In ternationa l Scholars Journa ls African Journal of Pig Farming ISSN 2375-0731 Vol. 8 (1), pp. 001-006, January, 2020. Available online at www.internationalscholarsjournals.org © International Scholars Journals Author(s) retain the copyright of this article. Full Length Research Paper The influence of the different content of protein fractions in sows’ milk in piglet rearing Ewa Skrzypczak1*, Karolina Szulc1, Monika Demkowicz2, Damian Knecht3, Anna Jankowska- Mąkosa3 , Janusz T. Buczyński1 and Marek Babicz4 1 Department of Pig Breeding and Production, Poznań University of Life Sciences, ul. Wołyńska 33, 60-637 Poznań, Poland. 2 Department of Cattle Breeding and Milk Production, The Institute of Animal Breeding, Wrocław University of Environmental and Life Sciences, ul. J. Chełmońskiego 38C, 51-630 Wrocław, Poland. 3 Department of Pig Breeding, The Institute of Animal Breeding, Wrocław University of Environmental and Life Sciences, ul. J. Chełmońskiego 38C, 51-630 Wrocław, Poland. 4 Department of Breeding and Technology of Pig Production, University of Life Sciences in Lublin, ul. Akademicka 13, 20- 950 Lublin, Poland. Accepted 15 June, 2019 The aim of this study was to investigate the influence of the percentage content of protein fractions in total protein of sow`s colostrums and milk and their influence on the traits related with piglet rearing. The animal specimens were 20 sows of the native Złotnicka White breed. Złotnicka pigs were subjected to the National Genetic Resources Conservation Programme. Colostrum and milk were collected between the 20th and 24th h after parturition and on the 2nd, 3rd, 7th, 14th and 21st day of lactation. A total of 120 samples (60 colostrum samples and 60 milk samples) were collected from all active mammary glands. Individual fractions of total protein were separated by means of electrophoresis on polyacrylamide gel in the presence of sodium dodecyl sulphate (SDS). The piglets’ body weights average daily gains and mortality were checked consecutively after 24 hours after parturition and on the day 7 th , 14 th , and 21 st of lactation. 207 piglets were examined. A highly significant correlation between the number of piglets, daily growths and protein fractions was observed. The most favourable rearing results were obtained at the highest level (III) of individual fractions. The study also proved most of the piglets are lost from the litters when the level of fractions is the lowest (I). Key words: Sows, milk, protein fractions, Złotnicka White, piglet rearing. INTRODUCTION Sows’ colostrum and milk are the main sources of necessary nutrients during the first days of piglets’ lives (Devillers et al., 2004; Skrzypczak et al., 2012a). Milk proteins are particularly significant due to the fact that they participate in all life processes (Wheeler et al., 2007; Stelwagen et al., 2009), they are *Corresponding author. E-mail: ewa.skrzypczak@op.pl. Abbreviations: SDS, Sodium dodecyl sulphate; LSM, least square means; SE, standard errors. an important building block and play an enormous role in the development of the immune system (Bernatowicz and Reklewska, 2003; Sangild, 2003; Lipiński, 2007; Płusa, 2009). The proteins whose role has only partly been recognised deserve attention. Lactoferrin and β- lactoglobulin exhibit the antineoplastic effect (McIntosch et al., 1998; Szulc, 2010), β-lactoglobulin and immuno- globulins G also exhibit the supportive effect in viral and bacterial infections (Pan et al., 2006, 2007; Płusa, 2009), α-lactalbumin is a Ca 2+ carrier and it is an anti- carcinogenic, antibacterial, immunological and supportive factor. In stressful situations, it may lower the blood pressure (Meisel, 1997; Zimecki and Arytm, 2005). Figure 