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HANUŠ, O., SOJKOVÁ, K., HANUŠOVÁ, K., SAMKOVÁ, E., HRONEK, M., HYŠPLER, R., KOPECKÝ, J., JEDELSKÁ, R.: An experimental comparison of methods for somatic cell count determination in milk of various species of mammals. Acta univ. agric. et silvic. Mendel. Brun., 2011, LIX, No. 1, pp. 67–82 Somatic cell count (SCC) is important foodstuff , hygienic and health indicator of milk and animal mammary gland. The goal of this paper was to evaluate an ability of chosen methods to reach the SCC reliable results in various biological kinds (species) of milk. The various methods of SCC determination were compared in cow (CM), goat (GM), sheep (SM) and human (HM) milk: direct microscopy (DM); fl uoro-opto-electronic (Fossomatic 90; Foss); fl uorescent (DCC; De Laval). Used methods had cow milk calibration basically. The DM, Foss and DCC result relations about SCC were very close, mostly > 0.92 (P < 0.001) for CM, GM and SM. In CM the regression equations between methods were near ideal form y = 1x + 0. The mean diff erences SCC data sets between mentioned methods were small for CM, larger for SM and HM and the largest for GM. It is possible to convert all DCC results in SM, HM and GM to DM or Foss method. The conversion equations were stated from DCC: to DM in cow milk y = 1.1293x − 5.5029; to Foss in goat milk y = 3.603x − 3171.4; to Foss in sheep milk y = 1.3805x − 18.149; to Foss in human milk y = 2.6246x + 158.63. Assesment of conversion equations should be individual laboratory event. Results had relatively good correspondence among DM, Foss and DCC for SCC determination in CM, GM, SM and HM for milk quality control. DCC had lower results in small ruminants as compared to Foss calibrated on CM using DM. DCC in HM had lower results as Foss adjusted by CM at good correlation (0.84; P < 0.001). cow, sheep, goat, human milk, somatic cell count (SCC), direct microscopy, fl uoro-opto-electronic method, fl uorescence method, relationship Somatic cell count (SCC) in milk is important food-hygienic (bulk samples) and also health (individual samples) indicator of animal mammary gland. Somatic cells are mostly leucocytes in milk (nucleus cells) which show on actual state of physiological and pathological response of animal defense system with respect to possible mammary gland infection. The SCC values show on milk secretion disor ders occurrence. Therefore the SCC determination serves over whole world to control of milk food chain quality in dairying and human nutrition from beginning of this succession. Critical quality values of SCC are limited by legislation for milk of some animal species. For cover of above mentioned facts more analytical methods are existing which are based on various principles, standardized and also unstandardized. Beside reference direct optical microscopy method (CSN EN ISO 13366–1 (DM)) there are also instrumental methods. Electronical counting of fi rm particules (somatic cells fi xed by formaldehyde) during their passage throught defi ned electrical fi eld in capillary was previous procedure (so called Coulter Counter method). Today other procedures such as fl uoro-opto-electronic counting (CSN EN ISO 68 O. Hanuš, K. Sojková, K. Hanušová, E. Samková, M. Hronek, R. Hyšpler, J. Kopecký, R. Jedelská 13366–2) of stained (ethidiumbromid) somatic cell nuclei (DNA) which emit red emission a er lighting which is registered electronically by disc rotation (DR, disc rotation; Fossomatic) method or more modern fl ow cytometry (FC, fl ow cytometry; for instance Fossomatic – Foss Electric, Denmark (Foss), Somacount – Bentley Instruments, USA or Somascope – Delta Instruments, The Netherlands) are used preferably. Also various modifi cations of viscosimetric methods (Wisconsin Mastitis test or Ruakura (New Zealand) rolling ball viscosimeter (Hanuš et al., 1993d)) on the principle of so called Schalm reaction (reaction among DNA, milk proteins and added detergent with creation of gel and its density increase at SCC rise; Hejlíček et al., 1987) are disposable for estimation of cell quantity as well. There are above all Schalm California mastitis test in modifi cation known as Mastitis tets NK in the Czech Republic. Recently a further instrumentation (Chemometec (2004), NucleoCounter SCC–100 and De Laval (2006), DCC cytometer, fl uorescent method principle (DCC)) is appearing, especially for direct advisory service purposes at fast and operative moni toring of udder health in animal stables (Castro et al., 2008; Sánchez–Macías et al., 2008). All these indirect instrumental methods of SCC determination should be calibrated or adjusted in other way according to direct microscop SCC determination results. Diff erences in result reliability regarding biological kind of milk can exist here. The row of papers was interested in methodical support of reference standard preparation for somatic cell count (SCC) determination in milk, especially in cows (Szijarto and Barnum, 1984; Lintner et al., 