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1.  Experimental bovine genital ureaplasmosis. I. Granular vulvitis following vulvar inoculation. 
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.
PMCID: PMC1320070  PMID: 7427772
2.  Effects of Fertility on Gene Expression and Function of the Bovine Endometrium 
PLoS ONE  2013;8(8):e69444.
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.
PMCID: PMC3734181  PMID: 23940519
3.  Effect of Heifer Frame Score on Growth, Fertility, and Economics 
A non-traditional forage-based protocol was employed to evaluate replacement heifer growth, fertility, and economics between small frame (SF, 3.50; n = 50) and large frame (LF, 5.56; n = 50) heifers using three increasing gain growth phases. Preceding an 85 d growing-breeding period (Phase 3; P3) the heifers were managed as a common group for Phases 1 and 2 (P1 and P2). During P1, heifers grazed common fields of unharvested corn and corn residue (total digestible nutrients [TDN] 56%) with supplemental hay. For P2, heifers grazed early spring crested wheatgrass pasture (CWG; TDN 62%) that was followed by the final P3 drylot growing and breeding period (TDN 68%). Small frame heifers were lighter at the end of P1 in May and at the start of P3 breeding in August (p = 0.0002). Percent of mature body weight (BW) at the end of P1 (209 d) was 48.7% and 46.8%, respectively, for the SF and LF heifers and the percent pubertal was lower for SF than for LF heifers (18.0% vs 40.0%; p = 0.02). At breeding initiation (P3), the percentage of mature BW was 57.8 and 57.2 and the percentage pubertal was 90.0 and 96.0 (p = 0.07) for the SF and LF heifers, respectively; a 5-fold increase for SF heifers. Breeding cycle pregnancy on days 21, 42, and 63, and total percent pregnant did not differ (p>0.10). In drylot, SF heifer dry matter intake (DMI) was 20.1% less (p = 0.001) and feed cost/d was 20.3% lower (p = 0.001), but feed cost/kg of gain did not differ between SF and LF heifers (p = 0.41). Economically important live animal measurements for muscling were measured in May and at the end of the study in October. SF heifers had greater L. dorsi muscle area per unit of BW than LF heifers (p = 0.03). Small frame heifer value was lower at weaning (p = 0.005) and the non-pregnant ending heifer value was lower for SF heifers than for the LF heifers (p = 0.005). However, the total development cost was lower for SF heifers (p = 0.001) and the net cost per pregnant heifer, after accounting for the sale of non-pregnant heifers, was lower for SF heifers (p = 0.004). These data suggest that high breeding efficiency can be attained among March-April born SF and LF virgin heifers when transitioned to a more favorable May-June calving period through the strategic use of grazed and harvested forages resulting in a lower net cost per pregnant SF heifer.
PMCID: PMC4283190  PMID: 25557677
Beef Heifer; Heifer Production Economics; Fertility; Frame Score; Increasing Energy Management; Percent Mature Body Weight
4.  Humoral and secretory antibodies to Ureaplasma diversum in heifers following subcutaneous vaccination and vaginal infection. 
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.
PMCID: PMC1263674  PMID: 8004534
5.  Characterization and Comparative Genital Tract Pathogenicity of Bovine Mycoplasmas 1 
Infection and Immunity  1970;2(1):101-104.
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.
PMCID: PMC415971  PMID: 16557786
6.  Isolation of Ureaplasma from bovine granular vulvitis. 
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.
PMCID: PMC1277608  PMID: 352491
7.  Bovine Granular Vulvitis Associated with Ureaplasma Infection 
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.
PMCID: PMC1789526  PMID: 427710
8.  Endometritis caused by Chlamydia trachomatis. 
Chlamydia trachomatis was found to be the aetiological agent of endometritis in three women with concomitant signs of salpingitis. All patients developed a significant antibody response to the organism. Chlamydia were recovered from aspirated uterine contents of two patients and darkfield examination of histological sections showed chlamydial inclusions in endometrial cells in one patient. Thus, C trachomatis can be recovered from the endometrium of patients in whom the cervical culture result is negative. In one patient curettage showed endometritis with a characteristic plasma-cell infiltration. The occurrence of chlamydial endometritis may explain why irregular bleeding is a common finding in patients with salpingitis. It also suggests a canalicular spread of chlamydia from the cervix to the Fallopian tubes.
