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Biomedical subjects

I M Sheldon

Publications and source records attributed to I M Sheldon.

At least 19 recordsLinked to original sources
PubMed2005

Use of the cow as a large animal model of uterine infection and immunity.

For most of the reproductive cycle in both humans and animals, the uterus is clear of pathogenic bacteria. However, it is readily contaminated with pathogens, such as Escherichia and Tritichomonas species, during sexual intercourse and after parturition. Uterine infection is particularly common after parturition in cattle (Bos taurus), causing clinical disease and infertility. The endocrine and immune responses to uterine infection in cattle have been investigated in vivo and using tissue culture. Cattle are of sufficient size to permit monitoring of reproductive and immune function throughout uterine infections, and primary cell cultures are readily established. In the whole animal, uterine infections suppress GnRH and LH secretion, and inhibit the growth of ovarian follicles and their estradiol secretion. The immune response is characterized by an influx of neutrophils into the uterus and increased concentrations of acute phase proteins in peripheral plasma. In vitro, the endometrial and ovarian cell function is modified by challenge with bacteria, their products such as lipopolysaccharide or pro-inflammatory cytokines. However, it is interesting to note that the susceptibility to uterine infection and the immune response are partially regulated by the ovarian steroid hormone mileu. In conclusion, the ease of working with cattle, the availability of tissues and the similarity of uterine infection between mammals, make Bos taurus a good model for studying uterine infection and immunity.

Animals↗
PubMed2004

Minimum inhibitory concentrations of some antimicrobial drugs against bacteria causing uterine infections in cattle.

The minimum inhibitory concentrations (MICs) of oxytetracycline, cephapirin, cephapirin/mecillinam, cefquinome, ceftiofur and enrofloxacin, candidate antibiotics for the principal bacteria associated with uterine infections: Escherichia coli, Arcanobacterium pyogenes and the anaerobic bacteria Fusobacterium necrophorum and Prevotella melaninogenicus, were determined by the agar dilution method. The bacteria were isolated from animals with clinical metritis and/or endometritis. For E coli, cefquinome and enrofloxacin had the lowest MIC90 and MIC50 values (< 0.06 microg/ml), and oxytetracycline and cephapirin had the highest values. For A pyogenes, oxytetracycline had the highest MIC50 value (16 microg/ml), but all the cephalosporins had values below 0.06 microg/ml. For the anaerobic bacteria, enrofloxacin and oxytetracycline had the highest MIC50 values but all the cephalosporins had values of 0.06 microg/ml or below.

Actinomycetaceae↗
PubMed2004

Association between postpartum pyrexia and uterine bacterial infection in dairy cattle.

The temperature of 90 dairy cattle was recorded for the first 10 days after parturition and the animals were categorised as either normal (< 39.7 degreesC) or pyrexic. Swabs were collected from the uterine lumen seven, 14, 21 and 28 days after parturition for aerobic and anaerobic culture; bacteria were identified and their growth was scored semiquantitatively. Blood samples were collected three times a week for the estimation of the concentrations of acute phase proteins. The cows' temperatures were often above the accepted normal range, but it was not a good indicator of the number of bacteria in the uterus. However, pyrexia was correlated with the presence of specific uterine pathogens (P < 0.05) and in particular with Prevotella species (P < 0.01). The pyrexic animals had a higher plasma concentration of the acute phase protein (alpha1-acid glycoprotein (P < 0.05). Although pyrexia is an indicator of postpartum inflammation, additional clinical signs are necessary to identify uterine bacterial infection.

Animals↗
PubMed2004

Effect of intrauterine administration of oestradiol on postpartum uterine bacterial infection in cattle.

