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Biomedical subjects

P J Hansen

Publications and source records attributed to P J Hansen.

At least 19 recordsLinked to original sources

Fertility of lactating dairy cows administered recombinant bovine somatotropin during heat stress.

Administration of recombinant bovine somatotropin (bST) to lactating dairy cows during heat stress increases milk yield, but it also can increase body temperature and may therefore compromise fertility. However, it is possible that bST treatment could increase fertility during heat stress because it has been reported to increase fertility in lactating cows. In addition, bST increases secretion of insulin-like growth factor-I (IGF-I) that promotes embryo survival. The purpose of this study was to determine effects of bST on reproductive function in lactating dairy cows during heat stress. The experiment was conducted in southern Georgia from July to November 2005 using lactating Holstein cows (n = 276 for reproductive traits). For first service timed artificial insemination (TAI), cows were presynchronized with 2 injections of PGF(2alpha) given 14 d apart followed by a modified Ovsynch protocol (GnRH and insemination at 72 h following PGF(2alpha) ). Pregnancy was diagnosed by using ultrasonography on d 29 and reconfirmed by palpation between d 45 and 80 post-TAI. Nonpregnant cows were resynchronized with the modified Ovsynch protocol and received a second TAI. Treatment with bST started 1 wk before the start of Ovsynch and continued at 2-wk intervals. Blood samples were collected from a subset of cows to determine IGF-I profiles immediately before the first bST injection, 1 wk later, and at d 35 of bST treatment. Rectal temperatures were assessed on d 29 of bST treatment. Pregnancy rates (d 45 to 80 post-TAI) did not differ between bST and control cows for first- (16.7 vs. 15.2%) or second-service TAI (14.8 vs. 17.2%). Plasma concentrations of IGF-I and milk yield were greater for bST-treated cows following the initiation of bST treatment and bST increased rectal and vaginal temperatures. Body condition score was less for bST-treated cows. In conclusion, treatment with bST during heat stress increased IGF-I concentrations, milk yield over time, and rectal and vaginal temperatures without affecting first- or second-service pregnancy rates. Thus, at least under certain housing conditions, bST can be used to improve milk yield during heat stress without compromising fertility.

Animals↗

Effectiveness of administration of gonadotropin-releasing hormone at Days 11, 14 or 15 after anticipated ovulation for increasing fertility of lactating dairy cows and non-lactating heifers.

One strategy for improving fertility in cattle is mid-cycle administration of GnRH to increase progesterone secretion and delay luteolysis. This strategy might be especially useful during hot weather because heat stress increases uterine prostaglandin release and reduces development of the elongating embryo. A series of experiments was conducted to test the efficacy of GnRH for increasing fertility. There was no effect of administration of 100 microg GnRH at Day 11 after anticipated ovulation on pregnancy rates in virgin heifers subjected to timed artificial insemination (TAI) during the summer. Similarly, there was no beneficial effect of administration of GnRH at Day 11 after anticipated ovulation on pregnancy rates of lactating cows subjected to TAI in summer and winter. Three experiments tested effects of injection of GnRH at Days 14 or 15 after anticipated ovulation on pregnancy rates of lactating cows. The first experiment used 477 lactating cows subjected to TAI. Cows receiving GnRH at Day 14 had higher pregnancy rates in both summer and winter than cows receiving vehicle (20.3 versus 12.7%, P<0.02). When this experiment was repeated during summer with 137 cows, there was a negative effect of GnRH treatment at Day 14 on pregnancy rate. In the third experiment, lactating cows during summer were inseminated at detected estrus and cows were assigned to treatment with either GnRH or vehicle at Days 14 or 15 after insemination. Pregnancy rates were 25.6% (32/125) for cows receiving vehicle, 20.7% (19/92) for cows receiving GnRH at Day 14, and 20.3% (16/79) for cows receiving GnRH at Day 15. In conclusion, GnRH administration at Days 11-15 after anticipated ovulation or estrus did not consistently increase pregnancy rates in either cool or warm seasons.

Animals↗

Effects of dietary unsaturated fatty acids on oocyte quality and follicular development in lactating dairy cows in summer.

