"Embryonic mortality in cattle: endocrine

and infectious determinants, economical

impact and therapeutic options".

Monika Ptaszynska, DVM, PhD

Intervet International
The Netherlands
 Contents

Definitions
Occurrence and economic impact
Physiological conditions for pregnancy recognition and
maintenance in cattle
Contributing factors
- suspected non-infectious causes
- most relevant infectious causes
Therapeutic directions
Conclusions
Pregnancy losses in cattle -
definitions
Embryonic mortality loss of pregnancy during
embryonic phase fertilization 42d

EEM early embryonic LEM- late embryonic

mortality mortality
- death of embryo until - death of embryo after
15-16d post insemination 15-16d post insemination

- before maternal - maternal recognition of

recognition of pregnancy pregnancy have taken
have taken place place

- short or normal luteal - prolonged luteal phase

phase no change in the return later than 25d post
length of cycle AI
 Fetal mortality loss of pregnancy during fetal
phase 42d to calving

Here we usually already are talking about

abortion:
- early abortion first trimester
- mid term second trimester
- late abortion second/last trimester

Stillbirth birth of dead fetus

(usually at a physiological time)
 Occurrence and economic impact
Humblot (2001) showed the following frequency of pregnancy
losses in cattle:

- Fertilization failure and early embryonic death: 20-45%

- Late embryonic/fetal loss: 8-17.5%
- Late abortion: 1-4% represent
 Economic consequences of early
embryonic losses
Prolonged (inter)calving interval = increased number of
days open
Difficult planning of reproductive season and milk
production
Increased service cost
Premature culling of genetically valuable cows

Rarely such losses are directly evaluated and defined but can
be considerable:
Je-In Lee and Ill-Hwa Kim. J Vet Sci 2007;8:283-288
 Embryonic mortality what is it really that
we are talking about?
 The embryonic journey
D0-d42 Embryonic phase

d0 oviduct d6 Uterus pre-recognition d8-12 Uterus post recognition d42

Fertilization Transfer Pregnancy Finished

to the uterus recognition organogensis

Most of the losses occur when:

- Embryo actually dies before signaling takes place (incorrect fertilization process, poor embryo
development) and luteolysis occurs as if no fertilization did not take place
- Embryos production of INT t is delayed or inadequate and luteolysis starts or is triggered by
some luteotoxic factor and this causes the death of embryo
 The most important factors deciding of pregnancy
maintenance during the first weeks

Adequate luteal function

that guarantees sufficient
progesterone levels
Progesterone
stimulates growth of
the embryo and
synthesis of INT t

Uterine environment supportive

Adequate and for embryonic development and
INT t inhibits
early production of luteolytic cascade maternal recognition
INT t
 Most important non-infectious factors proposed
as causes of early embryonic mortality
Factors leading to low
quality of oocyte or
incorrect fertilisation
Factors leading to
insufficient luteal function
and precocious
luteolysis
Factors impairing embryo
development and INT t
production
Inadequate LH support for
growing DF
Inadequate ovulatory LH
stimulation
Inadequate LH support
Direct luteotoxic effect of
induction of luteolysis
Inadequate support for
development
Direct harmful effect on the
embryo
Nutritional deficiencies (negative energy
balance)
High ambient temperature (heat stress)
Delayed ovulation and follicular persistency
Nutritional deficiencies (negative energy
balance)
High ambient temperature (heat stress)
Inflammation processes, fever
Phytotoxins (esp. mycotoxins) in the feed
Inadequate progesterone levels
Suboptimal condition of endometrium
High ambient temperature (heat stress)
Inflammation processes, fever
 Main infectious factors proposed as causes
of early embryonic mortality

