AÑO 1, NÚMERO 12, AGOSTO 2008

LA TASA DE PREÑEZ
Existen diferentes parámetros para evaluar la fertilidad en el hato lechero y cada parámetro
permite identificar problemas diferentes. Así, el porcentaje de concepción se refiere a la
proporción de vacas gestantes del total inseminado, mientras que la tasa de preñez
representa la proporción de vacas que gestan del total elegible para que presenten estro y
que pudieran ser inseminadas durante un periodo equivalente a un ciclo estral (21 días). El porcentaje
de concepción permite identificar problemas relacionados con el momento del servicio, la técnica de
inseminación y los factores asociados con la muerte embrionaria temprana. La tasa de preñez es un
parámetro resultante de dos aspectos: la eficiencia en la detección de estros y el porcentaje de
concepción. La tasa de preñez se calcula multiplicando la eficiencia en la detección de estros por el
porcentaje de concepción, dividido entre 100. De tal modo que, en un hato con una eficiencia en la
detección de estros de 50% y con un porcentaje de concepción de 30, se obtiene una tasa de preñez de
15%. Este número indica que, de las vacas elegibles para que muestren estro y sean inseminadas en un
periodo de 21 días, sólo 15% queda gestante. La tasa de preñez permite identificar problemas
relacionados con la falla en la concepción y aquellos asociados con la eficiencia en la detección de estros.
La mayor aspiración de un productor o del veterinario es lograr que la tasa de preñez sea igual al
porcentaje de concepción, esto indicaría que de las vacas elegibles para ser detectadas en estro todas
son inseminadas (100% de eficiencia en la detección de estros). En los hatos lecheros nacionales, una
meta posible de alcanzar en la tasa de preñez es de 20%, cabe destacar que en los hatos de Estados
Unidos la tasa de preñez promedio alcanzada es de 14%. Dado que, el aumento en la fertilidad global
del hato mediante un incremento en el porcentaje de concepción, es realmente difícil de lograr, una
posibilidad para mejorar la fertilidad ―probablemente la única― es mediante el aumento de la tasa de
preñez. Es decir, con la misma proporción de vacas gestantes, después de cada servicio podemos
aumentar el número de vacas gestantes por mes, aumentando únicamente el número de vacas
inseminadas.

Dr. Joel Hernández Cerón y M.C. Álvaro Ortega León

Compiladores
Departamento de Reproducción

Resúmenes
Theriogenology 70 (2008) 682–691
Sperm Morphology and Fertility of Progeny-Tested AI
Dairy Bulls in Sweden
A. Al-Makhzoomia, N. Lundeheimb, M. Håårdc, H. Rodríguez-
a
Martíneza
Division of Reproduction, Department of Clinical Sciences, Faculty of
Veterinary Medicine and Animal Science, Swedish University of
Agricultural Sciences (SLU), Uppsala, Sweden. bDepartment of Animal
Breeding and Genetics, Faculty of Veterinary Medicine and Animal
Science, Swedish University of Agricultural Sciences (SLU), Uppsala,
Sweden. cSvensk Avel ek för, Örnsro, 532 94 Skara, Sweden.
Use of bull semen with high levels of sperm
abnormalities, reflecting genital dysfunction, is not
recommended for artificial insemination (AI) since
it would most likely lead to subfertility. Sperm
quality, including sperm morphology, may
deteriorate with increasing age of the bull thus
becoming a source of concern when using older,
progeny-tested AI bull sires. Although a
relationship between sperm morphology and
fertility after AI in progeny-tested bull sires has
been reported, it is yet unclear which sperm
abnormalities are most critical. This constituted
the core aim of a 22-month long retrospective
study in proven (aged 60–84 months at the start
of the study) AI sires of the Swedish Red (SR, n =
8) and Swedish Holstein (SLB, n = 4) breeds
where their semen (107 freezing batches in total,
built by a single ejaculate (n = 3) or pooling two
consecutive ejaculates (n = 104) collected at 1–3
months interval), were subjected to detailed
morphological examinations on wet- and dry,
stained smears. Attention was paid to between
and within-bull variations with regard to presence
and level of sperm abnormalities. Sperm
morphology differed significantly between sires
and ejaculates, with 6/12 sires having ejaculates
containing >10% of morphologically deviating
sperm head shapes, a commonly used threshold
for young AI bulls in Sweden. However, with the
exception of pear-shaped or narrow-at-the-base
anomalies, the mean values for individual defects
were always within the limits expected for a
normal bull sire, and were therefore considered
acceptable. The percentage of morphologically
normal spermatozoa was positively related to
fertility, whose output differed significantly among
bulls. Among sperm abnormalities, the proportion
of morphologically deviating sperm head shapes
were negatively correlated with fertility, pear-
Noticias de Reproducción Bovina AGOSTO 2008
shaped sperm heads in particular. In conclusion,
the relationship between sperm morphology and
fertility after AI calls for frequent (2–3 months
interval) detailed assessments of sperm
morphology in AI stud bull sires.
heriberto.rodriguez@kv.slu.se
Animal Reproduction Science 107 (2008) 36–47
Effect of Progesterone (P4) Intravaginal Device (CIDR)
to Reduce Embryonic Loss and to Synchronize Return
to Oestrus of Previously Timed Inseminated Lactating
Dairy Cows
Mufeed A. Alnimer, Wadie F. Lubbadeh
Department of Animal Production, Faculty of Agriculture, University
of Jordan, Amman 11942, Jordan.
A study on postpartum dairy cows was done to
evaluate the effect of using progesterone
intravaginal device (CIDR) to synchronize return
to oestrus of previously timed inseminated (TAI)
cows and to evaluate embryo survival and
pregnancy rate in the return to oestrus cows. All
cows were subjected to TAI program. On Day 14
after AI, cows were assigned randomly into two
groups: treated group (CTAI; n = 126) and
control group (TAI; n = 172). Every cow in the
CTAI group received CIDR device for 7 days.
Cows were observed for oestrus after CIDR
removal in the first period (Days 21–28). Cows,
which did not show oestrus, were retreated with
CIDR on Day 28 for 7 days and they were
observed for oestrus after CIDR removal in the
second period (Days 35–42). Pregnancy was
diagnosed on Days 28, 45 and 90 after AI. In the
first period, more cows in the CTAI group showed
oestrus within 2.4 days after CIDR removal
compared to cows within 4 days in the control. In
the second period, more cows in the CTAI group
showed oestrus within 4.5 days compared to cows
within 3.7 days in the control. Pregnancy rates on
Day 28 were not different between both groups.
Pregnancy rates on Days 45 and 90 were higher
for cows in the CTAI (42.1±5.3%, P = 0.028 and
38.9±5.2%, P = 0.020) compared to that in the
TAI group (35.5±3.9% and 31.9±3.8%).
Moreover, pregnancy rates were lower for
multiparous cows (34.2±4.1, P = 0.017 and
30.2±3.9%, P = 0.008) compared to primiparous
cows (42.7±5.2% and 39.9±5.1%) on Days 45
2

and 90, respectively. Pregnancy losses between
Days 28–45 and 45–90 tended to be lower for the
CTAI group and primiparous cows compared to
the TAI group and multiparous cows. Pregnancy
rates of first (P = 0.020), second, third and up to
fourth AI were higher (P = 0.001) for the CTAI
group (38.9±0.05%, 78.6±0.05%, 92.1±0.04%,
93.6±3.71%) compared to that for the TAI group
(31.9±0.03%, 54.1±0.04%, 68.0±0.03%,
81.4±2.75%, respectively). Days open for
pregnant cows were similar in both groups, but
lower (P = 0.025) for primiparous cows (81.6±4
day) compared to multiparous cows (93.8±3 day).
Number of services for pregnant cows was lower
(P = 0.002) for the CTAI group (1.70±0.10)
compared to that for the TAI group (2.14±0.09),
whereas it was similar for primiparous and
multiparous cows. Results indicate that the CIDR
device improved synchronization to return to
oestrus and increased pregnancy rate to first,
second, third and fourth AI by reducing embryonic
and fetal losses.
Animal Reproduction Science 107 (2008) 48–61
Kinetics of Fertilization and Development, and Sex Ratio
of Bovine Embryos Produced Using the Semen of
Different Bulls
M. Alomara,b, H. Tasiauxa, S. Remaclea, F. Georgea, D. Paula, I.
a
Donnaya
UCL, Institut des Sciences de la Vie, Unité des Sciences vétérinaires,
Place Croix du sud 5 Boîte 10, B-1348 Louvain-la-Neuve, Belgium.
b
Division of Animal Production, Department of Agriculture, Atomic
Energy Commission, P.B. Box 6091, Damascus, Syria.
The between bulls variation in in vitro fertility and
the shift of sex ratio towards male embryos are
two problems affecting the in vitro production
(IVP) of bovine embryos. Our objective was to
evaluate the kinetics of fertilization, embryo
development and the sex ratio of the resulting
embryos using the frozen/thawed semen of four
different bulls. In a first experiment, the kinetics
of pronucleus (PN) formation was evaluated at 8,
12 and 18 h post-insemination (hpi). Based upon
the pronuclei sizes and the distance between the
two pronuclei, inseminated oocytes were classified
in three PN stages. Differences between bulls
were observed at each time point, but were more
important at 12 hpi. At 8 and 12 hpi bull III
showed a significantly faster PN evolution by
comparison with the three other bulls (P < 0.05),
while at 18 hpi, the proportion of the three PN
stages was similar to those of bulls I and IV, bull
II being delayed. In a second experiment, the
Noticias de Reproducción Bovina AGOSTO 2008
kinetics of in vitro embryo development was
compared using time-lapse cinematography. The
analysis of embryos reaching the blastocyst stage
revealed significant differences in the mean time
of first cleavage (range of 22.7–25.6 h, P < 0.05),
while the lengths of the subsequent three cell
cycles did not differ between bulls. The early
mean time of first cleavage with bull III was
associated with an early blastulation and a high
blastocyst rate at Day 7, in opposition to what
was observed with bull II showing a later timing
of first cleavage (first cleavage 22.1 hpi versus
25.5 hpi; blastulation 140.4 hpi versus 152.5 hpi;
D7 blastocyst rates: 31.3% versus 21.9%; P <
0.05). In a third experiment, 65–76 Day 8
blastocysts per bull were sexed by PCR. Only
blastocysts obtained with bull III showed a shift in
sex ratio towards male embryos (76% male
embryos; P < 0.05). Such shift was already
observed at the 2-cell and morula stages. In
conclusion, the bull influences the kinetics of PN
formation, of embryo development and the sex
ratio of the embryos. Moreover, those parameters
might be related.
donnay@vete.ucl.ac.be
Animal (2008), 2:8, pp 1158–1167
Expression Profiles of Genes Regulating Dairy Cow
Fertility: Recent Findings, Ongoing Activities and Future
Possibilities
B. Beerda1, J. Wyszynska-Koko2, M. F. W. te Pas1, A. A. C. de
Wit1 and R. F. Veerkamp1
1
Animal Breeding and Genomics Centre, Animal Sciences Group of
Wageningen UR, 8200 AB Lelystad, The Netherlands; 2Institute of
Genetics and Animal Breeding, Polish Academy of Sciences,
Jastrzebiec, 05-552 Wolka Kosowska, Poland.
Subfertility has negative effects for dairy farm
profitability, animal welfare and sustainability of
animal production. Increasing herd sizes and
economic pressures restrict the amount of time
that farmers can spend on counteractive
management. Genetic improvement will become
increasingly important to restore reproductive
performance. Complementary to traditional
breeding value estimation procedures, genomic
selection based on genome-wide information will
become more widely applied. Functional
genomics, including transcriptomics (gene
expression profiling), produces the information to
understand the consequences of selection as it
helps to unravel physiological mechanisms
underlying female fertility traits. Insight into the
latter is needed to develop new effective
3

