Treatment of Repeat breeding syndrome

1. 1
POST GRADUATE CREDIT
SEMINAR ON
PRACTICAL APPROACHES FOR TREATMENT OF
REPEAT BREEDING SYNDROME IN DAIRY CATTLE
Major Advisor:
Dr. C. T. Khasatiya
Associate Professor & Head
Department of Veterinary
Gynaecology & Obstetrics
Speaker:
Yede Amol Baban
Reg. No :2040418016
M. V. Sc. Scholar
Minor Advisor:
Dr. V. R. Patel
Assistant Professor
Department of Animal
Nutrition
VANBANDHU COLLEGE OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY,
NAVSARI AGRICULTURAL UNIVERSITY
NAVSARI- 396 450
VOG - 691

2. 2
INTRODUCTION
INCIDENCE
ETIOLOGY
DIAGNOSISTREATMENT
CONCLUSIONS
FUTURE
PROSPECTS
CONTENTS

3. 3
INTRODUCTION
(Annual Report 2018-19)
Cows are raised for many reasons
including: milk, cheese, meat, leather
hide, other dairy products
(India livestock population in million)
(20th Livestock Census)
 Livestock plays an important role in Indian economy.
 About 20.5 million people depend upon livestock for their livelihood.
 Livestock contributed 16% to the income of small farmers.
 Livestock provides employment to about 8.8 % of the population in India.
 Livestock sector contributes 4.11% GDP and 25.6% of total Agriculture GDP.

4. 4
(3yr study on 576 dairy cattle)
(Khan et al., 2016)(Annual Report 2018-19)

5. 5
Repeat breeding syndrome (RBS) is a major problem in dairy cattle leading to large
economic loss for the dairy producer due to more inseminations per conception,
increased calving interval and culling rate
Repeat breeder cow is define as cow with
• Normal Oestrous cycle
• Normal Oestrous period
• Naturally Bred or artificially inseminated three or more
times continuously with fertile bull or semen of fertile
bull.
• But failed to conceive.
 RB cow return to oestrus after a mating or artificial insemination (AI), could
be caused by either fertilization failure or embryonic death.
(Perez-Marin et al., 2012)
Total early pregnancy loss of close to 40% during the first 21 days post AI.
(Roche et al., 1981)
(Lafi et al., 1992)

6. 6
INCIDENCES
Usually about 9 to12% cows are expected to be
repeat breeder in a herd with normal fertility and
with 50-55% conception rates.
Higher incidence of RBS reported in crossbred
cow than local breed.
(Nishi et al., 2017)
(Mahto and Sinha, 2019)

7. 7
0
10
20
30
40
50
Heifer 1st 2nd 3rd 4th
30.76
42.85
38.46 37.56
18.33
Percentage
Parity wise RBS %
0
5
10
15
20
25
JAN
FEB
MARCH
APRIL
MAY
JUNE
JULY
AUG
SEPT
OCT
NOV
DEC
Percentage
MONTH
 The highest incidence (23.52%) was recorded in the month of February and the
lowest (11.90%) during month of June.
 First calvers showed maximum incidence (42.85%) and fourth calvers the lowest
(18.33%).
(Mahto and Sinha, 2019)

8. 43%
10%7%
40%
Distribution of Embryonic mortality
Early embryo mortality
Fertilization failure
Late embryo mortality
Successful conception
8
ETIOLOGY
Etiology of RBS is multifactorial
Early embryonic death
Failure of fertilization

9. 9
Etiology of RBS
A) Maternal factors:
1. Influence of maternal age
2. Genetic factors
3. Uterine infection
4. Anatomical defects of the
genital tract
5. Hormonal dysfunctions
6. Nutrition causes
B) Bull factors:
1. Influence of bull fertility
and semen quality
2. Site of semen deposition
3. Time of semen deposition
4. Immunological
incompatibility
C) Environmental and Management factors:
1. Season
2. Estrus detection problem
3. Unhygienic at artificial insemination
and parturition
4. Stress
(Perez-Marin et al., 2012)

