The present invention refers generally to the preparation of milk samples for preparing the milk for further analysis with respect to the quality of the milk, especially the number and kind of somatic cells in the milk. More precisely, the present invention refers to a method for preparing a milk sample according to the preamble of claim 1. The invention also refers to a device configured to be used when preparing a milk sample.
In the prior art, it is known to estimate the number of somatic cells in milk extracted from an animal in order to establish whether the animal suffers from any disease, such as mastitis. For obtaining such an estimate, a milk sample is taken from the extracted milk. The milk sample is prepared before being analysed in order to facilitate counting of somatic cells in the milk.
U.S. Pat. No. 6,979,550 discloses a method for detection and prediction of mastitis. The method comprises the steps of determining the value of mastitis indicators and comparing the values with predetermined standards, wherein deviation form the standards provides a measure of mastitis.
In the milk sample, containing fat and casein, it is difficult to identify and observe somatic cells, especially in an optical manner through a microscope. According to the prior art, the preparation therefore comprises the addition of various chemicals to the milk sample to react with the somatic cells in order to make the somatic cells observable. The addition of chemicals, make the preparation difficult and time consuming.
These kind of chemicals can also affect the somatic cells, making it difficult to identify which types of somatic cell are contained in the milk sample. Moreover, the milk sample has to be taken care of after the analysis, which is a problem when the milk sample is contaminated with chemicals of various kind.
The object of the present invention is to provide an improved method and an improved device to be used when preparing a milk sample for being analysed. In particular, it is aimed at a method and a device enabling preparation of a milk sample without addition of chemicals reacting with somatic cells in the milk sample.
This object is achieved by the method initially defined which is characterised by the further steps of:
The invention permits to obtain a sample mixture, which comprises the solution and the remaining sample part, e.g. the first sample part or the second sample part. In case the remaining sample part is formed by the second sample part, the sample mixture constitutes a representative quantity of the milk extracted from the animal with respect to the content of the second constituent, such as somatic cells, wherein the solution is mixed with the second sample part. In other words, the invention permits to obtain a milk sample which contains the same percentage of somatic cells as the milk extracted from the animal. The sample mixture so obtained is thus suitable for being discharged and forwarded to an analysing equipment for analysing the milk, especially for optical counting of the number of somatic cells in a microscope. Furthermore, the invention permits somatic cells of various types in the sample mixture to be recognised and counted optically in a microscope. The somatic cells have not been negatively affected by any chemicals, which possibly could have prevented identification of the different types of somatic cells contained in the extracted milk and in the milk sample. Thus, the invention enables identification of each type of somatic cells, such as lymphocytes, epithelial cells, neutrophils, monocytes, basophils, macrophages, etc.
The invention also permits to obtain a sample mixture which constitutes a representative quantity of the milk extracted from the animal with respect to the content of the first constituent, such as fat, wherein the solution is mixed with the first sample part being the remaining sample part. The sample mixture so obtained is suitable for being discharged to an analysing equipment for analysing and/or counting the fat cells of the milk.
According to an aspect of the invention, the solution comprises or consists of a biologically acceptable saline solution. In particular, the solution may comprise or consist of a biological acceptable NaCl-solution. The salt content may then correspond to the salt content of the body liquid of the animal. Such a saline solution will not affect the somatic cells.
According to a further aspect of the invention, the first volume is greater than the second volume. Advantageously, the second volume may be less the 10% of the first volume. Moreover, the second volume may be more than 1% of the first volume.
According to a further aspect of the invention, the step of providing the milk sample is followed by a step of isolating the milk sample in a space formed by a first fill chamber, defining the first volume, and a second fill chamber, defining the second volume.
According to a further aspect of the invention, the step of providing the solution is followed by a step of isolating the solution in a space formed by a first mixing chamber, defining a volume equal to the first volume, and a second mixing chamber, defining a volume equal to the second volume, wherein the step of removing one of the sample parts comprises displacing at least one of the second fill chamber and the first mixing camber to communicate with each other. Advantageously, the second fill chamber and the first mixing chamber are then align with each other so that the second sample part in the second fill chamber may flow into the first mixing chamber, and the solution in the first mixing chamber may flow into the second fill chamber. The mixing step may then be efficiently performed.
