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Reprod Dom Anim 44, 384–389 (2009); doi: 10.1111/j.1439-0531.2007.01021.

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ISSN 0936-6768

Formation of Corpora Lutea and Central Luteal Cavities and Their Relationship with
Plasma Progesterone Levels and Other Metabolic Parameters in Dairy Cattle
C Perez-Marin
Medicina y Cirugia Animal, Universidad de Cordoba, Espan˜a

Contents
The corpus luteum (CL) may be looked upon as a compact or
cavitary structure. A number of papers have addressed the
relationship between CL type and parameters such as fertility
or progesterone levels. The present study assessed the morphological and functional sequence observed in cows with
different CL types, comparing pre-ovulatory follicle size,
progesterone levels, luteal tissue formation and some blood
biochemical parameters (calcium, albumin, inorganic phosphorus, glucose, magnesium, copper and zinc), oestrus cycle
length and oestrus expression, as a function of CL type.
Twenty-eight lactating dairy cows from two commercial dairy
farms in southern Spain were studied. Oestrus detection was
performed by monitoring daily oestrus behaviour, and artificial insemination (AI) was perfomed using the AM ⁄ PM rule.
Ovaries and uterus were sonographically examined and blood
samples were collected to measure progesterone and various
biochemical parameters. There was a slight tendency towards
the appearance of luteal cavities when pre-ovulatory follicles
were larger (1.9 ± 0.2 vs 1.7 ± 03; p = 0.074). Fertility was
not affected by cavity presence (cavity = 42.9% and compact = 57.1%; n.s.). Luteal tissue and function were not
modified as a function of CL type. Cows with luteal cavities
displayed significantly higher levels of albumin, suggesting a
possible metabolic influence on the formation of these structures, although specific research is required to confirm this
observation.

Introduction
The literature contains descriptions of luteal cavities
based on the examination of excised ovaries from
slaughterhouses or on rectal palpation (Hammond
1927; McEntee 1958; Morrow et al. 1966; Donaldson
and Hansel 1968). However, it was not until the advent
of ultrasonography that more detailed knowledge of
luteal development and function could be acquired, or
the different types of luteal gland described (Pierson and
Ginther 1984; Kito et al. 1986; Kastelic and Ginther
1989; Kastelic et al. 1990a,b; Grygar et al. 1997;
Battocchio et al. 1999; Veronesi et al. 2002).
Cavitary corpus luteums (CL) were first reported by
Pierson and Ginther (1984), but cavities were not fully
described until 3 years later (Pierson and Ginther 1987).
Kastelic et al. (1990a,b) subsequently provided an ultrasonographic description of CLs and studied the correlation between the presence of luteal cavities and plasma
progesterone levels. Interesting results were reported by
Kito et al. (1986) from ultrasonographic assessment of
luteal cavities on days 5, 10, 16 and 21 and every 3 days
until their disappearance. These authors asserted that
the luteal cavity should not be seen as a pathological
structure, as had hitherto been the case. More recently,
Grygar et al. (1997) provided more detailed data

acquired by monitoring ovaries on days 4, 9, 20, 25,
30 and 40. Garcı´ a and Salaheddine (2000) have suggested that neither the presence nor the size of luteal
cavities affect the success and maintenance of gestation
in cows in an embryo-transfer program.
Researchers agree that the presence luteal cavity has
no detrimental effect on fertility. However, Grygar et al.
(1997) reported that cows with luteal cavities displayed
higher plasma progesterone levels, a finding not
reported by other authors. Although no previous results
confirm the direct involvement of biochemical parameters in luteal cavity formation, it has been suggested that
energy deficiency or uterine disorders (together with
associated chemical disorders) could contribute to CL
formation (Gabor et al. 2004). On this assumption, it
could be hypothesized that variations in blood biochemical parameters may modify the function and also
the morphology of the developing CL.
The present study assessed morphological and functional sequence in CLs, using ovarian US and progesterone RIA, respectively, in pregnant and non-pregnant
cows during natural oestrous cycles. The following were
measured: pre-ovulatory follicle size, progesterone levels, luteal tissue formation and metabolic parameters in
all cows, as well as oestrous cycle length and oestrous
expression in non-pregnant cows; results for different
CL types (compact vs cavitary) were compared.
The aim of the study was to monitor changes in luteal
morphology and simultaneous changes in circulating
progesterone levels, pre-ovulatory follicle size, oestrous
duration and metabolic parameter in cycling and
pregnant cows with cavitary or compact CLs.

