Calm Merino ewes have a higher ovulation rate and more multiple pregnancies than nervous ewes.

In 1990, two selection lines of Merino sheep were established for low and high behavioural reactivity (calm and nervous temperament) at the University of Western Australia. Breeding records consistently showed that calm ewes weaned 10% to 19% more lambs than the nervous ewes. We hypothesise that calm ewes could have a higher ovulation rate than nervous ewes and/or calm ewes could have a lower rate of embryo mortality than nervous ewes. We tested these hypotheses by comparing the ovulation rate and the rate of embryo mortality between the calm and nervous lines before and after synchronisation and artificial insemination. Merino ewes from the temperament selection lines (calm, n=100; nervous, n=100) were synchronised (early breeding season) for artificial insemination (day 0) (intravaginal sponges containing fluogestone acetate and eCG immediately after sponge withdrawal). On day-17 and 11 ovarian cyclicity and corpora lutea, and on days 30 and 74 pregnancies and embryos/foetuses were determined by ultrasound. Progesterone, insulin and leptin concentrations were determined in blood plasma samples from days 5, 12 and 17. Ovarian cyclicity before and after oestrus synchronisation did not differ between the lines, but ovulation rate did (day-17: calm 1.63; nervous 1.26; P<0.01; day 11: calm 1.83; nervous 1.57; P<0.05). Ovulation rate on day 11 in nervous ewes was higher than on day-17. Loss of embryos by day 30 was high (calm: 71/150; nervous: 68/130); but nervous ewes had a lower proportion (15/47) of multiple pregnancies compared with calm ewes (30/46; P<0.01). Reproductive loss between days 30 and 74 represented 7.3% of the overall loss. Temperament did not affect concentrations of progesterone, but nervous ewes had higher insulin (32.0 pmol/l±1.17 SEM; P=0.013) and lower leptin (1.18 µg/l±0.04 SEM; P=0.002) concentrations than calm ewes (insulin: 27.8 pmol/l±1.17 SEM; leptin: 1.35 µg/l±0.04 SEM). The differences in reproductive outcomes between the calm and nervous ewes were mainly due to a higher ovulation rate in calm ewes. We suggest that reproduction in nervous ewes is compromised by factors leading up to ovulation and conception, or the uterine environment during early pregnancy, that reflect differences in energy utilisation.

Dynamics of sperm DNA fragmentation in raw boar semen and fertility.

The aims were to evaluate sperm DNA fragmentation (SDF) in boars through the dispersion of their chromatin in raw semen samples, quantifying the extent of SDF, and to assess dynamic aspects of sperm DNA damage after incubation to obtain the rate of sperm DNA fragmentation (rSDF) under thermal conditions similar to the uterus (37°C) over a period of up to 24 hr and to correlate the reproductive outcome of the sows with the SDF of the boars at ejaculation. The study was performed on a pig-breeding farm in southern Uruguay. Sixty-one ejaculates from five of the most frequently used hybrid boars were evaluated. Semen was collected weekly from each of the boars, using the gloved-hand technique and discarding the jelly-like fraction of the ejaculate. Fresh semen was kept in a water bath at 37°C and protected from light, and was thereafter processed with Sperm-Sus-Halomax(®) to evaluate SDF. The smears for time 0 (T0) were made on farm, and thereafter smears were made at the laboratory at 4 hr of obtaining the semen (T4), then every 2 hr (T6, T8, T10, T12) and a final fixation at 24 hr (T24). Differences in SDF were observed among exposure times for all boars (p < .05), but not between T10 and T12 (p = .7751) nor T4 and T24 (p = .9113). In none of the T24 samples, sperm heads could be seen with chromatin dispersion halos. Furthermore, there were differences among boars when comparing sperm rSDF (p < .05). Farrowing rate was not affected by SDF at T0 (r = .38, p = .75), nor was litter size (r = .16, p = .70). With the present experimental conditions, we have not been able to show a relationship between sperm DNA fragmentation at ejaculation and reproductive performance. However, this could be a result of the low number of ejaculates and boars used.

Temperament and its heritability in Corriedale and Merino lambs.