Dietary sources of fatty acids were evaluated for their influence on oocyte quality and follicular development using 54 lactating cows in summer. Fat supplements were 1) sunflower oil (80% cis 18:1), 2) Ca salt of transoctadecenoic acids (57% trans 18:1), 3) Ca salt of vegetable oils (30% 18:2), and 4) linseed oil (56% 18:3 and 16% 18:2). Fats were fed at 1.35% of dietary dry matter beginning at 5 wk prior to expected calving date and at 1.5% (oils) and 1.75% (Ca salts) of dietary dry matter for 15 wk after parturition. Four days following a programmed induced ovulation, 5 transvaginal oocyte aspirations were performed 3 or 4 d apart. Three days after the last aspiration, PGF2alpha was injected, followed 3 d later by a GnRH injection and a timed artificial insemination (d 0) 16 to 20 h later. For the first 4 aspirations, oocytes grading 1 or 2 were used for in vitro embryo production. Total cell number and the proportion of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive blastomeres were analyzed at d 8. At the fifth aspiration, the occurrence of metaphase II, group II caspase activity, and TUNEL labeling were determined after oocyte maturation. A total of 1,011 oocytes were collected. The proportion of oocytes with high caspase activity was greater for grade 3 compared with grades 1 and 2 (37.5 vs. 1.54 and 1.61%). Feeding polyunsaturated fatty acids, as compared with monosaturated fatty acids, failed to affect oocyte quality, as demonstrated by subsequent embryo development. Cows fed 18:2- or 18:3-enriched diets had a larger preovulatory follicle at insemination and subsequent volume of the corpus luteum compared with those fed cis 18:1 or trans 18:1 diets (16.8, 16.2 vs. 15.0, 14.9 +/- 0.7 mm; 7,323, 8,208 vs. 6,033, 5,495 +/- 644 mm3, respectively). The previously documented benefits of polyunsaturated fatty acids on reproductive performance appear to reflect actions at alternative biological windows in lactating dairy cows.

Animals↗

Effect of transfer of one or two in vitro-produced embryos and post-transfer administration of gonadotropin releasing hormone on pregnancy rates of heat-stressed dairy cattle.

Pregnancy rates following transfer of an in vitro-produced (IVP) embryo are often lower than those obtained following transfer of an embryo produced by superovulation. The purpose of the current pair of experiments was to examine two strategies for increasing pregnancy rates in heat stressed, dairy recipients receiving an IVP embryo. One method was to transfer two embryos into the uterine horn ipsilateral to the CL, whereas the other method involved injection of GnRH at Day 11 after the anticipated day of ovulation. In Experiment 1, 32 virgin crossbred heifers and 26 lactating crossbred cows were prepared for timed embryo transfer by being subjected to a timed ovulation protocol. Those having a palpable CL were randomly selected to receive one (n = 31 recipients) or two (n = 27 recipients) embryos on Day 7 after anticipated ovulation. At Day 64 of gestation, the pregnancy rate tended to be higher (P = 0.07) for cows than for heifers. Heifers that received one embryo tended to have a higher pregnancy rate than those that received two embryos (41% versus 20%, respectively) while there was no difference in pregnancy rate for cows that received one or two embryos (57% versus 50%, respectively). Pregnancy loss between Day 64 and 127 only occurred for cows that received two embryos (pregnancy rate at Day 127=17%). Between Day 127 and term, one animal (a cow with a single embryo) lost its pregnancy. There was no difference in pregnancy rates at Day 127 or calving rates between cows and heifers, but females that received two embryos had lower Day-127 pregnancy rates and calving rates than females that received one embryo (P < 0.03). Of the females receiving two embryos that calved, 2 of 5 gave birth to twins. For Experiment 2, 87 multiparous, late lactation, nonpregnant Holstein cows were synchronized for timed embryo transfer as in Experiment 1. Cows received a single embryo in the uterine horn ipsilateral to the ovary containing the CL and received either 100 microg GnRH or vehicle at Day 11 after anticipated ovulation (i.e. 4 days after embryo transfer). There was no difference in pregnancy rate for cows that received the GnRH or vehicle treatment (18% versus 17%, respectively). In conclusion, neither unilateral transfer of two embryos nor administration of GnRH at Day 11 after anticipated ovulation improved pregnancy rates of dairy cattle exposed to heat stress.