10. 10
Maternal factors:
1. Influence of maternal age
Age Rate of RBS Fertility
(Hewett, 1968)
• 1.31ng/ml
Heifers
• 2.19 ng/ml
Cow at (3-5th )
Lactation
• 0.94 ng/ml
Cow after 5th
Lactation
LH level at AI
(Santana et al., 2000)
 Alteration of HPG
axis and ovarian
response.
 Low oocyte
viability

11. 11
2. Genetic factors
 Genetic abnormalities of parents
 Genetic abnormalities increase due
to high inbreeding or aged gametes
(Snijders et al., 2000)
X- Trisomy

12. 12
3. Uterine infection
E.coli, Staphylococcus aureus, Bacillus spp.,
Proteus spp., Enterobacter spp.,
Corynebacterium spp. and Pseudomonas
aeruginosa.
Endometritis
in RB cow
Clinical endometritis-
12%
Sub-clinical endometritis
29.7%
(Sharma et al., 2009)
Bacteria isolates from
uterus of RB cow
Spp. of Geotrichum, Cladosporium, Mucor,
Verticillium, Chrysosporium , Rhodotorula,
Cephalosporium, Candida albicans, Candida
tropicalis, Aspergillus fumigates, Aspergillus
ochraceous, Aspergillus niger and Rhizopus.
(Sharma and Singh, 2012)
Spp. of fungus isolates from
uterus of RB cow
Fungal
endometritis
(15.5%)
(Kumar et al., 2018)

13. 13
Congenital defects of the genital tract:Acquired defects of genital tract
 Oviduct obstruction:
• Salpingitis
• Hydrosalpinx
• Pyosalpinx
• Oviduct adhesions
 Cystic uterine tube
 Uterine tumor
 Cervical stenosis
• Aplasia of uterine tube
• Segmental aplasia of
uterine tube (unicornis)
• Accessory uterine tube
• Ovarian hypoplasia,
aplasia
• Double vagina
• Pneumovagina
• Hard & kinked
cervix
• Double cervix
• Adhesions of ovary,
fallopian tube or
ovarian bursa
(Shakya et al., 2012)
4. Defects of the genital tract

14. 5. Hormonal dysfunction
1) Anovulation 2) Delayed ovulation
3) Luteal insufficiency
Anovulation
Anovulation was reported 2-16% in
repeat breeder cows.
14(Wiltbank et al., 2002)
1
Nutrition deficiency & lactating cow
Mechanism of Normal ovulation

15. 15
Insensitivity of hypothalamus to positive
feedback effect of estradiol
2
High level of supra basal progesterone
(Wiltbank et al., 2002)
Anovulation + delayed ovulation
Incomplete luteal regression3

16. 16
CL-Corpus Luteum
P4 – Progesterone
OTr- Oxytocin receptor
LH-Luteinizing hormone
MRP-Maternal
recognition of pregnancy

17. 17
6. Nutritional causes:
Reduce food intake or
imbalance diet
Nutritional deficiency
Loss of body weight and
body score condition
Endocrine imbalances
RBS
Nutritional deficiency
during post partum
Delay uterine
involution
Increase open days
Uterine infection
RBS
(Purohit, 2008)
 Negative energy balance(NEB)
 Excess of dietary protein
 Deficiency of calcium, phosphorus,
iodine, cobalt, copper, zinc,
magnesium and selenium
 Deficiency of vitamin A and E
 Deficiency or excess of body
metabolite such as glucose, urea,
albumin, globulin and non
esterified fatty acid etc.
NEB
Reduce
GnRH/LH
Low
estradiol RBS
1 2

18. 18
Bull factors :
1. Influence of bull fertility and semen quality
Lower sexual behavioral and libido of bull
Requirement for successful AI
Post thaw total motile sperm More than 5 million/dose
Post thaw progressive motility % More than 50%
Post thaw live sperm count % More than 40%
Post thaw normal sperm More than 70%

