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Relationships between milk fatty acids composition in early lactation and subsequent reproductive performance in Czech Fleckvieh cows
Animal Reproduction Science 155 (2015) 75–79
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Animal Reproduction Science journal homepage: www.elsevier.com/locate/anireprosci
Relationships between milk fatty acids composition in early lactation and subsequent reproductive performance in Czech Fleckvieh cows L. Stádník, J. Ducháˇcek, J. Beran ∗ , R. Touˇsová, M. Ptáˇcek Department of Animal Husbandry, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kam´ ycká 129, 16521 Prague 6 – Suchdol, Czech Republic
a r t i c l e
i n f o
Article history: Received 16 July 2014 Received in revised form 30 January 2015 Accepted 2 February 2015 Available online 14 February 2015 Keywords: Negative energy balance Milk composition Fatty acids Reproduction
a b s t r a c t Increase of milk yield after calving causes changes in milk fatty acids (FA) composition and simultaneously corresponds with reproduction performance decrease. The objective of this study was to evaluate the relationships between milk FA group composition (SFA, saturated fatty acids; MUFA, monounsaturated fatty acids; and PUFA, polyunsaturated fatty acids) during the first 5 lactation weeks and subsequent reproductive results (INT, calving to first service interval; NUM, number of services per conception, and DO, days open) in Czech Fleckvieh cows. A total of 1231 individual milk samples from 382 cows were collected and subsequently analyzed. Simultaneously, body condition score (BCS) was weekly evaluated as well. Software SAS 9.1 was used for statistical analysis. Daily milk yields increased whereas BCS, milk fat and protein contents decreased during period observed. The reduction of basic milk components (% of fat, % of protein) was associated with increased SFA and decreased MUFA, respectively PUFA contents. Significant (P < 0.01–0.05 days) increase in NUM (+0.15 to +0.29 AI dose) and DO (+8.16 to 15.44 days) were detected in cows with the lowest SFA content. On the contrary, cows with the highest content of MUFA presented significantly (P < 0.01–0.05) higher values of NUM (+0.13 to +0.30) and DO (+7.26 to +15.35 days). Milk FA groups composition in early lactation potentially used as NEB indicators, especially SFA and MUFA proportion, affected subsequent reproductive results of Czech Fleckvieh cows. Therefore, its post-partum values could serve as predictors of potential fertility of dairy cows. © 2015 Elsevier B.V. All rights reserved.
1. Introduction Peripartal period is the most important in herd management (Adamski et al., 2011; Lane et al., 2013). The biggest changes in intensity of dairy cows’ metabolism are reflected mainly in the first part of lactation (Ducháˇcek et al., 2012). This period is characterised by the reduction of feed intake, insufficient for maintenance requirements,
∗ Corresponding author. Tel.: +420 224 383 070. E-mail address: [email protected] (J. Beran). http://dx.doi.org/10.1016/j.anireprosci.2015.02.002 0378-4320/© 2015 Elsevier B.V. All rights reserved.
milk production, and reproductive performance (Reist et al., 2000). As a result, cows enter the stage of negative energy balance (NEB) (Walsh et al., 2011). According to Rossi et al. (2008), the depth and duration of NEB is the main factor influencing the decline of reproductive efficiency in high-yielding dairy cows. The influence of NEB on reproductive parameters was documented in a number of studies, however mainly in Holstein cows only (Wathes et al., 2007; Tamadon et al., 2011; Esposito et al., 2014). The utilization of energy reserves is reflected in the content of milk fat (F) (Bauman et al., 2006), respectively in fatty acids (FA) composition and mutual ratio between
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individual FA groups proportion in milk (Ducháˇcek et al., 2014). These FA proportions are represented by saturated (SFA) (above 73%), unsaturated (UFA) (above 26%), from that monounsaturated (MUFA) (above 23%), polyunsaturated (PUFA) (above 3%) and other groups of FA as is volatile (VFA) or branched (BFA) (Peˇsek et al., 2006; Stoop et al., 2009; Stádník et al., 2013; Ducháˇcek et al., 2013). The composition of these FA groups in milk depends on breed, lactation stage, depth of NEB, individuality, and other factors (Kaylegian and Lindsay, 1995; Hanuˇs et al., 2010). Fatty acids are hormones precursors important for metabolism and reproduction (Gulliver et al., 2012), for example prostaglandins (Petit et al., 2001). Bastin et al. (2011a) confirmed the relationships between FA groups and reproduction, respectively days open length in Holstein cattle. Milk F varies throughout lactation and is affected by many factors (Petit et al., 2001), energy status among them ˇ (Cejna and Chládek, 2005). Milk F and FA are also easily and widely determined milk characteristics in practice. Studies by Berry et al. (2006), Soyeurt et al. (2006), and Bastin et al. (2011b) detected fatty acid composition in milk for predicting energy status and also fertility in Holstein dairy cows using MIR (mid infrared). Also, we can hypothesis that FA groups composition during the first lactation period influence reproductive parameters of Czech dual-purpos cattle breed. Therefore, the objective of the study was to evaluate the effect of milk FA group composition in milk during the first 5-week post partum on subsequent reproductive traits in Czech Fleckvieh cows.
