Granular vulvitis was reproduced in ten virgin heifers following vulvar inoculation with strains of ureaplasma previously isolated from natural cases. The disease appeared one to three days postinoculation and was characterized by vulvar swabs but not from the upper mucopurulent discharge. At necropsy 13 to 41 days later, ureaplasmas were recovered consistently from vulvar swabs but not from the upper reproductive tract. It was concluded that some strains of ureaplasma are pathogenic and should be viewed as a cause of bovine granular vulvitis.
Sixteen bovine genital mycoplasmal isolates obtained from semen and prepuce of bulls and from aborted fetuses were compared physiologically and serologically with the Donetta strain (tentatively Mycoplasma agalactiae var. bovis), a known pathogen. All isolates were distinct from the Donetta organism. Four appeared to be saprophytes, and the remainder were placed in one group which could not be further separated by the biochemical or serological methods used. Two of the organisms in the latter group have been subsequently identified as M. bovigenitalium. Uterine infusion of broth cultures of four isolates into virgin heifers failed to produce clinical evidence of disease, and significant lesions were not present at necropsy. The mycoplasmas were recovered from cervicovaginal mucus of only three heifers, and never for more than 3 days postinfusion. Since the organisms were not recovered from any organs at necropsy, it appears that the mycoplasmas were incapable of surviving in the clinically normal virgin female reproductive tract.
A granular vulvitis syndrome associated with ureaplasma infection was first recognized in Ontario dairy herds in 1972.
The acute form of the disease was characterized by a purulent vulvar discharge, an inflamed hyperemic vulvar mucosa and varying degrees of granularity. In the chronic form, there was an absence of a purulent discharge and a gradual decline in the severity of the hyperemia and granularity. Epithelial inclusion cysts were observed in the vulvar epithelium of approximately 10% of affected cows.
A seasonal variation in the incidence of the disease was observed. Herd morbidities during the summer months reached a low of 37% and increased to 75% during the winter months with constant housing.
When widespread in herds, the acute form of the disease had a significant effect on fertility. In four herds examined, first service conceptions dropped on average by 27%.
The chronic form of the disease had a less detrimental effect on fertility with first service conceptions being reduced on average by 13%.
Intrauterine infusions of a tetracycline 24 hours postbreeding were found to be of value in improving conception rates in acutely affected herds.
Cultures for mycoplasmatales, viruses and bacteria were made from bovine vulvar swabs to determine whether ureaplasma was associated with a clinical granular vulvitis observed in 16 Ontario dairy herds. Ureaplasma was isolated from 23.5% of 34 clinically normal cows, 74% of 27 cows with mild to moderate vulvar hyperemia but no discharge and 100% of 20 cows with acute vulvar hyperemia accompanied by purulent discharge. There were statistically significant differences in rates of isolation among clinical groups. Mycoplasma bovigenitalium was isolated from 7.7% and 20% of cows with moderate or acute vulvitis respectively but not from normal cows. Haemophilus somnus was isolated from 25% of cows with acute vulvitis. There were no significant differences in isolations of Escherichia coli, Corynebacterium pyogenes and alpha-hemolytic streptococcus between normal and clinically affected animals. Cultures of 135 repeat samples from 33 cows revealed that ureaplasma persisted in some animals for at least three months. No viruses were isolated from any of the animals in this study.
Two field isolates of Ureaplasma diversum spp. were used to infect heifers at the time of insemination in a preliminary study to observe the effect of infection on early pregnancy. M84-14c-1 was a field isolate from a bull's prepuce typed by immunofluorescence to be similar to U. diversum strain T-44 (Group C). M84-477c-4 was a field isolate from bovine semen typed by immunofluorescence to be similar to U. diversum strain T-288 (Group A). All three heifers infected with M84-477c-4 had a mild granular vulvitis at some time during the trial. None was pregnant when slaughtered 27 days after infection. The result of infection with M84-14c-1, a preputial isolate, was not consistent. One heifer had no infection and a normal pregnancy, one heifer was infected with an abnormal pregnancy, and one heifer was open but ureaplasmas were not detected until day 17 of the trial.