19. 19
2. Site of semen deposition
 Defective inseminations (Deposition of
semen at the entrance of the cervix or vagina )
are responsible for RBS.
 Higher conception rates are obtained when
sperm is deposited into the uterus.
(Gwazdauskas et al., 1986)
3. Time of semen deposition
 Estrus behavior could last between 13-17 hrs on average
 Ovulation will take place approximately 30 hrs after standing the heat
 It must be ensured that semen is deposited in the female tract as close as
possible to that time
 Mismatch between time of ovulation and AI responsible for RBS in cow
(Perez-Marin et al., 2012)
Site of AI

20. 20
Environment and Management factors
 Lactational stress
 Size of livestock
 Milk yield
(Hewett, 1968)
1. Season
2. Estrus detection failure
The poor heat detection is one of the
main cause of RBS in cattle
• Mis-timed inseminations
• AI without evident estrus
(Lafi and Kaneene, 1988)
Summer month
• Heat stress
• Depress endocrine function
• Reduced intensity of estrus
• Decreased preovulatory LH peak
• Decrease viability of embryo
Predisposing factors

21. 21
3. Unhygiene at Artificial insemination (AI) or at Parturition
Unhygienic AI
Improper Care during peripartum period
4. Stress
Stress
Increase
cortisol level
From adrenal
gland
Secretion
Adrenal
progesterone
Increase
supra
basal P4
(Bage, 2002)
(Erb et al., 1985)

22. 22
DIAGNOSIS
Diagnostic methods of RB
1. Record analysis
2. Visual
3. Recto genital palpation
4. Vaginoscopy
5. Test to evaluate uterine
health
 Uterine pH
 Uterine microbiology
 Uterine biopsy and
cytology
6. Metabolic profiles
7. In vivo imaging techniques
8. Immunological tests
9. Endoscopy
10.Tubal patency testing
11.Hormone assays
1. Record analysis
 The number of actual inseminations done
or number of times fail to conceive.
 Previous periparturient diseases.
 Management strategies
(Purohit, 2008)

23. 23
2.Visual
Visual observations of inseminators
during AI
 Colour of cervico-vaginal mucus
 Consistency of cervico-vaginal
mucus
 Clarity of cervico-vaginal mucus
 Vulvar Oedema
 Vaginal congestion
 Service without any ejaculatory thrust
 Urine pool of vagina contaminates the semen
(De Araujo et al., 2003)
Clear/ transparent Purulent discharge Vaginal mucous exam

24. 24
3. Recto –genital palpation
For estimation of ovulation, palpations must
be done every 12 hrs from AI till the finding
of an ovulation depression.
Evaluating the early or late corpus
luteum (CL) to rule out luteal
insufficiency.
(Bruyas et al., 1993)
 Examination of both ovaries(for
any abnormality/ies)
 Examination of uterine horn
 Os of cervix
 Irregularly shaped cervix
 Size of ovulatory follicles
 Uterine tone
 Ovarobursal adhesions
Per rectal examination of reproductive
organs before AI or Natural Service include:
Corpus Luteum
Follicular cyst

25. 4. Vaginoscopy diagnosis
Small quantities of infected material inside cervix and
vagina determine by Vaginoscopy to diagnose reproductive
tract infection (Miller et al., 1980)
Limitation
 Discharge is absent when cervix is close
 Absence of uterine discharge does not indicate
absence of uterine infection (Gilbert et al., 2005)
Metricheck device was more sensitive in detecting
endometritis compared with Vaginoscopy
40mm hemisphere of silicon
attached to a 500mm long stainless
steel rod
Vaginoscopy
(Mcdougall et al., 2007)
Metricheck with Pus 25

