Detection of bacteria in bovine ovarian follicular fluid.

The aim of this study was to examine the sterility of follicular fluid in large ovarian follicles in dairy cows. In all, 17 samples of paired follicular fluids and uterine contents collected from post-slaughtered dairy cows were cultured to detect aerobic and facultative anaerobic bacteria. Furthermore, the origin of the bacterial isolates from samples of follicular fluid and the uteri was also investigated using PFGE analysis. Follicular fluid concentrations of lipopolysaccharides were also determined. Of 17 uterine samples, 15 (88%) were detected as contaminated. In total, nine different bacterial genera and species were identified in the uterine and follicular fluid samples. Escherichia coli was the most prevalent bacterial species isolated from the uterine samples. Out of seven isolates of Staphylococcus aureus from the uterine samples, 6 (85%) were coagulase positive. Six isolates of Staphylococcus spp. were identified in 6 out of 17 follicular fluid samples (35%). Two out of six isolates were identified as Staphylococcus aureus (33%). Our results show that ovarian follicular fluid is not sterile in the bovine. The presence of Staphylococcus aureus in follicular fluid may partly explain the occurrence of infertility in some dairy cows. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of the present study show that ovarian follicular fluid is not sterile in bovines. The presence of Staphylococcus aureus in follicular fluid may partly explain the occurrence of infertility in some dairy cows.

A confident source of hard X-rays: radiation from a tokamak applicable for runaway electrons diagnosis.

In a tokamak with a toroidal electric field, electrons that exceed the critical velocity are freely accelerated and can reach very high energies. These so-called `runaway electrons' can cause severe damage to the vacuum vessel and are a dangerous source of hard X-rays. Here the effect of toroidal electric and magnetic field changes on the characteristics of runaway electrons is reported. A possible technique for runaways diagnosis is the detection of hard X-ray radiation; for this purpose, a scintillator (NaI) was used. Because of the high loop voltage at the beginning of a plasma, this investigation was carried out on toroidal electric field changes in the first 5 ms interval from the beginning of the plasma. In addition, the toroidal magnetic field was monitored for the whole discharge time. The results indicate that with increasing toroidal electric field the mean energy of runaway electrons rises, and also an increase in the toroidal magnetic field can result in a decrease in intensity of magnetohydrodynamic oscillations which means that for both conditions more of these high-energy electrons will be generated.

Cumulus cell expansion and first polar body extrusion during in vitro oocyte maturation in relation to morphological and morphometric characteristics of the dromedary camel ovary.

The morphological and morphometric characteristics of the ovary are fundamental properties for in vitro oocyte maturation. Nuclear maturation, including first polar body (1PB) extrusion, cytoplasmic maturation and cumulus cell (CC) expansion are the criteria for in vitro maturation (IVM) of oocyte. This study was designed to determine the effect of morphological and morphometric features of the ovary on CC expansion and 1PB extrusion during IVM of oocyte in the adult female dromedary camel. The weight, volume and three dimensions of ovaries from slaughtered dromedary camels and oocytes inside zona diameter and zona pellucida thickness were measured. The follicles were classified in regard to the size and oocytes according to their ooplasm appearance and CC compactness. Aspirated cumulus oocyte complexes (COCs) were incubated for 48 hr (with a 6-hr interval) in Hams-F10, and CC expansion and 1PB extrusion were assessed. Significant differences were seen in the shape, weight, volume and three dimensions of the ovaries between ≤4-year-old and >4-year-old dromedary camel (p < .5). Approximately, 95.82% of follicles were 2-4 mm in diameter. The mean (±SD) of inside zona diameter of the oocyte and zona pellucida thickness was 132.22 ± 13.8 and 14.64 ± 2.24 µm, respectively, in >4-year-old dromedary camel. The CC expansion and 1PB extrusion were seen in 86% and 21.88% of COCs, respectively. Age and sexual conditions of dromedary camel influence the morphological and morphometric characteristics of the ovary. Most COCs retrieved from 2-6 mm follicles are cultivable. The most slaughterhouse-derived COCs retrieved from 2-6 mm follicles of non-pregnant dromedary camels are excellent and good and yielding a most favourable diameter to achieve the developmental competence for IVM in an optimal time of 24-30 hr; the optimal time for CC expansion is 24-30 hr in this species. However, the CC expansion is a prerequisite process, but not sufficient for IVM.

