Method for cytological diagnostics of puerperal pathology in cows

FIELD: veterinary medicine.

SUBSTANCE: the present innovation deals with sampling a cow's vaginal smear followed by its staining and microscopic testing the cells. During microscopic testing one should count the cells of vaginal epithelium (surface, parabasal and basal) . In case of prevailing quantity of surface cells (49-60%) one should diagnose normal flow of puerperal period, in case of prevailing parabasal epithelial cells one should diagnose complicated puerperal period, moreover, if the quantity of surface and basal cells is similar or he difference is insignificant (15-20% against 15%, correspondingly) it is possible to diagnose uterine subinvolution, and at considerable difference in the quantity of surface and basal cells (0.5-11% against 20-22%) one should diagnose acute puerperal endometritis. The innovation enables to analyze and detect puerperal pathology in earlier terms.

EFFECT: higher accuracy of diagnostics.

1 dwg, 2 tbl

 

The invention relates to veterinary medicine, namely to veterinary obstetrics and gynecology.

There is a method of cytological diagnosis of postpartum pathology (Annacchino, 1974). To perform this method, perform the following manipulations:

1) take a swab from the cervix;

2) paint it Pappenheim;

3) count the number of neutrophils, the nature and degree of degenerative changes;

4) the number and status of neutrophils assess the condition of the sexual apparatus.

The method chosen for the prototype.

But it has serious disadvantages, namely, that:

1) a swab taken from the cervix, which makes study time-consuming, requires additional tools, skills veterinary specialist;

2) study material, you can get not earlier than 6-8 days after birth, because by that time the fence material is impossible because of pain research and the high likelihood of diagnostic error;

3) smear stained by the method of Pappenheim that restricts the use of the method in connection with the high cost of reagents for its implementation;

The objective of the invention is to develop a cheaper and more simple in execution of the diagnostic technique used to predict and identify postpartum pathology at an earlier date.

Those who practical result is the extension of the functionality of the method, expressed in the following:

1) the emergence of possible predictions of postpartum complications;

2) the possibility of early diagnosis and predictions, starting with the first day after calving;

3) ability to conduct research not only nagaragawa cows, but in heifers.

In addition, the technical result is to reduce the complexity and reduce the cost method.

To solve this problem, a method of cytological diagnosis of postpartum pathology in cows, as a prototype, is in taking a swab from the genital tract of the cow with its subsequent staining and microscopic examination of the cells, they will judge the nature of the flow of the postpartum period. Unlike the prototype, a swab is taken from the vagina, colouring make Romanovsky-Giemsa, and microscopic examination is considered cells of the epithelium of the vagina (surface, parabasal, basal, additionally considering the state of their nuclei) and their ratio determines the state of the reproductive system.

In General, the method is as follows. Material for cytological study is vaginal fluid, which is taken from the upper wall of the vagina to the rotational motion using a cotton buds soaked in sterile isotonic solution of sodium chloride. Received mater what al put on a skim glass slide dried in the air for 10-15 min, then fixed in 96% ethanol for 30 minutes Coloring carried out according to the method Romanovsky-Gimza. In one smear count 100 epithelial cells 25 cells in 4 fields of view.

Cytological composition of the vaginal mucus is determined by two indicators in percentage:

1) the number of surface, parabasal and basal epithelial cells;

2) nuclei of the above cells.

As a concrete example, we give the results of the research method proposed in 175 head of cattle black-motley breed at the age of 3-6 years, weighing 350-450 kg with an average milk yield of 3500 kg, divided into 3 groups according to the nature of the flow of the postnatal period (group 1 with normal during the postpartum period, 50 goals, group 2 with subinvolution of the uterus - 64 heads, group 3 with postpartum endometritis - 61 head).

The research results are summarized in the charts on the attached figure. It shows the percentage of epithelial cells in 3 groups: the first chart represents the ratio of the surface of epithelial cells, second - parabasal cells and third - basal epithelial cells. Black color indicates the columns reflecting the ratio of cells in subinvolution of the uterus, dark grey color - normalisation postpartum and white with postpartum endometritis.

As seen in the drawing, the percentage of epithelial cells in 3 different groups.

Thus, the surface of the cells in the group with normal during the postpartum period will be 49-60% of the total number of epithelial cells in the group of subinvolution of the uterus is 15-20%, and at endometritis - 0.5 to 11%.

Parabasal cells in normal course of smears there 28-38% of the total number of epithelial cells of the vagina, subinvolution of the uterus will be 53-63%, and at the postpartum endometritis - 40-55%.