management strategies to combat subfertility.
Here, the importance of functional genomics for
dairy cow reproduction so far and in the near
future is evaluated. Recent gene profiling studies
in the field of dairy cow fertility are reviewed and
new data are presented on genes that are
expressed in the brains of dairy cows and that are
involved in dairy cow oestrus (behaviour). Fast-
developing new research areas in the field of
functional genomics, such as epigenetics, RNA
interference, variable copy numbers and
nutrigenomics, are discussed including their
promising future value for dairy cow fertility.
bonne.beerda@wur.nl
Reproduction, Fertility and Development, 2008, 20,
784–788
Technical Note
Effect of Density Gradient Centrifugation With Trypsin
on the in vivo Fertilising Capability of Bovine
Spermatozoa
Brock A. BlevinsA,C, Morne de la ReyB and Naida M. LoskutoffA
A
The Bill and Berniece Grewcock Center for Conservation and
Research, Henry Doorly Zoo, Omaha, NB 68107, USA. BEmbrio Plus,
Brits, NorthWest 0250, RSA.
The present study investigated the effect of a
novel density gradient centrifugation (DGC)
treatment using recombinant trypsin on the in
vivo fertilising capability of bovine spermatozoa
compared with a standard method. In Trial 1,
semen collected from Boran and Ankole (Bos
indicus) bulls was treated either with a silane-
coated silica particle colloid formulated for
humans with a recombinant trypsin or processed
using a standard method (dilution in an egg yolk
based diluent). Semen processed by the two
methods was used to artificially inseminate (AI)
superovulated cattle. Day 7 embryos were flushed
and assessed for fertilisation rates and embryo
quality.Trial 2 used a trypsinised silane-coated
silica particle colloid formulated specifically for
bovine semen. Trial 1 resulted in significantly
higher fertilisation rates using the trypsinised
human DGC treatment than cows inseminated
using the standard method (75.2% v. 67%,
respectively; P <0.01), but the numbers of
transferable-quality Day 7 embryos did not differ
between the two groups (P >0.05). Results for
Trial 2 indicated that cows inseminated with the
trypsinised bovine DGC treatment had significantly
increased fertilisation rates compared with the
standard method (88.4% v. 63.1%, respectively;
Noticias de Reproducción Bovina AGOSTO 2008
P <0.01) and had significantly higher numbers of
transferable-quality embryos (70.3% v. 51.8%,
respectively; P <0.01). In summary, bovine sperm
treatment before AI by DGC and recombinant
trypsin increases fertilisation rates and can result
in more transferable-quality embryos compared
with standard methods.
repro@omahazoo.com
Theriogenology 70 (2008) 485–494
The Effect of Nutrition on Sexual Development of Bulls
A.D. Barth, L.F.C. Brito, J.P. Kastelic
Department of Large Animal Clinical Sciences, Western College of
Veterinary Medicine, University of Saskatchewan, Saskatoon,
Saskatchewan, Canada S7N 5B4.
Most bulls that are managed for sale as yearlings
are fed high-energy diets in the post-weaning
period to maximize rates of gain in body weight.
High-energy diets with adequate protein, vitamins
and minerals result in a larger scrotal
circumference at 1 y of age, however, part of this
increase in size is likely due to scrotal fat. It is
unclear whether testis size and spermatogenesis
is significantly affected by nutritional intake in the
post-weaning period. There are indications of an
effect of calfhood nutrition on age at puberty and
testis size. Scrotal circumference was smaller in
yearling bulls raised by first-parity dams,
compared to those raised by older dams. This
may have been due to lower milk production by
first-parity dams, an in utero effect, or both. The
effect of reduced calfhood nutrition may be
mediated through gonadotropin secretion. Calves
destined to become later maturing bulls with
smaller testes had lower amounts of LH secretion
during the period of the early gonadotropin rise
(8–16 wk of age). Furthermore, augmenting
circulating LH concentrations at this time by
treating calves with GnRH hastened pubertal
development. In addition, FSH treatments in
calfhood also increased scrotal circumference and
hastened spermatogenesis. In that regard, FSH
has been considered a main driver of Sertoli cell
proliferation in prepubertal animals. Since Sertoli
cell multiplication ceases at 20–25 wk of age in
bulls, final testis size in bulls is likely determined
in calfhood. Four experiments were done to
investigate the effects of calfhood nutrition on
pubertal development. These studies confirmed
that superior calfhood nutrition augmented
gonadotropin secretion (which is probably
mediated by metabolic hormones); this resulted in
4

larger testes at 1 y of age and an earlier onset of
spermatogenesis.
albert.barth@usask.ca
Animal Reproduction Science 107 (2008) 9–19
Effect of Progesterone Administration on the Ovarian
Response to Superovulatory Treatments in Cattle
S. Callejasa, R. Alberiob, J. Cabodevilaa, J. Allerb, R. Catalanoa,
M. Teruela, F. Duloutc
a
Area de Reproducción (FISFARVET), Facultad de Ciencias
Veterinarias, Universidad Nacional del Centro de la Provincia de
Buenos Aires, Campus Universitario, Paraje Arroyo Seco S/N,
B7000GHG Tandil, Buenos Aires, Argentina. bIntituto Nacional de
Tecnología Agropecuaria Balcarce, 7620 Balcarce, Buenos Aires,
Argentina. cCIGEBA, Facultad de Ciencias Veterinarias, Universidad
Nacional de La Plata, 1900 La Plata, Argentina.
To evaluate ovarian response in Angus cows
previously treated with progesterone (P4),
animals were randomly assigned to two groups:
T600 group (n = 14), 600 mg of P4/day. P4 was
injected from days 3 to 7 of the estrous cycle. On
day 7, superovulatory treatments began. The
control group (n = 12) was given vehicle only.
The superovulatory treatments in the control
group began on days 7–9 of the estrous cycle.
The superovulatory total treatment dose of 400
mg NIH FSH P1 was given twice a day over a 4-
day period. Ultrasonography of the ovaries was
conducted 3 days preceding the initiation of
superovulatory treatment, every 24 h. In both
groups, an additional ultrasonographic evaluation
was made at 24 h after the end of superovulatory
treatment. Blood samples were collected 4 days
preceding the initiation of superovulatory
treatment, every 24 h. Additional samples were
taken from the P600 group for 12 day after of
initiation of superovulatory treatment every 24 h,
except on the fifth day after the initiation of
superovulatory treatment. In the P600 group, P4
concentrations were greater than in the control
group (P < 0.01) and remained over 1 ng/ml up
to day 11 after beginning of superovulatory
treatment. The diameter of the dominant follicle
was larger in the animals of the control group (P
< 0.01). Cows of the P600 group had a greater
number of Class I (3–4 mm) follicles (P < 0.01). A
significant day and treatment effect (P < 0.01)
were observed in Class II (5–9 mm) follicles.
Effects due to treatment on the number of Class
III follicles (P < 0.05) were observed. In the P600
group, no estrous post-superovulatory was
observed and there were no ovulations that
occurred. Conversely, 100% of the cows of the
control group showed estrous. In the P600 group,
Noticias de Reproducción Bovina AGOSTO 2008
there were a greater number of Class III follicles
(P < 0.01) and a lesser number of Class II follicles
(P < 0.05) at 24 h after the end of
superovulatory. In the control group, 66.7% of
the cows responded to superovulatory treatments.
In conclusion, the daily administration of 600 mg
of P4, from days 3 to 7 of the estrous cycle,
produces an increase of plasma concentrations of
this hormone from day 4, resulting in changes in
follicular dynamics (absence of follicles greater
than 10mm of diameter and an increase of the
population of Class I follicles). As to the ovarian
stimulation using Folltropin V® in animals
receiving a daily injection of 600 mg of P4 from
days 3 to 7 of the estrous cycle, a greater
population of follicles≥10mm developed by 24 h
after superovulatory treatments were completed.
callejas@vet.unicen.edu.ar
Theriogenology 70 (2008) 469–472
Bull Breeding Soundness Evaluation: A Practitioner’s
Perspective
J.H. Alexander
Alexander Veterinary Services, 3135 Buck Reed Road, Edwards, MS
39066, United States.
It is well established that the bull breeding
soundness evaluation (BBSE) is an easily
performed, relatively inexpensive, and extremely
useful tool for the cow–calf operation. However,
the BBSE can only achieve its full potential if it is
performed properly by the practitioner in the field.
A properly performed BBSE should include the use
of the standards established by the Society for
Theriogenology and a systematic protocol by the
practitioner. This paper will review the Society for
Theriogenology BBSE standards and the author’s
protocol.
jayacims@tecinfo.com.
Theriogenology 70 (2008) 573–575
The Development of Beef Breeding Bulls
T.J. Engelken
Veterinary Diagnostic and Production Animal Medicine, Iowa State
University, College of Veterinary Medicine, Ames, IA 50011, United
States.
Management of the bull battery will have a
dramatic impact on profitability of the cow/calf
enterprise. It is critical that young bulls be
selected and developed to maximize longevity and
productivity for the eventual buyer. Bulls must be
structurally sound, healthy, and have adequate
libido in order to service the required number of
females. Once bulls complete their first breeding
5

season, special care must be taken in order to
ensure that they recover and regain needed body
condition and pass a bull breeding soundness
examination (BBSE). Mature bulls that have
reached their genetic potential for growth require
less intensive management, but the health
program and annual BBSE cannot be overlooked.
Mature bulls are also more likely to carry venereal
disease and should be screened according to local
disease incidence and state regulations. All bulls,
regardless of age, should be observed early
during the breeding season to ensure that they
are physically capable of mounting and servicing
females. The establishment of a complete
management program, especially for young bulls,
is essential to ensure that ranch resources are
used efficiently, including maintenance of a high
level of reproductive performance of the cow
herd.
engelken@iastate.edu.
Animal (2008), 2:8, pp 1104–1111
Why is it Getting More Difficult to Successfully
Artificially Inseminate Dairy Cows?
H. Dobson, S. L. Walkera, M. J. Morris, J. E. Routly and R. F.
Smith
Department of Veterinary Clinical Science, University of Liverpool,
Leahurst, Neston, Wirral CH64 7TE, UK.
Successfully using artificial insemination (AI) is
defined as getting cows pregnant when the
farmer wants them in-calf and making the best
use of appropriate genetic potential. Over the past
30 to 50 years, the percentage of animals in
oestrus that stand-tobe-mounted (STBM) has
declined from 80% to 50%, and the duration of
STBM from 15 h to 5 h; both in parallel with a
reduction in first-service-pregnancy-rate from
70% to 40%. Meanwhile, the incidence of
lameness and mastitis has not decreased; and it
takes more than an extra 40 and 18 days,
respectively, to get a lame or mastitic cow in-calf
compared to healthy herd-mates. The intensity of
oestrus is 50% lower in severely lame cows, and
fewer lame cows ovulate. Luteal phase milk
progesterone concentrations are also 50% lower
in lame cows, and follicular phase oestradiol is
also lower in non-ovulating lame cows compared
to ovulating animals. Furthermore, lame cows that
do not ovulate do not have an LH surge, and the
LH pulse frequency in their late follicular phase is
lower (0.53 v. 0.76 pulses/h). Thus, we suggest
that the stress of lameness reduces LH pulsatility
Noticias de Reproducción Bovina AGOSTO 2008
required to drive oestradiol production by the
dominant follicle. The consequent low oestradiol
results in less-intense oestrus behaviour and
failure to initiate an LH surge; hence there is no
ovulation. A series of experimental studies
substantiate our hypothesis that events activating
the hypothalamus–pituitary–adrenal axis interfere
at both the hypothalamus and the pituitary level
to disrupt LH and oestradiol secretion, and thus
the expression of oestrus behaviour. Our inability
to keep stress at a minimum by appropriately
feeding and housing high-production cows is
leading to a failure to meet genetic potential for
yield and fertility. We must provide realistic
solutions soon, if we want to successfully use AI
to maintain a sustainable dairy industry for the
future.
hdobson@liv.ac.uk
Animal (2008), 2:8, pp 1128–1134
Developmental Programming of Reproduction and
Fertility: What is the Evidence?
D. S. Gardner1, R. G. Lea1 and K. D. Sinclair2
1
School of Veterinary Medicine and Science, Sutton Bonington
Campus, University of Nottingham, Loughborough LE12 5RA, UK;
2
School of Biosciences, Sutton Bonington Campus, University of
Nottingham, Loughborough LE12 5RA, UK.
The concept of the foetal/developmental origins of
adult disease has been around for 20 years and
from the original epidemiological studies in human
populations much more evidence has accumulated
from the many studies in animal models. The
majority of these have focused upon the role of
early dietary intake before conception, through
gestation and/or lactation and subsequent
interactions with the postnatal environment, e.g.
dietary and physical activity exposures. Whilst a
number of theoretical models have been proposed
to place the experimental data into a biological
context, the underlying phenomena remain the
same; developmental deficits (of single (micro)
nutrients) during critical or sensitive periods of
tissue growth alter the developmental pathway to
ultimately constrain later functional capacity when
the individual is adult. Ageing, without exception,
exacerbates any programmed sequelae. Thus,
adult phenotypes that have been relatively easy to
characterise (e.g. blood pressure, insulin
sensitivity, body fat mass) have received most
attention in the literature. To date, relatively few
studies have considered the effect of differential
early environmental exposures on reproductive
6