Cell chip-based monitoring of toxic effects of cosmetic compounds on skin fibroblast cells.

The present study estimated the efficacy of electrochemical detection of imidazolidinyl urea-induced cell toxicity in skin human fibroblast cells (HFF cells). The gold nanopunct structures were fabricated through a nanoporous alumina mask, and the structural formations were confirmed via scanning electron microscopy. The HFF cells were allowed to attach to RGD (Arg-Gly-Asp) peptide nanopatterned surfaces, and electrochemical tools were applied to skin cells attached to the chip surface. The HFF cells evidenced inflammation responses to allergens such as imidazolidinyl urea. The cells were subsequently treated with different concentrations of imidazolidinyl urea for 24 h in culture, which induced a change in the cyclic voltammetry (CV) current peak. Treatment with imidazolidinyl urea induced a loss of cell viability and accelerated inflammation in a concentration-dependent manner. The expression level of inflammation-related proteins such as IL-1 beta were increased in imidazolidinyl urea-treated cells. The CV results demonstrated that imidazolidinyl urea significantly reduced the current peaks in a dose-dependent manner. The results showed that the current peak was reduced in accordance with the increases in imidazolidinyl urea-induced inflammation. In conclusion, the results of this study suggest that the electrochemical-based chip provides crucial information for improvements to a cell chip system for drug screening applications.

Relationship between different concentrations of the plasma progesterone at the time of FSH-P treatment and the superovulatory response in Holstein dairy heifers.

The aim of this study was to evaluate the influence of two different concentrations of plasma progesterone at the time of FSH-P treatment on the superovulatory response in dairy heifers. Sixteen reproductively sound Holstein heifers (13-15 months of age) were used in this study. Superovulatory treatment was commenced at mid-dioestrus (Day 10 ± 2 of the oestrous cycle) of the synchronized (using two injections of PGF2α, 11 days apart) oestrous cycles. Blood samples were collected on the day and the day after commencing FSH-P treatment and at oestrus for plasma progesterone determination. Heifers were grouped based on two levels of plasma progesterone; Group low progesterone (LP; ranging from 2 to 4.5 ng /ml; n = 7) and Group high progesterone (HP; ≥ 4.6 ng /ml; n = 8) at the beginning of FSH-P treatment (one heifer was excluded from the statistical analysis because of the abnormal progesterone level at oestrus). The superovulatory response in terms of mean numbers of palpable corpora lutea (ovulation rate) was significantly higher (p < 0.05) in group LP than group HP. Ovulation rate was negatively correlated (r = -0.51) with the progesterone concentration at the time of commencing FSH-P treatment (p < 0.05). Data suggest that varying concentrations of plasma progesterone at the time of FSH-P treatment may have a different effect on the outcome of superovulatory response in dairy heifers.

Cell chip with nano-scale peptide layer to detect dopamine secretion from neuronal cells.

A cell chip with a nano-scaled thin film of cysteine modified synthetic oligopeptide C(RGD)4 was fabricated to detect dopamine secretion from neuronal cells. Thin C(RGD)4 peptide layer was fabricated on chip surface for increasing the binding affinity of cells to gold electrode surface, which is essential for the electrochemical detection of dopamine released from PC12 cells. The structural formation of the peptide thin film was confirmed by both atomic force microscopy (AFM) and scanning electron microscopy (SEM). Redox characteristics of chemical dopamine were firstly characterized by voltammetric tool to compare the dopamine released from PC12 cells. Cells grown on the chip were then subjected to cyclic voltammetric (CV) analysis after 48 hours of incubation. The intensities of reduction peaks were found to be increased with increasing the concentrations of PC12 cells. In addition, the electrochemical redox signal increased more in the cells treated with glucose and potassium compared to the control group. Hence, the developed cell chip can be used to determine the effects of drugs on living cells electrochemically.

Induction of superovulation in mature mice and rats using serum of spayed female dogs.