2. Material and methods 2.1. Animals A total of 382 Czech Fleckvieh cows from a single farm were included in the analysis (103, 113, 84, 83 cows in the 1st, 2nd, 3rd, and 4th and subsequent lactations, respectively). Milk samples (n = 1231) were collected weekly during the first 5 weeks of lactation. Lower number of milk samples was caused by practical conditions as mastitis incidences and precocious transfer of cows to other production groups/stable with different feeding ratio composition. The BCS of cows was determined in accordance with the methodology for Czech Fleckvieh dairy cows (Hanuˇs et al., 2004; Kuˇcera, 2009) by the only one technician weekly as well. A body condition index (a 5-point scale with 0.25 point increments) was used to evaluate BCS level. The cows calved within the period of 6 months, from October to March. The cows were loose housed in a cubicle straw-bedded barn and fed a total mixed ratio (TMR). The ingredient composition of the diet corresponded to the level of daily milk yields (MY). Thus the TMR was same for all the animals through the entire period (5 weeks of lactation). Therefore the detailed proportion of FA content in TMR was not objective of this study. TMR was consisted of maize and alfalfa silage (51%), grass and alfalfa hay (20%), brewery draff (6%), bakery waste (5.95%), molasses commercial concentrates (7%), barley (10%) and mineral supplements (0.05%).
2.2. Collection of samples and analyses Two aliquot milk samples from each cow were collected in accordance with the official methodology of the milk performance recording system (ICAR, 2012). The first sample with a preservative was heated to 39 ± 1 ◦ C and used to determine basic milk components (% of fat, % of protein) using Milkoscan 133B (N. Foss Electric; Denmark). The second sample without a preservative was used for the extraction of F and determination of FA composition. Standard method for F extraction in accordance ˇ with CSN EN ISO 1211 (570534) was applied. The extract was obtained using a water-based-solution of ammonia, ethanol, diethylether and petrolether. FA methyl esters were prepared by the potassium hydroxide catalysed methylation and extracted into heptane. Gas chromatography (GC) of FA methyl esters was performed using the Master GC (DANI Instruments S.p.A.; Italy) (split regime, FID detector) on a column with polyethylene glycol stationary phase (FameWax – 30 mm × 0.32 mm × 0.25 m). Helium was used as the carrier gas at a flow rate of 5 ml/min. The temperature programme used for GC was as follows: 50 ◦ C (2 min), after which the temperature was increased to 230 ◦ C at 10 ◦ C/min (8 min), the temperature of the detector being 220 ◦ C. Content (mg.100 g−1 ) and proportion (%) of 34 individual FA subsequently divided to FA groups (SFA, MUFA, PUFA) were determined from individual milk samples. Data characterizing cows´ı reproductive performance (INT, calving to first service interval; NUM, number of services per conception; and DO, days open) were obtained from the farm records software. Cows were examined sonographically between 60 and 74 days after parturition. Cows with corpus luteum detected on ovary were subsequently inseminated at natural heat. If sonographic examination detected luteal or follicular cysts, the cow was included to OVSYNCH program as biotechnology generally applied in dairy farms (Alkar et al., 2011). 2.3. Data handling and statistical analysis The data were analysed with statistical software SAS 9.1 (SAS/STAT® 9.1. 2004). The procedures MEANS and UNIVARIATE were used to calculate basic statistics. The REG procedure (STEPWISE option) was used to develop a model including effects of parity, FA group, and regression on days in milk for the evaluation of reproductive parameters in Czech Fleckvieh cows. The normal distribution of dataset was expected according to Salkind and Rasmussen (2007). To obtain balanced data, cows were divided into four classes for parity (1st − n = 103, 2nd − n = 113, 3rd − n = 84, and 4th and subsequent lactations − n = 83). FA groups were divided into three classes. Limit criteria were determined as the mean ± standard deviation (<¯x − 1/2 s; x¯ − 1/2 s to x¯ + 1/2 s; >¯x + 1/2 s) from the basic statistics of the whole set of samples collected during 5 weeks of observation due to obtaining balanced groups analysed. Each individual sample of milk was subsequently allocated to one of three groups according to limit values stated. The distribution of milk samples to groups
L. Stádník et al. / Animal Reproduction Science 155 (2015) 75–79 Table 1 The sorting of FA groups. FA groups
% In milk
n (Samples)
n (Cows)
SFA
<67.93 67.93–73.57 >73.57 <22.89 22.89–28.23 >28.23 <3.2 3.2–4.11 >4.11
390 447 394 394 454 383 402 495 334
241 263 202 229 252 191 248 271 210
MUFA
PUFA