function and fecundity in predominantly mono-
ovular species such as the sheep, cow and
human. The available evidence suggests that
prenatal insults, undernutrition for example, have
little effect on lifetime reproductive capacity
despite subtle effects on the hypothalamic–
pituitary–gonadal axis and gonadal progenitor cell
complement. The postnatal environment is clearly
important, however, since neonatal/adolescent
growth acceleration (itself not independent from
prenatal experience) has been shown to
significantly influence fecundity in farm animals.
The present paper will expand these interesting
areas of investigation and review the available
evidence regarding developmental programming
of reproduction and fertility. However, it appears
there is little strong evidence to indicate that
offspring fertility and reproductive senescence in
the human and in farm animal species are overtly
affected by prenatal nutrient exposure.
Nevertheless, it is clear that the developing gonad
is sensitive to its immediate environment but
more detailed investigation is required to
specifically test the long-term consequences of
nutritional perturbations during pregnancy on
adult reproductive well-being.
david.gardner@nottingham.ac.uk or kevin.sinclair@nottingham.ac.uk
Theriogenology 70 (2008) 622–630
Increased Pre-implantation Development of Cloned
Bovine Embryos Treated With 5-aza-20-Deoxycytidine
and Trichostatin A
X. Ding, Y. Wang, D. Zhang, Y. Wang, Z. Guo, Y. Zhang
Institute of Bioengineering, Northwest A&F University, Yangling,
Shaanxi 712100, China.
Limited success of somatic cell nuclear transfer is
attributed to incomplete reprogramming of
transferred nuclei. The objective was to determine
if 5-aza-20-deoxycytidine (5-aza-dC) and
trichostatin A (TSA) promoted reprogramming and
improved development. Relative to untreated
controls, treatment of donor cells, cloned
embryos, and continuous treatment of both donor
cells and cloned embryos with a combination of
0.01 µM 5-aza-dC and 0.05 µM TSA significantly
increased the blastocyst rate (11.9% vs 31.7%,
12.4% vs 25.6%, and 13.3% vs 38.4%,
respectively) and total cell number (73.2 vs 91.1,
75.2 vs 93.7, and 74.6 vs 96.7). Moreover,
blastocyst rate and inner cell mass (ICM) cell
number of embryos continuously exposed to both
reagents were significantly higher than that of a
Noticias de Reproducción Bovina AGOSTO 2008
TSA-treated group (38.4% vs 23.9% and 27.4 vs
18.2). The DNA methylation level of 2-cell
embryos was decreased significantly, whereas the
histone acetylation level increased dramatically
after donor cell treatment and continuous
treatment with both reagents. However, these
epigenetic features of cloned blastocysts were not
significantly different than the untreated control
group. Following embryo treatment, DNA
methylation and histone acetylation levels of
cloned blastocysts were unchanged, except for
the group given 0.5 µMTSA (acetylation level was
significantly increased, but development potential
was reduced). In conclusion, development of
cloned bovine embryos was enhanced by 5-aza-
dC and TSA; furthermore, the combination was
more effective than either one alone.
zh_yong@yahoo.com.cn; yongzhang06@163.com
Journal of Dairy Science 91:3025–3033, 2008
Pregnancy Incidence in Norwegian Red Cows Using
Nonreturn to Estrus, Rectal Palpation, Pregnancy-
Associated Glycoproteins, and Progesterone
R. T. Garmo,* A. O. Refsdal,† K. Karlberg,* E. Ropstad,* A.
Waldmann,‡ J. F. Beckers,§ and O. Reksen*
*Department of Production Animal Clinical Sciences, Norwegian
School of Veterinary Science, PO Box 8146, NO-0033 Oslo, Norway.
†Geno Breeding and AI Association, Holsetgata 22, 2326 Hamar,
Norway. ‡Estonian University of Life Sciences, 62 Kreutzwaldi Street,
51014 Tartu, Estonia. §University of Liege, Faculty of Veterinary
Medicine, B 4000 Sart-Tilman, Belgium.
The objectives of the study were to estimate
pregnancy incidence and calving rate after first
artificial insemination (AI) in Norwegian Red cows
undergoing spontaneous estrus, to assess the
relationship between pregnancy and management
factors at herd or cow level, to evaluate
differences between 60-d nonreturn rate
(NRR60d) and pregnancy incidence, and to
compare the accuracy of pregnancy diagnosis by
rectal palpation and plasma pregnancy-associated
glycoproteins (PAG) analysis supported by
progesterone measurements. In total, 829
animals (n = 229 heifers, 234 first lactation, 173
second-lactation, and 193 >second-lactation
cows) were included. Milk samples for
progesterone analysis were collected both at AI
and 3 wk later. Cows with progesterone
concentrations <3 ng/mL at AI were considered in
estrus or having non active ovaries, whereas cows
with progesterone concentrations >7 ng/mL 3 wk
later were considered pregnant. Blood sampling
for PAG analysis and pregnancy diagnosis by
7

rectal palpation were conducted 57.6 ± 0.92 d
after AI. Pregnancy-associated glycoprotein
concentrations equal to 2.5 ng/mL gave the
greatest sensitivity (94.3%) and specificity
(94.6%) in the assessment of pregnancy. The
number of days from calving to first AI was 85.3
± 1.71. Overall NRR60d after first AI was 72.5%.
The corresponding values for heifers, first-
lactation, second lactation, and >second-lactation
cows were 76.9, 67.1, 69.9, and 76.2%. Overall
pregnancy incidence after first AI was 63.7%. The
corresponding values for heifers, first-lactation,
second-lactation, and >second-lactation cows
were 70.0, 58.2, 61.6, and 64.9%. Overall calving
rate to first AI was 57.2%. The corresponding
values for heifers, first-lactation, second-lactation,
and >second lactation cows were 64.9, 54.3,
54.7, and 53.9%. The overall difference between
NRR60d and pregnancy incidence was 8.8%,
whereas the parity-specific differences were 6.9,
8.9, 8.3, and 11.3% for heifers, first-lactation,
second-lactation, and >second-lactation cows,
respectively. Eight animals with PAG <2.5ng/mL
and classified as pregnant by rectal palpation
calved, whereas 5 animals with PAG ≥2.5 ng/mL
and classified as nonpregnant by rectal palpation
also calved. The study showed that Norwegian
Red cows have relatively high reproductive
performance. Breeding for fertility traits over 35
yr is probably an important reason for such high
fertility.
randi.garmo@veths.no
Animal (2008), 2:8, pp 1144–1152
Integration of Physiological Mechanisms That Influence
Fertility in Dairy Cows
P. C. Garnsworthy, K. D. Sinclair and R. Webb
University of Nottingham School of Biosciences, Sutton Bonington
Campus, Loughborough LE12 5RD, UK.
Fertility in dairy cows has been declining for the
past three decades. Genetic selection for
increased milk production has been associated
with changes in key metabolic hormones (growth
hormone, insulin, IGF and leptin) that regulate
metabolism by homoeostasis and homeorhesis.
These metabolic hormones, particularly insulin,
provide signals to the reproductive system so that
regulation of ovarian function is coordinated with
changes in metabolic status. Studies have shown,
for example, that increasing circulating insulin
concentrations during the early post partum
period can advance the resumption of oestrous
Noticias de Reproducción Bovina AGOSTO 2008
cycles by enhancing follicular growth. However,
high concentrations of insulin can be detrimental
to the developmental competence of oocytes,
which is also influenced by the supply of fatty
acids at the systemic level and at the ovarian
level. Insulin status is also associated with the
incidence and characteristics of abnormal ovarian
cycles. These changes can occur without
significant variation in circulating gonadotrophin
concentrations. This suggests that additional
factors, such as peripheral metabolites, metabolic
hormones and locally produced growth factors,
may have a modulating role. Recent evidence has
demonstrated that ovarian responses to metabolic
signals and nutrient profile vary according to the
stage of the reproductive cycle. Improved
understanding of this multifactorial process
enables nutrition to be matched to genotype and
milk production, with a positive impact on
pregnancy rate.
Phil.Garnsworthy@nottingham.ac.uk
Livestock Science 117 (2008) 88–92
Effects of GnRH in Combination With PGF2α on the
Dynamics of Follicular and Luteal Cells in Post-pubertal
Holstein Heifers
R. Harpera, W.A. Bennettb, E.J. Cuadraa, C.F. Vaughna, N.S.
Whitworthb
a
Alcorn State University, 1000 ASU Drive, Box 750, Alcorn State, MS
39096, United States. bDepartment of Obstetrics and Gynecology,
University Medical Center, 2500 North State Street, Jackson, MS
39216, United States.
Holstein heifers were randomly allotted by weight,
age and body condition score to one of three
treatments to test the hypothesis that GnRH
administration concurrent with PGF2α injection
would advance follicle or corpus luteum (CL)
development parallel to an induced luteolysis of
the pre-existing CL. Heifers in the control group
(n=14) received two treatments of PGF2α (25 mg,
im) given 10 days apart. Groups 2 (n=14) and 3
(n=14) received an additional treatment of GnRH
(100 µg, im) after the first and second PGF2α
respectively. Estrus detection began immediately
after PGF2α and continued for 80 h. Blood
sampling was initiated 7 days prior to the first
PGF2α (day −7) and continued on days 0, 7, 10
(prior to the second PGF2α), 17 and 24. Heifers
were artificially inseminated after the second
PGF2α and pregnancy diagnosed at 60 days.
There was a trend (Pb.10) toward a lower estrus
response in group 3 when compared to the other
groups. Pregnant heifers in group 2 had lower
8

