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In the present study 795 teats of slaughtered cows were examined endoscopically. Teats which showed lesions were examined histologically. Abnormalities were detected endoscopically in 37 teats. In five cases red discolorations were observed. Histologically these alterations proved to be fresh bleedings within the fibromuscular layer of the teat cistern. Thirteen teats showed yellowish discolorations of the cistern surface. Three of these cases could not be cleared up histologically. In the other cases moderate or severe blood pigment deposition (n = 10) or severe fibrosis (n = 3) of the fibromuscular layer were found. Neither fresh bleeding nor severe blood pigment deposition was found in teats with normal coloured epithelium of the cistern. Cistern surface was smooth in 18 cases, whereas in 19 cases nodules were found. Histologically all nodules consisted of fibrovascular tissue ("granulation tissue") covered with cisternal epithelium. Detailed histological examinations, however, revealed considerable morphological variations regarding the overlying epithelium, the extent and kind of cellular infiltration, the extent of angioplasia and fibroplasia as well as fresh bleedings or deposition of blood pigment in the underlying fibrovascular tissue in some cases. It was concluded, that teat endoscopy enables a differentiation between fresh and old bleedings according to the coloration of the cisternal surface. Nodular proliferations of the cistern surface generally consists of fibrovascular ("granulation") tissue. Further histological details, however, and thus possible information about the pathogenesis of the lesions cannot be assessed endoscopically.

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In the present study endoscopically diagnosed changes of the Fürstenberg's rosette were examined histologically. For this purpose the teats of 200 slaughtered cows were inspected. In 19 cases proliferations in the region of the Fürstenberg's rosette were found. Additionally, tissue samples were taken from the Fürstenberg's rosette of 26 cows which showed milk flow disorders due to stenoses of this part of the teat. Teats of slaughtered cows as well as biopsy specimens were fixed in 5% formaldehyde and embedded in paraffin and in plastic. All specimens were stained according to H.E., Giemsa and Turnbull, respectively. Histologically, the proliferations of the Fürstenberg's rosette consisted of fibrovascular tissue (granulation tissue) covered by keratinized squamous epithelium (teat canal epithelium) in eleven cases. In the other cases double-layered cuboid epithelium (cisternal epithelium) was found additionally. The formation of granulation tissue and deposition of blood pigment was observed exclusively in areas covered by stratified epithelium. Therefore a traumatic lesion of the teat canal was supposed to be the cause of the histological findings. In contrast to several reports in the literature, however, no indications of an inversion of the teat canal epithelium were found. On the basis of these findings, the hitherto assumed idea of pathogenesis of proliferations of the Fürstenberg's rosette was modified and a new pathogenesis pattern was developed.

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In previous studies, transrectal color Doppler sonography was used to demonstrate an increase in genital blood flow resistance in subfertile mares. The objectives of the present study were to determine the effects of an anticoagulant (acetylsalicylic acid) and a vasodilator (captopril) on uterine and ovarian perfusion and plasma progesterone concentrations in cycling mares. From Day 1 to 11 of an estrous cycle (Day 0=day of ovulation following prostaglandin-induced luteolysis), five Trotter mares were given 2500 mg lactose, 2500 mg ASA, or 50 mg captopril twice daily in their feed (one compound per cycle, in random order). Transrectal color Doppler sonography was used to examine both uterine arteries and the ovarian artery ipsilateral to the corpus luteum once daily, immediately prior to administration of the drug. Blood flow resistance was determined semiquantitatively using the pulsatility index (PI) and plasma progesterone concentrations were determined with an enzyme immunoassay. Compared to the placebo, both ASA and captopril decreased mean PI values of both uterine arteries of all mares. On average, ASA decreased the PI of the uterine arteries by 25%; this was more (P<0.05) than the average decrease (13%) caused by captopril. Both drugs decreased (P<0.05) blood flow resistance in the ovarian arteries, although there was no difference (P<0.05) in their efficacy. In addition, both ASA and captopril increased (P<0.0001) plasma progesterone concentrations (18 and 17%, respectively). In conclusion, either ASA or captopril improved uterine and ovarian perfusion; however the effects on fertility were not determined.