Material and Methods
Animals and protocol
Twenty-eight lactating Holstein-Friesian cows from two
commercial dairy farms in southern Spain (from 38N
to 4W) were housed in open-air stalls with sheltered
areas, and fed with a balanced diet twice daily following
National Research Council Nutritional Guidelines. A
physical examination was conducted during spontaneous oestrous cycles to rule out congenital and acquired
defects of the reproductive organs. All cows included in
the study, of 3–7 years of age, had calved at least once
and presented good body condition. They were tested
for being free of tuberculosis and brucellosis. All cows
were milked twice daily, at 6:00 and 18:00 h. Average
annual milk yield was 7500 kg per animal. Oestrous
detection was performed by monitoring daily oestrous
behaviour three times a day for 20 min. The main 

2008 The Author. Journal compilation  2008 Blackwell Verlag

The SPSS 11. each animal had been examined a total of nine times. USA) was used for statistical analysis.0 days after AI.2 cm2). If a cow returned to oestrus before pregnancy diagnosis. anti-progesterone serum and 3H-progesterone. Journal compilation  2008 Blackwell Verlag . the area under the curve (AUC) was calculated as (ng · ml) · time. Similar statistical analyses were carried out to establish differences between different reproductive statuses. Blood was drawn from a coccygeal vessel into vacuum tubes containing lithium heparin. For extraction. luteal tissue area and metabolic levels. Ovine anti-progesterone serum was supplied by Dr Butcher of West Virginia University. Chi-squared analysis indicated no significant difference in luteal cavity presentation between groups. Spain) was used to measure metabolic parameters. 1.6%) exhibited a CL containing a central cavity 4 days after oestrous detection. inorganic phosphorus. with the aid of commercial kits (Spinreact. If the CL was cavitary. Japan) equipped with a 5. 100% 80% 60% 40% 20% 0% 4 8 12 16 20 Days after insemination 24 28 32 Fig. approximately 12 h. Barcelona.03–0. A functional CL was considered to be present if plasma progesterone levels were above 1 ng ⁄ ml.03 cm2). To acquire more detailed data on this issue. Differences were considered statistically significant at p £ 0. using the trapezium rule. and large cavity (>0. sampling was continued until confirmation of pregnancy. Biochemical parameters Plasma biochemical parameters were measured to determine metabolic status. By the end of the 32-day study period. some cows were sampled for more than one cycle. IL. oestrous expression. Ovaries were scanned several times in lateromedial and dorsoventral planes to determine the position and the diameter (height and width) of follicles.4% (n = 8) and 13. glucose. A BTS 310 photometer (Biosystem. Samples were immediately centrifuged at 1000 · g for 20 min. Pregnancy diagnosis was made on day 32 by transrectal ultrasonography of the uterus. the reproductive tract of each cow was examined ultrasonographically every 4 days. Chicago. Ovarian ultrasound examination From day 0 (onset of oestrus). CL was monitored by ultrasonography and was characterized as intra or projected ovarian structures with thickened wall.8 cm2). and incubated at 4C for Presence and description of cavities in pregnant and nonpregnant cows Luteal cavities were recognized in 42.and inter-assay variation coefficients were 10. cavity area was not taken into account in the measurement. a study was carried out to determine calcium. Spain).21– 0. copper and zinc levels over 32 days following AI. 3 ml of hexane were mixed with 200 ll of plasma. medium cavity (0. CL was considered absent or non-functional. Tokyo. after the oestrus was observed. The separated plasma was then refrigerated for 3–6 h and subsequently frozen at )20C until assayed.05. Eight of 28 cows (28. Barcelona. To minimize bias.6% (n = 6) respectively. 385 24 h.8 cm2). data for daily progesterone levels. To chart progesterone availability over time as an indicator of hormone production. Separation of free. followed by 14 ⁄ 28 (50%) on day 8 after oestrous detection and two cows (7%) on day 32 (Fig.0 MHz linear-array transducer. all cows were inseminated by the same technician. Cavities were detected for the first time 5. Examinations were performed using an ultrasound scanner (Aloka SSD-210 DX II. corpora lutea and other structures. intra.1% (8 ⁄ 14) of pregnant and non-pregnant cows respectively. Results Plasma progesterone levels Plasma progesterone levels were measured by radioimmunoassay (RIA) using liquid-phase extraction. The progesterone dry residue obtained was mixed with buffer.0 package (SPSS. 1). magnesium. Four groups were established based on luteal cavity area (Kastelic et al. (1990b). All data are presented as arithmetic means (mean ± SD). If progesterone levels were below 1 ng ⁄ ml. small cavity (0. 1990b): no cavity (<0. with or without a hypoechogenic central fluid-filled cavity at the site of the former pre-ovulatory follicle.9% (6 ⁄ 14) and 57. Incidence of corpora lutea containing central fluid-filled cavities on day of estrous detection  2008 The Author. and no statistically significant % Cows with CL containing central luteal cavity Blood sampling Blood samples were collected every 4 days from day 0 (=AI) to day 32. Diagrams were made to chart changes in these structures on subsequent monitoring.7 ± 2.from bound steroid was accomplished using dextran-coated charcoal. Assay sensitivity was 16 pg ⁄ ml. These cows were analysed to determine whether the pre-ovulatory follicle was involved in cavity formation and whether cows usually presented cavities. Statistical analysis Several ANOVA tests were performed to determine the possible relationship between the CL cavity and other variables such as pre-ovulatory follicle size. total progesterone production. Cross-sectional area (also termed total luteal tissue) was calculated as reported by Kastelic et al. and confirmed by the subsequent visualization of a CL.Luteal Cavities in Dairy Cows criterion for oestrous detection was the observed cow’s acceptance of mounting by other cows. Ovulation was defined as the disappearance of a large follicle (>9 mm) identified by ultrasonography at the previous examination. Therefore. using high-quality frozen-thawed semen.