progesterone (0.44±0.09 vs. 1.72±0.56 ng/ml) a
week after the second PGF2α than the non-
pregnant animals in that group (Pb.05). Similar
results were observed in the control group but
only within the responding heifers (0.61±0.08 vs.
0.93±0.03 ng/ml; Pb.05). Progesterone in heifers
in group 2 remained high on day 0, 7, and 10
(1.48±0.37, 1.23±0.39, 1.96±0.36 ng/ml) in spite
of the treatment with PGF2α. This data suggest
that administration of GnRH following PGF2α
alters bovine luteal and/or follicular cell function.
cuadra@alcorn.edu
Theriogenology 70 (2008) 270–285
Infectious Causes of Embryonic and Fetal Mortality
a
a,b
M. Daniel Givens , M.S.D. Marley b
Department of Clinical Sciences, College of Veterinary Medicine,
Auburn University, Auburn, AL 36849, United States. bDepartment of
Pathobiology, College of Veterinary Medicine, Auburn University,
Auburn, AL 36849, United States.
The purpose of this review is to summarize
bacterial, fungal, protozoan, and viral causes of
reproductive dysgenesis in cattle, sheep, goats,
pigs, horses, dogs, and cats. The clinical
presentations of disease due to reproductive
pathogens are emphasized, with a focus on
assisting development of complete lists of causes
that result in abortion and infertility in these
species. Clinicians are encouraged to assess
clinical presentation, create complete lists of
differential diagnoses, obtain appropriate
diagnostic samples, maximize diagnostic
laboratory support, and avoid zoonotic infections
resulting from reproductive pathogens of animals.
The foundation of an accurate diagnosis of
reproductive loss due to infectious pathogens
facilitates the prudent use of immunization and
biosecurity to minimize reproductive losses.
givenmd@vetmed.auburn.edu
Theriogenology 70 (2008) 504–507
Pathogens that cause infertility of bulls or
transmission via semen
M. Daniel Givensa, M.S.D. Marleyb
a
Departments of Pathobiology and Clinical Sciences, College of
Veterinary Medicine, Auburn University, Auburn, AL 36849, United
States. bDepartment of Pathobiology, College of Veterinary Medicine,
Auburn University, Auburn, AL 36849, United States.
The purpose of this paper is to review scientific
evidence regarding pathogens that cause
infertility of bulls or that could be transmitted via
bovine semen. Although several pathogens can
cause male infertility and potentially be
Noticias de Reproducción Bovina AGOSTO 2008
transmitted via semen, adhering to disease
control recommendations provided by Certified
Semen Services (CSS) and the World Organization
for Animal Health (OIE) can prevent infectious
male infertility and ensure that the risk of
pathogen transmission via semen is negligible.
Regarding bulls to be used for natural breeding,
quarantine prior to herd introduction and
appropriate diagnostic testing during quarantine
will commonly prevent introduction of pathogens
that adversely affect reproduction.
givenmd@vetmed.auburn.edu
Animal (2008), 2:8, pp 1120–1127
The Consequences of Metabolic Changes in High-
Yielding Dairy Cows on Oocyte and Embryo Quality
J. L. M. R. Leroy1, A. Van Soom2, G. Opsomer2 and P. E. J.
Bols1
1
Laboratory for Veterinary Physiology, Department of Veterinary
Sciences, Faculty of Biomedical, Pharmaceutical and Veterinary
Sciences, University of Antwerp, Universiteitsplein 1, B-2610 Wilrijk,
Belgium; 2Department of Reproduction, Fertility and Herd Health;
Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133,
B-9820 Merelbeke, Belgium.
Unsatisfactory reproductive performance in dairy
cows, such as reduced conception rates, in
addition to an increased incidence of early
embryonic mortality, is reported worldwide and
has been associated with a period of negative
energy balance (NEB) early post partum.
Typically, NEB is associated with biochemical
changes such as high non-esterified fatty acid
(NEFA), high b-hydroxybutyrate (b-OHB) and low
glucose concentrations. The concentrations of
these and other metabolites in the follicular fluid
(FF) of high-yielding dairy cows during NEB were
determined and extensively analyzed, and then
were replicated in in vitro maturation models to
investigate their effect on oocyte quality. The
results showed that typical metabolic changes
during NEB are well reflected in the FF of the
dominant follicle. However, the oocyte seems to
be relatively isolated from extremely elevated
NEFA or very low glucose concentrations in the
blood. Nevertheless, the in vitro maturation
models revealed that NEB-associated high NEFA
and low glucose levels in the FF are indeed toxic
to the oocyte, resulting in deficient oocyte
maturation and developmental competence.
Induced apoptosis and necrosis in the cumulus
cells was particularly obvious. Furthermore,
maturation in saturated free fatty acid-rich media
had a carry-over effect on embryo quality, leading
9

to reduced cryotolerance of day 7 embryos. Only
b-OHB showed an additive toxic effect in
moderately hypoglycemic maturation conditions.
These in vitro maturation models, based on in
vivo observations, suggest that a period of NEB
may hamper the fertility of high-yielding dairy
cows through increased NEFA and decreased
glucose concentrations in the FF directly affecting
oocyte quality. In addition to oocyte quality, these
results also demonstrate that embryo quality is
reduced following an NEB episode. This important
observation may be linked to the typical diet
provided to stimulate milk yield, or to
physiological adaptations sustaining the high milk
production. Research into this phenomenon is
ongoing.
jo.leroy@ua.ac.be
Reprod Dom Anim 43, 415–421 (2008)
Immunohistochemical Detection of Receptors for
Oestrogen and Progesterone in Endometrial Glands and
Stroma during the Oestrous Cycle in Nelore (Bos
taurus indicus) Cows
I Martin1, R Torres Neto2, E Oba1, J Buratini Jr3, M Binelli4, R
Laufer-Amorim2 and JCP Ferreira1
1
Department of Animal Reproduction and Veterinary Radiology,
School of Veterinary Medicine and Animal Science, UNESP;
2
Department of Veterinary Clinic, School of Veterinary Medicine and
Animal Science, UNESP; 3Department of Physiology, Biosciences
Institute, UNESP, Botucatu; 4Department of Animal Reproduction,
School of Veterinary Medicine and Animal Science, USP,
Pirassununga, Brazil.
The aim of the present study was to monitor
endometrial distribution and concentrations of
oestrogen receptors a (ERa) and progesterone
receptors (PR) by immunohistochemistry in Nelore
cows (Bos taurus indicus) during the oestrous
cycle. Blood samples were collected for
progesterone measurement and endometrial
samples were taken from the uterine horn contra
lateral to the corpus luteum in 16 cows at days 0
(ovulation), 5, 9, 13 and 19 of the oestrous cycle.
Immunostaining evaluation for ERa and PR in the
glandular epithelium and uterine stroma was
performed by two methods: positive nuclei
counting and staining intensity of the nuclei.
Specific positive staining reactions for both
receptors were limited to cell nuclei and they were
not identified in the cytoplasm. The proportion of
ERa positive nuclei had a temporal variation
throughout the oestrous cycle in both cell types
evaluated and was higher in uterine stroma than
the glandular epithelium (p < 0.05). The greatest
Noticias de Reproducción Bovina AGOSTO 2008
proportion of ERa stained nuclei was observed at
oestrus and during the initial and mid luteal phase
(days 5, 9 and 13) (p < 0.05) in the glandular
epithelium and at days 0, 5 and 9 in the uterine
stroma (p < 0.01). The proportion of PR positive
nuclei remained constant throughout the entire
oestrous cycle for both cell types evaluated (p >
0.05). A higher proportion of PR positive nuclei
was measured in the uterine stroma compared
with the glandular epithelium (p < 0.05). Intensity
of staining for ERa and PR varied throughout the
oestrous cycle (p < 0.01). There was a higher
staining intensity at days 0 and 5 in the stroma
for ERa (p < 0.01) and PR (p < 0.01) and in the
glandular epithelium at days 0, 5, 9 and 13 for Era
(p < 0.01) and at days 0, 5 and 9 for PR (p <
0.01) when compared with the other evaluated
days. These data demonstrate that ERa and PR
expression varied throughout the oestrous cycle in
Nelore cows, in general with highest
concentrations at oestrus and the lowest during
the luteal phase. This is similar to patterns
observed in Bos taurus taurus.
Reprod Dom Anim 43, 464–472 (2008)
Luteolytic Effect of Prostaglandin F2α on Bovine Corpus
Luteum Depends on Cell Composition and Contact
AJ Korzekwa1, JJ Jaroszewski2, I Woclawek-Potocka1, MM
Bah1 and DJ Skarzynski1
1
Division of Reproductive Endocrinology and Pathophysiology,
Institute of Animal Reproduction and Food Research, PAS;
2
Department of Pathology and Pharmacology, Faculty of Veterinary
Medicine, University of Warmia and Mazury in Olsztyn, Olsztyn,
Poland.
Prostaglandin F2α (PGF2α) is a main luteolytic
factor in vivo; however, its direct luteolytic
influence on steroidogenic cells of bovine corpus
luteum (CL) is controversial and not fully
understood. The aim of the study was to clarify
PGF2α action on bovine CL in different in vivo and
in vitro conditions and to examine whether the
contact among all main types of CL cells is
necessary for luteolytic PGF2α action. In
experiment 1, the bovine CL (day 15 of the
oestrous cycle) was perfused using in vivo
microdialysis system with dinoprost (an analogue
of PGF2α) for 0.5 h. Dinoprost caused a short-
time increase in progesterone (P4), whose
concentration decreased there after (at 6-, 10-,
12- and 24-h after treatment). In experiment 2,
the direct effect of PGF2α on P4 accumulation in
CL steroidogenic cells cultured in monolayer (day
15 of the cycle) was determined. PGF2α after 24 h
10

of incubation increased P4 accumulation in
steroidogenic CL cells. In experiment 3
steroidogenic, endothelial CL and immune cells
(day 15 of the cycle) were incubated with PGF2α
in cocultures for 24 h in glass tubes and the levels
of P4, stable metabolites of nitric oxide (NO) and
leukotriene (LT) C4 were determined. Although
PGF2α treatment increased P4 secretion in
homogeneous steroidogenic CL cell culture, the
decrease in P4 secretion in cocultures of all types
of CL cells was observed. The secretion of NO and
LTC4 increased after the treatment of PGF2α both
in pure cultures of CL cells and in cocultures. The
interactions between endothelial and immune cells
with steroidogenic CL cells are needed for
luteolytic PGF2α action within the bovine CL. Our
results indicate that the cell coculture model,
including the main types of CL cells, is the most
approximate to study PGF2α role in vitro.
Theriogenology 70 (2008) 569–572
Developing Replacement Beef Heifers
T.J. Engelken
Veterinary Diagnostic and Production Animal Medicine, Iowa State
University, College of Veterinary Medicine, Ames, IA 50011, United
States.
The replacement heifer represents the next
generation of genetic progress for the cow herd.
Producers invest a substantial amount of capital in
these females, even if they fail to become
pregnant. In order to get a return on this
investment, it is imperative that these heifers
become pregnant early in the first breeding
season, calve with a minimum of dystocia, breed
back in a timely fashion, and then continue to be
productive for a number of years. Practitioners
working with heifer development programs need
to emphasize a systemic approach that evaluates
these females at critical times. These programs
need to address such areas as weaning and
nutritional management, genetic selection,
prebreeding evaluation, the breeding season, and
heifer management from pregnancy examination
through the end of their first calving season. This
increased level of scrutiny should not end until the
heifer weans her first calf and is determined to be
pregnant the second time. This type of program
will ensure optimal reproductive rates, female
longevity, and a positive return on the producer’s
investment.
engelken@iastate.edu.
Noticias de Reproducción Bovina AGOSTO 2008
Reprod Dom Anim 43, 477–480 (2008)
Oocyte Recovery by Ovum Pick Up and Embryo
Production in River Buffaloes (Bubalus bubalis)
BM Manjunatha2, JP Ravindra1, PSP Gupta1, M Devaraj2 and
1
S Nandi1
National Institute of Animal Nutrition and Physiology (NIANP), ICAR,
Bangalore, India; 2Department of Animal Reproduction, Gynaecology
and Obstetrics, Veterinary College, Karnataka Veterinary, Animal and
Fishery Sciences University (KVAFSU), Bangalore, India.
Ovum pick up (OPU) was conducted twice a week
for 12 weeks in six cycling, non-descriptive (local
breed), Indian buffaloes to study the efficiency of
OPU on recovery of oocytes for embryo
production. OPU was performed using an
ultrasound equipment with a 5-MHz transvaginal
transducer, a single-lumen, 18-gauge, 55-cm-long
needle and a constant vacuum pressure of 110
mmHg. The number and size of follicles were
determined before puncture. The recovered
oocytes were graded, washed, matured for 24 h
and then fertilized with frozen–thawed semen,
followed by embryo culture on the oviductal
monolayer. The mean number of follicles
observed per animal per session did not differ
between animals or between puncture sessions. A
mean number of 3.62 ± 0.32 mm follicles were
observed, 2.90 ± 0.15 mm follicles were
punctured and 1.21 ± 0.07 oocytes were
recovered per animal per session, with an average
recovery rate of 42%. Of the total oocytes
recovered, 64% were suitable for in vitro embryo
production (grade A + B) whereas 36% were
classified to be of grades C + D. A mean number
of 0.25 ± 0.2 transferable embryos was produced
in vitro per buffalo per session with a transferable
embryo production rate of 32%. In conclusion,
this study demonstrated that twice-aweek OPU
could be applied repeatedly, without any adverse
effects on the follicular growth and oocyte
recovery and that recovered oocytes could be
used for in vitro embryo production in buffaloes.
Theriogenology 70 (2008) 565–568
Epidemiology and disease control in everyday beef
practice
R.L. Larson
College of Veterinary Medicine, Kansas State University, 111B Mosier
Hall, Manhattan, KS 66506, USA.
It is important for food animal veterinarians to
understand the interaction among animals,
pathogens, and the environment, in order to
implement herd-specific biosecurity plans. Animal
factors such as the number of immunologically
11