After parturition fewer first dominant follicles are selected in the ovary ipsilateral to the previously gravid uterine horn in cattle. However, the presence of a large oestradiol-secreting follicle in the ipsilateral ovary is a predictor of fertility, possibly due to a localised effect of oestradiol which increases the rate of elimination of the ubiquitous uterine bacterial contamination that occurs after calving. The present study tested the hypothesis that oestradiol reduces uterine bacterial contamination when administered into the uterine lumen around the expected time for selection of the first postpartum dominant follicle. Animals were infused with saline (n=15) or 10mg oestradiol benzoate (n=15) into the previously gravid uterine horn on Days 7 and 10 postpartum. Peripheral coccygeal blood samples were collected daily and oestradiol concentrations measured by radioimmunoassay (RIA). Uterine lumen swabs were collected 7, 14 and 21 days postpartum for aerobic and anaerobic culture, bacteria were identified and growth scored semi-quantitatively. Plasma oestradiol concentrations were higher for treated animals between Days 7 and 14 (1.4+/-0.1 versus 2.0+/-0.2 pg/ml, P<0.05). Control animals had a similar bacterial growth score on Days 7 and 14, with a lower value on Day 21 (5.7+/-1.0 and 6.1+/-0.7 versus 0.3+/-0.1, P<0.05). However, treated animals had a surprising higher bacterial load on Day 14, than on Days 7 or 21 (7.1+/-0.9 versus 4.0+/-0.6 or 3.6+/-0.6, P<0.05). The increased score was attributable to more pathogens associated with endometritis on Day 14 than Day 7 (5.1+/-1.0 versus 2.5+/-0.5, P<0.05), in particular Prevotella melaninogenicus (1.5+/-0.5 versus 0.7+/-0.2, P<0.05) and Fusobacterium necrophorum (1.5+/-0.4 versus 0.3+/-0.2, P<0.05). In conclusion, administration of oestradiol into the uterine lumen surprisingly increased uterine pathogenic anaerobic bacterial contamination. Thus, it is unlikely that increased fertility associated with a first dominant follicle in the ipsilateral ovary is a consequence of the elimination of bacterial contamination by ovarian oestradiol.

Animals↗
PubMed2004

Postpartum uterine health in cattle.

Uterine health is often compromised in cattle because postpartum contamination of the uterine lumen by bacteria is ubiquitous, and pathogenic bacteria frequently persist causing clinical disease. The subfertility associated with uterine infection involves perturbation of the hypothalamus, pituitary and ovary, in addition to the direct effects on the uterus, and appears to persist even after clinical resolution of the disease. Absorption of bacterial components from the uterus can prevent the follicular phase LH surge and ovulation. In addition, the first postpartum dominant follicle has a slower growth rate and secretes less estradiol at the end of the growth phase. There are also localised ovarian effects of high uterine bacterial growth density, because fewer first dominant follicles are selected in the ovary ipsilateral than contralateral to the previously gravid uterine horn. Thus, it is important to diagnose and treat uterine disease promptly and effectively. Examination of the contents of the vagina for the presence of pus is the most useful method for diagnosis of endometritis. The character and odor of the vaginal mucus can be scored and this endometritis score is correlated with the growth density of pathogenic bacteria in the uterus, and is prognostic for the likely success of treatment. The challenge for the future is to design prevention and control programs to reduce the incidence of disease, and understand how the immune and endocrine systems are integrated.

Animals↗
PubMed2004

Influence of Griseofulvin treatment on semen quality in the dog.

Griseofulvin is used to treat dermatomycosis in many species and causes oligospermia in supra-pharmacological doses. The aim of the present study was to evaluate the effect of Griseofulvin administered at therapeutic doses upon semen quality in dogs. Four dogs were treated with Griseofulvin (25 mg/kg per day) for 30 days. Semen collections and analyses were performed before, during and for 100 days after treatment for the Griseofulvin group and 10 untreated control dogs. Semen analyses included mean percentage of forward progressively motile sperm, total sperm output, normal live sperm and normal dead sperm. There was no significant difference between control and treated dogs for each of the semen quality parameters. Therapeutic dosage of Griseofulvin had no deleterious effect upon semen quality in dogs, although this does not preclude potential embryotoxic and teratogenic effects.

Animals↗
PubMed2003

The effect of oestradiol on postpartum uterine involution in sheep.