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composition is probably more practical and could present potential of reproduction abilities prediction as well. The average daily MY in individual 5 weeks observed ranged from 25.57 to 29.17 L with standard deviations from 6.13 to 7.2 L. The content of F decreased during the entire period, while P content was reduced only to week 4, and then slightly increased. Daily MY increased during the first 4 weeks of lactation. Changes in MY and milk components proportions could be explained by lipomobilization during NEB (Ducháˇcek et al., 2014). The average contents of FA groups in observed 5 weeks of lactation were 70.78%, 25.54%, and 3.65% for SFA, MUFA and PUFA, respectively. Whereas SFA contents continuously increased, BCS and UFA decreased. The average values of INT and DO were 71.57 and 100.73 days, respectively. The average NUM of observed Czech Fleckvieh cows was 1.75. Mentioned relationships already have been widely documented by ˇ Komprda et al. (2005), Roche et al. (2009), and Rehák et al. (2012). Influence of parity on basic reproductive parameters of cows was observed in first part of evaluation. However, no significant (P > 0.05) and relatively small differences were computed for INT (+1.04 to +4.08 days), NUM (+0.02 to +0.25), and DO (+3.21 to +8.51 days) as well. The effects of low, medium, and high SFA in milk on reproductive parameters are presented in Table 2. Division of cows was selected due to elimination of cow’s metabolism individuality and necessarily to evaluate effect of repeated FA groups proportion on dairy cows reproductive capabilities. No relationship was determined between INT and SFA groups, probably due to breeding decisions associated with the system of herd reproductive management commonly used in Czech dairy farms. Management of dairy cows´ı reproduction usually starts using OVSYNCH or other hormonal treatment program (Alkar et al., 2011) from 60 to 70 days in milk. The NUM tended to decrease (from 1.9 to 1.61) with increasing SFA contents associated with the recovery from NEB confirmed by BCS increase. A significant difference (−0.29 AI dose; P < 0.01) in the value of NUM was found between the groups with the lowest (below 67.93%) and the highest (above 73.57%) SFA proportion. This might be explained by the increased content of SFA over lactation weeks (Komprda et al., 2005), when the high concentration of SFA group is associated with the recovery from NEB. Similarly to NUM, the length
SFA, saturated fatty acids; MUFA, monounsaturated fatty acids; PUFA, polyunsaturated fatty acids; n, number of observation.
according to SFA, MUFA, and PUFA proportion is shown in more detail in Table 1. The model equation used for the reproductive parameters evaluation was as follows: Yijkl = + PARITYi + FAi + b.DAYk + eijkl Yijkl = + PARITYi + FAj + b.DAYk + eijkl , Yijkl = dependent variable (INT, NUM, DO), = mean value of dependent variable, PARITYi = fixed effect of ith parity of lactation (j = 1 − 1st lactation, n = 103; 2 − 2nd lactation, n = 113; 3 − 3rd lactation, n = 84; 4 − 4th and subsequent lactations, n = 83), FAj = fixed effect of jth FA group (SFA – <67.93%, n = 390; 67.93–73.57%, n = 447; >73.57%, n = 394), or (MUFA – <22.89%, n = 394; 22.89–28.23%, n = 454; >28.23%, n = 383), or (PUFA – <3.2%, n = 402; 3.2–4.11%, n = 495; >4.11%, n = 334), b.DAYk = regression on days in milk, eijkl = random error. Significance levels P < 0.05, P < 0.01, and P < 0.001 were used to evaluate the differences between groups. 3. Results and discussion Milk components (F content, F to P ratio and, milk FA) are ˇ commonly recommended (Cejna and Chládek, 2005; Gross et al., 2011) for determination of NEB level. On the other hand, these indicators are not always entirely applicable for prediction precise state of metabolism and metabolic disorders in NEB period. For example, NEFA (non-esterified fatty acids) concentration in blood could be a more valuable indicator (González et al., 2011). However, sampling and analysis of milk is simpler, more accessible and cheaper. Therefore, determination of milk FA content, respectively Table 2 The effect of milk SFA, MUFA and PUFA content on reproduction results of cows. FA groups
Categories (%)
INT LSM ± SE
SFA
<67.93 67.93–73.57 >73.57 <22.89 22.89–28.23 >28.23 <3.2 3.2–4.11 >4.11
72.62 71.39 71.58 71.51 71.71 72.33 71.83 71.95 71.73
MUFA
PUFA
± ± ± ± ± ± ± ± ±
NUM LSM ± SE 0.681 0.621 0.698 0.696 0.617 0.684 0.672 0.584 0.73
1.9 1.75 1.61 1.61 1.74 1.91 1.7 1.73 1.85
± ± ± ± ± ± ± ± ±
0.058a 0.052 0.06b 0.059a 0.052 0.058b 0.057 0.049 0.062
DO LSM ± SE 108.68 100.52 93.24 93.32 100.58 108.67 98.02 100.64 104.94
± ± ± ± ± ± ± ± ±
2.508A,a 2.267B 2.589b 2.581a 2.253A 2.514b,B 2.496 2.153 2.715
SFA, saturated fatty acids; MUFA, monounsaturated fatty acids; PUFA, polyunsaturated fatty acids; INT, calving the first service interval; NUM, number of services per conception; DO, days open; PUFA, polyunsaturated fatty acids; a,b reps. A,B – different superscript letters confirm statistical significance of difference among within the effect rows at the P < 0.05; resp. P < 0.01 level.