Temperament can be defined as the fearfulness and reactivity of an animal in response to humans and strange, novel or threatening environments. The productive performance of an animal is affected by its temperament, and selection of calm animals might improve their adaptation to the farming environment and handling, as well as improve productivity. The temperament was measured in lambs of two breeds of sheep in Uruguay. The effects of dam's age, type of birth, age of the lamb and contemporary group (CG; lambs belonging to the same year, flock, sex and rearing group) on the temperament of the lambs and the heritability of temperament were estimated with a Bayesian analysis using Gibbs sampling. Overall, 4962 Corriedale lambs and 2952 Merino lambs from 13 farms were tested. Temperament was measured using the isolation box test, isolating a lamb inside the box for 30 s, and recording the vibrations produced by its movements. The average temperament score (±s.e.m.) of the Corriedale lambs was 24.7 (±0.23) and that of the Merino was 36.8 (±0.45). Temperament was not associated with dam's age, type of birth or lamb's age. There were no relevant differences in the agitation score between lambs born in 2010 and 2011. The mean of the distribution of possible values of heritability (±s.d.) was 0.18 (±0.05) for the Corriedale and 0.31 (±0.06) for the Merino. The likelihood of heritability values to be greater than 0.15 exceeded 70% in the Corriedale and 90% in the Merino. The temperament of Merino and Corriedale sheep in Uruguay is moderately heritable. It is not related to dam's age, type of birth or age of the lambs; however, it is affected by some aspect of the CG.

Sex steroid modulation of cortisol secretion in sheep.

There is strong evidence that the gonads modulate the hypothalamic-pituitary-adrenal axis. To investigate these sex differences at the adrenal glands of sheep we compared the cortisol response to ACTH (experiment 1) and measured the relative expression of oestrogen receptor alpha (ERS1), androgen receptor (AR), melanocortin 2 receptor (MC2R) and steroid acute regulatory protein (STAR) mRNA in adrenal glands (experiment 2) of gonadectomised rams and ewes either with or without sex steroid replacement. In experiment 1 six castrated adult rams and four ovariectomised adult ewes were used in two ACTH trials. On each trial blood samples were taken every 15 min for 4 h through an indwelling jugular catheter and each animal received 0.5 mg of an ACTH analogue i.v., immediately after the sample at 1 h from the beginning of the trial. Four days after the first trial the males received 100 mg of Testosterone Cyclopentilpropionate (TC) i.m. and the females received 2.5 mg of Oestradiol Benzoate (EB) i.m. At 72 h after TC or EB administration the second trial was performed. In experiment 2 the adrenal glands were obtained from gonadectomised adult rams (n=8) and adult ewes (n=8). Four rams received 100 mg of TC i.m. and four females received 0.5 mg of EB i.m. Blood samples were taken at 0, 12, 24, 48 and 72 h relative to steroid replacement and the animals were thereafter slaughtered. Cortisol, testosterone and 17ß-oestradiol were determined by radioimmunoanalysis. The transcripts of ERS1, AR, MC2R and STAR were determined by real-time reverse transcription PCR in adrenal tissue. Cortisol secretion was higher in female sheep than in male sheep, and higher in EB-treated than non-treated ewes. No difference in cortisol secretion was observed between TC-treated and non-treated rams. Gonadectomised rams treated with TC presented greater AR mRNA and MC2R mRNA expression than males without the steroid replacement. Gonadectomised ewes treated with EB tended to present lower AR mRNA than the ones without steroid replacement. Gonadectomised rams with TC also had greater AR mRNA, ERS1 mRNA and MC2R mRNA expression than ewes treated with EB. The relative amount of STAR transcript was not different among the different groups. The results confirm sex differences in ACTH-induced cortisol secretion in sheep, as well as in the expression of the receptor proteins for both 17ß-oestradiol and testosterone in the sheep adrenal gland. However, the underlying mechanisms for sex steroid modulation remain unresolved.

The burden of 2009 pandemic influenza A(H1N1) in the Netherlands.