26. 26
5. Tests to evaluate uterine health
 Uterine pH
 Uterine microbiology
 Uterine biopsy and
cytology
1 2 3 4 5 6 7 8 9 10
7.35 7.32 7.28 7.27 7.26 7.32 7.32 7.32 7.33 7.22
Ovulation 10th Day
(Mather, 1975)
Reduction of pH from7.2 to 6.9 at estrus compromise
embryonic development (Ocon and Hansen, 2003)
pH in case of metritis-8.23 to 8.80
(Boitor et al., 1980)
A. Uterine pH
Vaginal
mucus pH
At estrus
Repeat
breeder
8.5±1.16
Normal
breeding
7.2±1.10
(Gupta et al., 1981)

27. 27
B. Uterine microbiology
White side test
1 ml of estrus cervical mucus+ 1 ml
of 5 to 10% sodium hydroxide
Heated up to boiling point
After cooling the intensity of color changes study
Normal (turbid or no color)
Mild infection (light yellow color)
Moderate infection (yellow color)
Severe infection (dark yellow color) (Popov, 1969)
Collect cervical mucus and evaluate for
isolation and identification of bacteria on
the basis of cultural, morphological,
colony characteristics, motility,
biochemical reactions etc.
(Cowen and Steel, 1970)
Bacterial culture:

28. Method of sample collection
 Flushing the uterus
 Cytobrush
28
C. Uterine biopsy and
cytology
 Increase percentage of
polymorphonuclear leucocytes (PMNs)
 Infiltration of lymphocytes
 Supra nuclear vacuolation
 Denudation of epithelial lining
 Inflammatory changes and fibrosis
(Bonnett et al.,1991)
Histological finding:
Cytological finding
(Gilbert et al., 1879) Cytobrush
Uterine cytology

29. 29
6. Metabolic profiles
Parameters
Value in
Normal cow
Value in Repeat
Breeding cow
Cholesterol (mg%) 83.0–249.22 77.8–182.37
Ca (mg%) 10.73 ± 0.31 8.49 ± 0.30
P (mg%) 4.22–8.19 3.37–8.03
Fe (mg/dl) 1.9–2.4 2.47–11.3
Mn (mg/ml) 0.46–0.58 0.17–0.19
Zn (mg/ml) 1.09–3.14 0.65–1.19
Cu (mg/m 0.65–1.14 0.22–0.99
Blood urea (mg%) 18.80 28.88
Vitamin A (mg/dl) 41.216 37.14
Co (mg/ml) 2.18–9.67 0.85
Attributes N.C RB
Serum
glucose(mg/dl)
49.2 42.8
Serum total protein
g/dl
5.71 5.52
Serum albumin g/dl 2.81 2.71
Serum globulin g/dl 2.89 2.82
Albumin : globulin 0.97 0.96
(Mondal and Paul, 2012)
(Purohit, 2008)
*N.C (Normal cow) value

30. 30
7. In vivo imaging techniques USG
 USG is useful for measuring
follicles and detecting CL(3-5
days after ovulation.
(Pierson et al., 1987)
Ovarian dysfunction that commonly
diagnose by USG
 Ovulation defects
 Luteal dysfunction
(Bage et al., 2002)
 Optimum size diameter of a
follicle at AI would be 1.5–2.0 cm
for dairy cows.
1
4
2
3
Graafian follicle Follicular cyst
Polycystic ovary Corpus luteum

31. 31
Performed experiment on Progesterone Level
on virgin heifers(VH) and repeat breeding
heifers(RBH) and concluded following
observations in RBH
Mean plasma
progesterone level at
the time of AI in
RBH
(nmol/l)
0.64
±0.8
<0.5
Fail to pregnant
Pregnant
8. Circulating hormone assays – P4
Higher supra basal progesterone
level during estrus
Low post ovulatory rise of P4
(Bage et al., 2000)

32. 32
9. Immunological tests
Antisperm antibody has sperm-immobilizing activity, leading to penetration
reduction of sperm in cervical mucus and resultant reduced fertility
The detection of Antisperm antibodies in serum or cervical mucus of cattle
10. Using endoscopy
 Direct hysteroscopy has been used to evaluate the uterus
 A paediatric gastroscope with air insufflation allow good visibility of both
uterine cornua
(Gerring, 1986)
(Kremer and Jager, 1992)
{Lander et al.(1990 ) and Deo and Roy. (1971)}