Animals↗

Realizing the promise of IVF in cattle--an overview.

The in vitro-produced embryo could play a central role in dairy and beef production systems because of its potential role in genetic selection strategies and crossbreeding schemes, and because it can be integrated into reproductive management strategies for improving pregnancy rates in herds with low fertility. The promise attendant upon use of in vitro-produced embryos is not being fully realized, however. Indeed, there are important technical limitations to their production that reduce the desirability of in vitro-produced embryos because of increased costs, sub-optimal embryonic and fetal survival, and offspring that are occasionally abnormal. Most technical problems associated with in vitro production of embryos can be overcome through research. Among the requirements for a successful research program will be renewed emphasis on conducting embryo transfer trials to determine effects of modifications to embryo production protocols on pregnancy rates and fetal development. Given the promise of in vitro embryo technologies, there is an urgent need for a concerted and sustained investment in research to improve these technologies. Developing a consensus that the study of embryo technology is one of the most important areas of agricultural research should be both an individual and organizational priority.

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Disruption of nuclear maturation and rearrangement of cytoskeletal elements in bovine oocytes exposed to heat shock during maturation.

Meiotic maturation in mammalian oocytes is a complex process which involves extensive rearrangement of microtubules, actin filaments and chromosomes. Since cytoskeletal elements are sensitive to disruption by heat shock, a series of experiments were performed to determine whether physiologically relevant heat shock disrupts the progression of the oocyte through meiosis, fertilization and zygote formation. Cumulus-oocyte complexes were cultured at 38.5, 40.0 or 41.0 degrees C for the first 12 h of maturation. Incubation during the last 10 h of maturation and 18 h after fertilization was at 38.5 degrees C and in 5% (v/v) CO2 for both treatments. Examination of the cytoskeleton and the chromosome organization in matured oocytes revealed that oocytes matured at 38.5 degrees C were mostly at metaphase II (MII) stage, while the majority of heat-shocked oocytes were blocked at the first metaphase (MI), first anaphase or first telophase stages. A subset of heat-shocked oocytes possessed misshapen MI spindles with disorganized microtubules and unaligned chromosomes. A higher percentage of TUNEL-positive oocytes was noted for oocytes matured at 41.0 degrees C. Addition of 50 nmol/l sphingosine 1-phosphate to maturation medium blocked the effect of heat shock on progression through meiosis and apoptosis and increased the proportion of oocytes matured at 41.0 degrees C that were at MII. Following insemination, a high percentage of heat-shocked oocytes were unfertilized, while the majority of the control zygotes were fertilized and had two visible pronuclei. In conclusion, heat shock disrupts nuclear maturation and induces apoptosis. These alterations are likely to be involved in the mechanism underlying heat-shock-induced disruption of oocyte capacity for fertilization and subsequent development.

Actin Cytoskeleton↗

Factors associated with early and mid-to-late fetal loss in lactating and nonlactating Holstein cattle in a hot climate.

The purpose of this study was to evaluate associations of lactation, somatic cell count score (SCCS) at breeding, milk yield, lactation number, interval from calving to breeding (days open), number of times inseminated, and season of breeding on fetal loss for lactating Holstein females (both first-parity and multiparous cows) and nonlactating Holstein heifers in a hot climate. Females were palpated between d 40 and 50 of gestation and again at d 70 to 80 to determine pregnancy status. Early fetal loss was defined as a loss that occurred after d 40 to 50 but before d 70 to 80. Mid-to-late fetal loss represented losses after d 70 to 80 but before expected calving. Lactating females had higher early (P = 0.055) and mid-to-late fetal loss (P < 0.05) than nonlactating heifers. Those lactating females with increased days open experienced greater early (P < 0.05) and mid-to-late fetal loss (P = 0.055), whereas lactating females with an elevated SCCS encountered greater mid-to-late fetal loss (P < 0.01). Milk yield, lactation number, number of times inseminated, and season were not associated with early or mid-to-late fetal loss. For nonlactating heifers, there were no associations between number of times inseminated, season, or age at breeding on early or mid-to-late fetal loss. In conclusion, lactating females were more likely to suffer early and mid-to-late fetal loss than nonlactating heifers. Also, days open and SCCS at breeding were related to ability of lactating females to maintain pregnancy, but there were no relationships between fetal loss and milk yield, lactation number, number of times inseminated, or season.

