbiotics in milk reduced cholesterol, glucose, ammonia nitrogen and high-density lipoproteins (P˂0.05). Feces score decreased significantly with increasing sybibiotic level in milk, so that at the level of 5 g of synbiotic, the lowest consistency score was observed and in the control treatment, the highest feces score was observed (P˂0.05). But numerically, the control treatment had the highest number of days of diarrhea (P>0.05). In general, according to the present results, the use of synbiotics in suckling lambs is recommended to improve the performance and health of lambs.
Conclusion: Overall, the results of this study showed that the use of Synbiotics in milk Increases dry matter intake and daily weight gain in lambs. Also, the uptake of immunoglobulins into the blood of lambs increased with increasing synibiotic levels in milk. Elevated levels of synbiotics cause elevated triglycerides, low-density lipoproteins, and very low-density lipoproteins, but decreased cholesterol, glucose, amniotic nitrogen, and lipids .Improvement of feces score and diarrhea was observed with increasing sybiotic level in milk.The use of synbiotics in suckling lambs is necessary to improve the performance and health of suckling lambs.

Keywords

References

Abaadi MH, Dehghan Bonadaki M and Zali A, 2013. The effect of feeding of bacterial probiotic in milk or starter on growth performance, health, blood and rumen parameters of suckling calves. Research on Animal Production 4: 57-69. (In Persian).
Abe F, Ishibashi N and Shimamura S, 1995. The effect of administration of bifidobacteria and lactic acid bacteria to newborn calves and piglets. Journal of Dairy Science 78: 2823-2826.
Adams MC, Luo J, Rayward D, Kingm S, Gibson R and Moghaddam GH, 2008. Selection of anovel directfed microbial to enhance weight gain in intensively reared calves. Animal Feed Science andTechnology 145: 41-52.
Alberda C, Gromlish L, Meddings J, Field C and McCargar L, 2007. Effects of probiotic therapy in critically ill patients: A randomized double-blind, placebocontrolled trial. The American Journal of Clinical Nutrition 85: 816-823.
Al-Saidy MY, 2010. Effect of bacteria on immunoglobulin G concentration and other blood components of newborn calves. Journal of Animal and Veterinary Advances 9: 604-609.
Antunovic Z, Speranda M, Amidzic D, Seric V, Steiner Z, Doma-Cinovic N and Boli F, 2006. Probiotic application in lambs' nutrition. Agriculture, Animal Sciences and Food Sciences 4: 175-180.
Antunovic Z, Speranda M, Liker B, Seric V, Sencic D, Domacinovic M and Sperandat T, 2005. Influence of feeding the probiotic PioneerPDFM® to growing lambs on performances and blood composition. Acta Veterinaria 55: 287-300.
AOAC, 1990. Official methods of analysis 15th edition. Association of official analytical chemist, Arlington, U.S.A.
Asadi M, Toghdory A and Ghoorchi T, 2018. Effect of Oral Administration and Injection of Selenium and Vitamin E on Performance, Blood Metabolites and Digestibility of Nutrients in Suckling Dalagh Lambs. Research on Animal Production 9(20):79-87. (In Persian).
Avita FA, Paulillo AC, Schucken-iturrino RP, Lucas FA, Orgaz A and Quntana JL, 1995. A comparative study of efficiency of probiotic and the K-99 and anti A-14 vaccines in the control of diarrhea in calves in Brazil. Veterinary immunology and immunopathology journal 48: 239-2243.
Azimzadeh V, Assadi-Alamouti A, Khadem AA, Bagheri-Varzaneh M and Mohammad Moradi J, 2015. Effects of Supplementation of a Symbiotic Product on Growth Performance and Health of Holstein Calves. Research on Animal Production 6(12):105-114. (In Persian).
Bomba A, Nemcova R, Mudronova D and Guba P, 2002. The possibilities of potentiating the efficacy of probiotics. Trends in Food Science & Technology, 13: 121-126.
Brakefield K, Godden S, Fetrow J, Rapnicki P, Jones C, Bey R and Haines D, 2010. Effect of feeding Bovamine® probiotic on passive transfer of immunoglobulin G in newborn calves. Proceeding of Minnesota Dairy Health Conference, University of Minnesota, USA, Pp: 103-104.
