Heinrich Bollwein

TRANSRECTAL DOPPLER SONOGRAPHY OF UTERINE BLOOD FLOW IN COWS DURING THE ESTROUS CYCLE

Transrectal Doppler ultrasound was used for the noninvasive investigation of uterine blood flow in cows. Both the left and right Aa. uterinae were scanned to obtain blood flow velocity waveforms over 2 consecutive estrous cycles. Blood flow was reflected by the resistance index (RI) and the time-averaged maximum velocity (TAMV). Intra-observer reproducibility of Doppler measurements was evaluated. The intra-class correlation coefficient (Intra-CC) was 0.97 for the RI and 0.95 for TAMV. While RI values did not differ between the left and right A. uterina (P > 0.05), differences in TAMV occurred between both vessels in 2 cows. These differences were not related to the ovary bearing the dominant follicle or to the corpus luteum (P 0.72; P 0.05). The highest RI and lowest TAMV values occurred on Day 0 (= day of ovulation) and Day 1, while the lowest RI and highest TAMV values were measured between Days -3 and -1 of the estrous cycle, respectively. There was a positive correlation between TAMV and estrogen concentrations and a negative correlation between RI and plasma estrogen levels. Plasma progesterone levels and TAMV were negatively correlated, but no correlation could be measured (P > 0.05) between RI values and plasma progesterone concentrations. While there were no differences in plasma concentrations of estrogens and progesterone between estrous cycles within cows, the levels of these hormones differed between cows. The results show that transrectal Doppler sonography is a useful, noninvasive method for examining uterine blood flows in cows. If there is an influence of uterine perfusion on fertility in cows its role needs further investigation.

Transrectal Doppler sonography of uterine blood flow in cows during pregnancy

Transrectal Doppler ultrasound was used for the noninvasive investigation of uterine blood flow in three cows during pregnancy. The uterine arteries ipsi and contralateral to the conceptus were scanned monthly. Blood flow was reflected by the following parameters: resistance index (RI), time-averaged maximum velocity (TAMV), diameter of the vessel (D) and the volume of blood flow (VOL). RI values were negatively correlated to all other blood flow parameters (P < 0.01). Positive correlations occurred between TAMV, D and VOL (P < 0.0001). While blood flow parameters did not differ between cows (P > 0.05), the month of gestation showed a positive effect on RI and negative effects on TAMV, D and VOL (P < 0.0001). The RI was lower and TAMV, D and VOL higher in the uterine artery ipsilateral to the conceptus (P < 0.05). RI values decreased continuously during the first 8 months of gestation and remained from then until birth at a relatively constant level. While TAMV increased especially in two-thirds of pregnancy, a relatively uniform rise of D was noticed. VOL increased exponentially with stage of gestation. The results show that transrectal Doppler sonography is a suitable, noninvasive method for the examination of uterine blood flow during pregnancy in cows. Using this technique it might be possible in the future to determine the role of uterine blood flow in cows at the risk of abortion.

Luteal blood flow is a more appropriate indicator for luteal function during the bovine estrous cycle than luteal size

The objective of this study was to assess the reliability of luteal blood flow (LBF) as recorded by color Doppler sonography to monitor luteal function during the estrous cycle of dairy cows and to compare the results with that for the established criterion luteal size (LS) as determined by B-mode sonography. In total, 14 consecutive sonographic examinations were carried out in 10 synchronized lactating Holstein-Friesian cows (Bos taurus) on Days 4, 5, 6, 7, 8, 10, 12, 14, 16, -5, -4, -3, -2, -1 of the estrous cycle (Day 1=ovulation). Plasma progesterone concentrations in venous blood (P(4)) were quantified by enzyme immunoassay. Luteal size was determined by sonographic measurement of the maximal cross-sectional area of the corpus luteum (CL). Luteal blood supply was estimated by calculating the maximum colored area of the CL from power Doppler sonographic images. Luteal size doubled during the luteal growth phase (until Day 7) and remained at this level during the luteal static phase (Day 8 to 16) before decreasing rather slowly during luteal regression (Days -5 to -1). Luteal blood flow doubled during the growth phase, doubled furthermore during the static phase, and decreased rapidly during luteal regression. Thus, LBF values represented highly reliable predictors of luteal status. Luteal blood flow predicted reliably a P(4)>1.0 ng/mL by reaching only 35% of the maximal values, whereas LS had to exceed 60% of the maximal values to indicate reliably a functional CL. It is concluded that LBF reflected luteal function better than LS specifically during luteal regression.

