Seasonal changes in somatic cell content in milk of Polish Holstein-Friesian cows Maciej Gierdziewicz The Faculty of Animal Sciences Agricultural University in Cracow Somatic cells in milk In cattle breeding, the number of somatic cells in milk (Somatic Cell Count - SCC) is a commonly accepted indicator of subclinical mastitis. Somatic cells in milk (2) • Healthy cow – SCC up to 100 000 /ml SCC depends on lactation number: • 1st lactation cow – up to 35 000 /ml • 2nd lactation cow – up to 50 000 /ml • 3rd lactation cow – up to 60 000 /ml • Cow with (sub)clinical mastitis – SCC 500 000 5 000 000 50 000 000 ? COUNCIL DIRECTIVE 92/46/EEC, 16 June 1992: Health rules for the production and placing on the market of raw milk, heat-treated milk and milk-based products „ [...] CHAPTER IV Standards to be met for collection of raw milk [...] A. Raw cow's milk [...]” COUNCIL DIRECTIVE 92/46/EEC 16 June 1992 (continued) „Somatic cell count (per ml) 400 000 (b) (b) Geometric average over a period of three months, with at least one sample a month, or, where production levels vary considerably according to season, method of calculating results to be adjusted [...]” Rozporządzenie Ministra Rolnictwa i Gospodarki Żywnościowej z dnia 29 września 1999 r. (Polish regulations) A.Wprowadzone w normie zmiany obejmują: [...] 2. Ograniczenie ilości klas jakościowych mleka. [...] klasa Ekstra, klasa I i klasa II Wymagania szczegółowe dla poszczególnych klas mleka: Ekstra: Komórki somatyczne - liczba w 1 ml (SCC/ml limit)→ 100 000 Transformation of SCC to SCS Somatic Cell Count is very variable: 0-1-10-100- ... 10 000 000 - 100 000 000 ... logarithmic transformation: SCS - Somatic Cell Score SCS = log2(SCC/100 000) + 3 SCC SCS 6 250 12 500 25 000 50 000 100 000 200 000 400 000 800 000 -1 0 1 2 3 4 5 6 ... Selected references I. General Svendsen M., B. Heringstad, 2006: Somatic cell count as an indicator of sub-clinical mastitis. Genetic parameters and correlations with clinical mastitis. (Interbull Bulletin) Correlation coefficients between clinical and sub-clinical mastitis: 0,3 - 0,6 Selected references II. Age, test month, % of HF genes Effect of age: Kennedy B.W. et al., 1982: Environmental Factors Influencing Test-Day Somatic Cell Counts in Holsteins (Journal of Dairy Science) slow decrease with age; steep increase in VI and XII Effect of HF genes percentage: Philipsson J., G. Ral, B. Berglund, 1995. Somatic cell count as a selection criterion for mastitis resistance in dairy cattle. (Livestock Production Science) — HF share may influence SCS; Selected references III. Days in milk (in Polish cattle) Strabel T., Jamrozik J. 2006. Genetic analysis of milk production traits of Polish Black and White cattle using large-scale random regression test-day models. (Journal of Dairy Science) Ptak E., Brzozowski P., Jagusiak W., Zdziarski K., 2007: Genetic parameters for somatic cell score for Polish Black-and-White cattle estimated with a random regression model. (Journal of Animal and Feed Sciences ) The goal The aim of the work was to apply nonlinear regressions on month of calving and on month of test to investigate seasonal changes in somatic cell content in milk