Chamoro M, 2009. Environment, dam, management: Factors influencing passive transfer of immunoglobulins to neonatal calves. Veterinary Quarterly Research Gate 12: 1-7.
Chigerwe M, Tyler JW, Schultz LG, Middelton JR, Steevens BJ and Spain JN, 2008. Effect of colostrum administration by use of oroesophageal intubation on serum IgG concentrations in Holstein bull calves. American Journal of Veterinary Research 69: 1158-1163.
Cruywagen CW, Jordaan I and Venter L, 1996. Effect of Lactobacillus acidophilus supplementation of milk replacer on preweaning performance of calves. Journal of Dairy Science 79: 483- 486.
Darrezereshkipour M, Parsaeimehr K, Hosseinzadeh S and Farhovand P, 2014. The effects of different levels of prebiotic (A-MAX) on digestibility, and blood biochemical parameters in West Azarbaijan kids. Journal of Veterinary Clinical Pathology 7(28): 314-321. (In Persian).
Dayani O, Dadvar P and Afsharmanesh M, 2011. Effect of dietary whole cottonseed and crude protein level on blood-parameters and performance of fattening lambs. Small Ruminant Research 97: 48-54.
De Valdez DF, Martos G, Taranto MP, Lorca GL, Oliver G and De Ruiz Holgado AP, 1997. Influence of bile on bgalactosidase activity and cell viability of Lactobacillus reuteri when subjected to freeze-drying. Journal of Dairy Science 80: 1955-1958.
Didarkhah M and Bashtani M, 2018. Effects of Probiotic and Peribiotic Supplementation in Milk on Performance and Nutrition Digestibility in Holstein Calves. Research on Animal Production 9(20): 70-78. (In Persian).
Elizondo-Salazar JA and Heinrichs AJ, 2009. Feeding heat-treated colostrum to neonatal dairy Heifers; effect on growth characteristics and blood parameters. Journal of Dairy Science 92: 3265-3273.
Ellinger DK, Muller LD and Glantz PJ, 1980. Influence of-feeding fermented colostrum and Lactobacillus acidophiluson fecal flora of dairy calves. Journal of Dairy Science 63: 478-482.
Fayed AM, El-Ashry MA, Youssef KM, Salem FA and Aziz HA, 2005. Effect of feeding falvomycin or yeast as feed supplement on ruminal fermentation and some blood constituents of sheep in Sinai. Egyptian Journal of Nutrition and Feeds 8: 619 – 634.
Fuller R, 1989. Probiotics in man and animal. Journal of Applied Bacteriology 66: 365-378.
Ghosh S, Mehla RK, Sirohi SK and Roy B, 2010. The effect of dietary garlic supplementation on body weight gain, feed intake, feed conversion efficiency, faecal score, faecal coliform count and feeding cost in crossbred dairy calves. Tropical Animal Health Production 42: 961-968.
Ghosh S, Mehla RK, Sirohi SK and Tomar SK, 2011. Performance of crossbred calves with dietary supplementation of garlic extract. Animal phisiology and Animal Nutrition 95: 449-455.
Gibson GR and Collins MD, 1999. Probiotics, prebiotics, and synbiotics: approaches for modulating the microbial ecology of the gut. American Journal of Clinical Nutrition 69(5):1052-1057.
Gibson GR and Roberfroid MB, 1995. Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. Journal of Nutrition 125: 1401–1412.
Hasunuma T, Kawaahima K, Nakayama H, Murakami T, Knaagawa H, Ishii T, Akiyama K, Yasuda K, Terada F and Kushibiki S, 2011. Effect of cellooligosaccharide or synbiotic feeding on growth performance, fecal condition and hormone concentrations in Holstein calves. Journal of Animal Science 82: 543–548.
Heinrichs AJ, Jones CM, Elizondo-Salazar JA and Terrill SJ, 2009. Effects of aprebiotic supplement on health of neonatal dairy calves. Livestock Science 125: 149-154.
Hillal H, El-Sayaad G and Abdella M, 2011. Effect of growth promoters (probiotics) supplementation on performance, ru-men activity and some blood constituents in growing lambs. Leibniz Institute for Farm Animal Biology 6: 607-617.