BACKGROUND: The disease burden of the 2009 influenza pandemic has been debated but reliable estimates are lacking. To guide future policy and control, these estimates are necessary. This study uses burden of disease measurements to assess the contribution of the pandemic influenza A(H1N1) virus to the overall burden of disease in the Netherlands. METHODS: The burden of disease caused by 2009 pandemic influenza was estimated by calculating Disability Adjusted Life Years (DALY), a composite measure that combines incidence, sequelae and mortality associated with a disease, taking duration and severity into account. Available influenza surveillance data sources (primary care sentinel surveillance, notification data on hospitalizations and deaths and death registries) were used. Besides a baseline scenario, five alternative scenarios were used to assess effects of changing values of input parameters. RESULTS: The baseline scenario showed a loss of 5800 DALY for the Netherlands (35 DALY per 100 000 population). This corresponds to 0.13% of the estimated annual disease burden in the Netherlands and is comparable to the estimated disease burden of seasonal influenza, despite a different age distribution in incidence and mortality of the pandemic compared to seasonal influenza. CONCLUSIONS: This disease burden estimate confirmed that, although there was a higher mortality observed among young people, the 2009 pandemic was overall a mild influenza epidemic. The disease burden of this pandemic was comparable to the burden of seasonal influenza in the Netherlands.

Displaying random variation in comparing hospital performance.

INTRODUCTION: The role of transparency in quality of care is becoming ever more important. Various indicators are used to assess hospital performance. Judging hospitals using rank order takes no account of disturbing factors such as random variation and case-mix differences. The purpose of this article is to compare displays for the influence of random variation on the apparent differences in the quality of care between the Dutch hospitals. METHOD: The authors analysed the official 2005 data of all 97 hospitals on the following performance indicators: pressure ulcer, cerebro-vascular accident and acute myocardial infarction. The authors calculated CIs of the point estimate and the simulated CIs of the ranks with bootstrap sampling, and visualised the influence of random variation with three modern graphical techniques: forest plot, funnel plot and rank plot. RESULTS: Statistically significant differences between hospitals were found for nearly all performance indicators (p<0.001). However, the CIs in the forest plot revealed that only a small number of hospitals performed significantly better or worse. The funnel plot provides a representation of differences between hospitals compared with a target value and allows for the uncertainty of these differences. The rank plot showed that ranking hospitals was very uncertain. CONCLUSION: Despite statistically significant differences between hospitals, random variation is a crucial factor that must be taken into account when judging individual hospitals. The funnel plot provides easily interpretable information on hospital performance, including the influence of random variation.

[Effects of ageing on infectious disease]. / Effecten van vergrijzing op infectieziekten.

Expectations are that 25% of the Dutch population will be aged 65 years or older in 2050 whilst in 2008 this proportion was only 15%. As a consequence the annual absolute number of new cases of specific infectious diseases will increase. Elderly people often have less clear symptoms of infection, making it harder for clinicians to recognize the disease. Morbidity and mortality caused by infections increase with age. Ageing will possibly cause an increase in levels of antibiotic resistance. There will be more elderly people in hospitals and nursing homes, where more antibiotics are used. This development will come in addition to other developments in the field of antibiotic resistance. There will be more elderly people with chronic HIV and viral hepatitis B and C infection and their complications because patients who are presently infected with those pathogens are ageing. Policy makers should more often consider introducing vaccination of older age groups against certain infectious diseases.

[High vaccination coverage of the National Immunization Programme in the Netherlands]. / Hoge vaccinatiegraad van het Rijksvaccinatieprogramma in Nederland.

OBJECTIVE: To describe vaccination coverage of the National Immunization Programme (NIP) in the Netherlands for the years 2006-2008. DESIGN: Descriptive, retrospective. METHOD: Vaccination coverage was determined per vaccination and birth cohort based on data from the vaccination-registration system 'Praeventis'. This new system enables more accurate testing for compatibility with NIP guidelines. Furthermore, it was determined on an individual level whether the desired vaccination status was reached before a certain target age (1, 2, 5 or 10 years). RESULTS: In the final year reported, all national vaccination percentages came up to the WHO standards. Vaccination coverage for babies ranged from 94.5% for diphtheria, whooping cough, tetanus and polio (DTaP-IPV) to 96.0% for measles, mumps and rubella (MMR). The coverage for toddlers and school-aged children was over 90% for each of the vaccinations. In 5 of the 12 Dutch provinces and 128 of the 443 municipalities, one or more vaccination percentages were below the standard of 90%. Vaccination coverage for the second MMR vaccination and for hepatitis B vaccination in children of whom one or both parents were born in a country where the incidence of hepatitis B is higher than average, was relatively low (92.5% and 90.7%, respectively). CONCLUSION: Vaccination coverage in the Netherlands is high. In general, vaccination coverage for toddlers and school-aged children requires extra attention. This applies in particular to the second MMR vaccination and to vaccination in children of whom one or both parents were born in a country where the incidence of hepatitis B is higher than average.