Age Factors↗

Timing of inhibitory actions of gossypol on cultured bovine embryos.

Culture of bovine preimplantation embryos with gossypol, a polyphenolic pigment in cottonseed, inhibits development. Neither stage at which embryos are most sensitive to gossypol, nor the mechanism by which development is blocked is known. Our objectives were to characterize stages at which gossypol inhibits embryonic development and evaluate involvement of apoptosis in actions of gossypol. When presumptive 1-cell embryos were cultured continuously in medium containing gossypol at concentrations of 0, 2.5, 5, and 10 microg/mL, cleavage rate was not reduced by any concentration of gossypol, but percentages of 1-cell embryos that became blastocysts 8 d after insemination was reduced by the 10 microg/mL dose of gossypol. Culture of presumptive 1-cell embryos with gossypol at 10 microg/mL for 24 h was not sufficient to block development. Furthermore, gossypol did not affect development to the blastocyst stage when 2-cell embryos were cultured with gossypol at 10 microg/mL for 24 h or 7 d. Culture of embryos > or =16 cells with gossypol at 10 microg/mL for 24 h failed to reduce cell number 24 h later or increase blastomere apoptosis. Results indicate that embryonic development can be disrupted by long-term exposure to gossypol at 10 microg/mL and that exposure at the 1-cell stage is required. Thus, it is likely that the deleterious effects of gossypol involve disruption of events at the 1-cell stage and such effects are reversible if gossypol is removed. After the 1-cell stage, gossypol does not affect development because the critical event that gossypol disrupts occurs at the 1-cell stage only or the embryo develops cytoprotective mechanisms after the 1-cell stage that limit actions of gossypol.

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Sphingosine 1-phosphate protects bovine oocytes from heat shock during maturation.

Sphingosine 1-phosphate (S1P) is a sphingolipid metabolite that can block apoptosis by counteracting the proapoptotic effects of ceramide. Experiments were performed to evaluate whether S1P blocks the disruption in oocyte developmental competence caused by heat shock. Cumulus-oocyte complexes (COCs) were placed in maturation medium and cultured at 38.5 or 41 degrees C for the first 12 h of maturation. Incubation during the last 10 h of maturation, fertilization, and embryonic development were performed at 38.5 degrees C. Heat shock during the first 12 h of maturation reduced cleavage rate, the number of oocytes developing to the blastocyst stage, and the percentage of cleaved embryo that subsequently developed to blastocysts. Addition of 50 nM S1P to maturation medium had no effect on oocytes matured at 38.5 degrees C but blocked effects of thermal stress on cleavage and subsequent development. The blastocysts formed at Day 8 did not differ between S1P and control groups in caspase activity, total cell number, or percentage of cells that were apoptotic. Blocking endogenous generation of S1P by addition of 50 nM N1N-dimethylsphingosine, a sphingosine kinase inhibitor, reduced or tended to reduce cleavage rate and blastocyst development regardless of whether maturation of COCs was at 38.5 or 41 degrees C. Results demonstrate that S1P protects oocytes from a physiologically relevant heat shock and affects oocyte maturation even in the absence of heat shock. The S1P-treated oocytes that survived heat shock and became blastocysts had a normal developmental potential as determined by caspase activity, total cell number, and percentage of apoptotic cells. Thus, modulation of developmental competence of oocytes using S1P may be a useful approach for enhancing fertility in situations where developmental competence of oocytes is compromised.

Animals↗

Involvement of apoptosis in disruption of developmental competence of bovine oocytes by heat shock during maturation.