The present study tested the hypothesis that ovarian oestradiol increases the rate of uterine involution after parturition in sheep. The day after parturition, ewes were randomly assigned as un-operated controls (n=5), or a 3 cm silastic implant containing oestradiol (n=8) or empty (n=7) was sutured within the bursa of the ovary ipsilateral to the previously gravid uterine horn. Blood samples were collected daily for measurement of PGFM and acute phase proteins until 17 days postpartum when the ewes were slaughtered and the genital tract was collected. There was no consistent effect of treatment group on uterine involution determined by the collagen density, dry matter content, width, length, or weight of the genital tract. Furthermore, there was no evidence of a localised effect of oestradiol on involution as there were no significant differences between the previously gravid and non-gravid uterine horns. However, oestradiol-treated ewes had lower plasma concentrations of 15-keto-13,14-dihydro-prostaglandin F2alpha (P<0.01), alpha1-acid glycoprotein (P<0.05) and ceruloplasmin (P<0.001); but, not haptoglobin. These observations could reflect a direct effect of oestradiol on inflammatory mediator synthesis or secretion because, in the absence of parallel physical measurements, it is unlikely that these observations reflect a beneficial effect of treatment on uterine health.

Animals↗
PubMed2003

Reproductive challenges facing the cattle industry at the beginning of the 21st century.

The aim of this review is to pinpoint the areas that require further research for greatest impact to improve the efficiency of dairy and beef production. Increased knowledge about the principal causes of reduced fertility is essential. Increases in milk yield have been at the expense of reduced fertility in dairy cows and although diet has a major impact, the precise interaction between nutrition and reproduction still needs to be characterized in both beef and dairy cows. Furthermore, during periods of inadequate nutrition or stress, the intensity of oestrus is reduced by inadequate exposure to oestradiol. However, it is still unclear which pheromones are involved at oestrus, how synthesis is controlled and how pheromones are detected in herd-mates. GnRH may be involved in behaviour but the brain centres that translate hormonal messages are unknown. Attempts to overcome poor oestrous detection include measurements of milk progesterone, telemetric pressure detectors and devices that record the extra activity at oestrus. Substitution of heat detection by 'hormone treatment remedies' has met strong consumer resistance in Europe. The creation of larger cattle units and increased movements world-wide render herds more susceptible to infectious agents, such as Neospora, Leptospira, Trypanosoma and Bovine Viral Diarrhoea virus (BVDv), but it is unclear how other clinical conditions, such as lameness or endometritis, also interfere with ovarian function. The future of dominant follicles selected within 14 days after parturition is crucial--normal ovulation, prolonged persistence or atresia. Calving carries the greatest risk in the reproductive life of a cow and yet little work has focused on reducing the frequency of this event. For dairy cows, a greater understanding about induced or extended lactation is required. For beef animals, precise induction of twinning and nutritional adjustments could produce two offspring per pregnancy. At the start of pregnancy, the trophoblast produces interferon to prevent luteolysis, but the immunological implications are unknown and it is not clear how the rest of pregnancy is maintained. Profiles of pregnancy specific protein B (PSPB) have increased understanding of embryonic death. However, 25% of cows in abattoirs are pregnant, even though 30% of involuntary cullings are 'for failing to conceive'. Clearly, this is an area of wastage that requires urgent resolution. It is unknown why undernutrition at the time of insemination or in early pregnancy leads to delayed births, low fetal weights and later adverse health. At the end of pregnancy, the fetus controls the onset of parturition, but very little is known about the biochemical control of cervical dilation and placental separation. On the male side, bulls are selected for optimal freezability of semen; however, there is as yet no reliable predictor for semen fertility. Methods for accurately pre-determining the sex of both semen and embryos will revolutionize the dairy and beef industries.

Animal Nutritional Physiological Phenomena↗
PubMed2002

Effect of postpartum manual examination of the vagina on uterine bacterial contamination in cows.