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of DO period was shortened with the increasing content of SFA. The longest DO (108.68 days) was observed in the cows with the lowest content of SFA (below 67.93%), whereas the shortes one (93.24 days) in the cows with the highest content of SFA (above 73.57%). The differences between SFA groups below 67.93% and 67.93–73.57% (−8.16 days) and above 73.57% (−15.44 days) were statistically significant (P < 0.05 and P < 0.01, respectively). These results simultaneously confirmed the same trend detected in INT. Above mentioned findings are also documented by cows recovery from NEB associated with higher SFA contents (Ducháˇcek et al., 2012). This statement was confirmed by Bastin et al. (2011a), who described a negative genetic correlation between SFA content and length of DO period in Holstein cows. As explained and confirmed above higher SFA content can be associated with later stages of lactation (Komprda et al., 2005) and/or NEB overcome (Ducháˇcek et al., 2012). Hence, Czech Fleckvieh cows with higher SFA proportion in milk during first 5 weeks of lactation presented higher robustness to NEB followed by significantly favourable level of reproductive indicators. The effect of milk MUFA content on reproductive parameters is shown in Table 2 as well. Opposite tendencies were observed in comparison with the content of SFA. The first evaluated reproductive indicator INT, tended to increase with increasing milk MUFA contents (from 71.51 to 72.33 days). Although, the differences detected were not significant (P > 0.05). However, the influence of the recovery from NEB on reproductive parameters was evident as findings of Podpeˇcan et al. (2008). Similarly NUM tended to increase together with increasing MUFA content. The values of NUM (1.61) in the group with the lowest MUFA (below 22.89%) significantly differed (−0.30; P < 0.01) compared to the highest one (above 28.23%). Also DO increased with increasing MUFA content. The DO differences (7.26–15.35) between MUFA groups below 22.89%, 22.89–28.23%, and above 28.23% were statistically significant (P < 0.01). However, the higher days in milk the lower MUFA proportion as the cows recovered from NEB (Ducháˇcek et al., 2012). This statement was also confirmed by positive correlations between MUFA contents and DO detected by Bastin et al. (2011a). In accordance with results of Gross et al. (2011), our findings indicated the content of MUFA was the highest in early lactation with subsequent gradual decrease corresponding to NEB reducing. Thus, the cows with lower MUFA contents in the period observed were not in severe NEB and exhibited improved reproductive parameters. The results of reproductive parameters as affected by the PUFA content in milk are described in Table 2. Unfortunately, these differences were completely no significant (P > 0.05). There were no tendencies in the length of INT. Again, NUM tended to increase with increasing PUFA contents, related to NEB intensity as well as in MUFA evaluation. This corresponds to the process of the recovery from NEB indicated by the decreasing content of PUFA in milk over lactation (Komprda et al., 2005; Ducháˇcek et al., 2012). Holstein cows with high PUFA contents suffer from a more severe NEB resulting in a longer period of DO (Bastin et al., 2011b). In opposite, a lot of authors (Dirandeh et al., 2013; Petit et al., 2002; Robinson et al., 2002) observed mostly
positive relationships between PUFA supplementation to feeding ration and reproductive performance. Generally, higher UFA proportion in milk in the beginning of lactation (especially MUFA) indicated possible negative effect on subsequent reproductive performance of Czech Fleckvieh cows. Tamadon et al. (2011) explained decrease of reproductive characteristics by inhibition of maturing follicles, ovulation, corpus luteum development, and prolong onset of ovulation activity after calving in extreme cows NEB corresponding to higher MUFA, resp. PUFA proportion in milk yield (Stádník et al., 2013). Moreover, the occurrence and severity of NEB prolong the period between calving and first ovulation documented by Wathes et al. (2007), increase embryonic mortality, and the risk of uterine diseases as stated by Vacek et al. (2007). Mentioned findings and relationships confirm importance of depth and duration of NEB in dual purpose Czech Fleckvieh cows in relation to reproduction as well as economy effectiveness of dairy cows husbandry as a whole. 4. Conclusions The results observed in Czech Fleckvieh cows in this study indicated significant relationships between SFA, respectively MUFA proportion in milk in the first 5 weeks of lactation and subsequent reproductive characteristics. The values of NUM and DO were more preferable (P < 0.05–0.01) in the cows with a higher milk SFA content documenting less deep NEB. On the contrary, less favourable values of NUM and DO (P < 0.05–0.01) were determined in the groups with a higher MUFA content corresponding with more severe NEB. The determination of FA composition in milk of Czech Fleckvieh cows during the first weeks after calving may be used as a pre-selection criterion for cows with subsequent reproductive problems. A special attention should be given to these cows by herd managers. Conflict of interest The authors declare that there are no conflict of interests. Acknowledgement This research was funded by “S” grant of MSMT CR, and by Project NAZVQI91A061. References ´ Adamski, M., Kupczynski, R., Chládek, G., Falta, D., 2011. Influence of propylene glycol and glycerin in Simmental cows in periparturient period on milk yield and metabolic changes. Arch. Tierz. Arch. Anim. Breed. 54, 238–248. Alkar, A., Tibary, A., Wenz, J.R., Nebel, R.L., Kasimanickam, R., 2011. Presynchronization with GnRH 7 days prior to resynchronization with CO-Synch did not improve pregnancy rate in lactating dairy cows. Theriogenology 76, 1036–1041. Bastin, C., Gengler, N., Soyeurt, H., 2011a. Genetic relationships between fertility and content of major fatty acids in milk for first-parity Walloon Holstein cows [online]. In: Joint Annual Meating 2011, New Orleans, USA [2014-04-12], J. Anim. Sci. 89, E-Suppl. 1/J. Dairy Sci. 94, E-Suppl.1, p. 705. Available from http://www.jtmtg.org/JAM/2011/abstracts/0705.PDF