The following experiments were designed to examine the effect of serum of spayed dogs on superovulation response in mice and rats. In Experiment 1, female mice at diestrus (n=30) were divided into three equal groups and superovulated with either administration of 5 IU pregnant mare serum gonadotropin (PMSG) or recombinant follicle stimulating hormone (rFSH) (reducing dose from 2.5 to 0.5 IU) and 5 IU human chorionic gonadotropin (hCG) administered 48h later. Serum of spayed dogs was administered intraperitoneally at a reduced dose from 0.1 to 0.025 mL in a 48 h period. In Experiment 2, female rats (n=30) at diestrus stage were divided into three equal groups. Superovulation was induced using either 30 IU PMSG, or a dose reduced from 5 to 1 IU rFSH and 25 IU hCG administered 48h later. Serum of spayed dogs was administered in a reduced dose from 0.6 to 0.1 mL in a 48 hour period. Female mice and rats were mated 24 h following hCG administration. On day 14 after mating, animals were euthanized and ovarian sections were fixed for histopathological evaluation and corpus luteum (CL) counting. No significant difference observed in mean (±SEM) number of CLs between the PMSG group and the mice that received serum of spayed dog (10.4±1.3 vs 9.2±1.0). Mean (±SEM) number of CLs tended to be lower in rats that received serum of spayed dog than those of rats which received either PMSG or rFSH (15.1±1.9 vs 23.6±3.1 and 23.1±2.9, P=0.06, respectively). In conclusion, serum of spayed dogs is able to induce a superovulatory response in mice and rats.

Inherent inferior quality of follicular fluid in repeat breeder heifers as evidenced by low rates of in vitro production of bovine embryos.

The aims of the present study were to determine the effect of follicular fluid obtained from the ovulatory follicle of repeat breeder heifers on in vitro oocyte maturation (Experiment 1), fertilization (Experiment 2) and production of bovine embryos (Experiment 3). Holstein virgin heifers (VH, n = 5) with normal fertility or repeat breeder syndrome (RBH, n = 5) were used in the present study. Follicular fluid of VH and RBH was aspirated from ovulatory follicles and used as maturation medium. Bovine oocytes were aspirated from follicles of slaughterhouse ovaries and randomly allocated in three groups; in Group 1, oocytes cultured in TCM-199 supplemented with 10% heat-treated fetal calf serum and hormones (5 IU/mL hCG plus 0.1 IU/mL rFSH); in Group 2, oocytes cultured in TCM-199 supplemented with 10% filtered follicular fluid of VH without hormones; in Group 3, oocytes cultured in TCM-199 supplemented with 10% filtered follicular fluid of RBH without hormones. The mean (±SEM) percentage of matured oocytes was different between VH and RBH groups (72.2 ± 4.0 vs 56.4 ± 4.6%; P < 0.05, respectively). Further, the mean (±SEM) percentage of normal oocyte fertilization was higher in the VH than the RBH group (49.3 ± 2.1 vs 32.0 ± 4.2; P < 0.05, respectively). The mean percentage of embryos developed to the blastocyst stage was higher in the VH than the RBH group (12.0 ± 1.3 vs 7.0 ± 1.6; respectively; P < 0.05). In conclusion, our findings support our hypothesis that the ovulatory follicle microenvironment of Holstein repeat breeder heifers places their oocytes at a developmental disadvantage compared with Holstein fertile virgin heifers and that this suggest the existence of an inherent inferior quality of the ovulatory follicle microenvironment in repeat breeding Holstein heifers.

Factors affecting the size of ovulatory follicles and conception rate in high-yielding dairy cows.

Two studies were designed to determine (1) the effects of Heatsynch and Ovsynch protocols versus spontaneous ovulation and (2) the effects of calving problems, clinical uterine infections, and clinical mastitis on the size of the ovulatory follicle, conception rate, and embryonic/fetal (E/F) death in high-yielding dairy cows. In study 1, cows without the history of calving problems, clinical uterine infections, and clinical mastitis were randomly allocated to either an Ovsynch (n = 45) or Heatsynch (n = 39) ovulation synchronization protocol or spontaneous ovulation (n = 43) groups. Blood samples were collected on the day of artificial insemination (AI) to measure progesterone (P4), estradiol-17ß, and insulin-like growth factor 1 (IGF-1) and 7 days later to measure P4. Study 2 consisted of cows (n = 351) with or without the history of calving problems, clinical uterine infections, and clinical mastitis which were artificially inseminated after a 55-day voluntary waiting period. Transrectal ultrasonography was performed at the time of AI to measure the ovulatory follicle size and on Days 30 and 68 after AI to diagnose pregnancy in both studies. In study 1, the mean (±standard error of the mean) diameter of the ovulatory follicle was greater (P = 0.0005) and E/F mortality was lower (P = 0.007) for the spontaneous ovulation group compared with Ovsynch and Heatsynch groups. Serum concentration of P4 on Day 7 after AI was correlated with the size of the ovulatory follicle (P = 0.007). Conception rate at Days 30 and 68 was not significantly different between the three experimental groups in study 1. Cows with serum IGF-1 concentrations greater than 55 ng/mL at AI had significantly higher Day 68 conception rate (50% vs. 24%) and lower E/F death (16.6% vs. 40%) compared to cows with serum IGF-1 concentrations lower than 56 ng/mL at AI. The conception rate on Days 30 and 68 for follicles of 10 to 14 mm in diameter (34% and 21.8%) was significantly lower than follicles of 14.1 to 19 mm in diameter (60% and 50%), respectively (P < 0.05). In study 2, the ovulatory follicle in cows with clinical uterine infections was smaller than that in cows without clinical uterine infections (16.4 vs. 17.1 mm; P = 0.04). In conclusion, the size of the ovulatory follicle is affected by ovulation synchronizing protocols and postpartum clinical uterine infections. In addition, cows with higher serum IGF-1 concentrations on the day of AI had higher Day 68 conception rate and lower E/F death.