33. 33
11. Tubal patency testing
Phenol sulphonphthalein (PSP)Test
Time (min) Result
>15 Both tube normal
15-45 Both tube normal
30 Both tube normal
60
Left/ Right oviduct
blocked
90
Bilateral oviduct
blocked
0.3g phenol red + 4.2 g of
anhydrous sodium
bicarbonate + 100 ml of
distilled Water (pH- 5.8-8.4)Infusion 20-50 ml of PSP dye in uterus
using catheter.
Urine samples are collected at
5,10,15,30,45,60,75, and 90 minutes
Alkalinized by adding a drops of
1M NaOH
Red colour
(positive response)

34. Uterine infection commonly occurs in repeat breeder cow falls under
the definition subclinical endometritis.
1. Treatment for uterine infection
Treatments
of RBS
Control
uterine
infection
Correcting
ovulatory
disturbance
Supplementing
for luteal
insufficiency
Improving
management
34
TREATMENTS

35. A) Intra-uterine infusion of 1%
Lugol's iodine
Role of iodine in RBS
 Initiation of follicular wave dynamic
 Release of best quality mature oocytes
 Enhance the healing and restore the
activities of endometrium and ovaries
 Improvement rate of service per
conception
Lugol’s iodine has a broad spectrum
bactericidal, fungicidal and anti
protozoa effect
(Sarkar, 2006)
(10% Potassium iodide,5% iodine and
85% D.W.)
(Sheldon et al., 2006)
35
Potent bactericidal effects
Adjust the pH of uterus
Hyperemia of the endometrium
Increase the defense mechanism
Lugol's iodine

36. B) Treatment with parental antimicrobials
Zarekar et al. (2019) conducted experiment on 36 crossbred repeat breeder cow
and concluded as follow:
Twenty-four infectious repeat breeder cows were treated with Enrofloxacin long
acting @ 7.5 mg/kg b. wt. and Metronidazole @ 15 mg/kg b. wt. by parental route on
first to fourth day of oestrus whereas twelve (12) cows were kept as control.
Groups Treatment Insemination Conception%
Group –I
(n=12)
Enrofloxacin +
Metronidazole
Subsequent
estrus
91.67%
Group-II
(n=12)
Enrofloxacin +
Metronidazole
Same oestrus 83.33%
Group –
III (n=12)
Kept control Same oestrus 00.00%
36

37. 0%
10%
20%
30%
40%
50%
Genta Enro Cepha Control
42%
50%
42%
25%
29%
33%
43%
22%
20%
25%
50%
29%
1st service 2nd service 3rd service
C) Treatment with intra- uterine antibiotics
Parikh et al., (2017) conducted an experiment on 48 repeat breeder Gir cows by using
different intra uterine antibiotics and concluded following observations.
Groups Treatment
(Intrauterine)
Conception
(overall)%
Group –I
(n=12)
Gentamicin (100
mg)
67%
Group –II
(n=12)
Enrofloxacin
( Enrogyl-35mg)
75%
Group –
III (n=12)
Cephalexin
(Lixen- 4g)
83%
Group –
IV (n=12)
Control 58%
37
Percentage
Antibiotics
Treatment given on the day of estrus and
service provided in Subsequent estrus

38. Conducted experiment using 30 ml Hydro-alcoholic / 30 ml hydro-acetonic extract of Neem
bark for intra uterine treatment of endometritis from 0 to 7 days of estrus and service given
subsequent standing estrus.
38
Groups
(n=8)
Treatment Conception
%
Group A 30 ml Hydro-
alcoholic
75%
Group B 30 ml hydro-
acetonic
50%
Group C 30 ml normal
saline
62.50%
D) Herbal therapy for endometritis
1 Using Neem extract
Neem has a potent immuno-modulatory action
and effectively use for treatment of endometritis
in RB cow.
(Kumar et al., 2013)