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Animal Reproduction Science journal homepage: www.elsevier.com/locate/anireprosci
Relationships between milk fatty acids composition in early lactation and subsequent reproductive performance in Czech Fleckvieh cows L. Stádník, J. Ducháˇcek, J. Beran ∗ , R. Touˇsová, M. Ptáˇcek Department of Animal Husbandry, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kam´ ycká 129, 16521 Prague 6 – Suchdol, Czech Republic
a r t i c l e
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Article history: Received 16 July 2014 Received in revised form 30 January 2015 Accepted 2 February 2015 Available online 14 February 2015 Keywords: Negative energy balance Milk composition Fatty acids Reproduction
a b s t r a c t Increase of milk yield after calving causes changes in milk fatty acids (FA) composition and simultaneously corresponds with reproduction performance decrease. The objective of this study was to evaluate the relationships between milk FA group composition (SFA, saturated fatty acids; MUFA, monounsaturated fatty acids; and PUFA, polyunsaturated fatty acids) during the first 5 lactation weeks and subsequent reproductive results (INT, calving to first service interval; NUM, number of services per conception, and DO, days open) in Czech Fleckvieh cows. A total of 1231 individual milk samples from 382 cows were collected and subsequently analyzed. Simultaneously, body condition score (BCS) was weekly evaluated as well. Software SAS 9.1 was used for statistical analysis. Daily milk yields increased whereas BCS, milk fat and protein contents decreased during period observed. The reduction of basic milk components (% of fat, % of protein) was associated with increased SFA and decreased MUFA, respectively PUFA contents. Significant (P < 0.01–0.05 days) increase in NUM (+0.15 to +0.29 AI dose) and DO (+8.16 to 15.44 days) were detected in cows with the lowest SFA content. On the contrary, cows with the highest content of MUFA presented significantly (P < 0.01–0.05) higher values of NUM (+0.13 to +0.30) and DO (+7.26 to +15.35 days). Milk FA groups composition in early lactation potentially used as NEB indicators, especially SFA and MUFA proportion, affected subsequent reproductive results of Czech Fleckvieh cows. Therefore, its post-partum values could serve as predictors of potential fertility of dairy cows. © 2015 Elsevier B.V. All rights reserved.
1. Introduction Peripartal period is the most important in herd management (Adamski et al., 2011; Lane et al., 2013). The biggest changes in intensity of dairy cows’ metabolism are reflected mainly in the first part of lactation (Ducháˇcek et al., 2012). This period is characterised by the reduction of feed intake, insufficient for maintenance requirements,
∗ Corresponding author. Tel.: +420 224 383 070. E-mail address: [email protected] (J. Beran). http://dx.doi.org/10.1016/j.anireprosci.2015.02.002 0378-4320/© 2015 Elsevier B.V. All rights reserved.
milk production, and reproductive performance (Reist et al., 2000). As a result, cows enter the stage of negative energy balance (NEB) (Walsh et al., 2011). According to Rossi et al. (2008), the depth and duration of NEB is the main factor influencing the decline of reproductive efficiency in high-yielding dairy cows. The influence of NEB on reproductive parameters was documented in a number of studies, however mainly in Holstein cows only (Wathes et al., 2007; Tamadon et al., 2011; Esposito et al., 2014). The utilization of energy reserves is reflected in the content of milk fat (F) (Bauman et al., 2006), respectively in fatty acids (FA) composition and mutual ratio between
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individual FA groups proportion in milk (Ducháˇcek et al., 2014). These FA proportions are represented by saturated (SFA) (above 73%), unsaturated (UFA) (above 26%), from that monounsaturated (MUFA) (above 23%), polyunsaturated (PUFA) (above 3%) and other groups of FA as is volatile (VFA) or branched (BFA) (Peˇsek et al., 2006; Stoop et al., 2009; Stádník et al., 2013; Ducháˇcek et al., 2013). The composition of these FA groups in milk depends on breed, lactation stage, depth of NEB, individuality, and other factors (Kaylegian and Lindsay, 1995; Hanuˇs et al., 2010). Fatty acids are hormones precursors important for metabolism and reproduction (Gulliver et al., 2012), for example prostaglandins (Petit et al., 2001). Bastin et al. (2011a) confirmed the relationships between FA groups and reproduction, respectively days open length in Holstein cattle. Milk F varies throughout lactation and is affected by many factors (Petit et al., 2001), energy status among them ˇ (Cejna and Chládek, 2005). Milk F and FA are also easily and widely determined milk characteristics in practice. Studies by Berry et al. (2006), Soyeurt et al. (2006), and Bastin et al. (2011b) detected fatty acid composition in milk for predicting energy status and also fertility in Holstein dairy cows using MIR (mid infrared). Also, we can hypothesis that FA groups composition during the first lactation period influence reproductive parameters of Czech dual-purpos cattle breed. Therefore, the objective of the study was to evaluate the effect of milk FA group composition in milk during the first 5-week post partum on subsequent reproductive traits in Czech Fleckvieh cows.