protected individuals influence the number of
individuals that a potential pathogen is able to
infect, as well as the speed of spread through a
population. Pathogens differ in their virulence and
contagiousness. In addition, pathogens have
various methods of transmission that impact how
they interact with a host population. A cattle
population’s environment includes its housing
type, animal density, air quality, and exposure to
mud or dust and other health antagonists such as
parasites and stress; these environmental factors
influence the innate immunity of a herd by their
impact on immunosuppression. In addition, a
herd’s environment also dictates the ‘‘animal flow’’
or contact and mixing patterns of potentially
infectious and susceptible animals. Biosecurity is
the attempt to keep infectious agents away from
a herd, state, or country, and to control the
spread of infectious agents within a herd.
Infectious agents (bacteria, viruses, or parasites)
alone are seldom able to cause disease in cattle
without contributing factors from other infectious
agents and/or the cattle’s environment. Therefore
to develop biosecurity plans for infectious disease
in cattle, veterinarians must consider the
pathogen, as well as environmental and animal
factors.
Rlarson@vet.ksu.edu.
Livestock Science 117 (2008) 70–78
Effect of Milking Frequency in Early Lactation on Energy
Metabolism, Milk Production and Reproductive
Performance of Dairy Cows
a,b a b b
S. McNamara , J.J. Murphy , F.P. O'Mara , M. Rath , J.F.
Meea
a
Teagasc, Moorepark Dairy Production Research Centre, Fermoy, Co.
Cork, Ireland. bUCD School of Agriculture, Food Science and
Veterinary Medicine, Belfield, Dublin 4, Ireland.
The objective of this study was to evaluate the
effect of imposing different milking frequencies for
the first 4 weeks of lactation on energy balance,
blood metabolic profiles and reproductive
performance. The milking frequencies imposed
were once daily for the first 4 weeks of lactation
and twice daily subsequently (X1), twice daily for
the entire lactation (X2) and three times daily for
the first 4 weeks of lactation and twice daily
subsequently (X3). Two weeks prior to the first
expected calving date, sixty-three multiparous,
Holstein-Friesian cows, were allocated to
treatments based on expected calving date, parity
(lactation number=2 and lactation numberN2),
Noticias de Reproducción Bovina AGOSTO 2008
BCS and BW. Mean DMI was lower (Pb0.05) on
X1 than X2 (15.7 and 17.1 kg/day) over the first 4
weeks of lactation, but DMI on X3 was not
different to either. Milk yield was lower (Pb0.05)
on X1 than on X2 and X3 (24.1, 31.9 and 31.4
kg/day) from weeks 1 to 4 and at week 10 of
lactation (30.9, 33.9 and 33.7 kg/day) but was
not different at week 20. Yield of milk constituents
followed the same pattern as milk yield. Milking
frequency had no significant effect on milk fat or
protein concentration but milk lactose
concentration was higher (Pb0.05) on X1 than on
X2. Cows on X1 experienced less (Pb0.05)
negative energy balance than those on X2 and X3
from weeks 1 to 4, with no difference between X2
and X3. Cows on X1 lost less BW (Pb0.05) from
weeks 1 to 4 and lost less BCS up to day 73 of
lactation than cows on X2 and X3. Plasma NEFA
and BHB concentrations tended to be lower and
plasma glucose tended to be higher on X1 than
on X2 and X3 from weeks 1 to 4, while plasma
insulin was higher on X1 than on X2 and X3 for
the first 2 weeks of lactation. There was no
difference between treatments in any
reproductive performance variable measured.
These results indicate that reduced milking
frequency decreases milk yield and improves
energy balance and metabolic status in early
lactation.
jmurphy@moorepark.teagasc.ie
Animal (2008), 2:8, pp 1112–1119
Effect of Progesterone on Embryo Survival
D. Morris and M. Diskin
Teagasc, Animal Production Research Centre, Mellows Campus,
Athenry, Co. Galway, Irelan.
Increased genetic selection over the past 40 years
has resulted in a dairy cow with an improved
biological efficiency for producing milk but with an
associated reduced fertility. Embryo loss is the
greatest factor contributing to the failure of a cow
to conceive. The extent and timing of embryo loss
indicates that 70% to 80% of this loss occurs in
the first 2 weeks after artificial insemination (AI).
This is the period when a number of critical
phases in embryo development occur and where
protein accretion, substrate utilization and embryo
metabolism increase dramatically. During this time
the early embryo is completely dependent on the
oviduct and uterine environment for its survival
and it is likely that the embryo requires an optimal
uterine environment to ensure normal growth and
12

viability. There is increasing evidence of an
association between the concentration of systemic
progesterone and early embryo loss and that
progesterone supplementation of cows,
particularly those with low progesterone, can
reduce this loss. While progesterone is known to
affect uterine function and embryo growth, little is
known about the uterus during the period of early
embryo loss and how this is affected by changes
in the concentration of systemic progesterone.
The expression of uterine genes encoding the
transport protein retinol binding protein (RBP) and
the gene for folate binding protein (FBP) appear
to be sensitive to changes in systemic
progesterone, particularly during the early luteal
phase of the cycle. Uterine concentrations of
proteins also seem to be regulated by stage of
cycle; however, their relationship with the
systemic concentration of progesterone is unclear.
There is an urgent need to characterize the
uterine environment from a functional perspective
during the early part of the luteal phase of the
cycle, particularly in the high-producing cow, in
order to understand the factors contributing to
early embryo loss and in order to devise strategies
to minimize or reduce this loss.
dermot.morris@teagasc.ie
Journal of Dairy Science 91:3045–3056, 2008
Effect of Synchronization Protocols on Follicular
Development and Estradiol and Progesterone
Concentrations of Dairy Heifers
J. L. Stevenson,* J. C. Dalton,† J. E. P. Santos,‡ R. Sartori,§
A. Ahmadzadeh,# and R. C. Chebel,*1,2
*Caine Veterinary Teaching Center, and †Research and Extension
Center, University of Idaho, Moscow 83844. ‡School of Veterinary
Medicine, University of California-Davis, Tulare 93274. §Embrapa
Recursos Genéticos e Biotecnológicos, Brasília, Distrito Federal,
Brazil. #Animal and Veterinary Science Department, University of
Idaho, Moscow 83844.
The objectives were to evaluate the effect of
synchronization protocols on follicular
development and estradiol 17-β (E2) and
progesterone (P4) concentrations in dairy heifers.
In experiment 1, 36 heifers were assigned to 1 of
6 synchronization protocols in a 3 × 2 factorial
design: presynchronization with GnRH on study d
−6 or −9 [study d 0 = initiation of the Cosynch +
CIDR (controlled internal drug releasing insert
containing P4) protocol] or no presynchronization
(control) and one injection of PGF2α or not on
study d 0. In experiment 2, 126 heifers were
assigned to 1 of 4 synchronization protocols in a 2
Noticias de Reproducción Bovina AGOSTO 2008
× 2 factorial arrangement: presynchronization or
not with GnRH on study d −6 and injection of
PGF2α or not on study d 0. In experiments 1 and
2, all heifers received a modified Cosynch protocol
with CIDR for 7 d starting on study d 0. After the
PGF2α of the Cosynch and removal of the CIDR,
heifers were detected in estrus and inseminated.
Those not inseminated by study d 10 received an
injection of GnRH and were timed-inseminated.
Ovaries were scanned by ultrasound on d 0, 2,
and 5, daily from d 7 to 14, and on d 16. Blood
samples collected on d 0, 2, 7, 9, and 16 were
analyzed for P4, and the blood sample collected
on d 9 was analyzed for E2. Pregnancy was
diagnosed at 28 and 40 ± 3 d after artificial
insemination. In experiment 1, there was a
tendency for the presynchronization protocol to
affect the proportion of heifers ovulating in
response to the first GnRH injection of the
Cosynch + CIDR protocol. In experiment 2, a
greater proportion of presynchronized heifers
ovulated in response to the first GnRH injection.
Although heifers receiving PGF2α had larger
ovulatory follicles on d 7 and before ovulation and
shorter intervals to estrus and ovulation, these
heifers tended to have decreased concentrations
of E2 during proestrus. Presynchronization of
dairy heifers with GnRH increased ovulation in
response to the first GnRH injection, and
treatment of heifers with PGF2α at initiation of the
Cosynch + CIDR protocol increased the size of the
ovulatory follicle and reduced the intervals to
estrus and ovulation.
Animal Reproduction Science 107 (2008) 20–29
Protein Kinase C (PKC) Role in Bovine Oocyte
Maturation and Early Embryo Development
R.G. Mondadoria, J.P. Nevesb, P.B.D. Gonçalvesc
a
Animal Reproduction Laboratory, Department of Veterinary
Medicine, UPIS Faculdades Integradas, SEPS 712/912, Brasilia, DF
70390-125, Brazil. bAgronomy and Veterinary College, University of
Brasilia, Brasilia, DF 70910-900, Brazil. cDepartment of Large
Animals, Federal University of Santa Maria, Santa Maria, RS 97105-
900, Brazil.
The aims of the present study were to determine
the role of protein kinase C (PKC) on meiotic
resumption and its effects on pronuclear
formation and cleavage in the bovine. Oocytes
were matured in the presence of 0, 1, 10 and 100
nM of phorbol 12-myristate 13-acetate (PMA), to
evaluate the percentage of germinal vesicle
breakdown. To study pronuclear formation and
13

