Title: IMPROVING THE EFFICIENCY OF THE A.I. BY NEW TECHNICAL AND DIAGNOSTIC METHODS IN CATTLE
1IMPROVING THE EFFICIENCY OF THE A.I. BY NEW
TECHNICAL AND DIAGNOSTIC METHODS IN CATTLE
- 1Gábor G, 1Tóth F, 2Szász F, 1Dept. Cattle
Breeding and Dept. Cell Biology , Res. Inst.
Anim. Breed. Herceghalom, Hungary, 2Androvet
Ltd., Budapest, Hungary
2INTRODUCTION
- Prediction of the males fertility is based on
morphological examinations of the testis or
scrotum and the evaluation of the semen quality.
Prediction of the semen production would help to
reject all those young bulls which certainly will
not produce good quality of semen. - Early pregnancy detection looks a useful tool to
check the real fertility of the breeding bulls.
3OBJECTIVES
- The aims of all our examinations were
- to improve the standard BSE (Breeding Soundness
Evaluation) by using of testicular tonometry and
testicular ultrasonic echotexture, -
- 2.checking semen quality by an objective
inexpensive instrument based method (CASPAR) -
- 3. checking the real bull fertility by early
pregnancy detection used ultrasound and/or PSPB
(Pregnancy Specific Protein B) ELISA test
(Biopryn, BioTracking, Moscow, ID, USA) .
4MATERIAL AND METHODS
- 1. Improvement of the standard BSE
- Holstein-Friesian AI bulls were examined
during the BSE. -
- Morphological examinations of the testis scrotal
circumference (SC used Coulter scrotal tape)
scrotal form digital camera (storing format
jpg) testicular consistency (TM, Consitest
electronic tonometer). The ultrasonic examination
of the testes (ET) was done with a B-mode scanner
(Pie Medical, Model 100 LC VET, Maastricht, The
Netherlands) with an 8 MHz linear-array
transducer as described earlier and the
ultrasound-image was stored on floppy disk.
TesTracto? software was used for image analysis.
5Coulter Scrotal Tape
6Scrotal forms
Far from the body longer ligaments
Closer to the body shorter ligaments
Oval form rounded
shape testicular torsion (above)
and tided scrotum
7Consitest electronic tonometer
- The spring loaded measuring heads use 1500 grams
of pressure at all times. Because of the
difference in dimension of the two heads (13),
the penetration to the observed tissue will be
different. The transmission changes from a linear
movement to a circular movement. A rotating code
disk interrupts the infrared light, which counts
the impulses for the microprocessor, which then
summarizes the results and presents it on the
display.
8Examination of the testicular echotexture
- The ultrasonic examination of the testes was done
with a B-mode scanner (Pie Medical, Model 100 LC
VET) with an 8 MHz linear-array transducer. Gain
was held constant for all examinations. - The transducer was aligned at the center of each
testis and perpendicular to the vertical axis. A
custom, electro-mechanical device for holding the
probe was used to assure that the same pressure
was applied by the probe against tissues upon
which liberal amounts of ultrasound coupling gel
had been applied (except with dogs). - At approximately 0.8 kg/cm2 pressure a
spring-loaded switch was compressed. At that
instance, the ultrasound image was frozen on the
scanner and the image was stored.
9Examination of the ET by the help of an
electronic (auto image freezing function)
transducers holder
10Evaluation of the echotexture (ET) by Scanner 100
left testis
right testis
- The ultrasonic images of left and right testicles
are recorded on disk. The average gray level were
characterized on a 256 gray scale 0 (dark) and
255 (light).
11TesTracto software (developed for the data base)
was used for the evaluation of the ultrasonic ET
12Distribution evaluation of the gray levels
13- Semen evaluation the total number of spermatozoa
per ejaculate (TNS), the percentage of the motile
and progressive motile spermatozoa were
determined. A Zeiss Laboval phase-contrast
microscope with heated stages and a Makler
chamber was used for preparing the images. All
the images were saved to the HDD of a computer
and image analysis was carried out by a custom
software (CASPAR?, Pictron Ltd., Budapest,
Hungary). Morphological evaluation was done after
Cerovskys staining.
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172. Early pregnancy detection by ultrasound and
PSPB test
- The aims of these experiments were checking the
bull fertility by early pregnancy detection. -
- In Experiment 1 sixty-nine dairy cows were
checked by a B-mode ultrasound scanner (Pie
Medical, Model 100 LC VET, Maastricht, The
Netherlands) with a 6 MHz linear-array
transducer, 29-55 days after AI in order to check
the pregnancy status of them. At the same time
blood samples were also taken from the tail vein
to check the PSPB serum level of these cows. - In a second experiment 4 bulls were selected
during the BSE as a semen donor for a fertility
experiment. After semen collection and freezing,
1742 artificial inseminations were carried out at
23 dairies with the frozen-thawed semen. Early
pregnancy diagnosis was carried out once a week
30-36 days after the AI. For checking serum PSPB
level, Biopryn test was used. Results were
re-checked by rectal palpation 60 days after AI.
18Ultrasonic appearance of the bovine fetus at
different stages of the pregnancy
19PSPB (Pregnancy Specific Protein B)
- PSPB is a novel protein that was first reported
by scientists at the University of Idaho. First a
double antibody radioimmunosassay (RIA) was
developed with rabbit antisera against a more
nearly pure form of PSPB. The assay was used to
measure PSPB in sera of pregnant cows and
provided an accurate, early serological test for
pregnancy. - PSPB is detectable in serum from 24 to 282
(parturition) days of gestation and can be
applied reliably in dairy cow herds at 28 to 30
days after breading. - PSPB is located in the giant binucleate cells of
the trophoblastic ectoderm of the placenta and
this indicated that is was either synthesized or
sequestered by those cells. - The PSPB was found in sera of some primiparous
cows as early as 15 days following conception and
in sera of most cows at 24 days following
conception. - Complete clearence of the protein from blood may
not occur until 80 to 100 days after parturition.
For this reason testing of blood for a new
pregnancy following parturition may give false
positive lests. - An ELISA test (BioPryn by BioTracking LLC)has
been developed for the examination of the PSPB in
cattle 30 days after AI (at least 90 days after
the previous parturition).
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24Results
Table 1 Data of bulls in the data base with
complete record
BULL (n25)
Age (mo) 14.0 2.7
SC (cm) 32.9 4.1
TM (mm) 4.9 1.2
ET (0-255) 44.7 18.0
Motile spermatozoa () 71.0 10.8
Linear progressive motility () 48.0 8.6
Speed (mm/s) 96.9 10.9
Usually around -0.50, significant
Usually its a positive, significant correlation
25Categories of diagnosis (just for bulls)
- Based on SC, ET and TM. Examination of the bulls
was performed at least 2 months prior to the
semen production. - Semen was measured for at least 2 months after
the beginning of the semen production SV, SD,
TSN - NP - No predicted semen production problem
- PP - Possible predicted semen production problem
- NUAI - Prediction not useful as a breeding bull
26Table 2 Prediction results in young breeding
bulls
27Possible reasons of the incorrect predictions
Calcification. Dg. PP normal semen production
6 months later
Small SC (20 cm at 8 months of age normal SC size
(32 cm) and semen production4 months later
28Possible reasons of the rejection
Cryptorchidism
Left testis orchitis right testis fibrosis
29Table 3 OD values in the first PSPB experiment
(n69) We had 100 agreement between the
ultrasonic and PSPB diagnosis
30Table 4 Result of the bulls fertility in the
field experiment
31Table 5 Embryonic loss in farms with different
milk production