Chronological and ultrastructural changes in camel (Camelus dromedarius) oocytes during in vitro maturation.

Cumulus-oocyte complexes (COCs) were collected from non-pregnant camels at a local slaughterhouse by aspiration from antral follicles (2-6 mm). In Experiment I, camel COCs (n=304) were matured in vitro in Hams-F10, fixed at different time intervals (6, 12, 18, 24, 30, 36, 42, or 48 h) and stained with 1% aceto-orcein to assess nuclear changes in culture. A majority of the oocytes (81.5%) underwent germinal vesicle break down (GVBD) between 6 and 12h. Forty-eight percent of the oocytes were observed at the metaphase I (M I) stage by 18 h culture. The percentage of matured oocytes (M II stage) at 30 and 42 h were 66.5 and 71% respectively, which were significantly (p<0.05) different to that observed at 24 h (42.5%). In Experiment II, after different periods of culture (12, 24, 36, or 48 h), the COCs (n=26) were processed for transmission electron microscopy. Expansion of both the cumulus and corona radiate cells occurred between 12 and 24 h in the majority of oocytes concomitant with enlargement of the cumulus cell process endings (CCPEs) in the developed perivitelline space. After 12 h of culture disruption of the junctions between CCPEs and the oolemma was observed together with and breakdown of the GV. For 24-36 h of culture cortical granules had spread and aligned along the oolemma. Signs of degeneration in the cytoplasmic organelles of the oocytes were also observed from less than 36 h. After 48 h of culture, larger vesicles and lipid droplets had appeared in the central part of the oocytes and showed uneven distribution throughout the ooplasm. Predominantly non-penetrating CCPEs were also observed in four oocytes by 48 h. In conclusion, based on both light and electron microscopic evaluations, the optimal culture time for the development of competent Camelus dromedarius oocytes in vitro appears to be 30 h using Hams-F10 medium.

Functional histology of the ovarian follicles as determined by follicular fluid concentrations of steroids and IGF-1 in Camelus dromedarius.

Ovaries were collected from sexually mature non-pregnant dromedary she-camels. Follicles 6 to 19 mm in diameter per pair of ovaries were randomly selected and classified into clear (n = 30), or opaque (n = 14) based on macroscopic examination of the follicle surface, and then were divided into four classes: clear follicles with 6- 9.9 and 10-19 mm diameter; opaque follicles with 6- 9.9 and 10-19 mm diameter. Follicular fluid (FF) was aspirated for measurement of estradiol-17ß, progesterone and IGF-I concentrations, and then a section of tissue through the exposed surface of the follicle wall was removed and fixed in and processed for histological examination. Mean (±SE) number of clear follicles observed on the ovaries that contained a large dominant follicle was less than that on the ovaries which contained a large atretic follicle (p < 0.05; 2.6 ± 1 vs 8.6 ± 0.6). In conclusion, the estrogenic large follicles have suppressive effects on the growth of other follicles.

Reproductive responses of dairy cows with ovarian cysts to simultaneous human chorionic gonadotropin or gonadotropin-releasing hormone and cloprostenol compared to gonadotropin-releasing hormone alone treatment.