According to a further aspect of the invention, the step of mixing is performed with the aid of a stirring member. According to one embodiment, the stirring member may be provided in the first mixing chamber, in the case when the second sample part, i.e. the second constituent, is to be mixed with the solution. The stirring member may then be movable in the first mixing chamber and into the second fill chamber. According to another embodiment, the stirring member may be provided in the second mixing chamber, in the case when the first sample part, i.e. the first constituent, is to be mixed with the solution. The stirring member may then be movable in the second mixing chamber and into the first fill chamber.
According to a further aspect of the invention, the discharging step comprises transferring a predetermined portion of the sample mixture to an analysing equipment configured for counting and analysing the cell content. The predetermined portion will contain the same percentage of somatic cell, or alternatively fat cells, as the milk sample, and thus as the milk extracted from the animal. In either case, the somatic cells or the fat cells are uniformly distributed in the predetermined portion.
According to a further aspect of the invention, the first constituent is fat and the second constituent is somatic cells. The milk may also contain a third constituent, which may be casein. In case the sample mixture contains somatic cells and the solution, the third constituent may together with the first constituent be contained in the first sample part. Correspondingly, in case the sample mixture contains fat cells and the solution, the third constituent may together with the second constituent be contained in the second sample part.
The object is also achieved by the device initially defined, which comprises a plurality of sections including at least
Such a device is suitable for being used when performing the method defined in claim 1. The sections being transversely movable may thus be moved to various positions enabling to obtain the sample mixture. Consequently, the device according to the invention is suitable for receiving a milk sample and for being used to produce a sample mixture, which constitutes a representative quantity of said milk extracted from the animal with respect to the content of somatic cells, i.e. a sample mixture that contains the same percentage of somatic cells as said milk extracted from the animal. In particular, the sample mixture may be achieved by using the method discussed above, especially by subjecting the device to the method steps of claim 1. As mentioned above, the sample mixture is suitable for being discharged and forwarded to an analysing equipment for analysing the milk, especially for counting the number of different somatic cells in an optical manner. According to the explanations given above, the device is also suitable for obtaining a sample mixture permitting analysis of the fat cells of the milk.
According to an embodiment of the invention the second section and the third section are individually movable transversely to the longitudinal axis to said relative positions, which define different communications possibilities between the first communication passage, the first fill chamber, the first mixing chamber, the second fill chamber, the second mixing chamber, and the second communication passage.
According to a further embodiment of the invention, the first section, the second section, the third section and the fourth section are configured to permit the following consecutive relative positions:
a first position, in which the first communication passage, the first fill chamber, the second fill chamber and the second communication passage communicate with each other,
a second position, in which the first fill chamber and the second fill chamber are communicating with each other and closed to the first communication passage and the second communication passage,
a third position, in which the first communication passage, the first mixing chamber, the second mixing chamber and the second communication passage communicate with each other, a fourth position, in which either the first mixing chamber and the second fill chamber or first fill chamber and the second mixing chamber are communicating with each other and closed to the first communication passage and the second communication passage, and
a fifth position, in which the first communication passage and the second communication passage communicate with each other and with either the first mixing chamber and the second fill chamber or first fill chamber and the second mixing chamber.
The first position is suitable for performing the step of providing a milk sample. The second position is suitable for performing the centrifuging of the milk sample. The third position is suitable for providing the solution. The fourth step is suitable for mixing the second sample part or the first sample part with the solution. The fifth step is suitable for discharging the sample mixture.
According to a further embodiment of the invention,
the first position is configured to permit supply of a milk sample containing at least a first constituent and a second constituent to the first fill chamber and the second fill chamber,
the second position is configured to permit centrifuging of the milk sample contained in the first fill chamber and the second fill chamber to obtain a first sample part of a first volume, in which first sample part the first constituent is contained, and a second sample part of a second volume, in which second sample part the second constituent is contained,
the third position is configured to permit supply of a solution to the first mixing chamber and the second mixing chamber,
the fourth position is configured to permit mixing of the solution and either the second sample part, in the first mixing chamber and the second fill chamber to form a sample mixture in which the second sample part is distributed, or the first sample part, in the first fill chamber and the second mixing chamber to form a sample mixture in which the first sample part is distributed, and the fifth position is configured to permit discharge of the sample mixture.
