Albanian j. agric. sci. ISSN: 2218-2020, (2012), (Special Edition) Copyright © Agricultural University of Tirana

METABOLIC PROFILE AND BODY CONDITION SCORE OF COWS DURING THE PUERPERAL PERIOD ILIR DOVA*, DORIANA BEQIRAJ, BUJAR MANE, ENKELEDA OZUNI Faculty of Veterinary Medicine, Agricultural University of Tirana, Tirana, ALBANIA * Author of correspondence; Email:dova_ilir@yahoo. com

Abstract: The study was carried out in 30 cows in three different economies and in four different physiological states. The results are statistical processing by calculating the coefficients of correlations and multiple regression analysis between the BCS and blood biochemical indicators. Correlations between BCS with total proteins and urea are strong and statistically verified in the period two week after calving and 30 days after calving. Correlation between BCS and alkaline reserve is strong only two months after calving, inorganic phosphorus level in plasma is related statistically with BCS only two week before calving. Glucose shows strong correlation to BCS in the period two week before calving and two week after calving, Correlation of calcemia to BCS are not proved statistically in any physiological state and magnesemia have correlation with BCS only two week before calving and two week after calving. Multiply regression equations to calculate statistical validity for three physiological states and are not proved in the period a months after calving. Keyword: BCS; blood biochemical indicators; correlation; regression equation

1. Introduction Body score condition (BCS) is a useful method for the management nutrition of cows [27] and the assessment of energy reserves in the body. This method is based on the visual and the palpation assessment of body parts, especially in the back, tail and in the pelvic region [9]. BCS was not used of the assessment of energy reserves till 1970, instead of was used the difference of body weight. This is a faintly indicator, because the energy as fat in internal organs can vary up to 40% of the live weight [6]. BCS changes mainly by mobilizing the energy reserves of subcutaneous adipose tissue and muscles, and less by the adipose tissue of internal organs [25]. Determination of the BCS is realized by a 9 degree system [9]. This method is proposed by [7]. Dairy cows have a negative energy balance in the first weeks after calving and in the early stage of lactation [5]. High milk production cows significantly increase the negative energy balance due to reduction of dry matter intake [3]. Those cows have the negative energy balance too by a severe biological stress. This stress can have an impact on immune processes and on the cows reproductive. The stress also affects the metabolic diseases, but an important fact is that it affects health odd dairy [1, 2]. Metabolic profile is widely used in the identification and evaluation of the cows health condition [30, 2, 17]. Through metabolic profile we International Conference 31 October 2012, Tirana

may determine the earliest stages of metabolic disorders, which leading to economic losses [21]. The negative energy balance is aggravated in the following lactation. Increasing of the metabolic needs for milk production is compensate with the mobilization of body reserves which leads to the reduction of live weight and live weight body weight [5]. Protein and energy needs grow up to 4-10 times. According to two periods, at the peak lactation [13; 28], cows mobilize up to 7 kg of raw material. Cows synthesize 2. 7-4 kg glucose for milk lactose synthesis. This leads to the intensification of glukoneogenesis by glucogenic and glucoketogenic amino acids [22]. Intensification of the lipolysis and proteolysis brings the restriction of cholesterol synthesis as well as the intensification of keton synthesis [22]. Determination of the BCS is performed during all their reproductive lifelong but it is more useful in the puerperal period [20]. The purpose of this study is to analyze the relationship between (BCS) and the metabolic profile indicators in four physiological condition, (2 weeks before calving, 2 weeks after calving, 1 month after calving and 2 months after calving). 2. Material and Methods 2. 1 Sample collection and preparation The study was performed from February 2011 to November 2011. The study was carried out in the