AIM: Bovine ovarian cysts are a common cause of economic loss in modern dairy herds. The objective of the present study was to evaluate the reproductive responses to three protocols using hCG, GnRH and cloprostenol when the definite diagnosis of the type of ovarian cyst is/is not possible in dairy cows. MATERIALS AND METHODS: A total of 144 lactating dairy cows with ovarian cysts were divided into three groups. At diagnosis (Day 0), cows in Group 1 (the conventional method, n=47) were injected with 0.02 mg of a GnRH analogue i.m. (Buserelin); cows in Group 2 (n=47) were intramuscularly treated with 0.02 mg Buserelin plus 500 µg cloprostenol; and cows in Group 3 (n=50) were intramuscularly treated with 1500 IU hCG plus 500 µg cloprostenol. All cows received 500 µg cloprostenol intramuscularly on Day 10. RESULTS: No statistically significant differences were found in the recovery time, interval to conception, conception rate at first AI, and pregnancy rates by Days 70 and 100 after treatment among the three groups. CONCLUSIONS: Simultaneous treatment of ovarian cysts with hCG or GnRH and cloprostenol appeared to have no advantage over the conventional method, GnRH alone, in dairy cows. Furthermore, hCG and GnRH have an equal therapeutic effect in cows with ovarian cysts.

Continuous versus step-by-step scanning mode of a novel 3D scanner for CyberKnife measurements.

The purpose of the study is to investigate the continuous versus step-by-step scanning mode of a commercial circular 3D scanner for commissioning measurements of a robotic stereotactic radiosurgery system. The 3D scanner was used for profile measurements in step-by-step and continuous modes with the intent of comparing the two scanning modes for consistency. The profile measurements of in-plane, cross-plane, 15 degree, and 105 degree were performed for both fixed cones and Iris collimators at depth of maximum dose and at 10cm depth. For CyberKnife field size, penumbra, flatness and symmetry analysis, it was observed that the measurements with continuous mode, which can be up to 6 times faster than step-by-step mode, are comparable and produce scans nearly identical to step-by-step mode. When compared with centered step-by-step mode data, a fully processed continuous mode data gives rise to maximum of 0.50% and 0.60% symmetry and flatness difference respectfully for all the fixed cones and Iris collimators studied.

Factors associated with variation in the superovulatory response of cattle.

Kafi, M., McGowan, M.R., 1997. Factors associated with variation in the superovulatory response of cattle. Anim. Reprod. Sci. Variability in the superovulatory response of cattle is still one of the major limiting factors in extensive usage of embryo transfer technology. A variety of approaches including recent attempts to eliminate the suppressive effect of the dominant follicle have been used to reduce the unpredictability of the superovulatory response of cattle. The development of techniques such as transrectal ultrasonography, have enabled a re-evaluation of ovarian dynamics during superovulation of cattle. In addition, advances in reproductive hormone assays have increased knowledge of the mechanisms controlling follicular development, ovulation and corpus luteum function. This review focuses on the current understanding of factors affecting the superovulatory response of cattle. Also, abnormalities of ovulation and endocrine disorders that may occur during superovulation are reviewed.

In vitro maturation and fertilization of bovine oocytes and in vitro culture of presumptive zygotes in the presence of bovine pestivirus.