Twenty-one days after they had calved, the vaginas of 34 cows were examined with a clean gloved hand, and 26 cows were left unexamined. Swabs were collected from the lumen of the uterine body of the cows on the same day and seven days later; bacteria were identified by aerobic and anaerobic culture, and bacterial growth was scored semi-quantitatively. On the same days, blood samples were collected and the concentrations of acute phase proteins were measured, and the diameters of the cows' uterine horns were measured by transrectal ultrasonography. The vaginal examinations did not result in uterine bacterial contamination or an acute phase protein response, and they did not affect the diameters of the uterine horns.

Animals↗
PubMed2002

Influence of uterine bacterial contamination after parturition on ovarian dominant follicle selection and follicle growth and function in cattle.

First postpartum dominant follicles are preferentially selected in the ovary contralateral to the previously gravid uterine horn. The aim of the present study was to test the hypothesis that uterine bacterial contamination alters the location of ovarian follicle emergence and selection, and inhibits follicle growth and function. Swabs were collected from the uterine body lumen of cattle on days 7, 14, 21 and 28 after parturition. Bacteria were identified by aerobic and anaerobic culture; bacterial growth was scored semiquantitatively and animals were categorized into standard or high bacterial contamination categories on the basis of the number of colonies detected. Follicular growth and function were monitored by daily transrectal ultrasonography, and estimation of plasma FSH, oestradiol and progesterone concentrations. There was no effect of bacterial contamination on plasma FSH concentration profiles or emergence of the ovarian follicle wave. When uterine bacterial growth scores were high on day 7 or day 21 after parturition, fewer first (1/20 versus 15/50; P < 0.05) or second (1/11 versus 13/32; P < 0.05) dominant follicles were selected in the ipsilateral compared with the contralateral ovary, respectively. The diameter of the first dominant follicle was smaller in animals with a high day 7 bacterial score (P < 0.001), dominant follicle growth was slower (P < 0.05) and oestradiol secretion was decreased (P < 0.05). The present study provides evidence for an effect of the uterus on the ovary after parturition, whereby uterine bacteria have a contemporaneous localized effect on ovarian follicle selection and subsequent growth and function, but not on initial emergence.

Animals↗
PubMed2001

Cryopreservation of epididymal dog sperm.

A method of cryopreservation was developed for sperm salvaged from the cauda epididymis and vas deferens of domestic dog testes. Four modifications of the glycerol concentration of a buffer used for cryopreservation of dog ejaculates and two freezing rates were assessed for their effect upon post-thaw spermatozoal motility and morphology. There was no statistical difference between the four glycerol concentrations or the two freezing rates and the buffer containing 6% glycerol and the freezing rate provided by 0.5 ml straws was chosen for further study. This method resulted in a significant reduction in the percentage of live spermatozoa detected with Hoechst staining and a reduction in the percentage of capacitated spermatozoa after freeze-thawing. However, there was no difference in the ability of frozen-thawed spermatozoa to penetrate homologous oocytes. This study demonstrates that cryopreservation of epididymal canine sperm can be performed using methods similar to those established for ejaculates of the same species, and that despite some damage, spermatozoa retain their functional ability.

Animals↗
PubMed2001

Acute phase protein responses to uterine bacterial contamination in cattle after calving.

Repeated ultrasonographic examinations and collections of blood samples and uterine lumenal swabs between seven and 28 days after calving were used to examine the relative effects of bacterial contamination and involution of the uterus on the concentrations of acute phase proteins in the blood of 26 dairy cows. The severity of bacterial contamination, as determined by the total bacterial growth score, was a significant variable for the concentrations of the acute phase proteins alpha1-acid glycoprotein (P < 0.0001), haptoglobin (P < 0.05) and ceruloplasmin (P < 0.0001). In addition, the concentrations of alpha1-acid glycoprotein and ceruloplasmin were increased in the cows from which Escherichia coli (P < 0.0001) and Arcanobacterium pyogenes (P < 0.05), respectively, were isolated from the uterine lumen. Uterine involution, as determined by the decreasing diameter of the previously gravid uterine horn, was associated with a decrease in the concentrations of alpha1-acid glycoprotein (P < 0.005), haptoglobin (P < 0.05) and ceruloplasmin (P < 0.01). However, the response of the acute phase proteins to bacterial contamination was independent of the day on which the samples were collected, indicating that their concentrations were increased by bacterial contamination in addition to the changes associated with uterine involution.