2. Material and methods 2.1. Animals A total of 382 Czech Fleckvieh cows from a single farm were included in the analysis (103, 113, 84, 83 cows in the 1st, 2nd, 3rd, and 4th and subsequent lactations, respectively). Milk samples (n = 1231) were collected weekly during the first 5 weeks of lactation. Lower number of milk samples was caused by practical conditions as mastitis incidences and precocious transfer of cows to other production groups/stable with different feeding ratio composition. The BCS of cows was determined in accordance with the methodology for Czech Fleckvieh dairy cows (Hanuˇs et al., 2004; Kuˇcera, 2009) by the only one technician weekly as well. A body condition index (a 5-point scale with 0.25 point increments) was used to evaluate BCS level. The cows calved within the period of 6 months, from October to March. The cows were loose housed in a cubicle straw-bedded barn and fed a total mixed ratio (TMR). The ingredient composition of the diet corresponded to the level of daily milk yields (MY). Thus the TMR was same for all the animals through the entire period (5 weeks of lactation). Therefore the detailed proportion of FA content in TMR was not objective of this study. TMR was consisted of maize and alfalfa silage (51%), grass and alfalfa hay (20%), brewery draff (6%), bakery waste (5.95%), molasses commercial concentrates (7%), barley (10%) and mineral supplements (0.05%).
2.2. Collection of samples and analyses Two aliquot milk samples from each cow were collected in accordance with the official methodology of the milk performance recording system (ICAR, 2012). The first sample with a preservative was heated to 39 ± 1 ◦ C and used to determine basic milk components (% of fat, % of protein) using Milkoscan 133B (N. Foss Electric; Denmark). The second sample without a preservative was used for the extraction of F and determination of FA composition. Standard method for F extraction in accordance ˇ with CSN EN ISO 1211 (570534) was applied. The extract was obtained using a water-based-solution of ammonia, ethanol, diethylether and petrolether. FA methyl esters were prepared by the potassium hydroxide catalysed methylation and extracted into heptane. Gas chromatography (GC) of FA methyl esters was performed using the Master GC (DANI Instruments S.p.A.; Italy) (split regime, FID detector) on a column with polyethylene glycol stationary phase (FameWax – 30 mm × 0.32 mm × 0.25 m). Helium was used as the carrier gas at a flow rate of 5 ml/min. The temperature programme used for GC was as follows: 50 ◦ C (2 min), after which the temperature was increased to 230 ◦ C at 10 ◦ C/min (8 min), the temperature of the detector being 220 ◦ C. Content (mg.100 g−1 ) and proportion (%) of 34 individual FA subsequently divided to FA groups (SFA, MUFA, PUFA) were determined from individual milk samples. Data characterizing cows´ı reproductive performance (INT, calving to first service interval; NUM, number of services per conception; and DO, days open) were obtained from the farm records software. Cows were examined sonographically between 60 and 74 days after parturition. Cows with corpus luteum detected on ovary were subsequently inseminated at natural heat. If sonographic examination detected luteal or follicular cysts, the cow was included to OVSYNCH program as biotechnology generally applied in dairy farms (Alkar et al., 2011). 2.3. Data handling and statistical analysis The data were analysed with statistical software SAS 9.1 (SAS/STAT® 9.1. 2004). The procedures MEANS and UNIVARIATE were used to calculate basic statistics. The REG procedure (STEPWISE option) was used to develop a model including effects of parity, FA group, and regression on days in milk for the evaluation of reproductive parameters in Czech Fleckvieh cows. The normal distribution of dataset was expected according to Salkind and Rasmussen (2007). To obtain balanced data, cows were divided into four classes for parity (1st − n = 103, 2nd − n = 113, 3rd − n = 84, and 4th and subsequent lactations − n = 83). FA groups were divided into three classes. Limit criteria were determined as the mean ± standard deviation (<¯x − 1/2 s; x¯ − 1/2 s to x¯ + 1/2 s; >¯x + 1/2 s) from the basic statistics of the whole set of samples collected during 5 weeks of observation due to obtaining balanced groups analysed. Each individual sample of milk was subsequently allocated to one of three groups according to limit values stated. The distribution of milk samples to groups
L. Stádník et al. / Animal Reproduction Science 155 (2015) 75–79 Table 1 The sorting of FA groups. FA groups
% In milk
n (Samples)
n (Cows)
SFA
<67.93 67.93–73.57 >73.57 <22.89 22.89–28.23 >28.23 <3.2 3.2–4.11 >4.11
390 447 394 394 454 383 402 495 334
241 263 202 229 252 191 248 271 210
MUFA
PUFA
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composition is probably more practical and could present potential of reproduction abilities prediction as well. The average daily MY in individual 5 weeks observed ranged from 25.57 to 29.17 L with standard deviations from 6.13 to 7.2 L. The content of F decreased during the entire period, while P content was reduced only to week 4, and then slightly increased. Daily MY increased during the first 4 weeks of lactation. Changes in MY and milk components proportions could be explained by lipomobilization during NEB (Ducháˇcek et al., 2014). The average contents of FA groups in observed 5 weeks of lactation were 70.78%, 25.54%, and 3.65% for SFA, MUFA and PUFA, respectively. Whereas SFA contents continuously increased, BCS and UFA decreased. The average values of INT and DO were 71.57 and 100.73 days, respectively. The average NUM of observed Czech Fleckvieh cows was 1.75. Mentioned relationships already have been widely documented by ˇ Komprda et al. (2005), Roche et al. (2009), and Rehák et al. (2012). Influence of parity on basic reproductive parameters of cows was observed in first part of evaluation. However, no significant (P > 0.05) and relatively small differences were computed for INT (+1.04 to +4.08 days), NUM (+0.02 to +0.25), and DO (+3.21 to +8.51 days) as well. The effects of low, medium, and high SFA in milk on reproductive parameters are presented in Table 2. Division of cows was selected due to elimination of cow’s metabolism individuality and necessarily to evaluate effect of repeated FA groups proportion on dairy cows reproductive capabilities. No relationship was determined between INT and SFA groups, probably due to breeding decisions associated with the system of herd reproductive management commonly used in Czech dairy farms. Management of dairy cows´ı reproduction usually starts using OVSYNCH or other hormonal treatment program (Alkar et al., 2011) from 60 to 70 days in milk. The NUM tended to decrease (from 1.9 to 1.61) with increasing SFA contents associated with the recovery from NEB confirmed by BCS increase. A significant difference (−0.29 AI dose; P < 0.01) in the value of NUM was found between the groups with the lowest (below 67.93%) and the highest (above 73.57%) SFA proportion. This might be explained by the increased content of SFA over lactation weeks (Komprda et al., 2005), when the high concentration of SFA group is associated with the recovery from NEB. Similarly to NUM, the length
SFA, saturated fatty acids; MUFA, monounsaturated fatty acids; PUFA, polyunsaturated fatty acids; n, number of observation.