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three farms (Ndroq, Kashar, Lushnjë), in four physiological state. The determination of the BCS was realized according to the method with 9 degrees recommending by [9]. Cows in this study were followed at 4 physiological conditions, starting from the drying period up to 2 months after calving. The cows were selected, in each farm, in order to create the possibility for a dynamic in 4 physiological conditions. There were selected 10 cows in any economy from the lactation 2-4. Blood sample was taken in each physiological state. Blood was collected in then, in sterile and absolutely chemically pure test tubes. The separation of serum from blood cellular elements was carried out 8-10 hours after taking blood samples. The serum was preserved in refrigerator at +4 ° C until the all analysis was carried out. We determined the total proteins, by the biuret method, standard bicarbonate (alkaline reserve) by titrimetric Tashiro indicator method, inorganic phosphorus by spectrophotometric method, glucose with Frank Kirnberger method, urea by spectrophotometric diacetil monoksim method, total calcium and magnesium by titrimetric Eriochrom black indicator method [31]. 2.

2

Analyses

and

determination

of

biochemical indices The analyses were performed in the laboratory of Physiology at the Faculty of Veterinary Medicine of Tirana, Albania. The determination of BCS was realized in each physiological condition with a visual and palpation method. 2. 3 Statistical evaluation The data of the study were processed statistically by ANOVA method and defined correlations between BCS and metabolic profile indicators and multiple regression equation, where in the role of dependent indicators is BCS (Y), while metabolic profile indicators constituted the independent variables (X).

2. Results and Discussion Correlative interactions between BCS and metabolic indicators obtained in this study are presented in Table 1. Total protein in the serum cows two weeks after calving and one month after calving have strong correlation with BCS (P >0. 05), while in the period two weeks before calving and two months after calving these correlations are weak and statistically not proved (P<0. 05). Alkaline reserve (standard bicarbonate) has a statistically proved correlation only in the period 2 months after calving (P>0. 05). The correlations in other physiological state are weak (P<0. 05). The inorganic phosphorus level (Pin) has a correlation with BCS only in the period 2 weeks after calving (P>0. 05). Correlation of glicemia to BCS statistically proved in the period two weeks before calving and 2 weeks after calving (P>0. 05). The level of urea in blood serum has a correlation to BCS in the period two weeks after calving and a month after calving, while in the period before calving and two months after calving the correlation cannot be confirmed statistically (P>0. 05) [13] indicate that the urea in plasma is mainly associated with the ammonium ions in the rumen and is a derivative of the amino acid catabolism and their biosynthesis by the rumenoreticular microorganisms. The cows negative energy balance leads to an increasing of aminoacids biosynthesis by ruminal microorganisms and the urogenesis decreased because of restricted of the passing of ammonium ions in the blood. Correlation of total calcium level in blood serum to BCS is weak and unproved statistically (P <0. 05). Magnesemia represents a strong correlation to BCS in the period two weeks before calving and 2 weeks after calving (P>0. 05). The correlation of magnesemia during the period one month and two months after calving, is not confirmed statistically (P<0. 05).

Table 1: Correlation between metabolic indicators and BCS in cows Period

BCS:Prot

-14 day +14 day +30 day +60 day

-0. 26 0. 65a 0. 46a 0. 02

BCS:Bicarb. standard 0. 18 0. 13 0. 04 0. 48a

BCS:Pin

BCS:Glucosis

BCS:Ure

BCS:Ca

BCS:Mg

0. 6a 0. 19 0. 32 0. 22

0. 51a 0. 39a 0. 2 -0. 03

-0. 22 0. 43a 0. 53a 0. 09

0. 24 0. 2 0. 31 0. 36

-0. 39a 0. 47a 0. 3 0. 21 P a>0. 05; b<0. 05

The metabolic indicators obtained in this study have a positive correlation in the physiological International Conference 31 October 2012, Tirana

conditions. But however most correlation do not have a statistical validity (Tab. 1) achieved similar