Future consequences and challenges for dairy cow production systems arising from climate change in Central Europe - a review

It is well documented that global warming is unequivocal. Dairy production systems are considered as important sources of greenhouse gas emissions; however, little is known about the sensitivity and vulnerability of these production systems themselves to climate warming. This review brings different aspects of dairy cow production in Central Europe into focus, with a holistic approach to emphasize potential future consequences and challenges arising from climate change. With the current understanding of the effects of climate change, it is expected that yield of forage per hectare will be influenced positively, whereas quality will mainly depend on water availability and soil characteristics. Thus, the botanical composition of future grassland should include species that are able to withstand the changing conditions (e.g. lucerne and birds foot trefoil). Changes in nutrient concentration of forage plants, elevated heat loads and altered feeding patterns of animals may influence rumen physiology. Several promising nutritional strategies are available to lower potential negative impacts of climate change on dairy cow nutrition and performance. Adjustment of feeding and drinking regimes, diet composition and additive supplementation can contribute to the maintenance of adequate dairy cow nutrition and performance. Provision of adequate shade and cooling will reduce the direct effects of heat stress. As estimated genetic parameters are promising, heat stress tolerance as a functional trait may be included into breeding programmes. Indirect effects of global warming on the health and welfare of animals seem to be more complicated and thus are less predictable. As the epidemiology of certain gastrointestinal nematodes and liver fluke is favourably influenced by increased temperature and humidity, relations between climate change and disease dynamics should be followed closely. Under current conditions, climate change associated economic impacts are estimated to be neutral if some form of adaptation is integrated. Therefore, it is essential to establish and adopt mitigation strategies covering available tools from management, nutrition, health and plant and animal breeding to cope with the future consequences of climate change on dairy farming.

Uterine and ovarian blood flow during the estrous cycle in mares

Uterine and ovarian blood flow was investigated in four mares during two consecutive estrous cycles using transrectal color Doppler sonography. The uterine and ovarian arteries of both sides were scanned to obtain waves of blood flow velocity. The pulsatility index (PI) reflected blood flow. There were significant time trends in PI values of all uterine and ovarian blood vessels during the estrous cycle (P < 0.05). PI values did not differ between the uterine arteries ipsi- and contralateral to the corpus luteum or the ovulatory follicle. PI values of the uterine arteries showed a wave shaped profile throughout the estrous cycle. The highest PI values occurred on Days 0 and 1 (Day 0 = ovulation) and around Day 11, and the lowest PI values were measured around Days 5 and -2 of the estrous cycle. During diestrus (Days 0-15) PI values of the ovarian artery ipsilateral to the corpus luteum were significantly lower than PI values of the contralateral ovarian artery (P < 0.0001). No differences (P > 0.05) in resistance to ovarian blood flow occurred between sides during estrus (Days -6 to -1). In this cycle stage PI values decreased in both ovarian vessels (P < 0.05). During diestrus, high PI values of the ovarian artery ipsilateral to the corpus luteum were measured between Days 0 and 2, followed by a decline until Day 6 (P < 0.05). From this time on, the resistance to blood flow increased continuously until Day 15 (P < 0.05). The cyclic blood flow pattern in the contralateral ovarian artery was similar to that in the uterine arteries (r = 0.68; P < 0.0001). No correlations occurred between the diameter of the corpus luteum and the PI values of the ipsilateral ovarian artery (P > 0.05) during diestrus. During estrus, there was a negative relationship between growth of the diameter of the ovulatory follicle and changes in PI values of the dominant ovarian artery (r = -0.41; P < 0.05). PI values of the uterine arteries and of the ovarian artery ipsilateral to the ovulatory follicle were negatively related to estrogen (E) levels in plasma during estrus (uterine arteries: r = -0.21; P < 0.05; dominant ovarian artery: r = -0.35; P < 0.05). In diestrus, PI values of the dominant ovarian artery were negatively related to plasma progesterone levels (r = -0.38; P < 0.0001), but not the PI values of the uterine arteries (P > 0.05). The findings of this study show that there are characteristic changes in blood supply of the uterus and the ovaries throughout the equine estrous cycle. There are negative correlations between resistance to blood flow in the uterine and ovarian arteries and the plasma estrogen levels during estrus. In diestrus, there is a negative relationship between the resistance to ovarian blood flow and the progesterone levels.