Relative basal cells following picture: at physiological course of the postpartum period and their number is not more than 11% of the total number of epithelial cells; subinvolution of the uterus - 15%, and at the postpartum endometritis even more 20-22%. Differences between cells are reliable in all groups.

Thus, in the normal course of the postpartum period in smears of the epithelium of the vagina of cows prevail surface of the cell.

When subinvolution of the uterus greater percent parabasal epithelial, and the number of surface and basal cells are almost equal.

With postpartum endometritis picture smear another: the number of parabasal cells prevails over other types of epithelial cells, but there is a significant difference between the surface and the base is ranked on the cells (0.5 to 11% vs. 20-22%).

CellNormaThe subinvolutionEndometritis
Surface49-60%15-20%0.5 to 11%
Parabasal28-38%53-63%40-55%
Basal11% (maximum)15%20-22%

Additional diagnostic indicator in predicting the course of the postpartum period on the first day after calving, is the condition of the cores of the above cells of the vaginal epithelium.

So, under normal during the postpartum period on the first day after birth the number of parabasal epithelial cells with normal kernel will be of 33.8% of the total number of epithelial cells, basal cells with a nucleus in a state of lysis in a smear is not marked.

About the possibility of slowing down the processes of involution of the uterus indicates the presence of smears on the first day postpartum superficial cells with normal kernel, their population is about 7-10% of the total number of epithelial cells, superficial cells with a nucleus in a state of picknose - 0.5 to 2%, of the surface cells with a nucleus in a state of rexis - 2-8%, parabasal cells of normal core - 12-17%, parabasal cells is to the kernel in a state of rexis - 7,5%.

When endometritis on the first day after birth registered parabasal cells with a nucleus in a state of lysis - 10% and 2% of basal cells with a nucleus in a state of rexis of the total number of epithelial cells.

NormaThe subinvolutionEndometritis
Parabasal cells with normal core of 33.8%Superficial. cells with a normal kernel - 7-10%Parabasal cells with a nucleus in a state of lysis -10%
Basal cell with a nucleus in a state of lysis - noSuperficial. cells with a nucleus in a state of picknose - 0,5-2%Basal cell with a nucleus in a state of rexis - 2%
Superficial. cells with a nucleus in a state of rexis - 2-8%
Parabasal cells of normal core - 12-17%
Parabasal cells with a nucleus in a state of rexis - 7,5%

Thus was established the following:

differences cytological composition of vaginal mucus in three groups according to the ratio of epithelial cells and nuclei of these cells are logged in the first day after birth, and that gives the opportunity to predict postpartum complications from the Directors on the first day after calving. The technique of the method, i.e. taking vaginal, and not cervical smear reduces the complexity of the method and allows to obtain material not only nagaragawa animals, but also in heifers. The study of epithelial cells of the vagina more easily performed, cheaper and more informative than the study of neutrophilic leukocytes in the method adopted for the prototype.

It is obvious that the study of the composition of the vaginal mucus can serve as a valuable way to predict the course of the postpartum period and differentially diagnose its complications.

1. Method of cytological diagnosis of postpartum pathology in cows, including taking a swab from the genital tract of the cow with its subsequent staining and microscopic examination of the cells, characterized in that the PAP smear is taken from the vagina, and microscopic examination is considered cells of the epithelium of the vagina (surface, parabasal and basal)and at the prevailing number of surface cells (49-60%) diagnosed normal during the postpartum period, when the predominance of parabasal epithelial cells diagnose the complication of the postpartum period, and if the number of surface and basal cells while the same or differs only slightly (15-20% versus 15% respectively), while the automotive technician strout the subinvolution of the uterus, but with considerable difference in the number of surface and basal cells (0.5 to 11% vs. 20-22%) diagnosed with acute postpartum endometritis.

2. Method of cytological diagnosis of postpartum pathology in cows according to claim 1, characterized in that the staining of the smear produced by Romanovsky-Giemsa and, in addition, examine the condition of the cores of the above cells of the vaginal epithelium, and when the number of parabasal cells with normal kernel and 33.8% of the total number of cells and the absence of basal cells with a nucleus in a state of lysis diagnose normal during the postpartum process, when the number of surface cells with normal kernel 7-10%superficial cells with a nucleus in a state of picknose 0.5 to 2%, of the surface cells with a nucleus in a state of rexis 2-8%, parabasal cells with normal kernel 12-17%and parabasal cells with a nucleus in a state of rexis 7.5% of diagnosed subinvolution of the uterus, and when the number of parabasal cells with a nucleus in a state of lysis 10%, and basal cells with a nucleus in a state of rexis 2% diagnose acute endometritis.



 

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