1. Polyacrylamide gel with milk protein fractions. Casein has above all anticoagulant properties and it inhibits thrombocyte aggregation and serotonin liberation (Kuczyńska, 2008; Płusa, 2009). Therefore, investigation of the correlation between sows’ milk protein fractions and results of piglet rearing is justified. MATERIALS AND METHODS Animals The animal materials were 20 sows of the native Złotnicka White breed, which were subject to the National Genetic Resources Conservation Programme ( Szulc et. al., 2012). The research was carried out from October, 2009 to September, 2010. All experimental sows were housed in identical conditions meeting all welfare requirements. The sows were housed in single farrowing crates from about the 10 th day before parturition to the 4 th week of lactation. They were fed individually with standard total mixed rations according to polish Nutrient Requirements for Pigs (1993). Feed in the amount of 2 kg in one dose was supplied twice a day with ad libitum access to water. The sows in the experiment were naturally mated according to the mating plan approved on the farm and the offspring came from one boar. Each farrowing it was supervised by personnel. The piglets’ body weight was checked consecutively after 24 h, on the 7th, 14th, and 21st day of lactation. Altogether 207 piglets from 20 sows were examined. Collection of materials for analysis and analytical methods Colostrum and milk were collected after earlier intramuscular injection of 2 to 4 ml of oxytocin. The amount of oxytocin administered depended on the day of lactation. Colostrum and milk were manually collected from all active mammary glands to test tubes with a preservative (MILKOSTAT). The samples were Serum albumin α-casein β-casein κ-casein α-lactalbumin collected between the 20th and 24th h after parturition and on the 2nd, 3rd, 7th, 14th and 21st day of lactation. Then the samples were cooled down to the temperature of -20°C. Altogether 120 samples were collected (60 colostrum samples and 60 milk samples). The electrophoretic separation of individual fractions of total protein, that is, serum albumin, α- casein, β- casein, κ- casein, immunoglobulins G and α- lactalbumin and other protein structures were carried out according to Laemmli’s method (1970) on polyacrylamide gel in the presence of sodium dodecyl sulphate (SDS). The qualitative and quantitative analysis was carried out according to the electrophoretic separations developed by Kim and Jimenez-Flores (1994). The percentage share of individual fractions of total protein was calculated by defining their sum as 100%. The electrophoretic separations were compared by means of a protein marker. The separations were archived by means of an optical scanner. Bio Rad 6 program was used for qualitative and quantitative analysis. Figure 1 shows the vertical electrophoresis of colostrum and milk proteins on polyacrylamide gel. Statistical analysis The obtained data was statistically processed with SAS (2007) ver. 8.11. Package with the use of the following methods: 1. Normality test (UNIVARIATE). 2. Multivariate analysis of variance with PROC GLM LSM. 3. The results were presented as least square means (LSM) and standard errors (SE). 4. For discrete random variables (the number of piglets aged 24 h, 7, 14 and 21 days) probit transformation described by Żuk (1989) and Lynch and Welsh, (1998) was applied, which enables transformation from discrete random variables to continuous random variables. 