1984; Arndt et al., 1991; Heeschen et al., 1994; Aebi and Bühlmann, 2000a, b; Baumgartner et al., 2000). Also whole row of papers was interested in statistical evaluation of milk analytical result reliability in laboratory networks on national and international level (Vines et al., 1986; Grappin, 1987, 1993; Valenberg, 1990; Arndt et al., 1991; Leray, 1993, 2006, 2010; Wood, 1994; Heeschen et al., 1994; Golc–Teger, 1997; Wood et al., 1998; Baumgartner et al., 2000; Fuchs, 2000; Coveney, 2001; Feinberg and Laurentie 2006; Hanuš et al., 2006, 2007; Hering et al., 2008; Říha et al., 2008 – profi ciency testing (PT)). SCC determination in other biological kinds of milk in other farm animals (sheep, goats, buff alos, camels, horses or asses), mammals respectively, than in cows has growing importance. The main reason is an increase of herds of various animal species (Nicolas et al., 2008; Pellegrino and Rosi, 2008; Marchand et al., 2008) and also spectrum of milk food (for instance functional food) and its market. It means spreading of milk market and growth of an importance of food safety and consumer health control. The aim of this work was to examine pertinence of chosen analytical procedures to product reliable SCC results in various kind of milk which can methodically extend possibilities of control of hygiene standards and quality in dairying but especially in an animal health state. MATERIAL AND METHODS Model sample fi les of various biological kinds of milk Human milk samples were analysed as unpreserved. Other mammals species milk samples (ruminants) were preserved (0.02%; Pettipher and Rodrigues, 1982; Ardö, 1982; Kroger, 1985; Hanuš et al., 1992a, b; Genčurová et al., 1993a, 1994; Benda, 1995) by peletted bronopol (2–brom, 2–nitro, 1, 3 propandiol; Control System D&F Microtabs, England). First and also second samples were analysed either a er cold transport and storage (< 6 oC; according to Szijarto et al., 1990 and Sojková et al., 2009) or frozen preservation (−21 oC), according to conditions, howe ver always in accordance with standard procedure. Previous papers (Hanuš et al., 2002, 2008; Baumgartner and Landgraf, 2004) demonstrated also possibility of milk sample frozen preservation as usable procedure before analyse for obtaining reliable SCC results. Experimental comparison covered 1.5 lactation period of small ruminants by realization time and in this way it took 1.5 year. I) cow milk (CM): reference (for PT), modifi ed, mixed bulk and individual milk samples (n = 20, in two tests A and B) species Bos primigenius f. taurus (L), breeds Czech Fleckvieh and Holstein. Native and frozen preserved samples, native samples were chemical preserved. The used methods of SCC determination were DM (profi ciency testing, Fig. 1 with good result), Foss and DCC. II) goat milk (GM): bulk milk samples (n = 30 samples, 3 tests A, B and C) from small count of animals (from 4 to 6 animals in one sample) species Capra aegagrus f. hircus (L), breed White short haired, from morning milking. Native milk samples were subsequently chemical preserved and also frozen preserved. Used methods of SCC determination were Foss and DCC. III) sheep milk (SM): bulk milk samples (n = 40 samples, 4 tests A, B, C and D) from small count of animals (from 4 to 6 animals in one sample), species Ovis aries (L), greed Tsigai, from morning milking. Native milk samples were subsequently chemical preserved and also frozen preserved. Used methods of SCC determination were Foss and DCC. IV) human milk (HM): individual milk samples (n = 54 samples, 1 test A), species Homo sapiens sapiens (L), originated from primipar mothers in age from 23 to 27 years. Samples were taken during 1.5 year, from 2nd to 47th lactation week, always in morning hours a er 12 hunger hours. Whole volume of milk was exhausted from breast which was not suckled last time. The sample obtained in this way was frozen preserved before analyse. Used methods of SCC determination were Foss and DCC. Examined analytical methods for somatic cell count determination in milk The direct microscopy method (DM) was used for counting of stained somatic cells (CSN EN ISO An experimental comparison of methods for somatic cell count determination in milk 69 13366–1) at SCC determination in CM referen ce standards for profi ciency testing (PT) purposes. Milk samples from all species (CM, GM, SM, HM) were analysed on SCC using fl uoro–opto– electro nic method in DR type (Foss) on Fossomatic 90 (Foss Electric, Denmark) apparatus with regard to previous methodical knowledge (Grappin and Jeunet, 1974, 1975; Coleman and Moos, 1989; Genčurová et al., 1993b; Hanuš et al., 1993a, b, c, 2002, 2007, 2009) and according to CSN EN ISO 13366–2. This apparatus was included in profi ciency testing (State Veterinary Institute, Prague) for SCC determination regularly three times a year. The results were regularly in order (Fig. 1). The extended combine result uncertainty (Suchánek et al., 1999) of measurement was estimated in accredited labo