according to SFA, MUFA, and PUFA proportion is shown in more detail in Table 1. The model equation used for the reproductive parameters evaluation was as follows: Yijkl = + PARITYi + FAi + b.DAYk + eijkl Yijkl = + PARITYi + FAj + b.DAYk + eijkl , Yijkl = dependent variable (INT, NUM, DO), = mean value of dependent variable, PARITYi = fixed effect of ith parity of lactation (j = 1 − 1st lactation, n = 103; 2 − 2nd lactation, n = 113; 3 − 3rd lactation, n = 84; 4 − 4th and subsequent lactations, n = 83), FAj = fixed effect of jth FA group (SFA – <67.93%, n = 390; 67.93–73.57%, n = 447; >73.57%, n = 394), or (MUFA – <22.89%, n = 394; 22.89–28.23%, n = 454; >28.23%, n = 383), or (PUFA – <3.2%, n = 402; 3.2–4.11%, n = 495; >4.11%, n = 334), b.DAYk = regression on days in milk, eijkl = random error. Significance levels P < 0.05, P < 0.01, and P < 0.001 were used to evaluate the differences between groups. 3. Results and discussion Milk components (F content, F to P ratio and, milk FA) are ˇ commonly recommended (Cejna and Chládek, 2005; Gross et al., 2011) for determination of NEB level. On the other hand, these indicators are not always entirely applicable for prediction precise state of metabolism and metabolic disorders in NEB period. For example, NEFA (non-esterified fatty acids) concentration in blood could be a more valuable indicator (González et al., 2011). However, sampling and analysis of milk is simpler, more accessible and cheaper. Therefore, determination of milk FA content, respectively Table 2 The effect of milk SFA, MUFA and PUFA content on reproduction results of cows. FA groups
Categories (%)
INT LSM ± SE
SFA
<67.93 67.93–73.57 >73.57 <22.89 22.89–28.23 >28.23 <3.2 3.2–4.11 >4.11
72.62 71.39 71.58 71.51 71.71 72.33 71.83 71.95 71.73
MUFA
PUFA
± ± ± ± ± ± ± ± ±
NUM LSM ± SE 0.681 0.621 0.698 0.696 0.617 0.684 0.672 0.584 0.73
1.9 1.75 1.61 1.61 1.74 1.91 1.7 1.73 1.85
± ± ± ± ± ± ± ± ±
0.058a 0.052 0.06b 0.059a 0.052 0.058b 0.057 0.049 0.062
DO LSM ± SE 108.68 100.52 93.24 93.32 100.58 108.67 98.02 100.64 104.94
± ± ± ± ± ± ± ± ±
2.508A,a 2.267B 2.589b 2.581a 2.253A 2.514b,B 2.496 2.153 2.715
SFA, saturated fatty acids; MUFA, monounsaturated fatty acids; PUFA, polyunsaturated fatty acids; INT, calving the first service interval; NUM, number of services per conception; DO, days open; PUFA, polyunsaturated fatty acids; a,b reps. A,B – different superscript letters confirm statistical significance of difference among within the effect rows at the P < 0.05; resp. P < 0.01 level.