82 cm respectively.3 ± 8.05). p > 0. no significant difference was found in average values for pregnant and non-pregnant cows (1. but no cavities were identified. 3. 2. although differences were not Fig. detection took place earlier when cavities reached higher diameters in both groups.8 days) than those with compact CL (20. Journal compilation  2008 Blackwell Verlag . Variations in the duration of luteal cavities as a function of size Oestrous cycle duration and luteal cavities Cows that failed to become pregnant were monitored (using clinical signs and ⁄ or baseline plasma progesterone levels) to determine oestrous cycle duration. However. 2).0 days.9 days in non-pregnant.1 ± 2. there was a slight tendency towards cavity presentation when preovulatory follicles were larger (1. p = 0.88 and 1.7. 3).4 days.0 ± 5. Nonpregnant cows with central luteal cavities displayed a significantly longer oestrous cycle (23.05) (Fig.2 duration was longer. Note the ovulatory follicle on day 0  2008 The Author. no increase was observed because of the larger amount of secretory luteal tissue. plasma progesterone levels remained within a normal range. with no significant difference as a function of reproductive status (11.4 0 0 4 8 12 16 20 24 Day after insemination 28 32 Fig.8 day vs 10.1 day in non-pregnant. p > 0. Non-pregnant cows reached the maximum diameter sooner than pregnant cows (8.386 C Perez-Marin differences were observed as a function of reproductive status (5.1 ± 7.0 day in pregnant vs 6. Large cavity 0. Double ovulation was observed in another pregnant cow. Ultrasonographic evolution of the luteal cavity in a pregnant cow.0 ± 1. In one pregnant cow. p > 0.6 Small cavity Medium cavity 1. The cavities displayed continuous growth.00 ± 2.014).074).7 ± 0. although not significantly so.0 ± 2. reaching their maximum size on day 9.8 0.9 days in pregnant and 11. However. cavity duration extended beyond 32 days after AI (Fig.5 ± 2.5 ± 1.3 ± 2.2 cm vs 1.9 ± 0.5). No association was observed between the day of maximum cavity diameter and luteal cavity classification. Average cavity duration was 11.3 cm. Total CL size and luteal tissue area were calculated by adding both CLs. Pre-ovulatory follicle and luteal cavities On oestrous day an ultrasonographic assessment was conducted to determine the pre-ovulatory follicle diameter. The cycle also lasted longer when the cavity was larger. Cavity 2 Cavity size 1. This accounts for recorded values being higher than in other animals. in CLs with large cavities. p > 0. p = 0.05).5 day. However.