Transrectal Doppler sonography of uterine and umbilical blood flow during pregnancy in mares

Transrectal color Doppler sonography was used to investigate uterine and umbilical blood flow during pregnancy (duration, 46-48 weeks) in four mares. The resistance index (RI) and blood flow volume (VOL) of the uterine arteries ipsilateral and contralateral to the conceptus, and the presence of an early diastolic notch in the Doppler wave, were evaluated every 4 week throughout pregnancy. Fetal blood flow was calculated semiquantitatively every 2 week (from 20 to 40 weeks), using the RI of the umbilical arteries. During the entire period of investigation, there were no significant individual variations in uterine RI and VOL nor differences between the two uterine arteries. Mean RI decreased by more than half during pregnancy from 0.89 +/- 0.01 to 0.39 +/- 0.03, and mean VOL increased almost 400-fold from 69 +/- 37 to 27,467 +/- 8851 ml/min. There were relationships (P<0.0001) between week of pregnancy (x) and RI as well as VOL. These were described by the equations RI=0.938-0.150 ln(x) and VOL (ml/min)=7.621x(2.157). Log transformed total estrogen (TE) were related to RI (r=-0.879; P<0.05) as well as to VOL (r=0.888; P<0.05). The notch in the Doppler wave of the uterine artery disappeared between 18 and 26 weeks. There was a correlation (P<0.0001) between week of gestation (x) and RI values of the umbilical arteries; this was described by the equation RI=1.763-0.071x+0.001x2. Further studies are needed to determine whether transrectal color Doppler sonography could be used to identify mares at risk of abortion.

Transrectal Doppler sonography of uterine blood flow during early pregnancy in mares

Transrectal color Doppler sonography was used for the noninvasive investigation of uterine blood flow in five mares. Both the left and right uterine arteries were scanned to obtain blood flow velocity waveforms during two consecutive estrous cycles and two early pregnancies in each mare. Blood flow was expressed as the time-averaged maximum velocity (TAMV) and the resistance index (RI). In all pregnancies the embryonic vesicle could be detected for the first time on Day 11 (day of ovulation: Day 0). No differences in mean TAMV and RI values of both uterine arteries were observed in comparison to the corresponding days of the estrous cycle until Day 11 of pregnancy (P>0.05). From Day 11 onwards, mean TAMV values were higher and mean RI values lower in pregnant mares than in cyclic mares (P<0.05). During the estrous cycle TAMV and RI values did not differ between the right and left uterine arteries (P>0.05). From Days 15 to 29 of pregnancy, TAMV values were consistently higher and RI values lower in the uterine artery ipsilateral to the conceptus and they had a more distinct rise and decline, respectively, compared to the contralateral uterine artery (P<0.05). The variance component estimates for the effect of mare on TAMV and RI values during pregnancy were 60 and 53%, respectively, and for the effect of day of pregnancy, they were 29 and 34%, respectively (P<0.0001). Within mares there were no significant differences between the two pregnancies with regard to blood flow (P>0.05). The results show that uterine blood supply increases in mares during the second week of pregnancy compared to cyclic mares. Furthermore there are individual variations in blood flow between mares.

Luteal blood flow during the estrous cycle in mares.