Infertility and subfertility are important and pervasive reproductive problems in both domestic animals and humans. The majority of embryonic loss occurs during the first three weeks of pregnancy in cattle and women due, in part, to inadequate endometrial receptivity for support of embryo implantation. To identify heifers of contrasting fertility, serial rounds of artificial insemination (AI) were conducted in 201 synchronized crossbred beef heifers. The heifers were then fertility classified based on number of pregnancies detected on day 35 in four AI opportunities. Heifers, classified as having high fertility, subfertility or infertility, were selected for further study. The fertility-classified heifers were superovulated and flushed, and the recovered embryos were graded and then transferred to synchronized recipients. Quantity of embryos recovered per flush, embryo quality, and subsequent recipient pregnancy rates did not differ by fertility classification. Two in vivo-produced bovine embryos (stage 4 or 5, grade 1 or 2) were then transferred into each heifer on day 7 post-estrus. Pregnancy rates were greater in high fertility than lower fertility heifers when heifers were used as embryo recipients. The reproductive tracts of the classified heifers were obtained on day 14 of the estrous cycle. No obvious morphological differences in reproductive tract structures and histology of the uterus were observed in the heifers. Microarray analysis revealed differences in the endometrial transcriptome based on fertility classification. A genome-wide association study, based on SNP genotyping, detected 7 moderate associations with fertility across 6 different chromosomes. Collectively, these studies support the idea that innate differences in uterine function underlie fertility and early pregnancy loss in ruminants. Cattle with defined early pregnancy success or loss is useful to elucidate the complex biological and genetic mechanisms governing endometrial receptivity and uterine competency for pregnancy.
We measured antibody levels in serum and cervicovaginal mucus (CVM) of four heifers vaccinated with two inoculations of killed Ureaplasma diversum strain 2312 in incomplete Freund's adjuvant (IFA) two weeks apart, and six heifers given a placebo. Two weeks later, the vaccinates and four placebo heifers, were challenged by intravaginal inoculation with 6.4 x 10(8) colony-forming units of the homologous U. diversum strain. The remaining two placebo heifers served as unvaccinated, unchallenged controls. Antibody levels in serum and CVM of all heifers were determined by an enzyme-linked immunosorbent assay (ELISA). Vaccination stimulated specific IgG1 and IgG2 responses in serum and CVM but only a slight IgM and no IgA response. In both vaccinate and placebo heifers, subsequent intravaginal challenge resulted in a granular vulvitis (GV) with a predominant IgA response in the CVM. The GV gradually subsided during the 35 day observation period but ureaplasmas were consistently demonstrated by culture. We concluded that subcutaneous vaccination stimulated a specific, albeit nonprotective, IgG response in serum and CVM. In contrast, vaginal infection primarily induced a mucosal IgA response.
Ureaplasma urealyticum respiratory tract colonization in preterm infants has been associated with a high incidence of pneumonia and the development of bronchopulmonary dysplasia. However, study of this human pathogen has been hampered by the absence of animal models. We have developed the first juvenile mouse model of Ureaplasma pneumonia and characterized the histopathology during the month following inoculation. C3H/HeN mice were inoculated intratracheally with a mouse-adapted clinical Ureaplasma isolate (biovar 2) or sham inoculated with 10B broth. Culture of lung homogenates and PCR of DNA from bronchoalveolar lavage fluid (BAL) confirmed the presence of Ureaplasma in 100% of inoculated animals at 1 day, 60% at 2 days, 50% at 3 days, and 25% at 7 and 14 days. Ureaplasma was undetectable 28 days postinoculation. There were marked changes in BAL and interstitial-cell composition with increased number of polymorphonuclear leukocytes 1 to 2 days and 14 days postinoculation and macrophages at 2 and 14 days postinoculation. The Ureaplasma infection caused a persistent focal loss of airway ciliated epithelium and a mild increase in interstitial cellularity. There were no differences in BAL protein concentration during the first 28 days, suggesting that pulmonary vascular endothelial barrier integrity remained intact. Comparison of BAL cytokine and chemokine concentrations revealed low levels of tumor necrosis factor alpha (TNF-α) at 3 days and monocyte chemoattractant protein 1 at 7 days in Ureaplasma-infected mice but a trend toward increased TNF-α at 14 days and increased granulocyte-macrophage colony-stimulating factor and interleukin-10 at 28 days. These data suggest that Ureaplasma alone may cause limited inflammation and minimal tissue injury in the early phase of infection but may promote a mild chronic inflammatory response in the later phase of infection (days 14 to 28), similar to the process that occurs in human newborns.