A pathogen which has been shown to commonly contaminate in vitro bovine embryo production system is bovine pestivirus (bovine viral diarrhea virus). Three experiments were designed to evaluate the in vitro maturation (experiment I), fertilization (experiment II) and embryo development (experiment III) of immature oocytes, inseminated oocytes and presumptive zygotes in the presence of a bovine pestivirus (non-cytopathic, nCP type 1). The virus inoculum used was derived from a persistently infected cow. In experiment I, follicular oocytes (n=1257) recovered from slaughterhouse derived ovaries were randomly assigned to either a control group (n=578) which did not become exposed to bovine pestivirus and a treatment group (n=679) which was inoculated with bovine pestivirus (2.20-3.69 log(10) TCID(50)/50 microl) at the time of commencement of in vitro maturation. Overall, there was no significant difference between the control and pestivirus inoculated oocytes in either the cumulus cell expansion rate (79+/-7.5% versus 74+/-10.7%) or the nuclear maturation rate (89+/-4.8% versus 85+/-7.4%), respectively. In experiment II, in vitro matured oocytes (n=607) were inseminated either in the absence (control; n=301) or the presence of bovine pestivirus (4-4.6 log(10) TCID(50)/50 microl; n=306). A significant (P<0.01) reduction in the overall number of fertilized oocytes with two well formed male and female pronuclei was observed in the treatment group compared to the control group (58.5+/-5.8% versus 73.3+/-3.6%, respectively). In experiment III, after in vitro maturation and fertilization, presumptive zygotes were randomly assigned to either a control group (n=139) which was not exposed to bovine pestivirus or a treatment group which was inoculated with bovine pestivirus (2.97-4.47 log(10) TCID(50)/30 microl; n=139). The zygotes were then cultured under mineral oil in an atmosphere of 88% N(2), 7% O(2) and 5% CO(2) at 39 degrees C. The morphologic appearance of the embryos was assessed 48 h after the commencement of culture, and then every 48 h up to days 7-8 after insemination. The 22% (31/139) and 3.6% (5/139) of the presumptive zygotes developed to the morula or blastocyst stage in the control and the bovine pestivirus inoculated groups, respectively (P<0.001). This study demonstrates that bovine pestivirus has a significant detrimental effect on in vitro fertilization and early in vitro embryo development.

Relationships among calving season, heat load, energy balance and postpartum ovulation of dairy cows in a subtropical environment.

The study was designed to examine the relationships among calving season, energy balance, temperature humidity index (THI), and postpartum ovulation in high producing cows in a subtropical environment. Holstein cows calving in a feedlot dairy in southeast Queensland during winter (n = 23) and summer (n = 21) were monitored during the first 9 weeks of lactation. Cows were weighed and blood samples collected twice weekly: plasma progesterone, plasma metabolites related to energy and mineral balance, and haematological measurements were performed. Milk production was measured, body condition score was estimated, and trans-rectal ultrasound examinations of the ovaries were each undertaken once a week. The interval between calving and first ovulation was significantly longer in cows calving in summer (22.8 vs. 17.6 days, P < 0.05). Interval from calving to the first postpartum ovulation (FOVL) was inversely related to the mean plasma glucose concentration for the first 9 weeks after calving (GLU): FOVL = 80.0-17.9GLU, (R2 = 0.25, P < 0.001). Plasma progesterone concentration during the life of the second corpus luteum after calving was negatively correlated with THI during the first 2 weeks after calving (r = 0.55, P < 0.001). Plasma glucose concentration (GLU) was negatively correlated with milk yield (MYD) and rectal temperature (RT), and positively correlated with plasma calcium concentration (Ca) according to the following regression equation. GLU = 33.1 - 0.02MYD + 0.91Ca - 0.48RT, (R2 = 0.58, P = 0.0001).

The effect of bovine pestivirus infection on the superovulatory response of Friesian heifers.

The pathogenesis of reproductive loss associated with bovine pestivirus infection during the preovulatory period was investigated using superovulated heifers. Twenty-five Friesian heifers were selected and randomly assigned to either a control group (n = 12) which did not become infected or to a treatment group (n = 13) which became infected following intranasal instillation of 2 ml of serum inoculum containing 5.5 log(10) TCID(50)/ml non-cytopathic virus, 9 d prior to artificial insemination (AI). Transrectal ultrasonography was used to monitor follicular development and ovulation during the superovulatory period. Animals were superovulated using a standard protocol of twice-daily injections of FSH-P and then were inseminated twice commencing 12 h after the onset of estrus. The intensity of expression of estrus was higher in the control heifers than in the pestivirus-infected heifers. Of 13 pestivirus-infected heifers, only 3 heifers displayed standing estrus compared with that in the control group, in which 10 of 12 heifers exhibited standing estrus. The mean number of ova/embryos recovered from the control group heifers was 5.75 +/-2.31, of which 4.00 +/- 0.72 were evaluated as transferable quality embryos. In comparison, heifers in the pestivirus-infected group yielded only a mean of 0.60 +/-0.34 ova/embryos, of which 0.23 +/- 0.22 were transferable quality embryos. Based on ultrasonographic examination, 24 h after the first AI 82% of the presumptive ovulatory follicles had ovulated in the control group compared with an ovulation rate of only 17% in the treated group. The results of this experiment demonstrated that bovine pestivirus infection during the preovulatory period could adversely affect ovulation, thus leading to a significant reduction in the number of palpable corpora lutea and in the number and quality of embryos recovered.