PMCID: PMC1045915  PMID: 7237083
9.  The relative economics of feeding open, aborted, pregnant feedlot heifers 
The Canadian Veterinary Journal  1991;32(10):613-617.
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.
PMCID: PMC1481075  PMID: 17423875
10.  Bovine Vibriosis: The Distribution and Specificity of Antibodies Induced by Vaccination and Infection and the Immunofluorescent Localization of the Organism in infected Heifers 
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.
PMCID: PMC1319587  PMID: 4260944
11.  Immune response of heifers to vaginal submucosal or subcutaneous vaccination and intravaginal challenge with Ureaplasma diversum. 
Twenty beef heifers were randomly assigned to five equal groups and vaccinated: Group 1--in vaginal submucosa (VM) with Ureaplasma diversum ultrasonicated whole cells (WC) in complete Freund's adjuvant (CFA); Group 2--in VM with U. diversum cell membranes (CM) in CFA; Group 3--subcutaneously (SC) with CM in CFA; Group 4--in VM with CM alone; and Group 5--in VM with phosphate buffered saline (PBS) in CFA. A second vaccination with the same antigens in incomplete Freund's adjuvant was given after four weeks, and three weeks later, all heifers were challenged intravaginally with 3.6 x 10(7) colony-forming units (CFU) of U. diversum strain 2312. Immunoglobulins that reacted with U. diversum were measured in serum and cervicovaginal mucus (CVM) by an enzyme-linked-immunosorbent assay. In groups 1 and 2, vaccination by the VM route with WC or CM antigens, stimulated high levels of U. diversum-reactive IgG1 and IgG2 antibodies in serum as well as CVM, but a low IgA response only in CVM. In group 4, VM vaccination with CM (no adjuvant) elicited a minimal IgG1 and IgG2 response in serum and CVM. In group 3, SC vaccination with CM antigen stimulated high IgG1 and IgG2 reactivity in both serum and CVM, but no IgA reactivity. Very little IgM reactivity was detected in the four vaccinated groups. Intravaginal challenge resulted in characteristic granular vulvitis in all vaccinated and control heifers, with all animals remaining culture-positive for the 35 day observation period. The infection stimulated a marked increase in the specific IgA response in CVM of the three groups vaccinated with either, adjuvanted antigen.(ABSTRACT TRUNCATED AT 250 WORDS)
PMCID: PMC1263675  PMID: 8004535
12.  Conceptus-derived prostaglandins regulate gene expression in the endometrium prior to pregnancy recognition in ruminants 
Reproduction (Cambridge, England)  2013;146(4):377-387.
In cattle, the blastocyst hatches from the zona pellucida on days 8 to 9 and then forms a conceptus that grows and elongates into an ovoid and then filamentous shape between days 9 and 16. The growing conceptus synthesizes and secretes prostaglandins and interferon tau. Our hypothesis was that the ovoid conceptus exerts a local effect on the endometrium prior to maternal recognition of pregnancy on day 16 in cattle. In Study One, synchronized cyclic heifers received nothing or 20 in vitro produced blastocysts on day 7, and uteri were collected on day 13. Interferon tau was not detected by radioimmunoassay in the uterine flush of pregnant heifers containing multiple ovoid conceptuses; however, total prostaglandin levels were higher in the uterine lumen of pregnant as compared to cyclic heifers. Microarray analysis revealed that 44 genes were increased in the endometrium of day 13 pregnant as compared to cyclic heifers, and many of those genes were classical Type I IFN-stimulated genes (ISGs). Studies Two and Three determined effects of infusing prostaglandins at the levels produced by the elongating day 14 conceptus into the uterine lumen of cyclic ewes on ISG expression in the endometrium. Results indicated that prostaglandin infusion increased the abundance of several ISGs in the endometrium. These studies support the hypothesis that the day 13 conceptus secretes prostaglandins that act locally in a paracrine manner to alter gene expression in the endometrium prior to pregnancy recognition in cattle.
PMCID: PMC3791335  PMID: 23966582
13.  Reinfection with Chlamydophila abortus by Uterine and Indirect Cohort Routes Reduces Fertility in Cattle Preexposed to Chlamydophila  
Infection and Immunity  2004;72(5):2538-2545.
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.