Various pathological stimuli such as radiation, environmental toxicants, oxidative stress, and heat shock can initiate apoptosis in mammalian oocytes. Experiments were performed to examine whether apoptosis mediated by group II caspases is the cause for disruption of oocyte function by heat shock applied during maturation in cattle. Bovine cumulus-oocyte complexes (COCs) were cultured at 38.5, 40, or 41 degrees C for the first 12 h of maturation. Incubation during the last 10 h of maturation, fertilization, and embryonic development were at 38.5 degrees C and 5% (v/v) CO2 for all treatments. In the first experiment, exposure of COCs to thermal stress during the first 12 h of maturation reduced cleavage rate and the number of oocytes developing to the blastocyst stage. In the second experiment, a higher percentage of TUNEL-positive oocytes was noted at the end of maturation for oocytes matured at 40 and 41 degrees C than for those at 38.5 degrees C. In addition, the distribution of oocytes classified as having high (>25 intensity units), medium (15-25 intensity units), and low (<15 intensity units) caspase activity was affected by treatment, with a greater proportion of heat-shocked oocytes having medium or high activity. In the third experiment, COCs were placed in maturation medium with vehicle (0.5% [v/v] DMSO) or 200 nM z-DEVD-fmk, an inhibitor of group II caspases. The COCs were matured at 38.5 or 41 degrees C, fertilized and cultured for 8 days. The inhibitor blocked the effect of heat shock on cleavage rate and the percentage of oocytes and cleaved embryos developing to the blastocyst stage. In conclusion, heat shock during oocyte maturation can promote an apoptotic response mediated by group II caspases, which, in turn, leads to disruption of the oocyte's capacity to support early embryonic development following fertilization.

Animals↗

Insulin-like growth factor-I as a survival factor for the bovine preimplantation embryo exposed to heat shock.

Insulin-like growth factor-I (IGF-I) is a survival factor for preimplantation mammalian embryos exposed to stress. One stress that compromises preimplantation embryonic development is elevated temperature (i.e., heat shock). Using bovine embryos produced in vitro as a model, it was hypothesized that IGF-I would protect preimplantation embryos by reducing the effects of heat shock on total cell number, the proportion of blastomeres that undergo apoptosis, and the percentage of embryos developing to the blastocyst stage. In experiment 1, embryos were cultured with or without IGF-I; on Day 5 after insemination, embryos >or=16 cells were cultured at 38.5 degrees C for 24 h or were subjected to 41 degrees C for 9 h followed by 38.5 degrees C for 15 h. Heat shock reduced the total cell number at 24 h after initiation of heat shock and increased the percentage of blastomeres that were apoptotic. Effects of heat shock were less for IGF-I-treated embryos. Experiment 2 was conducted similarly except that embryos were allowed to develop to Day 8 after insemination. The percentage reduction in blastocyst development for heat-shocked embryos compared with those maintained at 38.5 degrees C was less for embryos cultured with IGF-I than for control embryos. Heat shock reduced the total cell number in blastocysts and increased the percentage of blastomeres that were apoptotic, whereas IGF-I-treated embryos had increased total cell number and a reduced percentage of apoptosis. Taken together, these results demonstrate that IGF-I can serve as a survival factor for preimplantation bovine embryos exposed to heat shock by reducing the effects of heat shock on development and apoptosis.

Animals↗

Physiological and cellular adaptations of zebu cattle to thermal stress.

During their separate evolution from Bos taurus, zebu cattle (Bos indicus) have acquired genes that confer thermotolerance at the physiological and cellular levels. Cattle from zebu breeds are better able to regulate body temperature in response to heat stress than are cattle from a variety of B. taurus breeds of European origin. Moreover, exposure to elevated temperature has less deleterious effects on cells from zebu cattle than on cells from European breeds. Superior ability for regulation of body temperature during heat stress is the result of lower metabolic rates as well as increased capacity for heat loss. As compared to European breeds, tissue resistance to heat flow from the body core to the skin is lower for zebu cattle while sweat glands are larger. Properties of the hair coat in zebu cattle enhance conductive and convective heat loss and reduce absorption of solar radiation. At the cellular level, genetic adaptations to resist deleterious effects of elevated temperature result in preimplantation embryos from zebu being less likely to be inhibited in development by elevated temperature than are embryos from European breeds. The zebu genotype has been utilized in crossbreeding systems to develop cattle for beef and dairy production systems in hot climates but success has been limited by other unfavorable genetic characteristics of these cattle. An alternative scheme is to incorporate specific thermotolerance genes from zebu cattle into European breeds while avoiding undesirable genes. Once specific genes responsible for thermotolerance in zebu have been identified or mapped, breeding strategies such as marker-assisted selection and transgenics can be applied to further the exploitation of the zebu genotype for cattle production systems.