Factors that impair follicular
growth and quality of the
oocyte
Factors that impair luteal
function
Indirectly through impaired LH
support
Directly through inflammation of
ovarian tissues
Indirectly through impaired LH
support or activation of
precocious luteolysis
Directly through inflammation of
ovarian tissues
BVD, IBR, generalised
infections, H. somnus (?)
BVD, IBR
BVD, IBR, mastitis,
endometritis, generalised
infections with fever
BVD, IBR

Factors directly damaging embryo or affecting placental BVD, IBR, Neospora

formation caninum (?), Trichomonas
foetus (?), Leptospira (?),
Campylobacter foetus
 EEM causative factors
The science is focusing at present on three levels:
Oocyte quality= factors
that in fact impair growth,
maturation and ovulation of
the DF
Inadequate LH support
during growth =poor quality
follicle (NEB, heat stress)
Inadequate LH
preovulatory peak = delayed
ovulation (NEB, heat stress)
Factors directly affecting
ovaries (BVD IBR)
Embryo quality = all
factors that either
decreased quality of the CL quality = all factors that
oocyte or directly damage lead to low P4 production: either
embryo creation of less efficient CL,
metabolism of steroids,
Inadequate LH support luteolysis
during follicular phase
Inadequate LH support
Inadequate P4 levels during CL formation
before recognition (high
metabolic rate) High metabolism of steroids
Direct influence of heat Inadequate embryo signaling
stress, uterine Luteotoxic factors (bacterial
environment, toxins, toxins, PGF)
infectious factors
 EEM causative factors
The science is focusing at present on three levels:
Oocyte quality= factors
that in fact impair growth,
maturation and ovulation of
the DF
Inadequate LH support
during growth =poor quality
follicle (NEB, heat stress)
Inadequate LH
preovulatory peak = delayed
ovulation (NEB, heat stress)
Factors directly affecting
ovaries (BVD IBR)
And proposes for the practice:
- The nutritional side plays the most important role
therefore the first step is to ensure adequate
nutrition level to avoid NEB and inhibition of
GnRH production
- As heat stress causes delayed ovulation -
minimize the influence of high ambient
temperature: fanning, spraying
- As the negative influence of BVDV and IBRV on
the ovarian function is confirmed ensure
protection from the infection through
vaccination that prevents free cell viraemia
- Provide protected follicular growth and
ovulation through application of Ovsynch-type
protocols or induce ovulation with GnRH at AI
Estimate of pregnancy rate as duration of dominance of the pre-ovulatory
follicle increases, established when a logistic model was fitted from four
different experiments (After Diskin at al., 2002)

100
90
80
70
60
50
40
30
20
10
0
1 2 3 4 5 6 7 8 9 10 11 12
 Why manage estrus with Ovsynch type
protocols?

Ovsynch type protocols provide additional GnRH/LH

support of follicular growth and induce ovulation:

- higher chance for a development of healthy DF

- elimination of delayed ovulation and follicular persistency

Higher chance for ovulation of an oocyte with high

developmental potential and for a correct fertilization
process
 What to chose?
Two aspects to be taken under consideration:
- feasibility
- luteotrophic ability
GnRH PGF GnRH
Recepta/Fertagyl 2.5ml Prosolvin/Cyclix Receptal/Fertagyl 2.5ml
Classical
7 days 48h 17-24 h Fixed time AI

Day 0 Day 7 Day 9

GnRH PGF GnRH

Easy Receptal/Fertagyl 2.5ml Prosolvin/Cyclix Receptal/Fertagyl 2.5ml

7 days 48h Fixed time AI

Day 0 Day 7
Day 9

New
GnRH PGF hCG
Receptal/Fertagyl 2.5ml Prosolvin/Cyclix Chorulon 1.500iu

7 days 48h 17-24 h Fixed time AI

Day 0 Day 7 Day 9

 Ovulation induction with GnRH

Well established and practiced

Usually gives significant improvement in herds with AI

performed at detected heat and where prolonged heat
signs are observed

In the meta-analysis of Morgan and Lean (1994) there was

a significant increase in pregnancy risk in cows treated
with GnRH analogue at first insemination post partum, at
second service after calving and in repeated breeder cows
treated with GnRH at the time of insemination