According to a further embodiment of the invention, the second communication passage comprises a primary channel and a secondary channel. Advantageously, the first communication passage, the first fill chamber and the second fill chamber may communicate with the primary channel in the first position for the supply of the milk sample to the first fill chamber and the second fill chamber. Moreover, the first communication passage, the first mixing chamber and the second mixing chamber may communicate with the primary channel in the third position for the supply of the solution to the first mixing chamber and the second mixing chamber. Still further, the first communication passage and either of the first mixing chamber and the second fill chamber or the first fill chamber and the second mixing chamber may communicate with the secondary channel in the fifth position for the discharge of the sample mixture.
According to a further embodiment of the invention, the first fill chamber and the second fill chamber extend in parallel with a longitudinal axis of the device, wherein the first mixing chamber and the second mixing chamber extend in parallel with said longitudinal axis.
According to a further embodiment of the invention, the first section, the second section and the third section are individually movable by being rotatable around the longitudinal axis. Alternatively, the first section, the second section and the third section are individually movable by being linearly displaceable transversely the longitudinal axis.
According to a further embodiment of the invention, a first actuator is provided for moving the first section, a second actuator is provided for moving the second section, and a third actuator is provided for moving the third section. Advantageously, the first, second and third actuators may be individually controlled by means of a controller.
According to a further embodiment of the invention, the plurality of sections comprises a fifth section provided between the second section and the third section, and forming a fifth fill chamber, defining a fifth volume, and a fifth mixing chamber, defining a volume equal to the fifth volume. Thanks to such fifth sections is possible to separate the any one of the first, second and third constituents of the milk. The invention thus enable analysis of the content of each of the three constituents.
According to a further embodiment of the invention, the sections, are configured to permit the following consecutive relative positions:
The present invention is now to be explained more closely through a description of various embodiments and with reference to the drawings attached hereto.
An amount of somatic cells may always be present in the milk, but in case the animal suffers from a disease, such as mastitis, the number of somatic cells in the milk may be significant. It is important to be able to count the number of somatic cells, i.e. to enable the performance of a so called Somatic Cell Count, SCC, in number of cells per micro litre, and to be able to determine the different types of somatic cells in the milk.
The device comprises a first section 1, a second section 2, a third section 3 and a fourth section 4. The device defines a longitudinal axis X, which, at least in the first position disclosed in
A first actuator 14 is provided for moving the first section 1. The first actuator 14 is attached to the frame 5 and connected to the first section 1 via a schematically indicated first transmission device 15 or transmission element, such as a piston, a gear rack, a pulley etc.
A second actuator 24 is provided for moving the second section 2. The second actuator 24 is attached to the frame 5 and connected to the second section 2 via a schematically indicated second transmission device 25 or transmission element, such as a piston, a gear rack, a pulley etc.
A third actuator 34 is provided for moving the third section 3. The third actuator 34 is attached to the frame 5 and connected to the third section 2 via a schematically indicated third transmission device 35 or transmission element, such as a piston, a gear rack, a pulley etc.
The surfaces of each section 1 to 4, facing and adjoining another section 1 to 4, have a fine surface roughness ensuring abutment, or tight abutment, against the adjoining surface. The surfaces will thus function as seal surfaces.
The first section 1 forms a first communication passage 11 extending through the first section 1. In the first embodiment disclosed, the first communication passage 11 extends in parallel with the longitudinal axis X.
The second section 2 forms a first fill chamber 21, defining a first volume, and a first mixing chamber 22, defining a volume equal to the first volume. The first fill chamber 21 and the first mixing chamber 22 both extend through the second section 2 in parallel with the longitudinal axis X.
The third section 3 forms a second fill chamber 31, defining a second volume, and a second mixing chamber 32, defining a volume equal to the second volume. The second fill chamber 31 and the second mixing chamber 32 both extend through the third section 3 in parallel with the longitudinal axis X. The first volume is greater than the second volume. The second volume may be less the 10% of the first volume, and more than 1% of the first volume.