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Transrectal color Doppler sonography was used to evaluate the effect of intrauterine infusion of skim milk semen extender, seminal plasma and raw semen on the endometrium and blood flow in the uterine and ovarian arteries in mares. Six Trotter mares (mean age: 12 years) were examined during estrus in three cycles. Each mare received an intrauterine infusion of 20 ml of skim milk semen extender, seminal plasma or raw semen during estrus in one of three cycles. Blood flow measurements in both uterine and ovarian arteries and the determination of intrauterine fluid via sonography were performed before each infusion and 1, 3, 6, 12, and 24 h after infusion. Forty-eight hours later, the intrauterine infusion and measurements were repeated using the same time intervals. Changes in blood flow were detected using transrectal color Doppler sonography and were evaluated using the mean time-averaged maximum velocity (TAMV) of the blood flow. Cytological and bacteriological examination of uterine swabs performed 48 h after the second infusion revealed less inflammation and bacterial growth in mares infused with skim milk semen extender than in those infused with seminal plasma or raw semen. There was an increase in intrauterine fluid as early as 1 h after infusion of any of the substances. The infusion of skim milk semen extender had no effect on uterine blood flow. Within 1 h after infusion of seminal plasma or raw semen, there was an increase in the TAMV values of both uterine arteries (P<0.05). In contrast, ovarian blood flow increased only in the artery ipsilateral to the preovulatory follicle and only after the infusion of raw semen (P<0.05). In conclusion, the changes in uterine perfusion observed after intrauterine infusion may be associated with endometrial inflammation and vasodilatory components in the seminal plasma, whereas the changes seen in ovarian blood flow are possibly attributable to the interaction between sperm and oviduct.

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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.

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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.

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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.

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During the occasional testing of Escherichia coli from faecal samples of young calves we observed multi-resistant isolates. Because of the significance of E. coli as an indicator bacterium for resistance trends we tested E. coli populations of young calves over a longer period. Here we present the results of a retrospective study comparing isolates from 1998 to 2000. Moreover, we compared, in a clinical study, the resistance rates of E. coli populations from 67 hospitalized calves both before and after hospitalization (with or without anti-microbial therapy), and with their anamnestic data of antibiotic usage. The highest resistance rates were found to be more than 80% for tetracyclines, ampicillin, sulfonamide/trimethoprim combinations, and chloramphenicol. A significant increase or decrease over the years was not observed. In analysing the data of hospitalized calves, an increase of resistance to some anti-microbials had to be registered that seemed to be connected with the selective pressure due to agents used in the clinic. In comparing anamnestic data and resistance rates it became obvious that reliable data are not easily available and that a number of potential anti-microbial influence factors have to be taken into account.

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Milk flow was recorded in 21 cows for three days after they were admitted to a large animal hospital. When the spontaneous flow of milk had stopped, a physiological dose (1 iu) of oxytocin was administered intravenously. Five of the cows were, in addition, treated with 0.35 mg of a long-acting analogue of oxytocin (carbetocin) one hour before the first milking after they were admitted. In the 16 cows not treated with carbetocin, only about 30 per cent of the total milk yield was released spontaneously on the first day, and the injection of 1 iu of oxytocin released approximately another 60 per cent of the total milk yield. On the second day, the proportion of the total milk yield released spontaneously increased and the fraction released after the injection of 1 iu oxytocin decreased. In contrast, the five cows treated with carbetocin released on average 94 per cent of the total milk yield spontaneously during the first milking.

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The importance of standardizing the procedures of sample and slide preparation for computer-assisted morphologic analysis has been emphasized in human and veterinary andrology. The purpose of this study was to optimize slide preparation (dilution grade and sperm washing), staining procedures and analysis conditions (colour of light source and objective magnification) for the morphometric analysis of bull spermatozoa using the Hamilton Thorne morphology analyzer integrated visual optical system (IVOS). For experiment 1, one ejaculate was collected from one bull and diluted to 200,000-300,000 spermatozoa/microl. Slides were prepared and stained using seven different procedures: rapid Papanicolaou (PAP), rapid Papanicolaou with prolonged staining times (PAP+). Diff-Quik (DIF), haematoxylin (HEM). Farelly (FAR), Spermac (SPER) and the modified GZIN (MGZIN) staining. All slides were analysed using a Hamilton Thorne Morphology Analyser IVOS equipped alternatively with a red, green or blue light source, and a 40x or 100x oil immersion objective. Recognition and digitization errors as well as morphometric parameters were determined. The IVOS was unable to detect DIF-stained spermatozoa. The GZIN and the SPER staining as well as the blue light source led to unsatisfactory results. Among the staining methods examined, the FAR, HEM, PAP+, and PAP staining, preferably in combination with the green light source, and the 40x objective yielded optimal results concerning sperm recognition and digitization. The 100x objective did not allow reliable analysis of the sperm heads because of a frequently appearing digitization error. For experiment 2, three ejaculates were collected from each of three bulls and diluted to five dilution grades (100 000-500 000 spermatozoa/microl). An aliquot of each dilution grade was washed additionally. The percentage of correctly digitized sperm heads decreased with increasing spermatozoal concentration. However, the evaluation speed increased. The range of 200 000-300 000 spermatozoa/microl appeared to be a reasonable compromise for both criteria. Sperm washing failed to further improve the analysis results. Sperm head dimensions were influenced significantly by all variations of the methods in both experiments. In conclusion, using the proposed methods, the IVOS allows precise and reliable morphometric analyses of bull spermatozoa. The consistent application of these procedures may lead to an inter-laboratory standardization and to further establishment of generally accepted morphometric criteria used in human andrology (e.g. World Health Organisation or strict criteria).