The burden of infectious diseases in Europe: a pilot study.

The main objectives of this pilot study were to test the potential use of the disease burden concept in the field of infectious diseases, including data quality and availability; to recommend future studies; and to stimulate a debate. The disease burden of seven infectious diseases (influenza, measles, HIV, campylobacteriosis, infection with enterohaemorrhagic Escherichia coli, salmonellosis and tuberculosis) in Europe was estimated by calculating Disability Adjusted Life Years (DALYs), a composite measure that attempts to combine mortality, incidence and sequelae, taking duration and severity into account. The results show that the relative burden of diseases as measured by DALYs differs from that only measured by incidence or mortality. Several limitations regarding data availability and quality have been identified, resulting in an underestimation of the true burden of disease in this pilot. Notwithstanding these, HIV-infection, tuberculosis (TB) and influenza are estimated to cause the highest burden in Europe among the selected diseases. The burden of foodborne diseases (campylobacteriosis, infection with enterohaemorrhagic Escherichia coli and salmonellosis) and in particular of measles is lower. A consideration of the relative comparison of burden between diseases can be useful when tackling the difficult, sensitive but necessary task of identifying priority actions. A low burden stresses the need for continued support for prevention and control whereas a high burden indicates the need for additional interventions. Following this pilot project, a generalised burden of disease study for infectious diseases in Europe is recommended. Such a study would benefit from an approach that identifies and combines several methods of investigation, including epidemiological modelling, and it should be done in collaboration with other international efforts in this field.

Influence of estrus of dairy goats on somatic cell count, milk traits, and sex steroid receptors in the mammary gland.

Two experiments were conducted to study the effect of the stage of a spontaneous estrus cycle on milk yield and constituents [somatic cell count (SCC), fat, protein, caseins, lactose, and urea content] and on estrogen receptor-alpha (ERalpha ) and progesterone receptor (PR) immunostaining in the mammary gland. In experiment I, the major components of milk and SCC were monitored weekly in 80 lactating Saanen goats for 6 wk, whereas detection of estrus was daily. In experiment II, milk samples were collected daily for SCC determination during 1 spontaneous estrus (d 0) until the second spontaneous estrus in 14 Saanen goats. The day of the estrous cycle was confirmed by plasma progesterone and 17beta-estradiol levels. Immunoreactivity of ERalpha and PR was analyzed in mammary gland samples of 8 Saanen goats (d 0, n = 4; d 10, n = 4) and the number of positive nuclei and intensity of the staining were evaluated in 1,000 cells. In experiment I, milk casein and protein percentages were significantly affected by the stage of estrous cycle; during proestrus and estrus, these variables were higher (3.32 +/- 0.06 and 4.44 +/- 0.08) than during metestrus (3.03 +/- 0.07 and 4.07 +/- 0.10), but not higher than during diestrus (3.23 +/- 0.06 and 4.35 +/- 0.09, respectively). In experiment II, daily measurement of SCC revealed higher levels at estrus (7,195 +/- 672 x 10(3) cells/mL) and a decline toward the luteal phase (1,694 +/- 672 +/- 10(3) cells/mL). Estrogen receptor-alpha and PR immunostaining were exclusively detected on epithelial cells. The percentage of positive nuclei to ERalpha was higher on d 0 than on d 10 (75.4 +/- 8.8 vs. 68.3 +/- 8.8%), but no change was observed for PR (4.0 +/- 0.3 vs. 3.5 +/- 0.4%). The average immunostaining intensity for both receptors was greater on d 0 than on d 10 (ERalpha : 1.44 +/- 0.02 vs. 1.35 +/- 0.02; PR: 0.079 +/- 0.008 vs. 0.057 +/- 0.008). The high SCC at estrus in experiment II was associated with high plasma estradiol and low progesterone, suggesting that the increased SCC could be brought about by the estrogen-induced proliferation and exfoliation of epithelial cells. In addition, this action may be supported by the higher sensitivity to estrogens (ERalpha content) found at d 0.