The fourth section 4 forms a second communication passage, which comprises a primary channel 41 and a secondary channel 42, which both extend through the fourth section 4. In the first embodiment disclosed, the primary channel 41 and a secondary channel 42 extend in parallel to the longitudinal axis X.
The first section 1, the second section 2, the third section 3 and the fourth section 4 are configured to permit the following consecutive relative positions.
In a first position, see
In the first position, the milk sample may thus be supplied to the first fill chamber 21 and the second fill chamber 31 via the primary channel 41. The primary channel 41 may be connected to an inlet conduit 44 and operate as an inlet to the first fill chamber 21 and the second fill chamber 31. The first communication passage 11 may be connected to an outlet conduit 16, so that the milk to be supplied may flow through the device, thereby ensuring that the first fill chamber 21 and the second fill chamber 31 are filled. The inlet conduit 44 may be connected to the milking equipment and receive milk from a milk collecting member (not disclosed) of the milking equipment.
During the supply of milk, the milk is thus flowing through the primary channel 41, the first fill chamber 21, the second fill chamber 31 and the first communication passage 11. When these are filled with milk and no gas bubbles are present, the second section 2 and the third section 3 are moved with the aid of the second actuator 24 and the third actuator 34 so that a second position is obtained. It is to be noted that the milk may flow also in the opposite direction during the supply of milk, i.e. in through the first communication passage 11 and out through the primary channel 41.
In the second position, see
In the second position, the milk sample contained in the first fill chamber 21 and the second fill chamber 31 may be centrifuged in order to obtain a first sample part of a volume equal to the first volume, and a second sample part of a volume equal to the second volume. The centrifuging may last for 15 to 60 s. The centrifugal force required is estimated to lie in the range 500 g to 1800 g. After the centrifuging, the first sample part contains all fat and casein, or substantially all fat and casein, of the milk in the milk sample. The second sample part contains all, or substantially all, somatic cells of the milk in the milk sample.
The centrifuging may be performed in a centrifuge (not disclosed) of any suitable kind. The device may be configured to be contained in the centrifuge during the centrifuging of the milk sample.
In a third position, see
The third position is configured to permit supply of a solution to the first mixing chamber 21 and the second mixing chamber 31. The solution comprises, or consists of, a biologically acceptable saline solution, and may comprise, or consist, of a biological acceptable NaCl-solution. The salt content may then correspond to the salt content of the body liquid of the animal.
During the supply of the solution, the solution is flowing through the first communication passage 11, the first fill chamber 21, the second fill chamber 31 and the primary channel 41. When these are filled with the solution and no gas bubbles are present, the second section 2 and the third section 3 are moved with the aid of the second actuator 24 and the third actuator 34 so that a fourth position is obtained, thereby removing one sample part, i.e. in this example the first sample part, and retaining a remaining sample part, i.e. in this example the second sample part.
In the fourth position, see
In the fourth position, the first volume of the solution and the remaining sample part, i.e. the second sample part, are mixed in the first mixing chamber 22 and the second fill chamber 31 to form a sample mixture in which the second sample part is uniformly distributed.
The mixing may be performed by means of a stirring member 26, such as a steel ball, contained in the first mixing chamber 22. The stirring member 26 may be moved reciprocally in the first mixing chamber 22 and the second fill chamber 21 by means of a stirring device 27 comprising a moving magnet or magnet field outside the first mixing chamber 22 and the second fill chamber or outside the device. The diameter of the first mixing chamber 22 and the second fill chamber 31 may be somewhat greater than the diameter of the first communication channel 11, the primary channel 41 and the secondary channel 42 in order to maintain the stirring member within the first mixing chamber 22 and the second fill chamber 31.
In a fifth position, see
It is to be noted that the abutment of the sections 1, 2, 3 and 4 against each other, which is discussed above, prevents any leakage of milk or the solution during the relative movement of the sections 1, 2, 3 and 4 to the different positions, and also when the sections 1, 2, 3 and 4 are positioned in the different positions.