of Polish HF cows. Data (1) over 12 600 000 test milking results about 880 000 cows first, second and third lactation 5 984 860, 4 139 104, 2 519 366 test results calving in 1997-2007, 1998-2007, 1999-2007 average SCS: 3.48, 3.92, 4.19 Data (2) average SCS by month of calving - lactations I - III mean I 4,4 mean II mean III 4,2 4 SCS 3,8 3,6 3,4 3,2 3 I II III IV V VI VII VIII IX Month of the year X XI XII Data (3) average SCS by month of test - lactations I - III mean I 4,4 mean II mean III 4,2 4,0 SCS 3,8 3,6 3,4 3,2 3,0 I II III IV V VI VII VIII IX Month of the year X XI XII Methods (1) Equation to fit to the data (each lactation separately): 2 y a 0 a 1 sin t a 2 error 12 y - SCS (dependent variable) t - time (independent variable); 0 ≤ t ≤ 12 time scale - months of the year (month of calving or month of test milking) a0 - lactation mean of SCS a1 – amplitude of seasonal changes of SCS a2 – phase shift in months Methods (2) Curve fitting - grid search phase Calving month: a0 = 3.0 to 4.5 by 0.1 a1 = -0.2 to 0.2 by 0.05 a2 = 1.0 to 4.0 by 0.5 Test month: a0 = 3.4 to 4.3 by 0.1 a1 = 0.0 to 0.20 by 0.05 a2 = -6 to 6 by 1; Curve fitting - Iterative phase Gauss-Newton method Hardware IBM BladeCenter HS21 cluster ("mars") • One of 56 nodes - containing: 2 Intel Xeon Dual Core processors (2.66 GHz) 8 GB memory hard disk 36.4 GB Software IBM BladeCenter HS21 cluster ("mars") • operating system : Linux RedHat • SAS® (Statistical Analysis System) ► version 9.1.3 • documentation: ►http://support.sas.com/documentation/onlinedoc/sas9doc.html Technical remarks The NLIN procedure of SAS was used. For examining the effect of the month of calving, grid search time was: 1h26’58’’, 1h00’02’’ and 0h35’13’’ for lactations 1, 2 and 3, respectively. For estimating the effect of test month: 0h56’19’’, 0h39’03’’ and 0h23’2’’ Without iterative phase, about 3’30’’ in both cases. Results for 1st lactation (1) 2,90E+07 2,80E+07 2,70E+07 2,60E+07 2,50E+07 2,40E+07 2,30E+07 2,20E+07 2,10E+07 2,00E+07 1,90E+07 1,80E+07 1,70E+07 1,60E+07 1,50E+07 1,40E+07 1,30E+07 1,20E+07 1,10E+07 1,00E+07 9,00E+06 8,00E+06 7,00E+06 6,00E+06 5,00E+06 4,00E+06 3,00E+06 2,00E+06 1,00E+06 0,00E+00 0,1 -0,0 AM PLITUDE 4,4 4,2 4 M EAN 3,8 3,6 3,4 3,2 3 RESIDUAL SUM OF SQUARES grid search - residual sum of squares for phase shift = +1month -0,2 28000000-29000000 27000000-28000000 26000000-27000000 25000000-26000000 24000000-25000000 23000000-24000000 22000000-23000000 21000000-22000000 20000000-21000000 19000000-20000000 18000000-19000000 17000000-18000000 16000000-17000000 15000000-16000000 14000000-15000000 13000000-14000000 12000000-13000000 11000000-12000000 10000000-11000000 9000000-10000000 8000000-9000000 7000000-8000000 6000000-7000000 5000000-6000000 4000000-5000000 3000000-4000000 2000000-3000000 1000000-2000000 0-1000000 Results for 1st lactation (2) 2,90E+07 2,80E+07 2,70E+07 2,60E+07 2,50E+07 2,40E+07 2,30E+07 2,20E+07 2,10E+07 2,00E+07 1,90E+07 1,80E+07 1,70E+07 1,60E+07 1,50E+07 1,40E+07 1,30E+07 1,20E+07 1,10E+07 1,00E+07 9,00E+06 8,00E+06 7,00E+06 6,00E+06 5,00E+06 4,00E+06 3,00E+06 2,00E+06 1,00E+06 0,00E+00 0,2 0,1 -0,1 4,4 4,2 4 3,6 0 3,8 MEAN 3,4 3,2 3 RESIDUAL SUM OF