5. The protein fractions were divided according to their levels in total protein (Table 1). In the statistical models applied for the analysis apart from the Table 1. Protein fractions and their various medium levels. Protein fraction Protein Levels (%) I (low) N = 40 II (medium) N = 40 III (high) N = 40 Serum albumin ≤ 0.645 0.646 - 0.963 ≥0.964 α- casein ≤ 0.673 0.674 - 0.858 ≥0.859 β- casein ≤ 0.665 0.666 - 0.884 ≥0.885 κ- casein ≤ 0.722 0.723 - 0.989 ≥ 0.990 α- lactalbumin ≤ 0.651 0.652 - 0.881 ≥0.882 Immunoglobulins G ≤ 0.879 0.880 - 1.212 ≥1.213 Other fractions ≤ 0.860 0.861 - 1.407 ≥1.408 Table 2. The influence of protein fractions on the average number of piglets Protein fraction Number of piglets (head) I II III Serum albumin LSM 10.31 9.00 a 10.54 b SE 0.68 0.48 0.42 α – casein LSM 10.21 9.00 a 10.64 b SE 0.66 0.47 0.43 β – casein LSM 9.53 9.36 10.66 SE 0.60 0.50 0.44 κ – casein LSM 10.25 9.06 a 10.56 b SE 0.71 0.48 0.42 α - lactalbumin LSM 10.36 9.07 a 10.54 b SE 0.74 0.47 0.42 Immunoglobulins G LSM 9.48 9.68 10.57 SE 0.55 0.50 0.48 Other protein structures LSM 9.92 10.59 10.47 SE 0.59 0.52 0.49 Means designated with small letters (a, b) statistically differ significantly at P ≤ 0.05. main effects under investigation, other effects were also taken into consideration: (i) Season of sample collection (autumn, winter, spring, summer). (ii) Sows’ lactation (according to the sow parity number) (iii) Colostrum and milk collection (time specified). (iv) Piglet’s sex (male, female). RESULTS Tables 2 and 3 show characteristics of the level- dependent influence of individual protein fractions on the number and growth of piglets in the litter. The average number of piglets and their growth rate was proved to be the most favourable when the level of fractions in total protein was the highest. Those differences were con- firmed at the level P ≤ 0.01 and P ≤ 0.05. The lowest level of protein fractions (I) was also found to be decisive for the highest piglet mortality (Table 4) and it was proved to affect the piglet’s mean body weight (Table 5). The trait was proved to have statistically significant differences. DISCUSSION In contrast to many other species of animals and Table 3. The influence of protein fractions on the growth of piglets Protein fraction Week's growth of piglets (kg) I II III Serum albumin LSM 0.36 a 0.52 a 0.81 b SE 0.14 0.10 0.09 α – casein LSM 0.44 a 0.51 a 0.80 b SE 0.14 0.10 0.09 β – casein LSM 0.43 a 0.53 a 0.81 b SE 0.13 0.10 0.09 κ – casein LSM 0.37 a 0.51 a 0.80 b SE 0.15 0.09 0.08 α - lactalbumin LSM 0.38 a 0.50 a 0.82 b SE 0.16 0.10 0.09 Immunoglobulins G LSM 0.47 a 0.52 a 0.84 b SE 0.11 0.10 0.10 Other protein structures LSM 0.51 A 0.53 A 1.03 B SE 0.15 0.13 0.12 Means designated with capital letters (A, B) differ statistically significantly at P ≤ 0.01. Means designated with small letters (a, b) statistically differ significantly at P ≤ 0.05. Table 4. The influence of protein fractions on the piglets mortality. Protein fraction Mortality of piglets (%) I II III Serum albumin LSM 21.00 a 9.00 b 11.75 SE 3.69 2.61 1.85 α – casein LSM 21.00 a 9.00 b 11.75 SE 3.69 2.61 1.85 β – casein LSM 21.00 a 9.00 b 11.75 SE 3.69 2.61 1.85 κ – casein LSM 21.00 a 9.00 b 11.75 SE 3.69 2.61 1.85 α - lactalbumin LSM 21.00 a 9.00 b 11.75 SE 3.69 2.61 1.85 Immunoglobulins G LSM 21.00 a 9.00 b 11.75 SE 3.69 2.61 1.85 Other protein structures LSM 9.0 10.0 12.33 SE 2.90 4.11 2.37 Means designated with small letters (a, b) statistically differ significantly at P ≤ 0.05. Table 5. The influence of protein fractions on the piglet`s body weight Protein fraction Average body weight of piglet (kg) I II III Serum albumin LSM 5.13 Aa 3.81 b 2.83 Ba SE 0.45 0.32 0.27 α – casein LSM 4.86 Aa 3.83 a 2.83 Bb SE 0.44 0.32 0.29 β – casein LSM 4.86 Aa 3.65 b 2.84 Bb SE 0.40 0.33 0.29 κ – casein LSM 5.19 Aa 3.84 ab 2.83 Bb SE 0.47 0.31 0.26 α - lactalbumin LSM 5.26 Aa 3.85 ab 2.80 Bb SE 0.49 0.31 0.27 Immunoglobulins G LSM 4.62 Aa 3.62 b 