In the present study. As progesterone is synthesized by luteal tissue.3 ± 1. The maximum cavity size was reached in around 8–12 days after AI.5 days. In a similar study. However. Cavitary CLs were found in 42. as reported by other authors (Kito et al. (1986). 5. It is reasonable to assume that if the refilled cavity is larger in size (i. magnesium or copper) 8 12 16 20 24 28 Time interval (days) after insemination 32 Fig.9% of pregnant cows.28 ± 3. showing that cavities have no negative effect on fertility. medium cavity: 22. this rate was lower than that reported by Kastelic et al. which is later than the span of time reported by Kastelic et al. Garcı´ a and Salaheddine (2000) reported that luteal cavity presentation was not associated with induced or spontaneous oestrus. Zinc levels tended to be slightly higher in cows with luteal cavities (156. Kastelic et al. 1986.26 lg ⁄ dl vs 137.93 ± 33. 1990b). subtracting the luteal cavity area. Discussion Fifty per cent of the cows studied displayed cavitary CL. There was no variation in luteal tissue as a function either of the presence or the size of luteal cavities. the luteal tissue takes more time to refill and constitute the luteal matrix.0 ± 2. p = 0. 5). Pregnant cows displayed significantly higher levels than non-pregnant cows from day 20 onwards. if the pre-ovulatory follicle is larger in size at ovulation time). p = 0. Kito et al.079).015). without hormonal synchronization treatment. as well as a cavity that is the remnant of the antrum of the follicle (Zheng et al. 4).8 days. No references have been found to any association between pre-ovulatory follicle diameter and luteal cavity presence. Journal compilation  2008 Blackwell Verlag .76 g ⁄ l vs 27.09 g ⁄ l. Corpus luteum size and luteal cavities Luteal ultrasonographic monitoring was carried out over 32 days following AI. However. Morrow et al. (1990b). Area under the curve of progesterone during the oestrous cycle in cows with or without central luteal cavity (data show means with bars for SD) showed no variation as a function of the presence of luteal cavities (Table 1). glucose. (1998).05). Pierson and Ginther (1988) and Viana et al. Pierson et al. there was no variation in total luteal size as a consequence of the presence or absence of luteal cavities (Fig. inorganic phosphorus. (1966). It has been suggested that cavitary CL formation is because of premature closing at the ovulation site (McEntee 1990). reproductive status differed significantly from day 20 onwards (p < 0. (1990b). Results show that the luteal cavity appears more frequently when pre-ovulatory follicles are larger. an early CL is formed by several tissue infoldings from the collapsed follicular wall. Average values obtained for the other parameters measured (calcium.5 and day 7. 1993).05). Luteal function and luteal cavities The presence of a luteal cavity did not influence progesterone production throughout the complete oestrous cycle (Fig. Quirk et al.67 lg ⁄ dl. The presentation of different classes of luteal cavities was  2008 The Author. Donaldson and Hansel (1968). but higher than those found by Rajakosky (1960). Chi-squared analysis showed that formation of luteal cavities was not related to silent or overt heat expression in the next oestrous cycle. However.90 ± 13. Corpus luteum size during the oestrous cycle in cows with or without central luteal cavity (data show means with bars for SD) significant (large cavity: 23. 1986.17 ± 2.0. located the maximum diameter between day 5.Luteal Cavities in Dairy Cows 387 200 9 Compact CL Cavitary CL 180 Progesterone AUC (ng/ml × time) CL size incows with or without CL 8 7 6 5 4 3 2 Compact CL Cavitary CL 160 140 120 100 80 60 40 20 1 0 0 4 0 4 8 12 16 20 24 Days after insemination 28 32 Fig. a slight tendency was noted towards higher progesterone levels in pregnant cows from day 8. Metabolic parameters and luteal cavities Albumin levels were significantly higher when luteal cavities were present (30. 1988. it was considered reasonable to analyse only the amount of luteal tissue.e. Results were similar to those obtained for total luteal size. cows were monitored during spontaneous cycles. 4. p > 0.