PMCID: PMC387841  PMID: 15102761
14.  Transmission of border disease virus from a persistently infected calf to seronegative heifers in early pregnancy 
This study describes the transmission of border disease virus (BDV) from a persistently infected calf to seronegative heifers in early pregnancy, resulting in persistently infected fetuses. On day 50 of pregnancy (= day 0 of the infection phase), six heifers were co-housed in a free stall with a bull calf persistently infected with BDV (pi BVD) for 60 days. The heifers underwent daily clinical examination, and blood samples were collected regularly for detection of pestiviral RNA and anti-pestivirus antibodies. After day 60 (= day 110 of pregnancy), the heifers were slaughtered, and the fetuses and placentae underwent post-mortem and immunohistochemical examination and RT-PCR for viral RNA detection.
Three heifers had mild viraemia from day 8 to day 14, and by day 40 all heifers had pestivirus antibodies identified as anti-BDV antibodies in the serum neutralisation test. The placenta of the three viraemic heifers had histological evidence of inflammation, and fetal organs from these heifers were positive for pestivirus antigen by immunohistochemical examination and for BD viral RNA by RT-PCR and sequencing. Thus, co-housing of heifers in early pregnancy with a pi-BDV calf led to seroconversion in all heifers and persistent fetal infection in three.
Considering that pi-BDV cattle can infect other cattle and lead to persistent infection of the fetus in pregnant cows, BDV should not be ignored in the context of the mandatory BVDV eradication and monitoring program. This strongly suggests that BDV should be taken into account in BVD eradication and control programs.
PMCID: PMC4336514
Cattle; Border disease; Early pregnancy; Persistent infection; BVDV; Pestivirus
15.  The effects of two Ureaplasma diversum strains on early pregnancy in heifers. 
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.
PMCID: PMC1255382  PMID: 3453276
16.  Effect of the estrous cycle on uterine infection induced by Escherichia coli. 
Infection and Immunity  1984;43(2):678-683.
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.
PMCID: PMC264353  PMID: 6363298
17.  Distribution and viability of spermatozoa in the canine female genital tract during post-ovulatory oocyte maturation 
Unlike other domestic mammals, in which metaphase-II oocytes are ovulated, canine ovulation is characterized by the release of primary oocytes, which may take 12 to up to 36 hours. Further 60 hours are needed for maturation to secondary oocytes which then remain fertile for about 48 hours. Oestrus takes 7 to 10 days on average and may start as early as a week before ovulation. This together with the prolonged process of post-ovulatory oocyte maturation requires an according longevity of spermatozoa in the female genital tract in order to provide a population of fertile sperm when oocytes have matured to fertilizability. Therefore the distribution and viability of spermatozoa in the bitch genital tract was examined during post-ovulatory oocyte maturation.
Thirteen beagle bitches were inseminated on the day of sonographically verified ovulation with pooled semen of two beagle dogs containing one billion progressively motile spermatozoa. Ovariohysterectomy was performed two days later (group 1, n = 6) and four days later (group 2, n = 7). The oviduct and uterine horn of one side were flushed separately and the flushing’s were checked for the presence of gametes. The oviducts including the utero-tubal junction and the uterine horns, both the flushed and unflushed, were histologically examined for sperm distribution.
The total number of spermatozoa recovered by flushing was low and evaluation of viability was limited. Prophase-I oocytes were collected from oviduct flushing in group 1, whereas unfertilized metaphase-II oocytes were detected in group 2. From day 2 to day 4 after ovulation a significant decrease in the percentage of glands containing sperm (P<0.05) and a marked reduction of the mean sperm number in uterine horn glands were observed. A concomitant diminution of spermatozoa was indicated in the utero-tubal junction accompanied by a slight increase in sperm numbers in the mid oviduct.
Oocyte maturation to metaphase-II stage is accompanied by a continuous sperm detachment and elimination in the uterine horns. Entrance of spermatozoa into the caudal oviduct seems to be steadily controlled by the utero-tubal junction thus providing a selected sperm population to be shifted towards the site of fertilization when oocyte maturation is completed.
PMCID: PMC3526419  PMID: 22932162
Dog; Sperm storage; Oviduct; Oocyte maturation; Fertilization
18.  Experimental Infection of the Genital Tract of Female Grivet Monkeys by Mycoplasma hominis: Effects of Different Routes of Infection 
Infection and Immunity  1979;26(3):1123-1128.