We report herein a survey in which cultures of bovine reproductive tracts for Ureaplasma diversum and mycoplasmas were carried out in order to better understand the role of these organisms in granular vulvitis (GV). Samples cultured were vulvar swabs from clinically normal cows or ones with GV, preputial swabs or raw semen from bulls, and abomasal contents of aborted fetuses.
Ureaplasma diversum was isolated from 104 (43.3%) of 240 dairy cows, 32 (27.1%) of 118 beef cows, 43 (47.2%) of 91 beef heifers, 23 (67.6%) of 34 beef bulls, and three (60%) of five dairy bulls. Mycoplasmas were isolated from 18 (7.5%) dairy cows, two (1.6%) beef cows, three (8.8%) beef bulls, and one dairy bull. No isolation was made from 97 aborted fetuses. For 65 dairy cows and 30 beef heifers with vulvar lesions, the isolation rates for ureaplasmas of 62.5% and 69.7%, respectively, were significantly higher (X2) than those for normal animals (37.5% and 30.3%). On immunofluorescent serotyping of 137 of the 205 isolates, there were 66 in serogroup C (strain T44), 18 in serogroup B (strain D48), eight in serogroup A (strain A417 or strain 2312), 14 cross-reacting, and 31 that were not identified. It was concluded that U. diversum is commonly present in the lower reproductive tract of beef/dairy cattle in Saskatchewan and is associated with granular vulvitis.
A 90-day finishing trial involving 144 feedlot heifers was conducted to compare the performance parameters and carcass characteristics of open heifers, therapeutically aborted heifers, and pregnant heifers. In the first 28 days of the trial, the aborted heifers had reduced (p < 0.05) feed intake (FI), average daily gain (ADG), and feed efficiency (FE) compared to pregnant and open heifers. Over the entire trial, on a live weight basis, the aborted group had reduced (p < 0.05) final weight, ADG, and FE compared to pregnant and open heifers. However, when the data were adjusted for total uterine weight, the aborted and open heifers had improved (p < 0.05) final weight, ADG, and FE compared to pregnant heifers. The aborted and open group had a higher (p < 0.05) carcass weight, rib eye area, dressing percentage, and cutability estimate compared to the pregnant heifers. The aborted group had lower (p < 0.05) carcass weight than the open heifers. Over the entire 90-day feeding period, there were no statistically significant differences among the groups with respect to feed intake (FI), average fat, grade fat, and carcass grades. Also, there were no significant health problems or mortality in any of the groups.
In the economic analysis, aborted heifers returned $26.41 per head more than pregnant heifers. Open heifers returned $39.94 per head more than aborted heifers, and $66.35 more than pregnant heifers. Thus, aborting feedlot heifers during the second trimester was determined to be a safe and cost effective management decision.
To determine the influence of Ureaplasma diversum on bovine fertility 11 uninfected virgin heifers with normal ovarian cyclic activity were randomly allocated to test or control groups. At a synchronized estrus, five test heifers were given an intrauterine broth inoculum containing 1.09 x 10(8) to 1.4 x 10(9) colony forming units of U. diversum and six control animals were infused with sterile ureaplasma broth medium. All animals were artificially inseminated within one hour of infusion. Pregnancy was diagnosed in one of five test heifers and all of six controls by serum progesterone concentrations measured to 25 days postinsemination. The difference in pregnancy rates between the two groups was statistically significant (p = 0.0152). It was concluded that under the conditions of this experiment U. diversum is capable of causing infertility in cattle.
Escherichia coli was inoculated into the uterine lumen of rats and rabbits at different estrous stages; one uterine horn of each animal was ligated at the cervical end. In rats, a large number of E. coli were retained in the ligated horns regardless of the estrous stage. E. coli inoculated at diestrus or pseudopregnancy induced purulent endometritis, but when inoculated at proestrus-estrus the organism caused asymptomatic infection. In nonligated horns, few E. coli were recovered, and marked histopathological changes were not observed. Large numbers of E. coli were retained in the nonligated horn at proestrus as a result of physiological constriction of the cervix. E. coli inoculated at proestrus never caused purulent endometritis in either the ligated horn or the nonligated horn. In rabbits, E. coli infused into ligated horns brought about purulent inflammation irrespective of ovarian states. The number of recoverable E. coli was reduced rapidly at the follicular phase as compared with the luteal phase. These results suggest that the stage of the estrous cycle when animals are inoculated with E. coli influences the course of the uterine infection.