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conclusions [24, 6]. The metabolic mechanism of this effect is related to the stimulation of insulin hormone synthesis as well as growth factors. It also inhibits the synthesis of GnRH. LH and FSH, a fact that leads in restriction of the development [24]. A typical characteristic of total proteins correlation to BCS is the result that in the before calving period we observe a negative correlation (r = 0. 26), while in the after calving periods we observe a positive correlation (r = 0. 65, r = 0. 46). This refers to the fact that in these periods, the BCS falls and as a consequence total protein level in the blood serum falls too. So, in the period 60 days after calving although the correlation is positive it becomes weaker (r = 0. 02). Those are accepted by many researchers [4, 29, 15] as the BCS declining inhibition period and the beginning of its growth. We notice a rising of total protein level before the growth of BCS, and most particularly a rising of the albumen fraction. Multiple regression equations, according to the physiological condition, presented below, show that the studied metabolic indicators provide strong connections or affinities in the three physiological conditions related to the BCS. These equations are statistically proved and they have a predictive value. The multi factorial regression equation in the period one month after calving has not a statistical proved, and consequently doesn’t have a predictive value. Multiple regression equation in cows 2 weeks before calving: Y = 1. 76 - 0. 42X1 + 0. 006X2 + 0. 45X3 + 0. 049X4 - 0. 05X5 - 0. 06X6 + 0. 17X7 R2 = 0. 69, Fllog = 6. 36 P = 0. 00051 BCS defining variables (Y), marked with an X present a metabolic profile indicator. X1 = total proteins; X2 = alkaline reserve; X3 = inorganic phosphorus; X4 = glucose; X5 = Urey; X6 = total calcium; X7 = Magnesium. The metabolic profile indicators affect the BCS taken in this study. This correlation is evidenced by the validity of the multiple regression equation for those cows in the two weeks before calving period. The equation has R2 = 0. 69 and P = 0. 00051. This validity degree shows that the regression equation can be used to predict the BCS. The increase of total proteins in the serum has an impact on the reduction of BCS in cows. This is related to the change of serum protein fractions. We notice, at this stage, a decrease of serum albumins until the minimum limit of the normal value, meanwhile the increase occurs mainly because the effect of globulins fraction [19, 26]. International Conference 31 October 2012, Tirana

The special metabolic indicators results in different size and in a different direction. The positive correlations with BCS have Standard bicabonate, inorganic phosphorus, glucose and magnesium levels, while the other metabolic indicators have a negative impact on the BCS level in cows. The positive influence of the standard bicarbonate relates to the acids – basic equilibrium condition located in the body and which is reflected in the standard bicarbonate level in the limits ± 276 mg %. The reduction or increase of this parameter brings negative effects [14, 8]. This fact is also supported by the positive role of phosphate ion. Because this ion are a component of phosphate buffered system and a component of kation anion difference (DCAD) along with magnesium ions [ 23, 8, 10], we can say they have a great importance. The participation of phosphate ions in glycolisis, is another positive role they have in cows BCS [22]. Multiple regression equation in cows 2 weeks after calving: Y = -3. 35 + 0. 337X1-0. 002X2 + 0. 07X3 + 0. 019X4 + 0. 03X5 + 0. 22X6 + 0. 445X R2 = 0. 65, F llog = 4. 5, P = 0. 0053 Two weeks after calving period the validity of multiple regression equation is reduced compared to its before calving period (R2 = 0. 65, Fllog = 3. 1 and P = 0. 0053). However, the equation is statistically true and can be used for predictive purposes. This condition indicates that in the animal organism has already started the mobilization of endogenous resources and especially the lipolysis intensification [24]. The urea level data demonstrated that proteolysis has not started, a fact that would increase the level of urea in the blood [18]. Activity of triacilgliceridlipase by adrenalin, increased triacylglicerol hydrolysis and decreased adipocit volume. As a consequence of these two actions BCS decreased [31, 24]. This is observed in the early stage of lactation in cows. The total protein level in blood serum has a positive impact on the BCS of cows. This phenomenon is related to the fact that the beginning of endogenous protein mobilization has led to the intensification of albumins biosynthesis [16]. The glucose needs in the first period of 21 days after calving increased from 1000 to 1100 g/day. The gluconeogenesis is significantly intensified [16]. This metabolic situation brings a positive impact of glicemia in the BCS, but this effect is lower than in the before calving period .