Hossein Abadi M, Dehghan Banadaki M and Zali A, 2013. Effect of adding probiotic bacteria in milk or initial feed on growth performance, health condition, blood and stomatal parameters of Holstein calves. Animal production research 4(8): 69-57. (In Persian).
Huang MK, Choi YJ, Houde RJ, Lee W, Lee BH and Zhao X, 2004. Effects of Lactobacilli and an acidophilic fungus on the production performance and immune responses in broiler chickens. Poultry Science 83: 788-795.
James RE, Polan CE and Cummins KA, 1981. Influence of administered indigenous microorganisms on uptake of [Iodine-125] {gamma}-globulin in vivo by intestinal segments of neonatal calves. Journal of Dairy Science 64: 52-61.
Jatkauskas J and Vrotniakiene V, 2010. Effects of probiotic dietary supplementation on diarrhea patterns, faecal microbiota and performance of early weaned calves. Veterinary Medicine 55: 494-503.
Jenny BF, Vandijk HJ and Collins JA, 1991. Performance and fecal flora of calves fed abacillus subtilis concentrate. Journal of Dairy Science 74: 1968-1973.
Kaur IP, Chopra K and Saini A, 2002. Probiotics: potential pharmaceutical applications. European Journal of Pharmaceutical Sciences 15: 1-9.
Kawakami S, Yamada T, Nakanishi N and CAI Y, 2010. Feeding of lactic acid bacteria and yeast affects fecal flora of Holstein calves. Journal of Animal and Veterinary Advances 10: 269-271.
Keithly JI, Kott RW, Berardinelli JG, Moreaux S and Hatfield PG, 2011. Thermogenesis, blood metabolites and hormones, and growth of lambs born to ewes supplemented with algae-derived docosahexaenoic acid. Journal of Animal Science 89: 4305-4313.
Khan MA, Lee HJ, Lee WS, Kim HS, Kim SB, Ki KS, Ha JK, Lee HG and Choi YJ, 2007. Pre- and post-weaning performance of Holstein female calves fed milk through step-down and conventional methods, Journal of Dairy Science 90: 876–885.
Khan MA, Weary DM and Keyserlingk MA, 2011. Invited review Effects of milk ration on solid feed intake, weaning, and performance in dairy heifers. Journal of Dairy Science 94:1071-1081.
Kong XF, Wu GY and Yin YL, 2011. Roles of phytochemicals in amino acid nutrition. Frontiers in Bioscience, 3: 372-384.
Krehbiel CR, Rust SR, Zhang G and Gilliland SE, 2003. Bacterial direct- fed microbials in ruminant diets: Perfomance response and mode of action. Journal of Dairy Science 81(E. Suppl. 2): E120- E132.
Kyriakis SC, Tsiloyiamnis VK, Velmmas K, Tsinas AC, Alexpopoulus C and Jansegres L, 1999. The effect of probiotic LSP 122 of control of post weaning diarrhea syndrome of piglets. Research in Veterinary Science Journal 67: 223-228.
Langford RM, Weary DM and Fisher L, 2003. Antibiotic resistance in gut bacteria from dairy calves: a dose response to the level of antibiotics fed in milk. Journal of Dairy Science 86: 3963-3966.
 Lehloenya KV, Krehbiel CR, Mertz KJ, Rehberger TG and Spicer L, 2008. Effect of propionibacteria and yeast culture fed to steer on nutrient intake and site and extent of digestion. Journal of Dairy Science 91: 653-662.
Lesmeister K, Heinrichs A and Gabler M, 2004. Effects of supplemental yeast Saccharomyces cerevisiae culture on rumen development, growth characteristics and blood parameters in neonatal dairy calves. Journal of Dairy Science 87: 1832-1839.
Lubbadeh W, Haddadin MSY, Al-Tamimi MA and Robinson RK, 1999. Effect on the cholesterol content of fresh lamb of supplementing the feed of Awassi ewes and lambs with Lactobacillus acidophilus. Meat Science 52:381–385.
Mahmudianfard HR, 1996. Composition of the colostrum of Golpayegani native cows and effect of nutrition on serum immunoglobulins concentrations of Holstein calves.M.Sc. thesis, Isfahan University of technology. (In Persian).