cows with cavitary CL were also found to have slightly. 71. Moreover. Grygar I. B = 2. Kudlac E. 4th edn.  2008 The Author. in which case fertility is negatively affected. Progesterone profiles and luteal tissue areas did not vary as a function of CL type. This metabolite has a short average lifetime. Journal compilation  2008 Blackwell Verlag . Gabai G. Reprod Dom Anim 35. phosphorus.78 ± 0. ruling out the presence of ovarian pathological structures. Rajamahendran et al. Hansel W. pp. it is reported that very large fluid-filled cavities may in fact be CLs with a large cavity. 5. but not significantly. Results showed that the progesterone AUC was not modified by the type of CL (cavitary or compact). even though the cavity was very large (>15 mm). Dolezel R. and can be used as a protein deficiency sensor for 1 or 2 months (Van Saun 1999).93 ± 33. Average levels of metabolic parameters over the course of the study Compact Cavitary Calcium (mg ⁄ dl) Albumin (g ⁄ l) Magnesium (mg ⁄ dl) Glucose (mg ⁄ dl) Phosphorus (mg ⁄ dl) Zinc (lg ⁄ dl) Copper (lg ⁄ dl) 6. 1989: Lipids metabolism and its disorder. as reported by Kastelic et al. pp.80 ± 23. 1997: Volume of luteal tissue and concentration of serum progesterone in cows bearing homogeneous corpus luteum or corpus luteum with cavity. Only one pregnant cow displayed a very large cavity (area = 5. Canada. Fertility was not affected by luteal cavity presence. (2004). Donaldson L.86 n.26 ** 82. 37th Annual Meeting of Society for the Study of Reproduction. 35–41.05. London. Grygar et al.17 ± 2. University of British Columbia.6 cm) that was maintained for longer than 28 days. although specific research is required to confirm this observation. 27. There was no difference in luteal areas between compact and cavitary CLs. (1997) report that cavitary CLs in pregnant cows contain greater luteal tissue volume and secretory activity than homogeneous CLs.27 n.59 ± 17. Mollo A. Clinical Biochemistry of Domestic Animals. Nedbalkova J. Hammond J.77 ± 0.34 n.28 ± 3. 107– 141. while large cavities were detected earlier (around day 4) and lasted longer than small or medium cavities (detected on day 8).). Albumin is an important protein and aminoacid transporter reservoir.99 n.16 ± 1. however.26 82.s. cavitary CLs were similar in size to compact CLs. as has been recently confirmed by Garcı´ a and Salaheddine (2000) in embryo transfer recipient cows. higher concentrations of zinc. Garcı´ a A..57 n. 113– 118. (1986). suggesting a possible metabolic influence on the formation of these structures. It is reasonable to suppose that that these biochemical variations may modify CL formation and govern the appearance of luteal cavities. Theriogenology 51.s. Cambridge University Press.76 * 2. beta-carotene levels. Similar findings were noted by Kito et al.89 ± 19.388 C Perez-Marin Table 1. the authors reported no correlation between cavity size and serum calcium.09 30. Salaheddine M. in terms of both total luteal size and luteal tissue volume. These results need to be carefully analysed and confirmed by further testing. similar in pregnant and non-pregnant cows.19 ± 0. 1968: Cystic corpora lutea and normal and cystic Graafian follicles in the cow.18 ± 0. London. Pierson and Ginther (1988). (1986) who found no decrease in pregnancy rates. Studies in cows and buffaloes suggest that a reduction in blood albumin concentrations may be prompted by liver dysfunction. Battocchio M. supporting the view that the cavity does not impede the normal growth of luteal tissue.66 7. Soldado F. **p < 0. suggesting that the presence of a luteal cavity does not infuence luteal function. A preliminary paper by Gabor et al. In conclusion. References Bartley JC. Bono G. a view also advanced by Kito et al. 2004: Preliminary comparison of luteal cavity size with some serum metabolic parameters in dairy cows. it is also involved in mobilizing fatty acids (Bartley 1989). the results obtained showed that the luteal cavity had a slight tendency to be present when the pre-ovulatory follicle was larger. (1990a). 1927: The Physiology of Reproduction in the Cow. Results obtained here agreed with those of Gabor et al.s.73 73. Cairoli F. 304–308. in which case reproduction could be negatively affected. nor replace it. Veronesi MC. The cavities were filled with luteal tissue up to the point of disappearance.s. In: Kaneko JJ (ed. (1990a. 137.67 156. Academy Press Inc. glucose. the cavity was visible in cows with higher levels of albumin. Spain. Toth F.04 ± 22.90 ± 13. Acknowledgements This research was supported by regional project PIR-9520 (Andalusian Regional Government). 1999: Agreement between ultrasonographic classification of the CL and plasma progesterone concentration in dairy cows. Aust Vet J 44. Mezes M. in that calcium. Plasma albumin levels are indicative of whether an animal is capturing or eliminating proteins. Gabor G.97 ± 0.b). Anim Reprod Sci 49.41 2. The preovulatory follicle and CL formation were monitored here. (2004) compared luteal cavity size with some serum metabolic parameters.97 5. as observed in the present study. In contrast to these observations. Numerous studies report major metabolic imbalances affecting reproduction. binding and transporting other proteins. the differences were not significant. as previously reported in the literature. However.s. urea. or may be luteinized follicles or luteal cysts.1.3 cm2. which was around 8–12 days after formation. Here. phosphorus and glucose levels showed no correlation with luteal cavity presence. In the present study. (1989) and Kastelic et al. Small luteal cavities were associated with more luteal tissues and increased luteal size. *p < 0. 77–82. Vancouver. or aminotransferase and lactate dehydrogenase activities. total protein. 2000: Ultrasonic morphology of the corpus lutea and central luteal cavities during selection of recipients for embryo transfer. 1059–1069.