Acute-Phase Proteins↗
PubMed2000

Peripheral and intrauterine neutrophil function in the cow: the influence of endogenous and exogenous sex steroid hormones.

It has been accepted for many years that the susceptibility of the genital tract to infection is reduced during the follicular phase compared with the luteal phase of the estrous cycle. Since the role of intrauterine neutrophils is paramount in the elimination of bacteria, it can be hypothesized that these differences in resistance to infection could be mediated by differences in uterine-derived neutrophil function. In order to test this hypothesis two groups of cows were used in this study. Group 1 cows (n=5) were studied at estrus, diestrus, after ovariectomy, after exogenous estradiol and after progesterone treatment, at which time they underwent intrauterine infusion with 1% oyster glycogen (OG) and a bacterial-free filtrate (BFF) of Actinomyces genes (BFF), the latter having been recovered from a clinical case of endometritis; neutrophils were harvested by flushing from the lumen 15 to 18 h later. A peripheral blood sample was collected at the time of flushing for the assay of estradiol and progesterone for a WBC and differential count and for the harvesting of neutrophils using a Percoll single-stage discontinuous gradient. After the recovery of the cells they were re-suspended in HBSS. Group 2 (n=4) were infused with BFF during during all reproductive states as Group 1, but with OG only after ovariectomy and after treatment with progesterone and estradiol. Neutrophil chemotaxis was assessed by measuring their migration using a modified Boyden chamber and Zymogen-activated serum as a chemoattractant. Phagocytic activity was measured by determining the number of Candida albicans ingested by each neutrophil after incubation. The percentage of kill was determined using a radiometric assay in which C. albicans was labeled with L-(5-3H) Proline. Peripheral WBC concentration was not influenced by the reproductive state of the cow; however, the mean neutrophil concentration was significantly different between the reproductive states (P<0.001) and between individual cows (P<0.001). In Group 1, there was little difference in the function of the peripheral and uterine neutrophils, and while there were differences in all 3 aspects of neutrophil function from both sources between reproductive states and individual cows, of which some were statistically significant, there was no consistent pattern. In Group 2, neutrophils recovered after the infusion of BFF had poorer function compared with those recovered after the infusion of OG. There was no consistent influence of the reproductive state or individual animal. The hypothesis that the influence of the reproductive state of the cow on the resistance of the uterus to infection is mediated by the inherent differences in either peripheral or intrauterine neutrophil function was not supported by this study.

Animals↗
PubMed2000

The influence of ovarian activity and uterine involution determined by ultrasonography on subsequent reproductive performance of dairy cows.

The objective of this study was to test the hypothesis that a follicle >8 mm diameter in the ovary ipsilateral to the previously gravid uterine horn (PGUH), between 14 and 28 days postpartum, improves subsequent reproductive performance. Lactating Holstein-Friesian cows (n=284) in 3 commercial herds were examined using transrectal ultrasonography once between 14 and 28 days postpartum to determine associations between uterine and ovarian measurements and subsequent fertility. There were fewer cows with a corpus luteum in the ovary ipsilateral to the PGUH compared with the contralateral ovary (16.9% vs. 37.0%; P<0.001). In addition, in the ovary ipsilateral to the PGUH there were fewer follicles >5 mm diameter (mean +/- SEM; 0.69 +/- 0.06 vs. 1.02 +/- 0.06; P<0.001) and fewer animals with a follicle >8 mm diameter (26.1% vs. 49.6%; P<0.001). These differences between the ovaries ipsilateral or contralateral to the PGUH declined with increasing time between 14 and 28 days postpartum. The presence of a purulent vaginal discharge decreased the number of animals with a corpus luteum in the ovary contralateral to the PGUH (45/143 vs. 60/141; P<0.05), but not in the ovary ipsilateral to the PGUH. The presence of a follicle >8 mm diameter in the ovary ipsilateral to the PGUH was associated with a shorter calving to conception interval compared with animals without such a follicle (99.0 +/- 5.6 days, n=74, vs. 112.8 +/- 4.4 days, n=210; P<0.05). These observations raise an intriguing issue: how does this follicle affect subsequent fertility--does the follicle exert a local influence on the uterus, or vice versa?