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Immunolocalization of heat shock protein 70 in bovine spermatozoa.

Heat shock protein 70 (HSP70) is part of a superfamily of molecular chaperones, which protect cells from chemical and heat shock. The objectives of this study were to determine the presence of HSP70 in bovine spermatozoa and its subcellular localization during different stages of spermatogenesis. Analysis of sperm proteins by Western blotting using a monoclonal antibody to the inducible form of HSP70 revealed a single immunoreactive band with an estimated molecular weight of 70 kDa in samples from 18 of 18 bulls. Using immunofluorescence microscopy and the same antibody, HSP70 was localized to the cytoplasm of prophase spermatocytes and elongating spermatids, to cytoplasmic droplets of caput epididymal spermatozoa, and to cytoplasmic droplets, acrosome, post-acrosomal region and middle piece of corpus and cauda epididymal spermatozoa. The pattern of distribution changed in freshly ejaculated spermatozoa as HSP70 was detected on the acrosome only. During capacitation and acrosome reaction, HSP70 was once again redistributed, and was localized to the equatorial segment, post-acrosomal region and middle piece. Thus, HSP70 is present in the spermatozoa of mature bulls and redistribution of the protein occurs during capacitation and the acrosome reaction.

Acrosome Reaction↗

Alteration in uterine contractility in mares with experimentally induced placentitis.

An experimental model of ascending placentitis was developed in the mare to characterize the uterine myoelectrical pattern in late gestation and determine how ascending placentitis altered this pattern. In experiment 1, myometrial electrical activity was analyzed during the early morning, late morning and evening hours in four mares in the last 15 days of gestation to identify patterns of activity. In experiment 2, nine mares received intra-cervical inoculations of Streptococcus equi subspecies zooepidemicus. Myoelectrical activity in the early morning and evening hours in these mares was compared with four control mares. In experiment 1, the number of spike burst clusters >30 s was greater in the evening than in the late morning hours (P < 0.04). Spike burst activity (number x duration) of mares in experiment 1 was similar during day and night recordings until the last 6 days of gestation when it gradually increased each evening until parturition (P < 0.05). In experiment 2, control mares experienced a gradual increase in the number of small spike burst clusters in the last 6 days (P = 0.008) and an increase in large and small spike burst clusters in the evening hours in the last 4 days of gestation (P = 0.03). Mares with experimentally induced placentitis never exhibited a rise in spike burst clusters but had an increase in the mean duration and activity index of large spike burst clusters in the 4 days before parturition (P < 0.04). In conclusion, control mares had a progressive, reversible rise in myoelectrical activity at night in the week preceding parturition. This was not observed in mares with experimentally induced placentitis. They exhibited an increase in the intensity and duration of large spike burst clusters possibly in response to local inflammation.

Animals↗

Differences in heat tolerance between preimplantation embryos from Brahman, Romosinuano, and Angus breeds.

Exposure to 41 degrees C reduces development of embryos of heat-sensitive breeds (Holstein and Angus) more than for embryos of the heat-tolerant Brahman breed. Here it was tested whether embryonic resistance to heat shock occurs for a thermotolerant breed of different genetic origin than the Brahman. In particular, the thermal sensitivity of in vitro produced embryos of the Romosinuano, a Bos taurus, Criollo-derived breed, was compared to that for in vitro produced Brahman and Angus embryos. At d 4 after insemination, embryos > or = 8 cells were randomly assigned to control (38.5 degrees C) or heat shock (41 degrees C for 6 h) treatments. Heat shock reduced the proportion of embryos that developed to the blastocyst stage on d 8 after insemination. At 38.5 degrees C, there were no significant differences in development between breeds. Among embryos exposed to 41 degrees C, however, development was lower for Angus embryos than for Brahman and Romosinuano embryos. Furthermore, an Angus vs. (Brahman + Romosinuano) x temperature interaction occurred because heat shock reduced development more in Angus (30.3 +/- 4.6% at 38.5 degrees C vs. 4.9 +/- 4.6% at 41 degrees C) than in Brahman (25.1 +/- 4.6% vs. 13.6 +/- 4.6%) and Romosinuano (28.3 +/- 4.1% vs. 17.5 +/- 4.1%). Results demonstrate that embryos from Brahman and Romosinuano breeds are more resistant to elevated temperature than embryos from Angus. Thus, the process of adaptation of Brahman and Romosinuano breeds to hot environments resulted in both cases in selection of genes controlling thermotolerance at the cellular level.