Although well known, still new data are generated but

targeted at heat stress
The effects of GnRH treatment at the time of AI and 12 days later on
reproductive performance of high producing dairy cows during the
warm season in northeastern Spain.
Lopez-Gatius et al., Theriogenology 2006;65:820830

Lactating HF cows, May-September

100mcg gonadorelin
GnRH-d0
GnRH-d0+d12
Control no treatment

Treatment group Conception rate

Control 20.6%

GnRH-d0 30.8%

GnRH-d0+d12 35.4%
Dose dependent effect of GnRH analogue on pregnancy
rate of repeat breeder crossbred cows.
S.D. Kharche and S.K. Srivastava. Anim Reprod Sci 2007;99:196201

137 crossbred dairy cows with a history of repeat breeding

Group 1- 20mcg buserelin (Receptal) at the time of AI
Group 2 10mcg buserelin (Receptal) at the time of AI
Group 3 physiological saline +AI
 EEM causative factors

The science is focusing at present on three levels:

Embryo quality = all
factors that either
decreased quality of the
oocyte or directly damage
embryo
Inadequate LH support
during follicular phase
Inadequate P4 levels
before recognition (high
metabolic rate)
Direct influence of heat
stress, uterine
environment, toxins,
infectious factors
And proposes for the practice:
- The nutritional side plays the most important role
therefore the first step is to ensure adequate
nutrition level to avoid NEB and inhibition of
GnRH production
- As heat stress causes delayed ovulation and has
a direct negative effect on early embryos -
minimize the influence of high ambient
temperature: fanning, spraying
- We can do really little to directly influence the
development of the embryo and efficacy of INT t
production = we must give the embryo the best
environment = healthy uterus and high P4 levels
 Healthy uterus what would that
mean?
Proper feeding promotes high immune competence of
endometrium making it more resistant to infections (works
by Zerbe and Sheldon)

Early detection and treatment of acute endometritis

prevents the development of chronic conditions around the
breeding time

Properly balanced protein/energy ratio prevents excessive

production of ammonia and unfavorable changes of uterine
environment
 Chances of the embryo to survive can be
markedly decreased by excessive ammonia
levels
McNeill et all.(2006)
Day 4 post ovulation Day 5 post ovulation

1 1

Probability of embryo
Probability of embryo

0.8 0.8
survival

survival
0.6 0.6
0.4 0.4
0.2 0.2
0 0
2 4 6 8 10
2 4 6 8 10 12
Progesterone in m ilk (ng/m l)
Progesterone in m ilk (ng/m l)

Day 6 post ovulation

1
Probability of embryo

0.8
survival

0.6
0.4

0.2

0
2 4 6 8 10 12 14 16
Progesterone in m ilk (ng/m l)
 We focus more and more on
progesterone and luteal function

The science is focusing at present on three levels:

CL quality = all factors that
lead to low P4 production: either
creation of less efficient CL,
metabolism of steroids,
luteolysis
Inadequate LH support
during CL formation
High metabolism of steroids
Inadequate embryo signaling
Luteotoxic factors (bacterial
toxins, PGF)
And proposes for the practice:
- Again nutrition is the key and the first step
- Direct stimulation of early luteal function and
formation of additional CLs through administration of
hCG at 4-6d post AI increase in P4 levels
- Indirect luteotrophic support prevention of
precocious luteolysis and induction of accessory CLs
with administration of GnRH at 10-12d post AI
 Improvement of fertility with hCG
at 4-6d post AI
hCG is increasingly used for improvement of
pregnancy rates in dairy cattle

It is believed to give better results than GnRH,

especially in case of repeat breeders who return
to heat >25 days post AI

New data generated and confirming its value

(Santo et al., 2001; Nishigai et al., 2002; Stevenson et al., 2007)
 Why?