Sex differences in oestrogen receptor levels in adrenal glands of sheep during the breeding season.

The concentrations of the oestrogen receptor (ER), and the mRNA levels of ERalpha, progesterone receptor (PR) and insulin-like growth factor I (IGF-I) were characterised in adrenal glands and uterine tissue of adult Corriedale sheep during the breeding season. The sheep were of different sex and gonadal status. Ewes had higher levels of cytosolic ER in the adrenals than the rams (mean+/-S.E.M.: 7.3+/-2.0 fmol/mg protein and 2.5+/-1.0 fmol/mg protein, respectively; P=0.0091) and gonadectomy increased ER (mean+/-S.E.M.: 2.9+/-1.2 fmol/mg protein and 8.6+/-2.3 fmol/mg protein, intact and gonadectomised sheep, respectively; P=0.0071). No differences could be observed in mRNA levels for ERalpha and IGF-I in the adrenal glands of all of the sheep. PR mRNA levels were reduced in ovariectomised ewes and enhanced in castrated rams (sex x gonadal status: P=0.009). PR mRNA levels tended to be higher in ewes in the follicular phase than in ovariectomised ewes and intact rams (P<0.1). All of the animals had positive nuclear staining for ERalpha in the adrenal cortex, but no differences were observed between the groups. In this study, we demonstrated the existence of ER in the adrenal gland of sheep and found varying sensitivity to oestrogens as the ER levels differed among sex and gonadal status. These findings indicate that oestrogens most likely affect steroidogenesis directly at the adrenal cortex and suggest that oestrogens are partly responsible for the sex differences in cortisol secretion in sheep.

Sex differences in cortisol secretion after administration of an ACTH analogue in sheep during the breeding and non-breeding season.

The aim of this study was to compare the response of cortisol in sheep of different sex and gonadal status to adrenal cortex stimulation by an ACTH analogue in the breeding and non-breeding season. Twenty-four adult Corriedale sheep were used in the non-breeding season, and 19 in the breeding season. Three weeks prior to the first trial (non-breeding season), six rams and six ewes were gonadectomised. In each trial, blood was obtained every 15min for 9h and the animals received 0.5mg of ACTH (Tetracosactid, Synacthen Depot i.m., after 1.5h of sampling. Sampling began at 10:00a.m. in the non-breeding season and at 9:00a.m. in the breeding season. Three main effects (sex, gonadal status and season) were evaluated, each with two levels (male and female, intact and gonadectomised, breeding and non-breeding season, respectively). In both seasons, the females showed higher cortisol levels after ACTH than males (P<0.001), though the difference seemed less marked in the non-breeding season. The cortisol response in the ewes was not affected by season. The rams, however, showed a lower response in the breeding season (P<0.03). Gonadectomy reduced the response in the ewes (P<0.001) but had no effect in the rams. Nevertheless, gonadectomy also eliminated the differences between the ewes and the rams, such that the intact rams had lower levels of cortisol compared to the intact females, with those of the gonadectomised animals of both sexes being intermediate between the gonad-intact groups. The results of this study confirm sex differences in ACTH induced cortisol secretion in intact sheep in vivo. Furthermore, by applying exogenous ACTH we have directly stimulated the adrenal cortex, indicating the existence of sex differences also at this level. The circulating gonadal steroids, which are responsible at least in part for the sex differences in the responses to stress, may influence cortisol secretion from the adrenal gland by direct action at the cortex.

Use of medroxyprogesterone acetate (MAP) in lactating Holstein cows within an Ovsynch protocol: follicular growth and hormonal patterns.