cleavage, oocytes were randomly distributed in
four groups and matured in modified TCM-199
with LH and FSH (negative control); 10% of
estrous cow serum (positive control); 100 Nm of
PMA (treatment); 100 nM of 4α-PDD (phorbol
ester control). Oocytes were also matured in
positive control medium, fertilized and transferred
to KSOM with increasing concentrations of a PKC
inhibitor. The protein profile and the presence of
PKC at the end of maturation period were
determined by SDS-PAGE followed by Silver Stain
and Western blot, respectively. PMA stimulated
meiotic resumption in a concentration-dependent
manner. PKC stimulation during oocyte maturation
caused an increase in pronuclear formation and
did not cause parthenogenetic activation. Inhibitor
of PKC (MyrPKC) inhibited cleavage in a dose-
dependent and irreversible manner. A protein
band around 74 kDa was not detected in PMA-
treated oocytes and PKC was not detected by
Western blot at the end of the maturation period.
In conclusion, meiotic resumption was accelerated
and the rate of oocytes with two pronuclei was
increased when PKC was activated during oocyte
maturation. Moreover, cleavage was inhibited in
the presence of PMA.
mondadori@upis.br
Animal Reproduction Science 107 (2008) 68–84
Effect of Reactive Oxygen Species on Capacitation and
Associated Protein Tyrosine Phosphorylation in Buffalo
(Bubalus bubalis) Spermatozoa
S.C. Roy, S.K. Atreja
Division of Animal Biochemistry, National Dairy Research Institute,
Karnal-132 001, India.
In the present study, the effect of two particular
reactive oxygen species (ROS), superoxide anion
(O2•−) and hydrogen peroxide (H2O2) on buffalo
(Bubalus bubalis) sperm capacitation and
associated protein tyrosine phosphorylation was
studied. Ejaculated buffalo spermatozoa were
suspended in sp-TALP medium at 50×106/mL and
incubated at 38.5 ◦C for 6 h with or without
heparin (10 (g/mL; a positive control), or xanthine
(X; 0.5 mM)-xanthine oxidase (XO; 0.05 U/mL)-
catalase (C; 2100 U/mL) system that generates
O2•− or NADPH (5 mM) that stimulates the
endogenous O2•− production or H2O2 (50_M).
The specific effect of O2•−, H2O2 and NADPH on
buffalo sperm capacitation and protein tyrosine
phosphorylation was assessed by the addition of
superoxide dismutase (SOD), catalase and
Noticias de Reproducción Bovina AGOSTO 2008
diphenylene iodonium (DPI), respectively, to the
incubation medium. Each of X+XO+C system,
NADPH and H2O2 induced a significantly higher
percentage (P < 0.05) of capacitation in buffalo
spermatozoa compared to control. However, DPI
inhibited this NADPH-induced capacitation and
protein tyrosine phosphorylation and suggested
for existence of an oxidase in buffalo
spermatozoa. Using immunoblotting technique, at
least seven tyrosine-phosphorylated proteins (20,
32, 38, 45, 49, 78 and 95 kDa) were detected in
capacitated buffalo spermatozoa. Out of these,
the tyrosine phosphorylation of p95 was induced
extensively by both O2•− as well as exogenous
source of H2O2 and using specific activators and
inhibitors of signaling pathways, it was found this
induction was regulated through a cAMP-
dependent PKA pathway. Further,
immunofluorescent localization study revealed
that these ROS-induced tyrosine-phosphorylated
proteins are mostly distributed in the midpiece
and principal piece regions of the flagellum of
capacitated spermatozoa and suggested for
increased molecular activity in flagellum during
capacitation. Thus, the study revealed that both
O2•− and H2O2 promote capacitation and
associated protein tyrosine phosphorylation in
buffalo spermatozoa and unlike human and
bovine, a different subset of sperm proteins were
tyrosine-phosphorylated during heparin- and ROS-
induced capacitation and regulation of these ROS-
induced processes were mediated through a
cAMP/PKA signaling pathway.
sudhircroy@rediffmail.com
Livestock Science 117 (2008) 34–42
Days to Calving in Artificially Inseminated Cattle:
Alternative Models and Analyses
D.L. Robinson
Beef Industry Centre, NSW Department of Primary Industries,
University of New England, Armidale, NSW 2351, Australia.
Investigations by Robinson [Robinson, D.L., 2007.
Days to calving in artificially inseminated cattle:
comparison of potential traits. Livestock Science
110, 174–180] concluded that the most useful
trait for assessing fertility of artificially
inseminated (AI) beef cows is AI days to calving
(AIDC), a trait that mimics days to calving for
naturally mated cows. Various fixed and random
effects were fitted to AIDC to determine the best
way of modelling lactation status of the cow, the
effect of service sire, using smaller contemporary
14

groups and lowering the penalty value for non-
calvers. Fitting the time interval between calving
and the start of mating either as a 10-level factor
or a cubic spline function explained considerably
more variation than fitting the standard 2- level
factor (wet or dry). Estimated permanent
environmental effects of the cow were
considerably reduced. This suggests that, if a cow
calves late in the season (less than 60 days before
she is inseminated), her fertility is reduced.
Models should therefore account for this effect. If
fitted, service sire explained 1.6% of phenotypic
variation, compared to a much larger
sire×contemporary group interaction (3% of
phenotypic variation). It is therefore important to
account for sire×contemporary group interactions.
When the fertility of service sires is not being
evaluated, service sires could be incorporated into
the definition of contemporary groups. Ideally,
breeders should be encouraged to formally record
contemporary (or mating) groups. Reducing the
size of contemporary groups (inferred from the
data) by limiting the time interval between first
and last inseminations from 120 to 60 days had
only a marginal effect as did reducing the penalty
for non-calvers from 21 to 10 days.
Dorothy.Robinson@dpi.nsw.gov.au.
Theriogenology 70 (2008) 495–503
Update on Treatment of Vesiculitis in Bulls
H. Rovay, A.D. Barth, M. Chirino-Trejo, M.F. Martínez
Western College of Veterinary Medicine, University of Saskatchewan,
Saskatoon, SK S7N 5B4, Canada.
Four experiments were done to determine: (1) the
effectiveness of early detection and treatment of
vesiculitis in bulls; (2) whether antibiotic
treatment at recommended dosages will result in
adequate vesicular gland tissue concentrations of
antibiotics to prevent in vitro bacterial growth; (3)
whether intraglandular injection of antibiotics can
be a successful alternative to systemic antibiotic
treatment; and (4) the effectiveness of tilmicosin
versus tulathromycin for treatment of clinical
vesiculitis. In Experiment 1, there was a high rate
of spontaneous remission from vesiculitis detected
at 9–12 mo of age. Furthermore, there was no
advantage for early antibiotic treatment versus no
treatment for bulls of this age. In Experiment 2,
bacteria on agar plates were exposed to fluid
extracted from vesicular gland biopsies after
antibiotic treatment of normal, healthy bulls.
Although inadequate concentrations of antibiotics
Noticias de Reproducción Bovina AGOSTO 2008
were achieved to inhibit bacterial growth when
recommended dosages of various antibiotics were
administered, doubling the antibiotic dosage
increased in vitro bacterial growth inhibition. In
Experiment 3, relatively nonirritating antibiotics
were injected directly into the glands of bulls with
clinical vesiculitis, demonstrating that
intraglandular injections of antibiotic could be
used as a successful alternative to systemic
antibiotic treatment. Experiment 4 was a clinical
field trial to compare the efficacy of tilmicosin
versus tulathromide at recommended dosages for
the treatment of clinical vesiculitis. Although the
results favored tulathromycin, both antibiotics
resulted in clinical cures of vesiculitis.
albert.barth@usask.ca
Domestic Animal Endocrinology 35 (2008) 190–197
Role For des-acyl Ghrelin in the Responsiveness of
Plasma Hormones and Metabolites to Ghrelin in
Holstein Steers
Hnin Thidar Myint, Hideto Kuwayama
Department of Animal Science, Obihiro University of Agriculture and
Veterinary Medicine, Nishi 1-11, Inada, Obihiro 080-8555, Japan.
Gastric-derived peptide hormone ghrelin is known
for its potent growth hormone (GH) stimulatory
effects. The acyl-modification on N-terminal Ser3
residue is reported to be important to stimulate
the ghrelin receptor, GH secretagogue-receptor
type1a (GHSR1a). However, major portion of
circulating ghrelin lacks in acylation, and some
biological properties of des-acyl ghrelin have been
reported in monogastric animals. In the present
study, the responsiveness of plasma hormones
and metabolites to ghrelin in steers was
characterized, and role for des-acyl ghrelin in
these changes was investigated. The repeated
intravenous administrations of bovine ghrelin (1.0
µg/kg BW) every 2 h for 8 h to Holstein steers
significantly increased the plasma acylated
ghrelin, total ghrelin, GH, insulin and NEFA levels.
The GH responses in peak values and area under
the curves (AUCs) were attenuated by repeated
injections of ghrelin, however, the responses of
plasma total ghrelin were similar. Plasma insulin
AUC decreased after fourth injection of ghrelin
while plasma NEFA AUCs gradually increased by
repeated injections of ghrelin. Pretreatment of
des-acyl ghrelin (10.0 µg/kg BW) 5 min prior to
the single injection of ghrelin (1.0 µg/kg BW) did
not affect the ghrelin-induced hormonal changes.
Moreover, the responses of plasma GH to bovine
15

and porcine ghrelin, which differ in C-terminal
amino acid residues, were similar in calves. These
data show that (1) GH release was attenuated by
repeated administration of ghrelin, (2) ghrelin
regulates glucose and fatty acid metabolism
probably via different pathway, and (3) des-acyl
ghrelin is unlikely the antagonist for ghrelin to
induce endocrine effects in Holstein steers.
fujino@obihiro.ac.jp
Reprod Dom Anim 43, 457–463 (2008)
Field Trial on Progesterone Cycles, Metabolic Profiles,
Body Condition Score and their Relation to Fertility in
Estonian Holstein Dairy Cows
J Samarütel1, K Ling1, A Waldmann1,2, H Jaakson1, T Kaart1
and A Leesmäe3
1
Institute of Veterinary Medicine and Animal Sciences, Estonian
University of Life Sciences; 2Institute of General and Molecular
Pathology, University of Tartu, Tartu; 3Piistaoja Experimental Station,
Piistaoja, Pärnu County, Estonia.
Resumption of luteal activity postpartum and
fertility were investigated in an Estonian Holstein
high milk production and good fertility dairy herd.
Body condition was scored after every 10 days in
54 multiparous dairy cows (71 lactations) calving
inside from December to March during 4-year
period. Blood samples were taken 1–14 days
before calving and 1–14, 28–42 and 63–77 days
after calving: analytes estimated were serum
aspartate aminotransferase (AST), glucose,
ketone bodies, total cholesterol, non-esterified
fatty acids and triglycerides. The general linear
mixed model was used to compare the data for
cows with different characteristics in luteal activity
postpartum based on their milk progesterone
profiles. Fortyfive per cent of cases had abnormal
profiles; delayed resumption of ovarian cyclicity
postpartum (DC) was the most prevalent
abnormality. There was no difference in body
condition scores between the groups. The DC and
prolonged luteal phase groups had higher serum
AST activity (p < 0.01) 1–14 days postpartum
compared with normal group. The DC group also
had higher cholesterol and triglyceride values (p <
0.05) 28–42 days postpartum and higher milk
fat/protein ratio (p < 0.01) on the first month of
lactation compared with normal profile group.
Despite long post-calving anoestrous period (71 ±
5.0 days; mean ± SEM) DC group had 64.7% first
service pregnancy rate (normal group 48.6% and
PLP group 37.5%). This study did not find any
detrimental effect of prolonged anovulatory period
postpartum on subsequent fertility.
Noticias de Reproducción Bovina AGOSTO 2008
Reproduction, Fertility and Development, 2008, 20,
741–749
A unique Method to Produce Transgenic Embryos in
Ovine, Porcine, Feline, Bovine and Equine Species
F. Pereyra-BonnetA, R. Fernández-MartínA, R. OliveraA, J.
JarazoA, G. VicheraA, A. GibbonsB and D. SalamoneA,C
A
Laboratorio de Biotecnología Animal, Facultad de Agronomía,
Universidad de Buenos Aires, Av. San Martín 4453, C1417 Buenos
Aires, Argentina. BLaboratorio de Reproducción de Rumiantes
Menores, Instituto Nacional de Tecnología Agropecuaria, Box 277,
EEA Bariloche, Argentina.
Transgenesis is an essential tool in many
biotechnological applications. Intracytoplasmic
sperm injection (ICSI)-mediated gene transfer is a
powerful technique to obtain transgenic pups;
however, most domestic animal embryos do not
develop properly after ICSI. An additional step in
the protocol, namely assistance by haploid
chemical activation, permits the use of ICSI-
mediated gene transfer to generate transgenic
preimplantation embryos in a wide range of
domestic species, including ovine, porcine, feline,
equine and bovine. In the present study,
spermatozoa from five species were coincubated
with pCX-EGFP plasmid and injected into
metaphase II oocytes. The chemical activation
protocol consisted of ionomycin plus 6-
dimethylaminopurine. We detected high
proportions of fluorescent EGFP embryos for all
five species (23–60%), but with a high frequency
of mosaic expression (range 60–85%). To our
knowledge, this is the first study to produce
exogenous DNA expression in feline and equine
embryos. Chemical activation reduces the lag
phase of egfp expression in ovine embryos. Our
results show that this unique method could be
used to obtain ovine, porcine, feline, bovine and
equine transgenic preimplantation embryos.
salamone@agro.uba.ar
Theriogenology 70 (2008) 479–484
Insemination Factors Related to Timed AI in Cattle
R.G. Saacke
Virginia Polytechnic Institute and State University, Department of
Dairy Science, Blacksburg, VA 240610315, United States.
Six-day-old bovine ova/embryos were recovered
non-surgically and used as biomonitors to
evaluate time of artificial insemination. These
embryos/ova provided information regarding
fertilization status and embryo quality, as well as
quantitative and qualitative data regarding
associated accessory sperm. Both sperm access to
the ovum (addressed by accessory sperm) and
fertilization status/embryo quality were important
16