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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 < 0.001). As in all cows, changes of RI and TAMV values between the left and right artery during the estrous cycle were correlated (correlation coefficient r > 0.72; P < 0.0001); the mean values of both sides were used for subsequent analyses. Variance component estimates for the effect of cow on RI and TAMV were 8 and 13% and for the influence of day of estrous cycle they were 70 and 47%, respectively (P <0.0001). Between estrous cycles no significant differences could be measured within cows (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.

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Estrus detection has a tremendous impact on the reproductive efficiency in dairy farms. Recently the systematic use of hormones for synchronization of estrus and ovulation has been propagated. These programs are designed to facilitate estrus detection and/or increase its efficiency. Prostaglandin programs are used to improve estrus detection and reproductive management in dairy operations. All cows are treated up to three times in weekly or biweekly intervals at the end of the voluntary waiting period. This should lead to groups of cows in estrus within two to four days after PG treatment. Since PG programs cannot completely eliminate the need for estrus detection procedures for fixed time artificial insemination (AI) were developed. Fixed time AI not only requires control of the luteal phase of the cow but also synchronization of the follicular development. Treatment with GnRH will induce a new follicular wave and provide a dominant follicle in a defined growth phase at the time of PG administration seven days later. A second injection of GnRH given 48 hours after PG results in ovulation approximately 24 to 32 hours after GnRH (OvSynch-procedure). The pros and cons of hormone programs in dairy farming are discussed in the light of field trials and experiments performed in our department.

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Color Doppler ultrasound was used transrectally in 6 mares to locate both the left and right Aa. uterinae and to obtain flow velocity waveforms at defined times (Days 0, 5, 10, 15 and 20) during 4 estrous cycles. Blood flow reflected by the resistance index (RI) was determined for both arteries on 120 occasions. As there was no significant difference and a high correlation in the RI values between the left and right arteries (paired Student's t-test, correlation coefficient r > 0.94; P < 0.05), the average RI value was used for subsequent analyses. There were correlations between RI values, mares and day of estrous cycle (P < 0.0001). The mean RI was higher (P < 0.05) in the 2 multiparous mares (age, 12 to 13 yr) than in the 4 younger maiden mares (age, 6 to 10 yr). During the estrous cycle mean RI values on Day 0 (day of ovulation) and Day 10 were higher (P < 0.05) than on Days 5, 15 and 20, whereas between estrous cycles within mares no differences (P > 0.05) could be measured. The results suggest that transrectal Color Doppler sonography is a noninvasive method for examining differences in impedance to uterine blood flow between different mares and cycle periods.

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The effects of luteolytic doses of PGF2 alpha (25 mg Dinoprost) and its synthetic analogues Cloprostenol (500 micrograms), Luprostiol (15 mg) and Tiaprost (525 micrograms) on bovine myometrial activity were investigated using a miniature pressure transducer placed in one uterine horn. The compounds were administered intravenously to 4 lactating cyclic cows at diestrus, proestrus, estrus and metestrus. Intrauterine pressure changes were assessed by computerized planimetry of the pressure tracings 30 minutes before and 60 minutes after treatment. Baseline intrauterine pressure was set at zero and treatment effects were expressed as percent change from an equivalent control period (= 100%). Following administration of Dinoprost there was a significant increase of uterine contractility in diestrus (515%), proestrus (198%) and metestrus (256%), but not in estrus. In comparison to PGF2 alpha the analogues Luprostiol and Tiaprost were less effective (Luprostiol: 195% and 154% in diestrus and proestrus resp., Tiaprost: 215% during diestrus), while Cloprostenol did not cause a significant change of intrauterine pressure in any stage of the estrous cycle. The results indicate that the myotonic effects which F2 alpha-prostaglandins exert on the uterus of cycling cows is affected both by the type of prostaglandin and the stage of the estrous cycle.

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In 1970 semen from a Haflinger-stallion was frozen by the pellet method. 18 years later semen samples were used to inseminate 4 mares. Inseminations were performed shortly after ovulation with a total number of motile spermatozoa between 150 and 636 x 10(6), the percentage of motile spermatozoa being 20% to 40%. Three mares conceived after a single insemination, one mare got pregnant after 4 inseminations during 3 oestrous periods. Meanwhile, 3 foals were born and one of the mares is still pregnant. The results demonstrate that long-term storage of frozen semen in liquid nitrogen does not impair its fertilizing capacity.

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