Animals↗
PubMed2000

Effect of administration of eCG to postpartum cows on folliculogenesis in the ovary ipsilateral to the previously gravid uterine horn and uterine involution.

This study tested the hypothesis that increased oestradiol secretion by large follicles in the ovary ipsilateral to the previously gravid uterine horn has a local effect to increase the rate of uterine involution. Cows were administered an i.m. water placebo (n = 19), 250 iu equine chorionic gonadotrophin (eCG) (n = 18) or 750 iu eCG (n = 20) 14 days post partum (day 0). Transrectal ultrasonography at the time of treatment and 2, 4 and 6 days later monitored uterine horn diameter and ovarian structures. Blood samples collected contemporaneously were assayed for 15-keto-13, 14-dihydro-prostaglandin F(2alpha) and oestradiol concentration. For control cows, accumulated diameter of the largest follicle in the ovary ipsilateral to the previously gravid uterine horn, compared with the contralateral ovary, was smaller on day 0 (P < 0.05) and days 2, 4 and 6 (P < 0.001). There were no significant differences in the eCG-treated animals. There were 12, 8 and 11 cows with a plasma oestradiol concentration < 1 pg ml(-1) on day 0 in the control, 250 and 750 iu eCG treatment groups, respectively. For control cows, the peripheral oestradiol concentrations were higher on day 6 compared with days 0, 2 and 4 (P < 0.05); for cows treated with 250 iu eCG, concentrations were higher on days 4 and 6 compared with day 0 (P < 0.05); and for cows treated with 750 iu eCG, concentrations were higher on days 2 and 4 compared with day 0 (P < 0.01). Treatment with eCG, or the presence of a follicle > 8 mm in diameter in the ovary ipsilateral to the previously gravid uterine horn, did not affect the rate of uterine involution or plasma 15-keto-13,14-dihydro-prostaglandin F(2alpha) concentration. In conclusion, administration of eCG to increase follicular growth and oestradiol production overcame the inhibition of follicular growth in the ovary ipsilateral to the previously gravid uterine horn, but did not affect uterine involution.

Analysis of Variance↗
PubMed1998

Comparison of three treatments for bovine endometritis.

Three commercial preparations for the treatment of bovine endometritis were compared: an intrauterine infusion of 1500 mg oxtytetracycline hydrochloride solution, an intramuscular injection of 500 micrograms cloprostenol (a synthetic analogue of prostaglandin F2 alpha), and an intramuscular injection of 3 mg oestradiol benzoate/500 kg estimated bodyweight. A total of 300 cases of endometritis were treated, of which 225 involved first, 67 involved second, and eight involved third or subsequent treatments. The overall success rate of treatment was 68 per cent. Oxytetracycline was successful in 73 per cent of cases, cloprostenol in 67 per cent and oestradiol in 63 per cent of cases. There was no significant difference between the success rates of the treatments, except for cows with mild endometritis in which oxytetracycline was more successful than oestradol (86 v 66 per cent, P < 0.05). Mild cases were treated more successfully than moderate cases (78 v 61 per cent, P < 0.01), and more successfully than severe cases (78 v 44 per cent, P < 0.001). Prostaglandin F2 alpha was more successful if the milk progesterone concentration was > 7 ng/ml at the time of treatment (P < 0.05). The presence of a smelly discharge at the time of treatment reduced the success rate by 17 per cent (P < 0.02). The treatment to conception interval for all successful treatments of endometritis by prostaglandin F2 alpha was 18.1 days shorter than for oestradiol (68.3 v 86.4 days, P < 0.02), and the interval for oxytetracycline was 16.2 days shorter than for oestradiol (70.2 v 86.4 days, P < 0.05).

Animal Husbandry↗