In a previous study we produced acute salpingitis and parametritis in grivet monkeys by inoculation of Mycoplasma hominis directly into the uterine tubes. With the purpose of examining in the same animal model the effect of more natural routes of infection, six female grivet monkeys were inoculated, two by two, with M. hominis by the following methods: (experiment A) into the uterine cavity through the cervical canal; (experiment B) into the uterine cavity by the same route, but after ligature of the isthmus of the uterine tubes had been performed and followed by curettage of the endometrium; and (experiment C) into the cervical epithelium. Whereas the animals in experiment A developed no or only very slight signs of infection, all monkeys in experiments B and C developed pronounced gross and microscopic inflammatory lesions of the uterine tubes and parametria, together with a significant antibody response. It is concluded that ascending M. hominis infection of the genital tract must be preceded by mechanical injury of the epithelial barrier, and that subsequent spread of the infection occurs via blood and lymph vessels rather than by the canalicular route. The inflammatory genital tract disease produced experimentally with M. hominis in grivet monkeys is very similar in its pathogenesis and pathology to the type of salpingitis caused in humans by microorganisms other than Neisseria gonorrhoeae and Chlamydia trachomatis.
PMCID: PMC414737  PMID: 118928
19.  Bovine vaginal antibody responses to immunoaffinity-purified surface antigen of Tritrichomonas foetus. 
Journal of Clinical Microbiology  1995;33(5):1158-1163.
Bovine trichomoniasis is a prevalent sexually transmitted disease of cattle caused by the protozoan Tritrichomonas foetus. Currently, diagnosis is most often made by culture. In order to provide a faster immunodiagnostic approach, a specific enzyme-linked immunosorbent assay (ELISA) was investigated. A protective surface antigen (TF1.17 antigen) of T. foetus was immunoaffinity purified and used in an ELISA to detect antibodies in vaginal mucus from heifers inoculated with T. foetus. In preliminary studies, antibodies of the immunoglobulin A (IgA) isotype were detected in mucus from all experimentally infected heifers which were tested at 6 weeks postinoculation, whereas IgG1 and IgG2 were not. In addition, IgA responses detected in postinoculation samples were all greater than those detected in preinoculation samples, unlike those detected by a whole-cell antigen ELISA. For these two reasons, IgA antibodies appeared to be useful diagnostically. Further investigation of IgA antibodies used vaginal mucus collected weekly from heifers inoculated intravaginally with 10(2), 10(4), or 10(6) T. foetus organisms. Heifers with positive cultures for T. foetus had similar IgA responses to TF1.17 antigen over the 10 weeks of infection regardless of the initial inoculum dose. This indicates that if the dose is sufficient to establish infection, the magnitude and duration of the immune response are no longer dependent on dose. All of the infected animals receiving all dosages responded with high absorbance values in the IgA anti-TF1.17 antigen ELISA by 6 weeks postinoculation, and all absorbance values remained high at 10 weeks. To determine the duration of the IgA response, four other heifers inoculated with 7 x 10(6) T. foetus organisms were studied through 24 weeks postinoculation.(ABSTRACT TRUNCATED AT 250 WORDS)
PMCID: PMC228123  PMID: 7615722
20.  Performance of Clinical and Laparoscopic Criteria for the Diagnosis of Upper Genital Tract Infection 
Objective: The purpose of this study was to validate the standard minimal clinical criteria and the laparoscopic triad of tubal edema, erythema, and purulent exudate used to diagnose acute upper genital tract infection.
Methods: Subjects included women who either met the Centers for Disease Control and Prevention's (CDC) minimal criteria for acute pelvic inflammatory disease or had other signs of upper genital tract infection (i.e., atypical pelvic pain, abnormal uterine bleeding, or cervicitis). The subjects were evaluated with a baseline interview comprehensive laboratory testing, and either an endometrial biopsy or laparoscopy with endometrial and fimbrial biopsies for definitive diagnosis of upper genital tract infection. Patients were considered positive for upper genital tract infection if they had any of the following findings: 1) histologic evidence of endometritis or salpingitis; 2) laparoscopic visualization of purulent exudate in the pelvis without another source; or 3) positive testing for Neisseria gonorrhoeae or Chlamydia trachomatis from the endometrium, fallopian tubes, or pelvis.