SQUARES grid search - residual sum of squares for phase shift = +1.5 months -0,2 AMPLITUDE 28000000-29000000 27000000-28000000 26000000-27000000 25000000-26000000 24000000-25000000 23000000-24000000 22000000-23000000 21000000-22000000 20000000-21000000 19000000-20000000 18000000-19000000 17000000-18000000 16000000-17000000 15000000-16000000 14000000-15000000 13000000-14000000 12000000-13000000 11000000-12000000 10000000-11000000 9000000-10000000 8000000-9000000 7000000-8000000 6000000-7000000 5000000-6000000 4000000-5000000 3000000-4000000 2000000-3000000 1000000-2000000 0-1000000 Results (3) - analysis of variance effect of month of test - 1st lactation Source DF Model 2 Error 5.98E6 Corrected Total 5.98E6 Sum of Squares 52158.3 21769945 21822104 Parameter A0 A1 A2 Std Error 0.000781 0.00111 0.0158 Estimate 3.4885 0.1327 5.0324 Mean Square 26079.1 3.6375 F Value 7169.51 Pr > F <.0001 95% Confidence Limits 3.4870 3.4900 0.1306 0.1349 5.0014 5.0633 Results (4) – SCS vs month of calving mean I mean III mean II (predicted) 4,4 mean II mean I (predicted) mean III (predicted) 4,2 4 SCS 3,8 3,6 3,4 3,2 3 I II III IV V VI VII VIII IX Month of the year X XI XII Results (5) – SCS vs month of test mean I mean III mean II (predicted) 4,4 mean II mean I (predicted) mean III (predicted) 4,2 4,0 SCS 3,8 3,6 3,4 3,2 3,0 I II III IV V VIof the VII year VIII Month IX X XI XII Results(6) - Curve fitting - Iterative phase effect of test month of SCS Sum of Lactation Iter 1 0 1 2 3 4 2 0 1 2 3 4 3 0 1 2 3 A0 3.5000 3.4885 3.4885 3.4885 3.4885 3.9000 3.9231 3.9231 3.9231 3.9231 4.2000 4.1975 4.1975 4.1975 A1 0.1000 0.1138 0.1314 0.1327 0.1327 -0.1000 -0.0979 -0.1261 -0.1269 -0.1269 0.1000 0.0841 0.1126 0.1127 A2 4.0000 5.3045 4.9882 5.0328 5.0324 1.0000 -0.5422 -0.2509 -0.3174 -0.3169 4.0000 5.4321 5.3767 5.3907 Squares 21785676 21771906 21769979 21769945 21769945 16078374 16065773 16063684 16063643 16063643 9917903 9911426 9910394 9910393 Results(7) – fitted curves SCS vs month of calving y = 3.49 + 0.14 sin [ 2/12 ( t – 0.56 ) ] (lactation I) y = 3.92 + 0.13 sin [ 2/12 ( t – 1.30 ) ] (lactation II) y = 4.20 + 0.10 sin [ 2/12 ( t – 1.26 ) ] (lactation III) Results(8) – fitted curves SCS vs month of test y = 3.49 + 0.13 sin [ 2/12 ( t – 5.03 ) ] (lactation I) y = 3.92 + 0.13 sin [ 2/12 ( t – 6.32 ) ] (lactation II) y = 4.20 + 0.11 sin [ 2/12 ( t – 5.39 ) ] (lactation III) Conclusions ○There were seasonal changes in SCS in milk. ○The amplitude of SCS changes varied from about 0.1 to about 0.2, ○The maximum SCS has been reached in cows calving in April, May or June. ○ The cows milked in August or September had maximum SCS in milk. ○Although the effect of season on SCS has been proved, the variation is not high, and therefore according to the EEC rules no special adjustment on season is needed. ○ Technical remark: for fitting simple curves to the data a good guess is better than initial grid search. Data source: research funding for grant G-1594/KGiMDZ/06-09 (head: dr hab. Ewa Ptak, University of Agriculture, Kraków) ”Genetic and environmental factors influencing the number of somatic cells in milk”; data were used in 2009; research project No. N31101131/3353