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of DO period was shortened with the increasing content of SFA. The longest DO (108.68 days) was observed in the cows with the lowest content of SFA (below 67.93%), whereas the shortes one (93.24 days) in the cows with the highest content of SFA (above 73.57%). The differences between SFA groups below 67.93% and 67.93–73.57% (−8.16 days) and above 73.57% (−15.44 days) were statistically significant (P < 0.05 and P < 0.01, respectively). These results simultaneously confirmed the same trend detected in INT. Above mentioned findings are also documented by cows recovery from NEB associated with higher SFA contents (Ducháˇcek et al., 2012). This statement was confirmed by Bastin et al. (2011a), who described a negative genetic correlation between SFA content and length of DO period in Holstein cows. As explained and confirmed above higher SFA content can be associated with later stages of lactation (Komprda et al., 2005) and/or NEB overcome (Ducháˇcek et al., 2012). Hence, Czech Fleckvieh cows with higher SFA proportion in milk during first 5 weeks of lactation presented higher robustness to NEB followed by significantly favourable level of reproductive indicators. The effect of milk MUFA content on reproductive parameters is shown in Table 2 as well. Opposite tendencies were observed in comparison with the content of SFA. The first evaluated reproductive indicator INT, tended to increase with increasing milk MUFA contents (from 71.51 to 72.33 days). Although, the differences detected were not significant (P > 0.05). However, the influence of the recovery from NEB on reproductive parameters was evident as findings of Podpeˇcan et al. (2008). Similarly NUM tended to increase together with increasing MUFA content. The values of NUM (1.61) in the group with the lowest MUFA (below 22.89%) significantly differed (−0.30; P < 0.01) compared to the highest one (above 28.23%). Also DO increased with increasing MUFA content. The DO differences (7.26–15.35) between MUFA groups below 22.89%, 22.89–28.23%, and above 28.23% were statistically significant (P < 0.01). However, the higher days in milk the lower MUFA proportion as the cows recovered from NEB (Ducháˇcek et al., 2012). This statement was also confirmed by positive correlations between MUFA contents and DO detected by Bastin et al. (2011a). In accordance with results of Gross et al. (2011), our findings indicated the content of MUFA was the highest in early lactation with subsequent gradual decrease corresponding to NEB reducing. Thus, the cows with lower MUFA contents in the period observed were not in severe NEB and exhibited improved reproductive parameters. The results of reproductive parameters as affected by the PUFA content in milk are described in Table 2. Unfortunately, these differences were completely no significant (P > 0.05). There were no tendencies in the length of INT. Again, NUM tended to increase with increasing PUFA contents, related to NEB intensity as well as in MUFA evaluation. This corresponds to the process of the recovery from NEB indicated by the decreasing content of PUFA in milk over lactation (Komprda et al., 2005; Ducháˇcek et al., 2012). Holstein cows with high PUFA contents suffer from a more severe NEB resulting in a longer period of DO (Bastin et al., 2011b). In opposite, a lot of authors (Dirandeh et al., 2013; Petit et al., 2002; Robinson et al., 2002) observed mostly
positive relationships between PUFA supplementation to feeding ration and reproductive performance. Generally, higher UFA proportion in milk in the beginning of lactation (especially MUFA) indicated possible negative effect on subsequent reproductive performance of Czech Fleckvieh cows. Tamadon et al. (2011) explained decrease of reproductive characteristics by inhibition of maturing follicles, ovulation, corpus luteum development, and prolong onset of ovulation activity after calving in extreme cows NEB corresponding to higher MUFA, resp. PUFA proportion in milk yield (Stádník et al., 2013). Moreover, the occurrence and severity of NEB prolong the period between calving and first ovulation documented by Wathes et al. (2007), increase embryonic mortality, and the risk of uterine diseases as stated by Vacek et al. (2007). Mentioned findings and relationships confirm importance of depth and duration of NEB in dual purpose Czech Fleckvieh cows in relation to reproduction as well as economy effectiveness of dairy cows husbandry as a whole. 4. Conclusions The results observed in Czech Fleckvieh cows in this study indicated significant relationships between SFA, respectively MUFA proportion in milk in the first 5 weeks of lactation and subsequent reproductive characteristics. The values of NUM and DO were more preferable (P < 0.05–0.01) in the cows with a higher milk SFA content documenting less deep NEB. On the contrary, less favourable values of NUM and DO (P < 0.05–0.01) were determined in the groups with a higher MUFA content corresponding with more severe NEB. The determination of FA composition in milk of Czech Fleckvieh cows during the first weeks after calving may be used as a pre-selection criterion for cows with subsequent reproductive problems. A special attention should be given to these cows by herd managers. Conflict of interest The authors declare that there are no conflict of interests. Acknowledgement This research was funded by “S” grant of MSMT CR, and by Project NAZVQI91A061. References ´ Adamski, M., Kupczynski, R., Chládek, G., Falta, D., 2011. Influence of propylene glycol and glycerin in Simmental cows in periparturient period on milk yield and metabolic changes. Arch. Tierz. Arch. Anim. Breed. 54, 238–248. Alkar, A., Tibary, A., Wenz, J.R., Nebel, R.L., Kasimanickam, R., 2011. Presynchronization with GnRH 7 days prior to resynchronization with CO-Synch did not improve pregnancy rate in lactating dairy cows. Theriogenology 76, 1036–1041. Bastin, C., Gengler, N., Soyeurt, H., 2011a. Genetic relationships between fertility and content of major fatty acids in milk for first-parity Walloon Holstein cows [online]. In: Joint Annual Meating 2011, New Orleans, USA [2014-04-12], J. Anim. Sci. 89, E-Suppl. 1/J. Dairy Sci. 94, E-Suppl.1, p. 705. Available from http://www.jtmtg.org/JAM/2011/abstracts/0705.PDF
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