Results: One hundred twenty-nine women with adequate endometrial samples were evaluated between August 1993 and September 1997, and 62 had complete laparoscopic evaluations. The sensitivities of the CDC's minimal clinical criteria for pelvic inflammatory disease and the laparoscopic triad of edema, erythema, and purulent exudate were 65% and 60%, respectively.
Conclusions: Commonly used minimal clinical criteria for pelvic inflammatory disease and the laparoscopic triad of tubal edema, erythema, and purulent exudate have limited sensitivity with correspondingly high false negative rates.
PMCID: PMC2364552  PMID: 18476154
21.  Effect of precalving intramammary treatment with pirlimycin in nulliparous Holstein heifers 
A clinical trial was conducted to determine whether prepartum intramammary pirlimycin reduces the proportion of nulliparous heifers with intramammary infection (IMI) during early lactation and improves milk production. Quarter milk samples were collected from 428 heifers, systematically allocated to treatment and control groups, 6 to 12 d before the expected calving date and 2 to 8 d after calving. At the prepartum visit, heifers in the treatment group (n = 219) received an infusion of pirlimycin hydrochloride in all 4 quarters; the control heifers (n = 209) received no infusions. Intramammary infection was detected in 69% of the heifers and 33% of the quarters before calving. After calving, the proportion of treated heifers with IMI was significantly lower than the proportion of control heifers (31% versus 45%). Staphylococcus aureus was isolated from 10% of the heifers and 3% of the quarters before calving. After calving, the proportion of IMIs due to S. aureus was significantly lower in the treated heifers than in the control heifers (5.6% versus 10%). Antibiotic treatment increased the percentage of cures and prevented new IMIs caused by gram-positive bacteria after calving. The incidence of new IMIs caused by gram-negative bacteria and yeast was higher among treated heifers than among control heifers. There was no overall effect of treatment on milk production, but there was a significant interaction effect of treatment and the interval between treatment and calving. An increase of 302 kg of milk was observed when antibiotic treatment was applied more than 1 wk before calving. Treatment did not affect the milk somatic cell count on the 1st 3 test days after calving.
PMCID: PMC1940276  PMID: 17955903
22.  Persistence, Immune Response, and Antigenic Variation of Mycoplasma genitalium in an Experimentally Infected Pig-Tailed Macaque (Macaca nemestrina) 
Infection and Immunity  2013;81(8):2938-2951.
Mycoplasma genitalium is a sexually transmitted pathogen associated with several acute and chronic reproductive tract disease syndromes in men and women. To evaluate the suitability of a pig-tailed macaque model of M. genitalium infection, we inoculated a pilot animal with M. genitalium strain G37 in the uterine cervix and in salpingeal pockets generated by transplanting autologous Fallopian tube tissue subcutaneously. Viable organisms were recovered throughout the 8-week experiment in cervicovaginal specimens and up to 2 weeks postinfection in salpingeal pockets. Humoral and cervicovaginal antibodies reacting to MgpB were induced postinoculation and persisted throughout the infection. The immunodominance of the MgpB adhesin and the accumulation of mgpB sequence diversity previously observed in persistent human infections prompted us to evaluate sequence variation in this animal model. We found that after 8 weeks of infection, sequences within mgpB variable region B were replaced by novel sequences generated by reciprocal recombination with an archived variant sequence located elsewhere on the chromosome. In contrast, mgpB region B of the same inoculum propagated for 8 weeks in vitro remained unchanged. Notably, serum IgG reacted strongly with a recombinant protein spanning MgpB region B of the inoculum, while reactivity to a recombinant protein representing the week 8 variant was reduced, suggesting that antibodies were involved in the clearance of bacteria expressing the original infecting sequence. Together these results suggest that the pig-tailed macaque is a suitable model to study M. genitalium pathogenesis, antibody-mediated selection of antigenic variants in vivo, and immune escape.