Mehrdad N, Chashnidel Y, Teimori Yansari A and Khorvash M, 2017. Effects of two kinds of probiotics on performance, blood and ruminal parameters in Holstein male calves. Journal of Ruminant Research 5(1): 23-44. (In Persian).
Moarrab A, Ghoorchi T, Ramezanpour S, Ganji F and Koochakzadeh AR, 2016. Effect of Synbiotic on Performance, Intestinal Morphology, Fecal Microbial Population and Blood Metabolites of Suckling Lambs. Iranian Journal of Applied Animal Science 6(3): 621-628.
Mohamadi P and Dabiri N, 2011. Effects of probiotic, prebiotic and synbiotic on performance and humoral immune response of female suckling calves. In proceeding of the 62th annual meeting of the European Association for Animal Production. Stavanger, Norway, p: 204.
Mohamadi P and Dabiri N, 2012. Effects of probiotic and prebiotic on average daily gain, fecal shedding of Escherichia Coli, and immune system status in newborn female calves.Asian-Australasian Journal of Animal Sciences 25: 1255-1261.
Moslemipur F, Moslemipur F and Mostafaloo Y, 2014. Effects of using probiotic and synbiotic in colostrum and milk on passive immunoglobulin transfer rate, growth and health parameters of calf. Journal of Ruminant Research 1 (4): 19-30. (In Persian).
Mwenya B, 2005. Effects of Yeast Culture and Galacto-Oligosaccharides on Ruminal Fermentation in Holstein Cows. Journal of Dairy Science 88(4): 1404-1412.
Mwenya B, Sntoso B, Pen C, Morikava R, Takaura K and Umetsu K, 2005. Effects of yeast Culture and galacto-oligosaccharides on luminal fermentation in Holstein cows. Journal of Dairy Science 88: 1404-1412.
Mwenya B, Zhou X, Santoso B, Sar C, Gamo Y and Kobyashi T, 2004. Effects of probiotic-vitacogen and ß (1-4) galacto-oligosaccharides supplementation on methanogenesis and energy and nitrogen utilization in dairy cows Asian-Australia. Journal of Animal Science 17: 349- 354.
Nasiri A H, Towhidi A, Shakeri M, Zhandi M and Dehghan Banadaki M, 2019. Effects of probiotic on milk production, feed intake and some metabolic blood profiles under the hot seasons in dairy cows. Journal of Ruminant Research 6(4): 77-88. (In Persian).
National Research Council. 2007. Nutrient Requirements of Small Ruminants: Sheep, Goats, Cervids, and New World Camelids. Washington, DC.
Nemati A, Tabatabaie SN, Davar Frouzandekey Shahraki A and Eghbal Sh, 2010. Comparison effect of Saccharomyces cerevisiae yeast and Protexin probiotic in starter on blood parameter, Immunity blood, behavior and fecal score in suckling calves. The 4th congress on Animal Science, Karaj, Iran, 2141-2144 pp. (In Persian).
Nikkhah A, Sadeghi AA and shoorang p, 2005. Development, Nutrition andManagement of Milking Calves. 1st ed., Publication of Tehran University, pp, 255-291. (In Persian).
Peric L, Milosevic N, Zikic D, Bjedov S, Cvetkovic D, Markov S, Mohnl M and Steiner T, 2010. Effects of probiotic and phytogenic products on performance, gut morphology and cecal microflora of broiler chickens. Archives of Animal Nutrition 53: 350- 359.
Piret Kalmus W, Toomas Orro E and Kalle Kask S, 2009. Effect of yeast culture on milk production and metabolic and reproductive performance of early dairy cow. Acta Veterinaria Scandinavica 51(32): 1-7.
Plata FP, Mendoza GD, Blrcena-Gama JR and Gonzalez S, 1994. Effect of a Yeast culture (Saccharomyces Cerevisia) on neutral detergent fiber digestion in steers fed oat straw based diets. Animal Feed Science 4: 203-210.
Quigley JD, Drewry JJ, Murray LM and Ivey SJ, 1997. Body weight gain, feed efficiency, and fecal scores of dairy calves in response to galactosyl-lactose or antibiotics in milk replacers. Journal of Dairy Science 80: 1751- 1754.
