Milk testing and quality control should be carried out at all stages of the dairy chain. Milk can be tested for quantity, organoleptic characteristic, compositional characteristic, physical and chemical characteristics, hygienic characteristics, adulteration or drug residues. The content of the major constituents of raw milk is important for milk payment system. Enzymes naturally present in the milk can change the chemical composition of raw milk. Also, enzymes secreted by bacteria or enzymes from somatic cells can degrade the raw milk composition. Products of these degradation reactions can have undesirable effects on milk structure, smell and taste. It is very important that farm-fresh raw milk be cooled immediately to not more than 8 °C in the case of daily collection, or not more than 6 °C if collection is not daily. During transport the cold chain must be maintained. An authorized person, properly trained in the appropriate technique, shall perform sampling of bulk milk in farm. Laboratory samples should be dispatched immediately after sampling to the dairy company and consequently to the testing laboratory. The time for dispatch of the samples to the testing laboratory should be as short as possible, preferably within 24 h. Laboratory samples shall be transported and stored at temperature 1 to 5 °C. Higher temperatures may adversely affect the composition of the laboratory sample and may cause disputes between the farmer, the dairy company and the laboratory. The effect of refrigerated storage at temperature 4 °C during 24 h on the composition of raw milk were investigated in this work, because we wanted to know how the milk composition will be changed and how the laboratory results will be affected. In many cases, the samples are not preserved with chemical preservants like azidiol, bronopol, potassium dichromate or Microtabs. We found, that the composition of raw cows' milk after 24 was changed significantly (p >0.005). We found an average decrease in the fat content of -0.04 g/100g, increase in the protein content of +0.02 g/100g, increase in the lactose content of +0.02 g/100g, increase in the solid-not-fat content of +0.02 g/100g and decrease in the total solid content of -0.02 g/100g. It is necessary to cool the raw cows' milk after the milking to decrease the changes in milk composition caused mainly due to the lipolytic activity of lipase.
ABSTRACT:The direct fluorescence microscopy method with ethidium bromide staining can be used for somatic cell counting in raw cow's milk. However, this method has some limitations that may influence the results of the analysis. We therefore aimed at improving the procedure of somatic cell nuclei staining. We tested the hypothesis that ethidium bromide can better penetrate into the DNA of cells with degraded somatic cell walls or into dead cells. Therefore, we increased the temperature of the sample to 100 °C in order to disrupt the somatic cell wall membrane and to improve ethidium bromide penetration to somatic cell nuclei. In all, 90 samples of raw cow's milk were analysed in this experiment. Three parallel measurements were performed using each of the microscopic methods and the routine flow cytometry method. In all, 810 microscopic smears were analysed. The somatic cells were counted using fluorescence microscopic methods and flow cytometry. The increased temperature during the sample preparation improved (P < 0.005) the penetration of ethidium bromide into the somatic cell nuclei. It is concluded that the direct fluorescence microscopy method is suitable for precise laboratory analysis of somatic cell in raw cow's milk.
Merle patterning in dogs, caused by the insertion of a short interspersed element (SINE) in the genetic structure of SILV gene, is characterized by patches of diluted pigment intermingled with normal melanin. Sequencing analyses of SINE element localized in the canine SILV gene discovered a variability of the poly (A)-tail length which is responsible for the different expression of merle pattern. The SINE element with the length of poly(A)-tail between 91 -101 nucleotides is responsible for the merle phenotype with all characters of merle pattern. On the contrary the dogs which have SINE element with the shorter length of poly(A) tail between 54-65 nucleotides are referred as cryptic merles without expresion of Merle pattern. The aim of this study was to improve molecular genetics method for the detection of cryptic allele for merle patterning in dogs. A total of 40 dogs of four breeds -Border collie, Shetland sheepdog, Australian Shepherd dog, and Chihuahua were used in this study. Canine genomic DNA was isolated from samples of whole blood and buccal cells by commercial column kit. Detection of merle (M), cryptic merle (Mc) and non-merle (m) alleles was done using M13-tailed primer protocol and two different allele-sizing methods for the verifi cation of the electrophoresis result. In the analyzed population of dogs were detected 20 dogs with non-merle genotype mm, 17 dogs with merle genotype Mm, 2 dogs with double merle genotype MM and one dog with merle phenotype but with the presence of cryptic merle allele Mc with the consequential genotype MMc.
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