in addressing pregnancy rate for specific intervals
from the onset of estrus to insemination. Based
on these biomonitors, early insemination failed to
achieve optimum pregnancy rate due to
inadequate access of sperm to the ovum (i.e., low
fertilization rate, manifested by low accessory
sperm numbers). However, embryo quality was
high in early inseminations, which favors
pregnancy. Late insemination failed to achieve
optimum pregnancy rate (due to reduced embryo
quality), however, sperm access to the ovum was
highest. Thus, the selection of an insemination
time to achieve optimum pregnancy rate
appeared to be a compromise between the two
extreme intervals. For timed-AI programs,
consideration of the time of ovulation (and its
variability) becomes important, in addition to
conventional considerations, such as semen
handling, site of insemination, and bull selection.
saacke@vt.edu.
Journal of Dairy Science 91:3092–3101, 2008
Maintenance of Pregnancy in Dairy Cattle After
Treatment with Human Chorionic Gonadotropin or
Gonadotropin-Releasing Hormone
J. S. Stevenson,* S. M. Tiffany,* and E. K. Inskeep†
*Department of Animal Sciences and Industry, Kansas State
University, Manhattan 66506-0201. †Division of Animal and
Nutritional Sciences, West Virginia University, Morgantown 26506-
6108.
The objectives were to determine whether a
single injection of either human chorionic
gonadotropin (hCG) or GnRH would: 1) increase
ancillary formation of new luteal structures, 2)
increase serum concentrations of progesterone,
and 3) increase pregnancy survival in dairy
females treated once between 26 and 71 d of
pregnancy. A total of 421 cows were enrolled
between January and November 2001, with 92,
106, and 223 females (included 68 nulliparous
heifers at 1 location) treated at the 3 locations.
Upon diagnosis of pregnancy, females were
allocated randomly to receive 100 µg of GnRH,
1,000 IU of hCG, or 2 mL of saline. Blood samples
were collected at 0, 1, 2, and 4 wk after
treatment, and pregnancy status was reassessed
at 1, 2, and 4 wk. New luteal structures were
formed in 23.8% of cattle, with hCG (50%) and
GnRH (26%) being more effective than saline
(7%). Treatment had no effect on the proportion
of females forming 2 new luteal structures
(7.6%), and 36.2% of all induced structures
Noticias de Reproducción Bovina AGOSTO 2008
regressed during the 4- wk study period.
Pregnancy losses were unaffected by treatment,
stage of pregnancy, or number of induced luteal
structures but were nearly 9-fold greater in
females in which induced luteal structures
regressed. No loss occurred in females having 2
new luteal structures. Pregnancy losses decreased
quadratically from 30 to 42 d. Serum
progesterone did not differ among treatments, but
among females forming new luteal structures,
progesterone was greater at 1 (7.2 ± 0.3 vs. 6.3
± 0.2 ng/mL) and 2 wk (7.0 ± 0.3 vs. 6.1 ± 0.2
ng/mL) after treatment. Progesterone at the first
pregnancy diagnosis was predictive of imminent
pregnancy loss; the lower the initial progesterone,
the sooner subsequent loss was observed. The
right ovary was dominant in the location of new
luteal structures. Regression of new luteal
structures occurred more often on the left ovary
and contra-lateral to the corpus luteum of
pregnancy (53.2 vs. 22%). In conclusion,
treatment of dairy cattle with either GnRH or hCG
failed to prevent pregnancy loss, but
concentrations of progesterone were predictive of
subsequent pregnancy loss.
ss@k-state.edu
Journal of Dairy Science 91:3015–3024, 2008
Gossypol Disrupts Embryo Development in Heifers
M. Villaseñor,* A. C. Coscioni,* K. N. Galvão,* R. C. Chebel,*
and J. E. P. Santos*†
*School of Veterinary Medicine, University of California–Davis, Tulare
93274. †Department of Animal Sciences, University of Florida,
Gainesville 32611.
Our objectives were to determine the effects of
dietary free gossypol (FG) intake on plasma and
uterine gossypol concentrations and embryo
development and viability before and after culture
with gossypol. Fifty postpubertal Holstein heifers
weighing (±SD) 406 ± 34.5 kg at 11.5 mo of age
were blocked by age and body weight (BW) and
randomly assigned to 1 of 3 isocaloric and
isonitrogenous diets differing in their FG content:
control (0 mg of FG/kg of BW), moderate (17.8
mg of FG/kg of BW), and high (36.8 mg of FG/kg
of BW). Heifers were fed the diets for 70 d before
superovulation and embryo collection.
Superovulated heifers were flushed on d 5 after
induction of ovulation, and early morulae were
either stained, to determine the number and
proportion of live and dead cells, or randomly
assigned to an in vitro culture for 96 h in media
containing either 0 or 10 µg/mL of gossypol acetic
17

acid. Plasma and uterine gossypol concentrations
increased with increasing gossypol intake. The
number of low-quality embryos-ova was greater
for the high than for the moderate and control
diets. Embryos collected from the high diet had
the least number of cells because of fewer live
cells, and were smaller in diameter. Greater
dietary gossypol reduced blastocyst development
and extended the time to reach the blastocyst
stage. Similarly, gossypol concentration at 10
µg/mL compromised in vitro development and
increased the proportion of degenerated embryos
at 96 h in culture. These findings provide in vivo
and in vitro evidence that intake of 36.8 mg of
FG/kg of BW per d and gossypol concentrations
>7 µg/mL in plasma, in uterine flush, or in vitro
compromise early embryo development, which
might explain some of the negative effects of
gossypol on the fertility of dairy cows.
jepsantos@ufl.edu
Animal (2008), 2:8, pp 1153–1157
Effect of Escherichia coli Infection of the Bovine Uterus
From the Whole Animal to the Cell
E. J. Williams1, S. Herath1, G. C. W. England2, H. Dobson3, C.
E. Bryant4 and I. M. Sheldon1
1
Department of Veterinary Clinical Sciences, Royal Veterinary College,
Royal College Street, London, NW1 0TU, UK; 2The School of
Veterinary Medicine and Science, University of Nottingham, Sutton
Bonington Campus, Sutton Bonington, Leicestershire, LE12 5RD, UK;
3
Department of Veterinary Clinical Science and Animal Husbandry,
Faculty of Veterinary Science, University of Liverpool, Leahurst,
Neston, CH64 7TE, UK; 4Centre for Veterinary Science, Department
of Clinical Veterinary Medicine, University of Cambridge, Cambridge,
CB3 0ES, UK.
Following parturition, contamination of the uterine
lumen by bacteria is ubiquitous, and uterine
health is impaired in cattle because infection
persists in 10% to 15% of animals as
endometritis. Endometritis causes infertility for the
duration of infection, and subfertility persists even
after apparent successful resolution of the
disease. Escherichia coli is the pathogenic
bacterium most frequently isolated from the post
partum uterus, and is associated with increased
concentrations of peripheral plasma acute phase
proteins and fetid vaginal mucus. The presence of
E. coli is also associated with slower growth of the
first post partum dominant follicle and perturbed
oestradiol secretion. Furthermore, in animals that
ovulate the first dominant follicle, the corpus
luteum is smaller and secretes less progesterone.
The endotoxin lipopolysaccharide (LPS), which is
released from E. coli, can pass from the uterine
Noticias de Reproducción Bovina AGOSTO 2008
lumen to the peripheral circulation and LPS
concentrations are increased in cows with uterine
infection. Infusion of E. coli LPS into the uterine
lumen suppresses the pre-ovulatory luteinising
hormone surge and disrupts ovulation in heifers.
In vitro, endometrial explants produce
prostaglandins in response to LPS. Addition of LPS
or E. coli to stromal or epithelial cells increases
cyclooxygenase-2 mRNA expression, and
stimulates the production of prostaglandin E2 and
prostaglandin F2a. Furthermore, uterine and
ovarian cells express mRNA of the molecules
required for recognition of LPS, Toll-like receptor-
4 and CD14. In summary, E. coli is a common
cause of infertility involving the perturbation of
the hypothalamus, pituitary and ovary in dairy
cows.
erin.williams@ucd.ie
Animal (2008), 2:8, pp 1135–1143
Factors Influencing Heifer Survival and Fertility on
Commercial Dairy Farms
D. C. Wathes, J. S. Brickell, N. E. Bourne, A. Swalia and Z.
Cheng
Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield,
Herts AL9 7TA, UK.
The average dairy cow survives only three
lactations, reducing the availability of replacement
heifers. Prenatal losses occur due to early
embryonic mortality (about 40%), later embryo
loss (up to 20% in high-yielding herds) or
abortion (about 5%). A recent survey of 19 UK
herds showed that 7.9% of calves were born dead
and 3.4% died within 1 month. During the rearing
phase, 6.7% of animals were lost before reaching
first service at 15 months due to disease or
accident and another 2.3% failed to conceive.
Many potential replacements therefore never
enter the milking herd. This severely limits
opportunities for on-farm selection of breeding
cows in addition to presenting a welfare issue and
causing economic loss. The most profitable
animals once lactation is reached combine good
milk production with a regular calving pattern.
Some aspects of performance are related to age
at first calving (AFC), which in turn is influenced
by heifer growth rates. Poorly growing animals
required more services to conceive, calved later
and subsequently performed badly. Optimum
fertility and maximum yield in the first lactation
were associated with an AFC of 24 to 25 months.
However, heifers calving at 22 to 23 months
18

performed best in terms of total milk yield and
survival over the first 5 years, partly because
good heifer fertility was associated with better
fertility later. We have investigated some possible
juvenile predictors of future performance. Low-
birth-weight calves were more likely to come from
either primiparous mothers or older dams (31
lactations) with higher peak milk yields,
suggesting that the uterine environment may limit
prenatal calf growth due to competition for
nutrients with maternal growth or milk production.
Linear trait classification scores for frame size
show genetic correlations with longevity. The
skeletal measures of height and crown rump
length in 1-month-old calves was correlated to
subsequent stature, and frame size was correlated
to weight at 15 months. It may thus be possible
to predict performance from simple size
measurements as juveniles. Neither endogenous
nor stimulated growth hormone (GH) release in 6-
month-old calves were related to milk yield in the
first three lactations, but size of a stimulated GH
peak was positively related to milk energy values
in the first lactation. Cows with delayed ovulation
(.45 days) in the first lactation had a higher GH
pulse amplitude and lower IGF-I as a juvenile.
Cows that partition excess energy into milk in
their first lactation may suffer reduced longevity.
dcwathes@rvc.ac.uk
Animal Reproduction Science 107 (2008) 62–67
Effect of Oestrus Synchronization Methods on Oestrus
Behaviour, Timing of Ovulation and Pregnancy Rate
During the Breeding and Low Breeding Seasons in Nili-
Ravi Buffaloes
Hassan Mahmood Warriach, Aijaz Ali Channa, Nasim Ahmad
Department of Theriogenology, Faculty of Veterinary Science,
University of Veterinary and Animal Sciences, Lahore, Pakistan.
The objective of this study was to determine the
effect of oestrous synchronization methods on
oestrous behaviour, timing of ovulation and
pregnancy rate during the breeding and low
breeding seasons in Nili-Ravi buffaloes. In
Experiment 1, oestrous behaviour and timing of
ovulation were determined from (n = 34)
oestruses. The mean (± S.E.M.) time of ovulation
after the onset of standing oestrus was greater (P
< 0.05) in PGF2α-induced luteolysis (30.6±1.5 h)
compared to Ovsynch buffaloes (15.0±0.8 h). In
Experiment 2, pregnancy rates were compared
between two methods of synchronization
(detected oestrus and Ovsynch protocol) during
Noticias de Reproducción Bovina AGOSTO 2008
the breeding and low breeding seasons.
Pregnancy rates of buffaloes bred at detected
oestrus (62.5%) or by the Ovsynch protocol
(36.3%) during the breeding season did not differ
significantly (P > 0.05) from those which were
inseminated during the low breeding season
(55.5%) and (30.4%), respectively. This study
demonstrates clearly that (1) timing of ovulation
in Nili-Ravi buffalo is about 30 h after the onset of
standing oestrus and (2) buffaloes can be
successfully synchronized with optimum fertility
using either PGF2α alone (detected oestrus) or
using (Ovsynch protocol) during low breeding
season, to calve during the period when milk
availability is short.
drnasim@yahoo.com
Animal Reproduction Science 107 (2008) 30–35
High-Pressure Steam Sterilization of Previously Used
CIDR Inserts Enhances the Magnitude of the Acute
Increase in Circulating Progesterone After Insertion in
Cows
J.F. Zuluagaa,b, G.L. Williamsa,b
a
Animal Reproduction Laboratory, Texas A&M University, Agricultural
Research Station, 3507 Hwy 59E, Beeville, TX 78102-8571, USA.
b
Center for Animal Biotechnology and Genomics, Department of
Animal Science, Texas A&M University, College Station, TX 77843,
USA.
Objectives were to compare serum concentrations
of progesterone (P4) in ovariectomized cows
receiving (1) new, (2) re-used disinfected (DIS),
and (3) re-used autoclaved (AC) controlled
internal drug release (CIDR) inserts. Five
ovariectomized (OVX) beef cows were used in a
replicated 3×3 Latin square design. Each
experimental period was 7 days, with at least 48 h
between periods. All re-used CIDR had been
inserted previously in beef cows for 7 days. Upon
removal, CIDR used for the DIS treatment were
washed thoroughly and soaked in a chlorhexidine
gluconate solution (0.03%) for 2 h, rinsed
thoroughly with water and air-dried. For the AC
treatment, CIDR were not soaked in disinfectant
but were steam sterilized at 121 ºC and 724mm
Hg for 20 min before use. Blood samples were
collected at 0, 10, 30, 60, 180, and 480 min
relative to time of insertion of CIDR, daily until
day 7, and at 30, 60, and 180 min relative to time
of removal for radioimmunoassay of P4. Mean
serum concentrations (ng/mL) of P4 during the 7-
day period of insertion were greater (P < 0.03)
for new (3.7±0.2) and AC (3.4±0.3) than for DIS
19