In the example discussed above, the sample mixture to be discharged from the device and analysed contains the second constituent, i.e. somatic cells. However, it is to be noted that the device is suitable also for isolating the first constituent, i.e. fat cells. In this case, the sections 1, 2, 3 and 4 are the same and are arranged in the same way in the first, second and third positions. However, in order to obtain a first sample part containing only the first constituent, it may be advantageous to adapt the length and the volume of the second fill chamber 31 and the second mixing chamber 32 to the total volume of the first and third constituents after the separation. The fourth position, in which the first fill chamber 21 and the second mixing chamber 32 are communicating with each other and closed to the first communication passage 11 and the second communication passage 43, permits mixing of the solution and the first sample part, in the first fill chamber 21 and the second mixing chamber 32 to form a sample mixture in which the first sample part is distributed. In the fifth position, the first communication passage 11 and the second communication passage 41 communicate with each other and with the first fill chamber 21 and the second mixing chamber 32 to permit discharge of the sample mixture.
It is to be noted that elements having the same or similar functions in the different embodiments have been allotted the same reference signs in all embodiments.
In the third embodiment, the first section 1, the second section 2, the third section 3 and the fourth section 4 are held together by means of a bolt 50 extending through a centre hole through all section 1 to 4 along the longitudinal axis X forming a centre axis of the bolt 50. The sections 1 to 4 are pre-tensioned against each other by means of a spring 51. The pre-tensioning force may be regulated by means of one or more nut 52 threaded onto the bolt 50.
No frame is shown in
In the same way as the first and second embodiments, also the third embodiment is suitable for isolating and discharging a sample mixture containing fat cells.
The sections 1, 2, 3, 4 and 6, are configured to permit the following consecutive relative positions.
In a first position, the first communication passage 11, the first fill chamber 21, the second fill chamber 31, the fifth fill chamber and the second communication passage 43 communicate with each other. The first position permits supply of a milk sample containing at least a first constituent, such as fat, a second constituent, such as somatic cells, and a third constituent, such as casein, to the first fill chamber 21, the second fill chamber 31 and the fifth fill chamber 61.
In a second position, the first fill chamber 21, the second fill chamber 31 and the fifth fill chamber 61 are communicating with each other and closed to the first communication passage 11 and the second communication passage 43. The second position permits centrifuging of the milk sample contained in the first fill chamber 21, the second fill chamber 31 and the fifth fill chamber 61 to obtain a first sample part of a first volume in the first fill chamber 21, in which first sample part the first constituent is contained, a second sample part of a second volume in the second fill chamber 31, in which second sample part the second constituent is contained, and a third sample part of a third volume in the fifth fill chamber 61, in which third sample part the third constituent is contained.
In a third position, the first communication passage 11, the first mixing chamber 22, the second mixing chamber 32, the fifth mixing chamber 62 and the second communication passage 43 communicate with each other. The third position permits supply of a solution to the first mixing chamber 21, the second mixing chamber 31 and the fifth mixing chamber 61.
In a fourth position, one of the first, second and fifth fill chambers 21, 31, 61 is communicating with two of the first, second and fifth mixing chambers 22, 32, 62, thereby forming a channel extending through the second, fifth and third sections 2, 6 and 3 and being closed to the first communication passage 11 and the second communication passage 43. The fourth position permits mixing of the solution and either first sample part, the second sample part or the third sample part to form a sample mixture in which this sample part is uniformly distributed.
In a fifth position, the first communication passage and the second communication passage communicate with each other and with said channel to permit discharge of the sample mixture.
It should be noted that not only the first embodiment but also the second and third embodiments may be modified to include five sections as explained above for the fourth embodiment.
The present invention is not limited to the embodiments disclosed but may be varied and modified within the scope of the following claims.
It is also to be noted that the stirring member 26 and the stirring device 27 shown may be replaced by any other stirring equipment. For instance the stirring may be performed by circulating the solution and the second sample part through external channels temporarily connected to the first mixing chamber 22 and the second fill chamber 31 when these are in the fourth position.
Number | Date | Country | Kind |
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1350917-9 | Jul 2013 | SE | national |
Filing Document | Filing Date | Country | Kind |
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PCT/SE2014/050901 | 7/22/2014 | WO | 00 |