39. 39
Using Garlic, Turmeric,
Ashwagandha
GROUPS TREATMENT (I/U) C.R(%)
A 30 ml Normal saline 0.0(0)
B 30 ml Levofloxacin 83.33(5)
C 30 ml Ashwagandha extract 33.3(2)
D 30 ml Garlic extract 50(3)
E 30 ml Turmeric extract 50(3)
F 30 ml (Ashwagandha +
Garlic +Turmeric) extract
66.6(4)
 Treatment given on day of estrus
 Service provide on next estrus
 Each group contains 6 animals
Garlic extract
Release of cytokines
(IL-2, IFN-α & IFN-γ )
Natural killer activity , phagocytic
activity by peritoneal macrophages
Turmeric (Curcumin)
Immunomodulators
Activation T cell, B cells, macrophages,
Neutrophils, N.K cells, and dendritic cells
(Rawat et al., 2006)
(Jagetia and Agarwal, 2007)
Combination of different herbal drug are
more effective due to synergic effect
(Kumar et al.,2018)
2
Conception rate(C. R)

40. E) Treatment using immunomodulators
Alternative therapy for antibiotics.
Intra uterine infusion of immunomodulators such
 E.coli lipopolysaccharides (endotoxin)
 Oyster glycogen
 Plasma or hyper immune serum
 Leukotriene B4
(Hussain, 1992)
PMNs,
Blood monocytes,
macrophages
Chemo
attractant
Phagocytosis
(Watson et al., 1990)
40
F) Treatment using Enzyme
and antioxidant
 Trypsin
 Chymotrypsin
 Papain
 Ascorbic acid
 Vitamin E
(Guney et al., 2007)

41. 2. Correction of ovarian dysfunctions
Anovulation
Delayed ovulation
Groups Treatment Pregnancy
Rate(%)
Group 1 Single AI (112/353)
32.1%
Group 2 Single AI + 100 μg
GnRH at AI
(165/406)
41.6%
Group 3 Double AI (2nd AI 12
to 16 hrs. after the
first AI )
(119/364)
33.5%
Group 4 Double AI plus
100 μg of GnRH at AI
(135/359)
37.5%GnRH with single AI increase
pregnancy rate of RB cow compare
to other treatment protocol.
41
A) Treatment using
GnRH
GnRH analogue
Buserelin acetate
(Stevenson et al.,1990)

42. Kharche and Srivastava, (2007) investigated the effect GnRH on RB cows using
different doses of GnRH analogue ( Buserelin acetate) given at a time AI.
Pregnancy rate increase with GnRH treatment
The higher dose of GnRH increased first service conception as well as overall pregnancy rate
0
50
100
20 μg 10 μg N. S
Conception%
Dose
42
B) GnRH treatment using different doses
Overall conception rate
Groups Treatment
Conception rate %
1st
service
2nd
service
3rd
service
1(n =55) 20 μg 45% 22% 20%
2(n =40) 10 μg 25% 20% 13%
3(n =42) Normal
saline
17% 10% 21%

43. Group 1: AI during natural estrus.
Group:2: Ovsynch protocol
C) Treatment using Ovsynch protocol
TAI
day day day
10 μg
GnRH
500 μg
PGF2α
10 μg
GnRH
10
Conception rate: control-29.41%, ovsynch-58.82%
Ovsynch increase conception
 Prevent delayed ovulation,
Anovulation
 Induction of luteolysis
 Fixed time AI
(Keskin et al., 2010)
(Santos et al., 2004) 43
If protocol start in mid diestrus (5-12th ) increase
conception due to ovulation of 1st wave.
9th7th
0

44. 44
0 7thday day
10 μg
GnRH
25 mg
PGF2α
0
7th 9th
day day
10 μg
GnRH
25 mg
PGF2α
10 μg
GnRH
12th day
5 day Estrus
detection
And
insemination
D) Select synch
day
Timed AI After GnRH
E) Cosynch