Riddell JB, Gallegos AJ, Harmon DL and Mcleod KR, 2010. Addition of a Bacillus based probiotic to the diet of preruminant calves: influence on growth, health, and blood parameters. International Journal of Applied Research in Veterinary Medicine 8: 78-85.
Rodriguez C, Castro N, Capote J, Morales-delanuez A, Moreno-Indias H, Sanchez-Macias D and Arguello A, 2009. Effect of colostrum immunoglobulin concentration on immunity in Majorera goat kid.American Dairy Science Association 92: 1696-1701.
Rust SR, Metz K and Ware DR, 2000. Effects of BovamineTM rumen culture on the performance and carcass characteristics of feedlot steers. Journal of Animal Science 2:78- 83.
Sadik MF, 1989. Effect of Lactobacillus concentrate (LBC) as a new growth promoter on the performance of growing buffalo heifers raised on milk replacer. MS Thesis. Ain Shams University, Cairo, Egypt.
Salama AAK, Caja G, Garin D, Albanell E, Such X and Casals R, 2002. Effects of adding a mixture of malate and yeasts culture (Saccharomyces cerevisiae) on milk production of Murciano-Granadina dairy goats. Animal Research 51: 295-303.
Sami N, Salminen S, Bylund G and Ouwehand A, 2001. Characterization of propertiesof human- and Dairy-derived probiotic for prevention of infectious disease in fish. Applied and Environmental Microbiology 67: 2430-2435.
Santoso B, Mwenya B, Sar C, Gamo Y, Kobayashi T and Morikawa R, 2004. Effects of supplementing galacto-oligosaccharids, Yucca schidigra or nisin on rumen metanogenesis, nitrogen and energy metabolism in sheep. Livestock Production Science 91: 209-217.
SAS Institute. 2003. SAS User‘s Guide. Version 9.1 ed. SAS Institute Inc, Cary, NC.
Schiffrin EJ and Blum S, 2002. Interactions between the microbial and the intestinal mucosa. European Journal of Clinical Nutrition 56(3): 60-64.
Siggers RH, Thymann T, Siggers JL, Schmidt M, Hansen AK and Sangilda PT, 2007. Bacterial colonization affects early organ and gastrointestinal growth in theneonate. Livestock Science 109: 14-18.
Timmerman HM, Konig CJ, Mulder L, Rombouts FM and Beynen AC, 2004. Monostrain, multistrain and multispecies probiotics. A comparsion of functionality and efficacy. International Journal of Food Microbiology 96: 219-233.
Timmerman HM, Mulder L, Everts H, van Espen DC, van der Wal E, Klaassen G, Rouwers SMG, Hartemink R, Rombouts FM and Beynen AC, 2005. Health and growth of veal calves fed milk replacers with or without probiotics. Journal of Dairy Science 88: 75:894-899.
Tripathi MK and Karim SA, 2011. Effect of yeast cultures supplementation on live weight change, rumen fermentation, ciliate protozoa population, microbial hydrolytic enzymes status and slaughtering performance of growing lamb. Livestock Science 135: 17-25.
Tripathi MK, Karim SA, Chaturvedi OH and Verma DL, 2007. Effect of different liquid cultures of live yeast strains on performance, ruminal fermentation and microbial protein synthesis in lambs. Journal of Animal Physiology and Animal Nutrition 92: 631-639.
Vakili-Saleh F, Moslemipur F, Mostafaloo Y and Ghorbani R, 2012. Study of effects of heating and antibiotic addition to colostrum on passive transfer of immunoglubolins, growth and health parameters in calves. M.Sc. thesis, University of Gonbad Kavoos. (InPersian).
Wallace RJ, 1994. Ruminal microbiology, biotechnology and ruminant nutrition: progress and problems. Journal of Dairy Science 72: 2992-3003.
Yakhkeshi S, Rahimi S and Gharib Naseri K, 2011. The effects of comparison of herbal extracts, antibiotic, probioticand or-ganic acid on serum lipids, immune response, GIT microbial population, intestinal morphology and performance of broilers. Journal of Medicinal Plants 37: 80-95.
Animal Science Research
Volume 32, Issue 2
October 2022
Pages 31-46

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History

  • Receive Date: 16 February 2021
  • Revise Date: 21 December 2021
  • Accept Date: 01 January 2022

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