Two groups of three Holstein heifers were immunized respectively with Vibrio fetus venerealis and Vibrio fetus intestinalis incorporated in Freund's complete adjuvant. Both serum and vaginal mucus agglutination titers increased following immunization. Vaginal mucus samples were more frequently positive when the homologous cells were used as antigen in the agglutination test.
Ten non-immunized heifers were inoculated with another strain of V. fetus venerealis and slaughtered at periods of 30 to 40 and 60 to 70 days post-inoculation (DPI). Agglutinating antibodies were present in the vaginal mucus of some infected individuals by five weeks post-inoculation. In the course of the experiment 11 vaginal mucus samples were obtained which agglutinated heated cells of the infecting strain; one aggglutinated whole cells. Precipitins toward homologous antigens could not be demonstrated in vaginal mucus but four of six samples tested precipitated a heat stable extract from an intestinal strain of the same O-serotype. Bacterial antigen was detected by immunofluorescence on the surface, as well as within and beneath the epithelium at all levels of the reproductive tract regardless of time of slaughter. Lesions in infected animals consisted of focal and diffuse lymphocytosis, plasmacytosis, and epithelial vacuolation. Diffuse neutrophilic infiltration of the oviducts was observed.
Agglutinins appeared in the serum of each of nine heifers immunized with whole cells of same venereal strain. Group mean serum titers for whole and heated cells were 1/28,000 and 1/1,300 respectively. Vaginal mucus samples agglutinated whole cells in 48% of tests while 6.3% reacted with heated cells. Serum, but not vaginal mucus, of immunized animals precipitated soluble antigens of the immunizing strain. The immunizing strain of V. fetus did not infect the reproductive tract of any of six immunized heifers upon challenge.
The serology of 27 heifers found to be positive to culture after inoculation with Brucella abortus strain 544, was studied. Eighteen heifers had previously been vaccinated with strain 19 or strain 45/20 and nine were unvaccinated. Post-infection serum samples were tested for Brucella antibodies by radioimmunoassay (RIA), complement fixation test (CFT), indirect haemolysis test (IHLT) and Rose Bengal plate test (RBPT). All of the unvaccinated heifers showed strong humoral responses to experimental infection in the RIA, CFT, IHLT and RBPT. The CFT and RBPT became positive sooner after infection than the other tests in the unvaccinated heifers. However, in vaccinated heifers the RIA was the most sensitive test early in infection and the results of the RBPT were variable. Three of the vaccinated heifers showed weak and inconsistent humoral responses and, in these animals, the RIA gave fewer false negative reactions than the other tests.
To assess the influence of hormones on uterine infections, Escherichia coli was infused into uterine lumens of ovariectomized or adrenoovariectomized rats receiving exogenous administration of various doses of ovarian hormones. Large numbers of E. coli were recovered from the rat uterine lumens, irrespective of hormonal influences. The number of leukocytes in the uterine flushings, representing the magnitude of purulent inflammation, differed significantly depending upon the hormonal regimen given to each host. Purulent endometritis was induced by E. coli in ovariectomized rats receiving progesterone or corn oil (hormone vehicle). Infections were asymptomatic in rats receiving estradiol, but promethazine-treated uterine horns were susceptible to infection. When progesterone was administered along with estradiol, purulent inflammation was caused by E. coli, but the number of leukocytes in the uterine lumens was significantly less than that obtained from the rats treated with progesterone or corn oil. These effects of ovarian hormones on uterine infections were observed in adrenoovariectomized rats as well as in ovariectomized rats. It is suggested that estradiol alters the nature of endometrial epithelium and prevent manifestation of purulent endometritis; progesterone antagonizes estradiol. Adrenal hormones appear not to participate in the pathogenesis of endometritis induced by E. coli.