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Metabolic profile and body condition score of cows during the puerperal period

The bicarbonate standard level in blood serum has negative effect on the BCS. This indicates that the metabolic effects in terms of mobilization of endogenous energy reserves has influenced in the acids-basic equilibrium of the whole organism [12]. The inorganic phosphorus level in the blood serum has a positive effect on BCS. But in this physiological condition the impact of phosphate ions is lower. This happens because it is affected by two metabolism aspects in the liver such as the inhibition of glycolysis and the promotion of gluconeogenesis. Another metabolism aspect is excretion of phosphate ions in urine that affects the preservation of acids - basic equilibrium [11]. Multiple regression equation in cows 1 month after calving: Y = 0. 05 + 0. 18X1 +0. 0009 X2 + 0. 07X3 + 0. 0007X4 + 0. 022X5 + 0. 054X6 - 0. 04X7 R2 = 0. 44, Fllog = 1. 9, P = 0. 13 The multiple regression equation, in the period of one month after calving cows, undergoes significant changes. The validity of this equation is not confirmed statistically. (R2 = 0. 44, Fllog = 1. 9 and P = 0. 13). All multiple regression equation coefficients have a positive affinity with the BCS, except of magnesium level in the serum which has a negative affinity. The source of this difference remains still to be elucidated. We emphasize the lipolysis by hormonal action and the under action as well of the parasympathic nervous system. The concentration of free fatty acids in blood plasma is increased significantly [4]. The proteolysis and the gluconeogenesis is significantly intensified in this physiological condition, mainly in muscle tissue and the BCS further drops [19]. The level of urea in blood serum increases, proving in this way the intensification of proteolysis associated with glukoneogenesis. Multiple regression equation in cows 2 months after calving: Y = 0. 62 - 0. 34X1 + 0. 01X2 - 0. 2X3 - 0. 0018X4 + 0. 014X5 + 0. 27X6 + 0. 415X7 R2 = 0. 55, Fllog = 3. 1, P = 0. 025 Two months after calving period the multiple regression equation between BCS (Y) and metabolic profile indicators has a high validity (R2 = 0. 55, Fllog = 3. 1 and P = 0. 025). This proves that even this equation can be used for predictive purposes. The protein level in blood serum negatively affects the BCS. This is related to the fact that the decline in BCS is associated with the total protein increased in the International Conference 31 October 2012, Tirana

serum. But this has come mainly from the increase of globulin fraction and the albumins fraction decrease. 3. Conclusions The correlative and regressive analysis between BCS and metabolic indicators allow us to conclude: • Two weeks before calving, correlations statistically are proved between BCS with the inorganic phosphorus, glucose and magnesemia in blood serum. • Two weeks after calving, correlations statistically are proved between BCS with the total protein, glucose, urea and magnesium in blood serum. • One month after calving, correlations statistically are proved between BCS with total protein and urea. • Two month after calving, correlations statistically are proved between BCS and standard bicarbonate. • The multi factorial regression equations is statistically confirmed in two weeks before calving period, two weeks after calving period and 2 months after calving period, but it is not statistically confirmed for the one month after calving period.