45. F) Treatment using progesterone
21 day estrus
cycle
Estrus
0
10
Ear implant
19
Inj. SMB
2 ml(i/m)
48 hrs after removing
ear implant
(subsequent estrus)
AI
Removeearimplant
0.98 mg PGF2α
Group 2
( Selvaraju and Veerapandian, 2010) 45
Groups 10th day 19th day
Group 1
(n=6)
Ear implant Remove ear implant
SMB -
Group 2
(n=6)
Ear implant Remove ear implant
SMB 0.98 mg PGF2α (i/m)
Group 3
(n=4)
No treatment regular AI
SMB(Synchromate B) (5 mg Oestradiol
valearte+3 mg Norgestomate)
Ear implant - 6 mg Norgestomate
Conception rate
Group1 43.75%
Group 2 37.50 %
control 18.75%

46. 46
Selvaraju et al.,(2002) studied the influence of insulin treatment on conception rate.
Groups Treatment 12th day
8th ,9th ,10th day after
estrus
Group 1
(n=11)
Bovine insulin at 0.2
IU/kg BW (s/c)
0.75 mg Tiaprost
(PGF2α)
Group 2
(n=10)
control
0.75 mg Tiaprost
(PGF2α)
Services were given Post-treatment and recorded
following observations :
Group 1st service 2nd service overall
Group 1 5/11(45.4%) 2/6(33.3%) 7/11(63.6%)
Group 2 2/10(20%) 2/8(25%) 4/10(40%)
G) Insulin treatment
Inj. Hypurin bovine insulin
100 IU/ml
(Dose 0.2 IU/kg body weight)

47. 3. Treatments for luteal insufficiency
(Kimura et al., 1987)
47
(Mann et al., 2001)
Early embryonic death
Insufficient P4 production (luteal phase)
Disturbance in luteotropic hormone
Luteinizing hormone (LH) stimulates c AMP
and protein kinase A (PKA) to activate proteins
that will supply cholesterol for progesterone
synthesis.
LH- Work as a Luteotropic hormone
P4
Increase
Endometrial
protein +
Growth factor
Enhance
secretion
antiluteolytic
IF-tau
Maintain
pregnancy

48. 21 day estrus
cycle0
21
AI
5-6th
day Group:1
GnRH (20 μg)
Group:2
1500IU(hCG)
Group:3
0-10 Day
(CIDR)
Treatment Pregnancy
rate
GnRH
(n=26)(Group1)
29.9%
hCG(Group2)
(n=25)
60%
CIDR(Group 3)
(n=25)
56.0%
Control (n=27) 29.6%
10
( Khoramian et al., 2011) 48
CIDR
remove
A) Treatment using hCG
Post insemination treatment with hCG or CIDR during the very early stages of pregnancy
improve conception rate in repeat-breeder cows
Low dose
Progesterone
CIDR

49. B) Treatment with hCG at different days after AI
Rajamahendran (1992) treated 34 HF cows with four treatments of hCG(1000 IU)
Post-insemination on day 0(n=8),7(n=9),14(n=9) and control(n=8)
 Rate of formation of accessory corpora lutea and
diameter of CL among cow treated on day 0,7,14
higher than control.
Treatment Pregnant
cow
Conception
rate
0 day 4/8 50.0%
7th day 7/9 77.7%
14th day 4/9 44%
control 3/8 37.5%
Treatment on 7th
day of AI with
hCG reduce
incidence of
early embryonic
death.
49
Inj. Chorulon (hCG)

50. C) Progesterone injection(500 mg)
i/m on day 5th post AI
(Devanathan et al., 1999)
D) Progestagen vaginal implant
from day 5th -12th of AI.
( Villarroel et al., 2004)
E) Chlormadinone daily oral feeding
(10 mg) from day 5th -12th of AI.
(Jochle et al.,1976) 50
C) Treatment using progesterone
supplements