Ureaplasma parvum, an opportunistic pathogen of the human urogenital tract, has been implicated in contributing to chorioamnionitis, fetal morbidity, and fetal mortality. It has been proposed that the host genetic background is a critical factor in adverse pregnancy outcome as sequela to U. parvum intra-amniotic infection. To test this hypothesis we assessed the impact of intrauterine U. parvum infection in the prototypical TH1/M1 C57BL/6 and TH2/M2 BALB/c mouse strain. Sterile medium or U. parvum was inoculated into each uterine horn and animals were evaluated for intra-amniotic infection, fetal infection, chorioamnionitis and fetal pathology at 72 hours post-inoculation. Disease outcome was assessed by microbial culture, in situ detection of U. parvum in fetal and utero-placental tissues, grading of chorioamnionitis, and placental gene expression of IL-1α, IL-1β, IL-6, TNF-α, S100A8, and S100A9. Placental infection and colonization rates were equivalent in both strains. The in situ distribution of U. parvum in placental tissues was also similar. However, a significantly greater proportion of BALB/c fetuses were infected (P<0.02). C57BL/6 infected animals predominantly exhibited mild to moderate chorioamnionitis (P<0.0001), and a significant reduction in placental expression of IL-1α, IL-1β, IL-6, TNF-α, S100A8, and S100A9 compared to sham controls (P<0.02). Conversely, severe protracted chorioamnionitis with cellular necrosis was the predominant lesion phenotype in BALB/c mice, which also exhibited a significant increase in placental expression of IL-1α, IL-1β, IL-6, TNF-α, S100A8, and S100A9 (P<0.01). Fetal pathology in BALB/c was multi-organ and included brain, lung, heart, liver, and intestine, whereas fetal pathology in C57BL/6 was only detected in the liver and intestines. These results confirm that the host genetic background is a major determinant in ureaplasmal induced chorioamnionitis with fetal infection and fetal inflammatory response.
Uterine bacterial infection after parturition causes endometritis, perturbs ovarian function and leads to infertility in cattle. Although endometritis is caused by mixed infections, endometrial pathology is associated with the presence of Arcanobacterium pyogenes. The aims of the present study were to determine the effects of A. pyogenes on endometrial function in vitro, and on uterine and ovarian function in vivo. Heat-killed A. pyogenes did not affect the production of prostaglandin F2α (PGF) or prostaglandin E2 (PGE) from endometrial explants, or purified populations of endometrial epithelial or stromal cells. However, the explants produced more PGF and PGE than controls when treated with a bacteria-free filtrate (BFF) cultured from A. pyogenes. Similarly, BFF stimulated PGF and PGE production by epithelial and stromal cells, respectively. So, BFF or control PBS was infused into the uterus of heifers (n = 7 per group) for 8 days, starting the day after estrus. Emergence of the follicle wave, dominant follicle or corpus luteum diameter, and peripheral plasma FSH, LH, estradiol, progesterone, PGFM, or acute phase protein concentrations were unaffected by the BFF infusion. In the live animal it is likely that the intact uterine mucosa limits the exposure of the endometrial cells to the exotoxin of A. pyogenes, whereas the cells are readily exposed to the toxin in vitro.
Inflammation; postpartum infection; reproductive immunology
This study investigated the effects of controlled reinfection on fertility of cattle naturally preexposed to Chlamydophila abortus. All animals had high prechallenge levels of immunoglobulin M (IgM), IgG, IgG1, and IgG2 serum antibodies against ruminant C. abortus in a chemiluminescent enzyme-linked immunosorbent assay. Twenty virgin heifers were estrus synchronized with prostaglandin F2, artificially inseminated 2 to 3 days later, and challenged immediately by intrauterine administration of 0, 104, 105, 106, or 108 inclusion-forming units (IFU) of C. abortus. Ten heifers were estrus synchronized, inseminated, and uterine challenged 2 weeks later. These animals were also indirectly exposed to C. abortus infection (cohort challenged) by contact with their previously challenged cohorts. Pregnancy was determined by rectal palpation 42 days after insemination. All anti-C. abortus antibody isotypes increased in heifers following uterine challenge with 108 IFU. A total of 11, 83, 50, 66, and 0% of heifers were pregnant after uterine challenge with 0, 104, 105, 106, and 108 IFU of C. abortus, respectively. A total of 50 and 65% of heifers were pregnant with and without cohort challenge, respectively. Uterine inoculum dose and cohort challenge (or, alternatively, a negative pregnancy outcome [infertility]) correlated highly significantly with a rise in postchallenge anti-C. abortus IgM levels over prechallenge levels. Logistic regression modeled fertility, with uterine challenge dose and cohort challenge or prechallenge IgM as predictors (P < 0.05). The models predict that the uterine C. abortus inoculum causing infertility is 8.5-fold higher for heifers without cohort exposure and 17-fold higher for heifers with high IgM levels than for heifers with cohort exposure or with low IgM levels.