To evaluate the effects of incorporating medroxyprogesterone acetate (MAP) in an Ovsynch protocol, cyclic lactating dairy cows were assigned randomly to two groups (control and MAP, n=8 each). Ovsynch treatment (Day 0: GnRH, Day 7: PG, Day 9: GnRH) was initiated at random stages of the estrous cycle (control) and an intravaginal polyurethane sponge impregnated with 300mg of MAP was inserted intravaginally in the MAP group at Day 0 and removed at Day 7 of the Ovsynch protocol (MAP treatment). Ovaries were scanned daily from Day 0 until the second GnRH treatment on Day 9 and from then every 6h for 36 h. Milk samples were collected three times weekly starting 17 days before the initiation of treatment to determine the stage of the cycle at the beginning of the Ovsynch protocol. Blood samples were collected to monitor estradiol (E2), progesterone (P4), LH, and 15-ketodihydro-PGF(2alpha) (PGFM) by RIA. Response to the first GnRH treatment varied with the stage of the cycle at the time of initiation of treatment, as cows in metestrous and late diestrous did not ovulate. In cows ovulating, growth rate of the new follicle was not affected by the addition of MAP. No treatment differences were found in E2 concentrations which reached a maximum at Day 9, consistent with the maximum follicular size. At Day 7, cows with luteal concentrations of P4 had increased concentrations of PGFM, but cows with basal P4 did not show an active release of prostaglandins. There were no treatment differences in the ovulatory response to the second GnRH-induced ovulation, with 11 of the 16 cows ovulating between 16 and 32 h. The addition of MAP to the Ovsynch protocol could not mimic the normal high progesterone levels needed to prevent premature ovulations in those cows with premature CL regression.

Repeatability and relationship with field mating performance of a serving capacity pen test in rams.

Two studies were conducted to analyse the repeatability of a serving capacity (SC) pen test and to evaluate the correlation of the results of such a test with field performance. The Laborde SC test, which measures SC by exposing an individual ram for 40min to two restrained non-oestrous ewes in a pen was used. In the first study, SC was measured on four and six occasions during the autumn of 1994 and 1995, respectively. The results indicated that this test is reliable in measuring SC of rams, since SC was repeatable within, and between, breeding season of consecutive years (p<0.05). In the second study conducted on two commercial farms, two groups of ewes synchronised with intravaginal sponges containing medroxyprogesterone acetate, were served by rams of high or low SC. On both the farms, the group of ewes served by rams of high SC had higher pregnancy rates than those served by low SC rams (p<0.05). The Laborde test proved to be a reliable test for the evaluation of SC in rams and that high SC rams had better field performance.

Effects of adrenocorticotrophin (ACTH) and progesterone on luteinising hormone (LH) secretion in recently castrated rams.

The goal of the present study was to determine whether ACTH and progesterone have any effect on LH secretion and pulse frequency in recently castrated rams. Six 2-year-old Corriedale rams were castrated in the winter. The day before castration, blood samples were taken in order to establish the precastration LH levels. The rams were divided into an untreated group (group U: n = 2) and a treated group (group T: n = 4). The first treatment consisted of the i.v. administration of 0.5 mg of ACTH on day 20 post-castration, immediately after the first sample had been taken. During the second treatment, subcutaneous progesterone implants were given to group T for 5 days. Control samplings were performed one week before each treatment. Prior to castration, the testosterone levels were low, while after castration they were below the detection limit of the assay. Cortisol and progesterone concentrations were basal before castration in all of the animals and after castration in group U and also in the control samplings for group T. ACTH treatment caused a significant increase in both cortisol and progesterone levels for 3 h (P < 0.001). Progesterone implants raised progesterone levels in group T, but cortisol levels remained basal. Before castration, all animals had low LH levels and hardly any pulse activity was seen. After castration, both the number of LH pulses and the mean LH production increased significantly in all of the animals (P < 0.01). During the ACTH trial, LH pulse frequency was significantly reduced for the first 4 h following ACTH administration (P = 0.013), however, no such differences occurred in the prior control period. No effect was seen on mean LH concentration during the ACTH treatment. Progesterone treatment did not have any effect on either the number of LH pulses nor on LH concentrations (P > 0.05).