325–333. 1998: Avaliac¸ao ultra-sonogra´fica do corpo luteo em novilhas mestic¸as utilizadas como receptoras de embriao. San Diego. Redmer DA. Ginther OJ. 61–68. 1960: The ovarian follicular system in sexually mature heifers with special reference to seasonal.Luteal Cavities in Dairy Cows Kastelic JP. 1177–1189. progesterone and luteinizing hormone levels. Van Saun RJ. Reynolds LP. Viana JHM. Pierson RA. Theriogenology 21. Theriogenology 29. Reproductive Pathology of Domestic Mammals. 21–37. 1989: Fate of conceptus and corpus luteum after induced embryonic loss in heifers. Fortune JE. milk yield. Morrow DA. Theriogenology 29. J Reprod Fert 77. Torres CAA. 929–936. Kastelic JR. Ginther OJ. 495–504. 1984: Ultrasonography of the bovine ovary. km. Leon. Bono G. 2002: Ultrasonographic appearance of tissue is a better indicator of CL function than CL diameter measurement in dairy cows. 1993: Vascular development and heparin-binding growth factors in the bovine corpus luteum at several stages of the estrous cycle. 1990: Cystic in and around the ovary. Fernandes CA. 42–47. Walton JS. 16–18 April. Sato K. Theriogenology 31. Gray HG. 140/4 Carodoba. Mollo A. Ginther OJ. Quirk SM. Departamento Medicina y Cirugia Animal. Madrid-Cadiz. Ginther OJ. Ferreira AM. Academic Press Inc. Theriogenology 58. 1–64. 1990a: Relationship between ultrasonic assessment of the corpus luteum and plasma progesterone concentration in heifers. Spain. Roberts SJ. Rajamahendran R. Miyazawa K. Pierson RA. 1596–1609. Desbottes S. Spain. Pierson RA. 1986: Study on the appearance of the cavity in the corpus luteum of cows by using ultrasonic scannig. 1990b: Ultrasonic morphology of corpora lutea and central cavitites during the estrous cycle and early pregnancy in heifers. Cairoli F. 1966: Postpartum ovarian activity and uterine involution in dairy cattle. 120–128. J Amer Vet Med Assoc 194. Theriogenology 34. Gabai G. 1999: Valoracio´n de los problemas de la vaca en transicio´n: perfiles metabo´licos modificados. Theriogenology 28. 1988: Ultrasonic imaging of the ovaries and uterus in cattle. Ginther OJ. Pierson RA. Hickey GJ. 1987: Reliability of diagnostic ultrasonography for identification and measurement of follicles and detecting the corpus luteum in heifers. Journal compilation  2008 Blackwell Verlag . McEntee K. J Am Med Assoc 149. McEntee K. Pierson RA. 3–20. Biol Reprod 47. Kastelic JP. Universidad de Cordoba. 3ª Jornadas ANEMBE de Medicina Bovina. pp. Int J Fertil 3. Archiv Reprod Anim 5. Submitted: 15 Aug 2007 Author’s address (for correspondence): Carlos Perez-Marin. Kito S. 211–219. Theriogenology 25. In: McEntee K (ed.es  2008 The Author. Veronesi MC. 922–928. Ginther OJ. 1988: Basic principles and techniques for transrectal ultrasonography in cattle and horses. body temperature. 1173–1182. Facultad de Veterinaria.). 1269–1278. E-mail: pv2pemac@uco. Battocchio M. Theriogenology 33. Ctra. Soldado F. and ovulation in dairy cows. Okuda K. Ginther OJ. cyclical and left-right variations. Acta Endocrinol 34. 1989: Temporal relationships among estrus. 52–68. Campus of Rabauales. Zheng J. Robinson J. Rajakosky E.. 487–498. Kastelic JP. 1986: Growth and regresion of ovarian follicles during the follicular phase of 389 the oestrus cycle in heifers undergoing spontaneous and PGF2alpha-induced luteolysis. McEntee K. 1958: Cystic corpora lutea in cattle. 396. Bergfelt DR.