Nine 2-year-old heifers having BAd3-neutralizing antibody titers between 1:120 and 1:1080 were individually exposed intranasally to an aerosol of 10(8) pfu of wild type (wt) bovine adenovirus type 3 (BAd3). Four animals were kept as non-inoculated controls. The heifers were examined daily for rectal temperature, weight gain/loss, nasal and ocular discharges, and other clinical signs for 10 d post-inoculation. None of the animals showed any sign of clinical disease. Virus excretion was observed in one animal only on Day 3 post-inoculation. All BAd3-inoculated heifers demonstrated a significant (P < 0.005, paired t-test) rise in BAd3-specific serum IgG, IgG1, or IgG2 ELISA titers and virus-neutralizing antibody titers compared to the titers before inoculation. All virus-inoculated animals demonstrated increased levels of BAd3-specific IgA ELISA titers in nasal secretions. These results suggest that in the presence of circulating BAd3-neutralizing antibodies, intranasal inoculation of cattle with wt BAd3 would result in inapparent infection.
Ureaplasmas isolated from the human genital tract and from the genital and respiratory tracts of cattle were grown in association with organ cultures of bovine oviduct (uterine tube). All strains of unreaplasmas multiplied in organ cultures, stopped ciliary activity, and caused histological lesions. Most strains grew well, and 10(8) to 10(9) color-changing units were determined 18 to 144 h after inoculation. Twenty-four to 144 h after inoculation with unreaplasmas, ciliostasis was complete. Ciliostasis was also caused by additions of nonviable cultures at pH 8.8 (or adjusted to 7.4) or washed disrupted cells (100 mug of protein/ml); it occurred in 48 to 96 h. The cilia-stopping effect of nonviable cultures was diminished by heating (56 C for 30 min) and was abolished by boiling. When added to fresh medium in amounts exceeding 25%, nonviable unreaplasmal cultures completely inhibited ureaplasmal growth. By light, scanning, and transmission electron microscopy, cilia-stopping effect was correlated with collapse and sloughing of the cilia (the initial lesion was "bent" cilia), with bulging and vacuolization of secretory and ciliated cells, and finally with disorganization of the epithelium, necrosis, and desquamation.
The efficacy of a morantel long-acting device in preventing parasitic gastroenteric infections throughout a whole year was evaluated in heifers and steer calves in south central Québec. Thirty-two calves, comprising nine Hereford steers, one Hereford heifer, fourteen Holstein crossbred steers and eight Holstein crossbred heifers, were allotted into two treatment groups and maintained throughout the grazing season on adjacent pastures resulting from equivalently dividing one original pasture. One morantel long-acting device was administered to each animal in one group at the time of turnout onto the pasture in the spring while the calves in the other group remained as nonmedicated controls. The parasitic infections incurred during the pasture season were primarily Ostertagia, Trichostrongylus, Cooperia and Nematodirus.
The following fall, after twelve calves were necropsied for worm counts, the twenty remaining ones were brought into the barn where they were kept throughout the winter with access to an outside yard. They received good quality hay and rolled barley (1 kg/head/day) up until the following May, at which time they were weighed and had fecal samples taken for egg counts. In contrast to the results observed among controls, the morantel long-acting device treatment group had an 87% reduction in fecal worm egg excretion and a 67.3 kg per calf increased weight gain after one year.
Morantel long-acting device; gastrointestinal parasitism; bovine
Survival to maturity and age at first calving were studied in heifer calves from 34 randomly selected Holstein dairy farms in southwestern Ontario. Calves were divided into cohorts on the basis of treatment for pneumonia, scours, other diseases, or no treatments, during the first 90 days of life. An effect of pneumonia and scours together, over and above the effects of each disease alone, was assessed by means on an interaction term in the statistical analyses. Heifers which had been treated for pneumonia during the first three months of life were 2.5 times more likely to die after 90 days of age than heifers which had not been treated for pneumonia, after controlling for the farm effect. Heifers with a calfhood history of being treated for scours were 2.5 times more likely to be sold for dairy purposes than other calves. Heifers which had been treated for scours were 2.9 times more likely to calve after 900 days of age than other heifers, after controlling for the farm effect.