CIDR (2.8±0.2). These effects were created
primarily by differences occurring during the first
8 h after CIDR insertion. Within this interval,
mean concentrations (ng/mL) differed (P < 0.05)
among all groups, but values for AC (6.0±0.7)
exceeded both new (4.6±0.5) and DIS (2.7±0.3)
markedly. Autoclaving may be the best option
when re-using CIDR inserts because it creates
greater concentrations of P4 immediately after
insertion and reduces maximally the risk of
disease transmission.
glwilliams@tamu.edu
Theriogenology 70 (2008) 550–559
Pregnancy-Associated Glycoproteins and Pregnancy
Wastage in Cattle
B.K. Whitlocka, H.S. Maxwellb
a
Department of Anatomy, Physiology & Pharmacology, College of
Veterinary Medicine, Auburn University, Auburn, AL 36849 USA.
b
Department of Clinical Sciences, College of Veterinary Medicine,
Auburn University, Auburn, AL 36849 USA.
Accurate diagnosis of non-pregnancy and prompt
re-enlistment of ‘‘non-pregnant’’ cattle into an
appropriate breeding protocol are essential
components of successful reproductive programs.
Various methods aimed at improving detection of
pregnancy and identification of non-pregnant
cows earlier and more accurately are the focus of
previous review articles and beyond the scope of
this manuscript. Recently, the ability to measure
pregnancy-associated glycoproteins (PAGs) in
cattle has changed how pregnancy and, more
importantly, non-pregnancy are detected. This
presentation provides an overview of current
research on the pregnancy associated
glycoprotein family, and how these glycoproteins
might be utilized as indicators of pregnancy
wastage in cattle.
whitlbk@vetmed.auburn.edu
Theriogenology 70 (2008) 535–549
Heritable Bovine Fetal Abnormalities
B.K. Whitlocka, L. Kaiserc, H.S. Maxwellb
a
Department of Anatomy, Physiology & Pharmacology, College of
Veterinary Medicine, Auburn University, Auburn, AL 36849, USA.
b
Department of Clinical Sciences, College of Veterinary Medicine,
Auburn University, AL 36849, USA. cDepartment of Medicine, College
of Human Medicine, Michigan State University, East Lansing, MI
48824, USA.
The etiologies for congenital bovine fetal
anomalies can be divided into heritable, toxic,
nutritional, and infectious categories. Although
uncommon in most herds, inherited congenital
anomalies are probably present in all breeds of
Noticias de Reproducción Bovina AGOSTO 2008
cattle and propagated as a result of specific trait
selection that inadvertently results in propagation
of the defect. In some herds, the occurrence of
inherited anomalies has become frequent, and
economically important. Anomalous traits can
affect animals in a range of ways, some being
lethal or requiring euthanasia on humane
grounds, others altering structure, function, or
performance of affected animals. Veterinary
practitioners should be aware of the potential for
inherited defects, and be prepared to investigate
and report animals exhibiting abnormal
characteristics. This review will discuss the
morphologic characteristics, mode of inheritance,
breeding lines affected, and the availability of
genetic testing for selected heritable bovine fetal
abnormalities.
whitlbk@vetmed.auburn.edu
Theriogenology 70 (2008) 631–637
Characterization of Estrus Detection, Conception and
Pregnancy Risk of Holstein Cattle From the Central
Area of Chile
P. Melendeza,c, M. Duchensb, A. Perezb, L. Moragab, L.
a
Archbaldc
College of Veterinary Medicine, University of Santo Tomas, Av.
Limonares 190, Vin˜a del Mar, Chile. bCollege of Veterinary Medicine,
University of Chile, Av. Sta. Rosa 11735, La Pintana, Santiago, Chile.
c
College of Veterinary Medicine, University of Florida, PO Box
100136, Gainesville, FL 32610-0136, USA.
The objectives were to characterize the estrus
detection risk (HDR), conception risk (CR), and
pregnancy rate (PR) of postpartum (pp) Holstein
cattle from the central area of Chile. The study
used records of 2269 lactations from six dairy
farms in central Chile (Mediterranean-type
climate) during 2004. Three 21-d periods for
estrus detection were considered (50–70, 71–91,
and 92–112 d pp). Estrus detection risk, CR, and
PR at the first, second, and third periods were
analyzed by logistic regression, whereas overall
PR at the end of the 63-d study (112 d pp) was
assessed with survival analysis. The overall HDR
was 51.1%. The HDR, CR, and PR were 48.4,
42.2, and 17.3%, respectively, during the first
period; 52.8, 41.8, and 20.5% during the second
period; and 52.9, 39.2, and 19.7% during the
third period. The HDR was lower during Period 1
than during Periods 2 and 3 (P ≤ 0.05).
Conception risks were not different among periods
(P > 0.05); however, PR was lower during Period
1 than during Periods 2 and 3 (P ≤ 0.05). Overall
PR over time differed among parities, but was not
20

significantly different among seasons. There were
no significant interactions among parity, season
and herd for HDR, CR and PR for the three 21-d
periods. Parity 1 had higher CR and PR than Parity
2 and 3+ during Period 3. Overall, survival curves
for the risk of non-pregnancy among parities (1,
2, 3 or greater) were different over time (P ≤
0.05). Cows of Parity 1 became pregnant earlier
than cows of Parity 2, and Parity 3 or greater.
Survival curves for the risk of non-pregnancy
among seasons (summer, fall, winter, and spring)
were not different over time (P > 0.05).
melendezp@vetmed.ufl.edu
Livestock Science 117 (2008) 79–87
Effect of banding or burdizzo castration on plasma
testosterone, acute-phase proteins, scrotal
circumference, growth, and health of bulls
a
W.Y. Panga,b, B. Earleya, V. Gathb, M.A. Croweb
Teagasc, Grange Beef Research Centre, Dunsany, Co. Meath,
Ireland. bSchool of Agriculture, Food Science & Veterinary Medicine,
University College Dublin, Belfield, Dublin 4, Ireland.
The objective was to assess the effect of banding
or burdizzo castration performed on farms on
plasma testosterone, acute-phase proteins, scrotal
circumferences, growth, and well-being of bulls.
243 Continental bulls (12 months; 399.2±5.72 kg)
from three different farms were allocated at
random, after stratification on weight within breed
type, to one of three treatment groups: banding
castration (BAND; n=80), burdizzo castration
(BURD; n=83), or controls (CON; n=80). The
castration methods were conducted under local
anaesthesia, and tetanus toxoid vaccine and
antibiotic were also injected at castration. BAND
and BURD castrates had lower (Pb0.001) plasma
testosterone concentration than control bulls, with
no difference between BAND and BURD castrates
on 28 d post-castration. From days 0 to 14 post-
castration, BAND (P=0.0002) and BURD
(Pb0.0001) castrates had lower average daily gain
Noticias de Reproducción Bovina AGOSTO 2008
(ADG) than CON bulls, no difference (P=0.46)
was found between BAND and BURD castrates.
From days 15 to 28, BAND castrates had lower
ADG compared with BURD castrates (P=0.03) and
CON bulls (P=0.01), while no difference (P=0.76)
was found between BURD and CON. From days
29 to 56, BAND (P=0.01) and BURD (P=0.002)
castrates had lower ADG than CON bulls, no
difference (P=0.55) was found between BAND
and BURD. From days 57 to 84, the ADG of BAND
castrates was not different compared with BURD
castrates (P=0.12) and CON bulls (P=0.38), while
BURD had lower (P=0.02) ADG compared with
CON. The integrated ADG from day 0 to 112 of
BAND (P=0.0001) and BURD (P=0.02) groups
were lower compared with CON, while there was
no difference (P=0.09) between BAND and BURD
castrates. On d 14 post-castration, BAND
castrates had lower scrotal temperature than
BURD (Pb0.0001) and CON (Pb0.0001), and BURD
castrates had greater (Pb0.006) scrotal
temperature than CON; BAND castrates had lower
scrotal latitudinal and longitudinal circumferences
than BURD castrates (Pb0.001) and CON bulls
(Pb0.001), and BURD castrates had greater
(Pb0.001) scrotal latitudinal and longitudinal
circumferences than CON bulls. BAND (Pb0.0001)
and BURD (P=0.01) castrates had greater glucose
concentration than CON bulls, and BAND castrates
had greater (P=0.04) glucose concentration than
BURD. In conclusion, BAND or BURD castration
significantly reduced plasma testosterone
concentration; reduced average daily weight gain
mainly during the first 2 weeks, which was not
compensated during the subsequent 16 weeks;
increased withdrawal of stored energy and
increased plasma protein concentration. BURD
showed an advantage over BAND in growth
during days 15 to 28 following castration.
bernadette.earley@teagasc.ie
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Noticias de Reproducción Bovina AGOSTO 2008

Fuente: Gaceta UNAM

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 Noticias de Reproducción Bovina AGOSTO 2008

Próximos eventos
XXI CONGRESO PANAMERICANO DE CIENCIAS
VETERINARIAS Y 16 REUNIÓN ANUAL DEL
CONASA
Asociación Panamericana de Ciencias Veterinarias
Consejo Técnico Consultivo Nacional de Sanidad
Animal
Fecha: 12 al 16 de octubre de 2008
Sede: Guadalajara, Jal
Informes: www.panvet2008.com
8º CONGRESO INTERNACIONAL DE MVZ
ESPECIALISTAS EN BOVINOS
Asociación de Médicos Veterinarios Especialistas
en Bovinos de la Laguna
Fecha: 6, 7 y 8 de Noviembre de 2008
Sede: Auditorio del Centro Cultural Universitario
“Braulio Fernández Aguirre” de la Universidad
Autónoma de Coahuila. Torreón, Coah, México
Informes: asociación@amveblaguna.com

Sitios de interés
Secretaría de Agricultura, Ganadería, Desarrollo Rural, Pesca y Alimentación
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www.fedmvz.com

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www.fao.org

Organización Panamericana de la Salud.

www.cinu.org.mx/onu/estructura/mexico/org/ops.htm

Revista Veterinaria México

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Dirección General de Salud Animal

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Noticias de Reproducción Bovina AGOSTO 2008

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