Transrectal color Doppler ultrasound was used for the noninvasive investigation of luteal blood flow during the estrous cycle in six mares. Color was displayed in Power-Mode, in which the number of color pixels on the ultrasound image is related to the number of moving blood cells. Three pictures with a maximum number of color pixels of the corpus luteum (CL) during an examination period of about 20 min were selected and digitized on a laptop equipped with an external frame grabber card. The intra-class correlation coefficient for the number of color pixels was 0.90. In all estrous cycles similar patterns of changes in (C), in the cross-sectional area of sectional planes of the CL (A), and in plasma progesterone levels (P) occurred. Variance component estimates for the effect of the mare on (C), (A) and (P) were 14, 23 and 4%, for the influence of day of estrous cycle they were 41, 5 and 58% and for the effect of estrous cycle they were 7, 5 and 5%, respectively. There were high positive correlations between cyclic changes in (C) and (P) (r = 0.58; P < 0.0001). The increase in (C) between Days 0 and 5 (Day 0: ovulation) remained at high levels until Day 7 and then decreased until Day 15. There were relationships between (C) and (A) (r = 0.37; P < 0.0001) and between (A) and (P) (r = 0.24; P < 0.05), but correlation coefficients were not as high as between (C) and (P). Differences in (C), (A) and (P) between estrous cycles within mares and between mares were not related to each other (P > 0.05). The results show that transrectal color Doppler sonography is a useful, noninvasive method for examining luteal blood flow in mares, and that there are cyclic changes and individual differences in the vascularization of the CL. The possible influence of luteal perfusion on fertility in mares needs to be investigated in further studies.

Vascular and immune regulation of corpus luteum development, maintenance, and regression in the cow

The bovine corpus luteum (CL) is a unique, transient organ with well-coordinated mechanisms by which its development, maintenance, and regression are effectively controlled. Angiogenic factors, such as vascular endothelial growth factor A and basic fibroblast growth factor, play an essential role in promoting progesterone secretion, cell proliferation, and angiogenesis. These processes are critically regulated, through both angiogenic and immune systems, by the specific immune cells, including macrophages, eosinophils, and neutrophils, that are recruited into the developing CL. The bovine luteolytic cascade appears to be similar to that of general acute inflammation in terms of time-dependent infiltration by immune cells (neutrophils, macrophages, and T lymphocytes) and drastic changes in vascular tonus and blood flow, which are regulated by luteal nitric oxide and the vasoconstrictive factors endothelin-1 and angiotensin II. Over the period of maternal recognition of pregnancy, the maternal immune system should be well controlled to accept the semiallograft fetus. The information on the presence of the developing embryo in the genital tract is suggested to be transmitted to the ovary by both the endocrine system and the circulating immune cells. In the bovine CL, the lymphatic system, but not the blood vascular system, is reconstituted during early pregnancy, and interferon tau from the embryo could trigger this novel phenomenon. Collectively, the angiogenic and vasoactive factors produced by luteal cells and the time-dependently recruited immune cells within the CL and their interactions appear to play critical roles in regulating luteal functions throughout the life span of the CL.

Chromatin-unstable boar spermatozoa have little chance of reaching oocytes in vivo

In the present study, the prevalence of chromatin instability in the fertilizing-competent sperm population in the porcine oviduct in vivo was examined through qualitative analysis of the chromatin structure status of accessory boar sperm found in in vivo-derived embryos. The binding of chromatin-unstable sperm to oviductal epithelium in vitro was also studied. To examine the sperm chromatin state, a modified fluorescence microscopic sperm chromatin structure assay was used. Among a population of 173 fertile boars, individuals were selected for according to their chromatin status: 25 animals showed more than 5% of chromatin-unstable sperm in their ejaculates, and 7 showed consistently elevated percentages of chromatin-unstable sperm in three successively collected semen samples. A positive correlation was found between incidence of chromatin instability and attached cytoplasmic droplets (r=0.44, P<0.01). Analyses of accessory spermatozoa from in vivo-derived embryos demonstrated that the proportion of chromatin-unstable sperm was significantly (P<0.05) reduced in the population of fertilizing-competent sperm in the oviduct compared with the inseminated sperm. Populations of sperm bound to the oviduct in vitro had significantly (P<0.05) lower percentages of chromatin instability than in the original diluted semen sample. In conclusion, numbers of sperm with unstable chromatin are reduced in the oviductal sperm reservoir, possibly because of associated changes in the plasma membrane that prevent sperm from binding to the oviductal epithelium. We conclude that in vivo the likelihood that sperm with unstable chromatin will reach the egg and fertilize it is low.