The object of this study was to develop a prefixation protein A gold labelling technique for Ureaplasma diversum and to apply this to bovine embryos. Sixteen hour cultures of Ureaplasma diversum strain 2312 were incubated with either specific antiserum or nonimmune serum, followed by exposure to protein A gold and negative staining. The ureaplasmas which were incubated with specific antiserum were labelled with gold particles while those ureaplasmas which were incubated with nonimmune serum were not labelled. Twenty-three unhatched, day 7 bovine embryos were then incubated in either embryo culture medium (ECM) alone, ECM with sterile ureaplasma broth added or ECM with 1.7 X 10(6) colony forming units of Ureaplasma diversum strain 2312 per embryo. After 16 hours, the embryos were washed twice and incubated with either specific antiserum or nonimmune serum. The embryos were then incubated with medium containing protein A gold and examined by electron microscopy. No ureaplasmas were identified on the zona pellucida of the control embryos. Ureaplasmas were identified on the outer surface of the zona pellucida of 13 of the 17 embryos which had been exposed to the organism. Of these, the embryos which were incubated with specific antiserum had labelled ureaplasmas while the embryos which were incubated with nonimmune serum had unlabelled ureaplasmas on the zona pellucida. It was concluded that the protein A gold method was a suitable technique for the identification of ureaplasmas in EM preparations. The presence of ureaplasmas on the outer surface of the bovine zona pellucida following in vitro exposure to the organism was confirmed.(ABSTRACT TRUNCATED AT 250 WORDS)
Transrectal Doppler sonography was used to evaluate uterine blood flow during the first two weeks after parturition in six primiparous Simmental cows. The uterine blood flow was evaluated on the day of parturition (Day 0), once daily from Days 1 to 8 and then every other day until Day 14. Blood flow was quantified by determining the diameter (D), the time-averaged maximum velocity (TAMV), the pulsatility index (PI) and the blood flow volume (BFV) of the uterine arteries ipsilateral and contralateral to the formerly pregnant uterine horn. During the first four days after calving D, TAMV and BFV declined (ipsilateral: TAMV 70%, BFV 87%, contralateral: D 47%, BFV 84%; p < 0.05), while PI increased (ipsilateral 158%, contralateral 100%; p < 0.05) distinctly. Between Days 4 and 14 only the ipsilateral D (12%) and the BFV of both arteries (ipsilateral 5%, contralateral 8%) decreased (p < 0.05). Blood flow variables were very strongly correlated with each other (r > ±0.75, p < 0.05), with negative correlations with PI and positive correlations with all other investigated factors. Overall, this study revealed characteristic changes in uterine perfusion during the first two weeks after parturition in cows that were pronounced during the first four days postpartum.
involution; puerperal; uterus
During 3 consecutive calving seasons, calving performance, placental characteristics and endocrine profiles of total 98 pregnancies of late pregnant Swedish Red and White (SRB) and Swedish Holstein (SLB) dairy heifers and cows, were investigated. Ninety-four singleton pregnancies and 4 sets of twins were recorded. In animals with singleton pregnancy, 8 stillbirths, 7 weak calves, 3 premature parturitions and 1 abortion were registered. In the SLB heifers, 19% of stillbirth (5/26) were observed, while 5% (2/42) were noted for the SRB heifers. One stillborn calf derived from the SRB cows and none was found from the SLB cows. In the heifers and cows delivering a normal living calf with unassisted parturition, the placentome thickness monitored by ultrasonography was constant towards the end of pregnancy. The numbers of foetal cotyledons varied individually between animals but in total, fewer cotyledons were found in the foetal membranes of the SRB animals than in the SLB animals (69 ± 19) vs. (88 ± 29) (p < 0.05). No morphological and numerical differences of the placentome thickness in animals delivering a stillborn or weak calf, compared to animals delivering a normal living calf, could be observed. In animals with unassisted parturition and without birth complications, the levels of progesterone (P4), PGF2α metabolite (PG-metabolite), cortisol, oestrone sulphate (E1SO4) and pregnancy associated glycoproteins (PAGs) were not different by breeds and parities. In animals carrying stillbirth, higher levels of E1SO4 were found in 3 SRB animals and 1 SLB heifer, whereas lower levels of E1SO4 were recorded in 3 SLB heifers during the last week of pregnancy, compared to the profiles found in animals with unassisted parturition. Additionally, the levels of PAGs remained low and constant in 1 SRB cow (delivering a stillborn calf), 1 SRB heifer (giving birth prematurely), 4 animals (carrying twins) and 1 aborting SRB cow. Our results show a very high rate of stillbirth in especially SLB heifers and deviating profiles of E1SO4 and PAGs in animals with impaired parturition were recorded.
Cattle-pregnancy; parturition; endocrine profiles; calving performance; stillbirth