Animals↗

Differences between Brahman and Holstein cows in response to estrus synchronization, superovulation and resistance of embryos to heat shock.

Embryos from Bos indicus are more resistant to elevated culture temperature (i.e. heat shock) than embryos from some Bos taurus breeds. The present experiment was designed to determine if Brahman embryos have greater resistance to heat shock than Holstein embryos at a stage in development before the embryonic genome was fully activated. A second objective was to test breed effects on estrus synchronization and superovulation responses. A total of 29 Brahman and 24 Holstein cows were subjected to estrus synchronization using gonadotropin releasing hormone (GnRH) and prostaglandin F2alpha (PGF2alpha) superovulation. Embryos were collected at 48 h and day 5 after insemination. There was a tendency for a lower proportion of Brahmans to be detected in standing estrus than Holsteins. There were no differences between breeds in the proportion of cows detected in estrus using both tailpaint and standing estrus as criteria or in interval from PGF2alpha to estrus. The degree of synchrony in estrus was greater for Brahmans. Superovulation response was generally similar between breeds. At 48 h after insemination, there was a tendency for a greater proportion of Brahman oocytes to have undergone cleavage. Uncleaved oocytes were cultured for an additional 24 h-at this time, cleavage rate was similar between breeds. When embryos reached the 2-4-cell stage, they were heat-shocked for 4.5 h at 41 degrees C. This heat shock reduced the proportion of embryos that developed to the blastocyst stage but there was no breedxtreatment interaction. At day 5 after insemination, the number of embryos recovered was too low to allow comparison of breed effects. In conclusion, genetic effects on cellular thermotolerance that make Brahman embryos more resistant to heat shock are not expressed at the 2-4-cell stage. There were few differences between Brahman and Holstein in response to estrus synchronization and superovulation. The fact that cleavage tended to occur earlier in Brahman than Holstein embryos suggests breed differences in timing of ovulation, fertilization or events leading to cleavage.

Animals↗

Identification of possible mediators of embryonic mortality caused by mastitis: actions of lipopolysaccharide, prostaglandin F2alpha, and the nitric oxide generator, sodium nitroprusside dihydrate, on oocyte maturation and embryonic development in cattle.

PROBLEM: Mastitis and immunization against constituents of organisms causing mastitis can reduce fertility of cattle and sheep, respectively. For the current experiments, it was hypothesized that these effects are mediated via actions of lipopolysaccharide (LPS), prostaglandin F2alpha (PGF2), and nitric oxide on oocyte maturation and embryonic development. METHOD OF STUDY: To evaluate effects on oocyte maturation, oocytes were matured with various concentrations of LPS, PGF2alpha, or the nitric oxide (NO) generator, sodium nitroprusside (SNP). Following maturation, oocytes were fertilized and cultured until day 8 after fertilization. To test effects on embryo growth, oocytes were matured and fertilized and cultured after fertilization with LPS, PGF2alpha, or SNP. RESULTS: Addition of 100 and 1000 ng/mL LPS and 50 and 100 ng/mL PGF2alpha to oocyte maturation medium reduced the proportion of oocytes that became blastocysts at day 8 after fertilization. When added after fertilization, in contrast, neither LPS nor PGF2alpha reduced development to the blastocyst stage. Unlike for LPS and PGF2alpha, addition of SNP during oocyte maturation was without effect on the proportion of oocytes that became blastocysts at day 8 after fertilization. However, addition of 10 microM SNP to culture medium after fertilization completely prevented development to the blastocyst stage while 0.1 and 1 microM SNP did not affect development. CONCLUSIONS: Results indicate that increased local concentrations of LPS, PGF2alpha, and NO can have deleterious consequences on oocyte function (LPS, PGF2alpha) and embryonic development (NO). Thus, these molecules are putative mediators of effects of infectious disease or inflammation, including mastitis, on fertility of cattle.

Animals↗