2.76 Bb SE 0.36 0.33 0.32 Other protein structures LSM 3.44 a 3.82 Aa 2.17 Bb SE 0.41 0.36 0.34 Means designated with capital letters (A, B) differ statistically significantly at P ≤ 0.01. Means designated with small letters (a, b) statistically differ significantly at P ≤ 0.05. humans, pigs are in practice born without any immu- nological preparation for life in outer environment due to their placenta, which is impermeable to immune bodies. Therefore, colostrum is the only possibility to obtain immunoglobulins from the mother (Butler and Kehrli, 2005; Hurle and Theil, 2011). Later, when the level of immunoglobulins decreases and they can no longer be passively absorbed, the essential role of milk proteins is to provide the necessary amino acids for the needs of existence, growth and development of newly born animals (Patureau-Mirandi et al., 1990; Xu, 1996; Blӓttler et al., 2001; Sauter et al., 2004; Senda et al., 2011). Therefore, obtaining appropriate growths has a considerable influence on piglets’ balanced weight when weaned and thus produces the best results of pig fattening. The authors’ own research proved a statistically signifi- cant correlation between individual protein fractions of colostrum and milk, and the number of piglets and their daily gains. The results are also confirmed by experi- ments on other species of animals. Wroński and Sosnowska (2008) proved that bulls fed with colostrum with higher specific weight and higher content of whey proteins and immunoglobulins gained higher body dimensions than those fed with colostrum with a lower content of those proteins. The experiment on deer calves made by Landete- Castillejos et al. (2001) confirms the fact that the body weight is strongly correlated with the content of protein in the milk (P ≤ 0.05). The authors also noted a high correlation between the protein and fat ratio and calf growth. Buczyński et al. (2008) proved that the mean weight of the piglet (2.82 kg) and growths were higher in the group of sows whose milk had the highest content of protein. Boruta et al. (2009) also arrived at similar conclusions in their studies on Polish Large White and Polish Landrace breeds and their crossbreeds. The authors proved that the piglets from the sows with the highest share of protein in their milk achieved better rearing results. They also noticed a correlation between piglet mortality and the share of whey proteins. A higher content of whey proteins increased piglets’ survival rate. The authors’ own studies confirm the fact that the highest mortality of piglets (up to 21%) was according to the lowest content of all protein fractions in their mothers milk (level I). The results were statistically confirmed. Those results were not confirmed in an analogical experiment made by Walkiewicz et al. (2004). They noted that crossbreeds piglets (Puławska x Duroc) from the sows with a higher level of protein grew worse than the piglets from the sows with a lower content of total protein. To sum up the importance of milk proteins in the processes of growth and development, it is necessary to add that they are not only decisive in piglets’ acquisition of immunity but they are also significant in piglet rearing. 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Państwowe Wydawnictwo Rolne i Leśne Warszawa. http://www.google.pl/url?sa=t&source=web&cd=1&ved=0CBcQFjAA&url=http%3A%2F%2Fwww.phmd.pl%2F&rct=j&q=Post%C4%99py%20Hig%20Med.%20Do%C5%9Bw.%20&ei=lhFBTojECcyq-gbOt-DYCQ&usg=AFQjCNGWaq51uZbMZJlMp_DX-GE9TSFl0w&cad=rja http://www.google.pl/url?sa=t&source=web&cd=1&ved=0CBcQFjAA&url=http%3A%2F%2Fwww.phmd.pl%2F&rct=j&q=Post%C4%99py%20Hig%20Med.%20Do%C5%9Bw.%20&ei=lhFBTojECcyq-gbOt-DYCQ&usg=AFQjCNGWaq51uZbMZJlMp_DX-GE9TSFl0w&cad=rja