51. 4. Management strategies
Nutritional management
Improve timing and techniques of AI
Control periparturient disease
Reducing stress
A) Improving nutritional
imbalance
Reduce
 Blood glucose
 Insulin
 IGF-1
 Low LH pulse
frequency
Poor nutrition during
dry period, prepartum
and post partum
Increase
 B-hydroxy
butyrate
 Non-esterified FA
 Negative energy
balance
Key point of feeding :
 Supplementation of carbohydrate,
vitamins and mineral
 Dry cow feed with low energy high
pefibre diet feeding
 Dietary anion and cations balance
 Feeding less than 10% rumen
degradable protein. More RDP rise
BUN which impair embryo survival
(Purohit, 2008)
51

52. B) Improve timing and techniques of AI
Treatment for Failure of estrus
detection
 Use Multiple insemination or
ovulation treatment protocol
 Use of heat detection aids (Vaginal
electric resistance, Pedometers,
KaMaR)
Key point of AI
 AI Done in AM/ PM procedure
 Repeated AI after 12 hrs. interval
 Site of insemination for cattle mid –cervix
(Hunte and Wilmut,1983)
 Proper storage of frozen/liquid semen straw
in LN2 at -196 °C with all safety precautions
 Maintain Hygienic condition during AI
 Proper training of AI for inseminator
(Boettcher et al., 2007).
Vaginal electric resistance
 Pedometers
52
KAMAR

53. C) Avoid periparturient diseases
 Mastitis
 Retained placenta
 Hypocalcaemia
 Ketosis
 Acidosis
 Dystocia
 Endometritis
}
 Feeding anionic salts in combination with calcium
and magnesium (Wilde, 2006)
 Feeding high pefibre low energy chopped diet during
dry period (Beever, 2006)
 Maintain hygiene condition on animal shed
 Avoid manually removing of placentas without
sanitary measure
100 ml daily oral feeding
(Post calving feeding)
53
VETBOLITE
anionic Salts With
Vitamin- D3, E & H
(Pre-calving supplement)

54. D) Reducing stress
 Provide sufficient space for each
animal
 Cooling of cow during hot summer
 Balance feeding to high yielders
 Regular deworming and vaccination
Upper limit of the thermo neutral zone in cattle -
70-80°F
54
Fogger system

55. Repeat Breeding Syndrome ( RBS ) is an unclear and multi- etiological reproductive
disorder of dairy cattle
The key players of this multifactorial syndrome are cow, bull, environment and
management which are overlapped and difficult to determine root cause
Systematic diagnostic modalities and potential therapeutic approach are the main arms to
combat and cop up the RBS
Adaption of Integrated Livestock management practices, strategies and breeding techniques
has positive and qualitative impact on reduction of RBS
Failure to detect estrus is the major constrain in the field for getting optimum
reproductive performance and subsequent leads to RBS
55
CONCLUSIONS

56. Apart from conventional guidelines, thorough clinical examination of ovary and uterus
by advanced technique like USG is the most effective tool for diagnosis of RBS
Intrauterine treatment with Lugol's iodine and combination of anti-microbial agents
or Herbal therapy boost up the defensive power and improve the overall
reproductive health parameters
Customization of the estrus synchronization protocol improves the efficiency of fixed
time AI, synchronizes ovulation , overcome the problem of heat detection and ovarian
dysfunction
Post- insemination treatment with hCG / GnRH / CIDR reduce the incidences of luteal
insufficiency
Balance ration during the periparturient period, minimizing stress factors and
improving overall reproductive hygiene to obtain high-fertility and reduce the chances
of RBS
56

57. 57
Future prospects
Stem cell based therapy can be a potential therapeutic approach to a wider array of
reproductive problems in animals for endometrial damage, vaginal atrophy and
infertility in near future.
Gene therapy can be used to eliminate genes which are harmful and cause
reproductive problems in dairy animals.
Nano biosensors integrated with nanomaterial's such as carbon nanotubes,
nanowires and Nano fibers need validation for the detection of pathogens, oestrus,
hormone level and metabolites in animals for reproductive management.

58. 58