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7. Dale AB, Rasby RJ, Rush IG, Connee R: Cow body condition scoring management tool for monitoring nutritional status. Beef cattle handbook, 2001. 8. Drennan MJ, Berry DP: Factors affecting body condition score, live weight and reproduction performance in spring-calving sucler cows. Irish Journal of Agricultural and Food Research, 2006 45:25-38. 9. Edmonson AJ, Lean IJ, Weaver LD, Farver T and Webster G: A body condition scoring chart of Holstein dairy Cows. J. Dairy Sci.. 1989, 72:6878. 10. Erfle J, Fischer LJ and Sauer FD: In relationship between blood metabolites and an evaluation of their use as criteria of energy status in cows in early lactation. Can. J. Anim. Sci.. 1974, 54, 293303. 11. Esther OC and Martinez GM: Starch digestion and glucose metabolism in the ruminant. INERCIENCA.. 2003, 28(7): 380-385. 12. Fŭrll M, Jatzke N, Bauer K, Stenhōfel I, Gottschalk J: Metaboli syndromein cows: TNF1 and fa metabolism in heifers with different body condition(BCS) at partium. XXV Jubilee Worl Buitarics Congress, 2008. 13. Gergazc Z, Szűcs E: Critical points in the feeding of high yielding dairy Cows. J. of Veterinary Epidemiology.. 2007, 10(1):15-20. 14. Garnsworthy PC and Trops JH: The effect of body condition of dairy cows at calving on their food intake and performance when given complet diets. Animal. Production.. 1982, 35:113. 15. Johnson CL: The effect of feeding in early lactation on food intake, yields of milk, fat and protein and on live weight change over one lactation in dairy cows. J. Agric. Sci. Camb.. 1984, 103:629. 16. Kathleen LO: Effects of method of delivery of glicerol on performance and metabolism of dairy cows during the transition period. Thesis to Master Sci. Cornell University, 2006. 17. Kirovsci D, Samanc H, Jovanovic M, Fratic N, Gvozdis D, Vujanac I, Dimitrijevic B: Fatty liver and serum concentration of insulin and glucose in dairy cows. XXV Jubilee World. Buitarics Congress, 2008. 18. Kumprechtova D, Illek J, Ballet N: The effect of organic and inorganic dietary selenium sources on selenium levels in blood, colostrum and milk and metabolic profile in dairy cows. XXV Jubilee World Buitarics Congress 2008. International Conference 31 October 2012, Tirana

19. Kuniaki S, Kanameda M, Tachibana SH, Ogawa T, Dang T T and Pfeiffer D: A monitoring study on cattle growth and body condition in smalholder dairy farming system in Northern Vietnam. Jour Vet Epidem.. 2006, 10(1): 15-20. 20. Stengärd L, Traven M, Emanuelson U, Holtenius K, Hultgrenand J, Niskanen R: Metabolic profiles in five hight -producing swedish dairy herds with a history of abomasal displacement and ketosis. ACTA Veterinaria Scandinavica.. 2008, 50:31 doi:10. 1186/1751-0147-50-31. 21. Macrae AI, Withaker DA, Burrough E, Dowell:Use of metabolic profiles for theassessment of dietary adequacy in UK dairy herds. Veterinary record. 2006, 159: 655-661. 22. Mane B, Beqiraj D: Biokimia. Botime Vllamasi. Tiranë, 2011. 23. Mc Donald : Animal Nutrition, 5th edition, 1995. 24. Mouffok C, Madani T. Smara L. Baitiche M. Allouche LF: Relationship between BCS, body weight, some nutritional metabolites changes in blood and reproduction in Algerian Montabeliad cows. Vet. World.. 2011, 4(10): 461-466. 25. National Research Council: Nutrient requirements of dairy cattle. Natl. Acad. Press, Washington, DC, 2001. 26. Pedron O, Cheli F, Santore E, Baroli D and Rizzi R: Effect of body condition score on performance, some blood parametres and milk fatty acid composiotion I dairy cows. J. dairy Sci.. 1993, 76:2528-2535. 27. Pushpakumara PG, Gardner NH, Reynolds CK, Beever DE, Wathes DC: Theirology Relationship betwen transition period diet, metabolic parameters and fertility in lactating dairy cows. Theriogenology. 2003, 60(6):11 6585 28. Reynolds CK and Beever DE: Energy requirements and responses: UK prespective, 1995, 31-4. 29. Santos J, Bisinotto RS, Ribeiro ES, Lima FS and Thatcher WW: Impacts of metabolism and nutrition during the transition period on fertility of dairy cows. High plains dairy conference. Amarillo, Texas, 2012. 30. Burkholder WJ : Use of body condition scores in clinical assessment of the provision of optimal nutrition. JAVMA.. 2000, 217, No 5. 650-654.

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