PMCID: PMC3719596  PMID: 23732170
23.  Evaluation of remote monitoring of parturition in dairy cattle as a new tool for calving management 
Proper calving management of dairy herds is a crucial aspect of the bovine life cycle, as it has profound effects on calf viability and on the post-partum course of the dam. The objectives of this study were to monitor the calving process through the use of a remote alarm system and to determine the impact of prompt emergency obstetric procedures in case of dystocia for the prevention of stillbirths and post-partum reproductive pathologies, and for improving herd fertility. Six groups of experimental animals were studied: monitored heifers (n = 60) and multiparous cows (n = 60) were compared with non-observed animals (n = 60 heifers and n = 60 multiparous) giving birth during the same time period and housed in the calving barn, and with unmonitored animals placed in a dry zone (n = 240 heifers and n = 112 multiparous cows).
The incidence of dystocia ranged from a minimum of 23.4% (monitored multiparous cows) to a maximum of 33.3% (monitored heifers), and there were no differences compared with control groups. However, the rate of stillbirth was higher in control groups than in the monitored groups (P < 0.01). Among both heifers and multiparous cows, the incidence of post-partum uterine infections was higher in the unmonitored animals both in the calving barn (P < 0.01) and in the dry zone (P < 0.05) compared with monitored animals. Among both heifers and multiparous cows, the control groups showed higher rates of foetal membrane retention than did the monitored groups (P < 0.001). The calving-to-conception interval was shorter; in particular, observed heifers showed a significant advantage of approximately 46 days compared with the unmonitored group (P < 0.001) and 32 days compared with the group in the calving barn (P < 0.05). Multiparous cows also had a reduction in the number of days open.
The remote alarm system used to monitor the calving process assured the prompt presence of personnel, improving both the cow’s reproductive efficiency and neonatal viability.
PMCID: PMC3850716  PMID: 24079910
Dairy cattle; Calving monitoring; Dystocia; Post-partum fertility
24.  Effects of ovarian hormones on manifestation of purulent endometritis in rat uteruses infected with Escherichia coli. 
Infection and Immunity  1985;47(1):311-317.
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.
PMCID: PMC261514  PMID: 3880725
25.  Associations among serum pro- and anti-inflammatory cytokines, metabolic mediators, body condition, and uterine disease in postpartum dairy cows 
Adipose tissue is an active endocrine organ which secretes a wide range of hormones and protein factors, collectively termed adipokines. Adipokines affect appetite and satiety, glucose and lipid metabolism, inflammation and immune functions. The objectives were to evaluate serum concentrations of adipokines (adiponectin, leptin, tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6) in lactating dairy cows with postpartum uterine inflammatory conditions (metritis, clinical endometritis or subclinical endometritis) and in cows experiencing loss of body condition, and to assess the relationship of adipokines and body condition loss in the establishment of persistent uterine inflammatory conditions.
Lactating multiparous Holstein cows (N = 40), with body condition scores (BCS) from 2 to 4 (eight cows for each 0.5 score increment) were enrolled. Body condition was monitored for all cows weekly for 7 weeks post calving; cows with uterine inflammatory conditions were also re-evaluated 2 weeks later. Blood samples were collected from 1 week prior to calving to 7 weeks after calving for determination of serum concentrations of adipokines, insulin and insulin like growth factor (IGF)-1.
Cows with metritis or clinical endometritis had higher serum concentrations of adiponectin, leptin, TNF-alpha, IL-1beta and IL-6 compared to normal cows (P < 0.05). Furthermore, serum leptin, TNF-alpha, IL-1beta and IL-6 were higher in cows with subclinical endometritis compared to normal cows (P < 0.05), and insulin and IGF-1 concentrations were lower in cows with metritis or clinical endometritis. Cows with low BCS (2 and 2.5) had significantly higher adiponectin, TNF-alpha, IL-1beta and IL-6 than those with high BCS (3 to 4). Cows with persistent uterine inflammatory conditions had higher adiponectin, leptin TNF-alpha, IL-1beta and IL-6 and insulin compared to normal and spontaneously recovered cows, except for IGF-1 (P < 0.05).
Serum concentrations of adipokines, insulin, and IGF-1 had significant associations with BCS categories (low vs. high) and postpartum uterine inflammatory conditions. Perhaps loss of body condition mediated increases in anti- and pro-inflammatory cytokines, whereas increased pro- and anti-inflammatory cytokines concentrations mediated body condition loss and thereby prolonged persistence of uterine inflammation in dairy cows.
PMCID: PMC3830449  PMID: 24209779
Adipokines; Insulin; IGF-1; Postpartum; Uterine inflammation; Body condition; Dairy cows

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