It seems that hCG thanks to its longer half life:

1. Induces secondary CLs of higher secretory

capacity
2. Can directly stimulate the secretory function
and development of CL verum
 Why?

Schmitt et al., J. Anim. Sci. 1996. 74:10741083

Responses to injection of hCG at d 3 support the concept that LH-

like exposure early in the development of the CL may stimulate CL
development.
Therefore, a d 5 injection of 3,000 IU of hCG may induce
modifications of the original CL and contribute to a greater
development of the induced CL.

Perhaps the extended half-life of hCG may improve maturation of

the d 5 dominant follicle and formation of a CL with a greater
steroidogenic capacity.
 Improvement of fertility with GnRH
post AI

Also well established

Extensively studied and meta-analysed by

Peters (2000) who suggested that in certain
circumstances it may produce significant benefits

Recent publications confirmed both possible

benefits and herd variability of response
(Sterry et al., (J Dairy Sci 2006); Franco et al., (Thriogenology
2006); Howard et al., (J Anim Sci 2006); Lopez Gatius et al.,
(Thriogenology 2006); Stevenson et al., (J Dairy Sci 2007)
 Cows with low/delayed early progesterone rise show lower
embryonic survival

Mean milk progesterone concentration in cows after AI

(Mann and Lamming 1999)
25

concentrations (ng/ml)
Milk progesterone
20

15
pregnant
10 non pregnat

0
d0 d3 d6 d9 d12 d15 d18 d21 d24 d27 d30
Days after AI
hCG GnRH

Early repeats Late repeats

(35-45d post AI)
(15-25d post AI)
(np. Receptal, 2.5ml)
(np. Chorulon, 1.500jm)
 Early embryonic losses in cattle
therapeutic possibilities

Progesterone?

Not really so far we do not know the dose and best timing!

Author Dose Administration Timing Results

route. post AI
Walton et al (1990) 200mg inj 5-11d -
1g IVD 5-12d Increase P4
Thuemmel et al (1992) 75mg/d. Inj. 6-10d Tend. +

Mann et al (2001) 0.95g IVD 10-17d -

Villarroel et al (2004) 1.55g IVD 5d -, repeat br.+

Lopez-Gatius et al (2004) 1.55g IVD 36-42d Tend +

Hanlon et al (2005) 1.56g IVD 4-7d -

Mann et al (2005) 0.95g IVD 5-9d +

0.95g IVD 12-16d -

Early embryonic losses in cattle therapeutic
possibilities

NSAIDs e.g. flunixin meglumine?

Theoretically yes, if we suspects premature release of PGF

to be the reason for the pregnancy loss, as they block
prostaglandin release.

So far the results are quite inconsistent.

FM given on 15th
and 16th day post AI
 The therapeutic options

Pharmacological measures exist without previous attempts

to address nutritional and managerial inadequacies

There is no single system that would always be effective

and solve all the problems EEM is a multifactorial
problem

In many cases a solution on measure have to be

developed

In fact, often the therapy is based on historical data and

prevention is rarely possible (with the exception of heat
stress)
 To summarize .. what to do in
practice?
Induce ovulation with GnRH in a correct dose
alone or within the Ovsynch protocol:
- when prolonged oestrus and persistency of DF are
observed/suspected
- in dairy herds during summer months
Support early LC with hCG at 4-6d post AI:
- in highly producing dairy cows
- when early repeat are observed
Induce accessory CLs at 10-12d post AI with GnRH
- when late repeats are observed
 Summary

Decreased fertility rates in cattle can be caused by

numerous factors including nutrition, production status, heat
stress and infectious diseases affecting directly or indirectly
follicular and luteal function as well as embryonic
developmental potential

Pharmacological induction of timely ovulation and support of

luteal function can be used to improve the pregnancy rates

Any pharmacological treatment adopted should be

proceeded by adequate corrections of feeding and
managerial conditions