European bat lyssaviruses: Distribution, prevalence and implications for conservation.

Worldwide, there are more than 1100 species of the Order Chiroptera, 45 of which are present in Europe, and 16 in the UK. Bats are reservoirs of, or can be infected by, several viral diseases, including rabies virus strains (in the Lyssavirus genus). Within this genus are bat variants that have been recorded in Europe; European bat lyssavirus 1 (EBLV-1), European bat lyssavirus 2 (EBLV-2) and, four currently unclassified isolates. Since 1977, 783 cases of EBLVs (by isolation of viral RNA) have been recorded in Europe. EBLV-1 or EBLV-2 has been identified in 12 bat species, with over 95% of EBLV-1 infections identified in Eptesicus serotinus. EBLV-2 is associated with Myotis species (Myotis daubentonii and Myotis dasycneme). A programme of passive surveillance in the United Kingdom between 1987 and 2004 tested 4871 bats for lyssaviruses. Of these, four M. daubentonii (3.57% of submitted M. daubentonii) were positive for EBLV-2. Potential bias in the passive surveillance includes possible over-representation of synanthropic species and regional biases caused by varying bat submission numbers from different parts of the UK. In 2003, active surveillance in the UK began, and has detected an antibody prevalence level of 1-5% of EBLV-2 in M. daubentonii (n = 350), and one bat with antibodies to EBLV-1 in E. serotinus (n = 52). No cases of live lyssavirus infection or lyssavirus viral RNA have been detected through active surveillance. Further research and monitoring regarding prevalence, transmission, pathogenesis and immunity is required to ensure that integrated bat conservation continues throughout Europe, whilst enabling informed policy decision regarding both human and wildlife health issues.

Matrilineal genetic structure and female-mediated gene flow in red grouse (Lagopus lagopus scoticus): an analysis using mitochondrial DNA.

DNA sequence variation at the hypervariable 5' end of the mitochondrial control region was examined in 247 individuals to detect genetic divergence among 14 populations of red grouse (Lagopus lagopus scoticus) in northeastern Scotland. Ten haplotypes were resolved, several of which were shared among populations. Analysis of molecular variance, Nei's gamma ST, and a cladistic estimate of the amount of gene flow indicated a lack of overall population differentiation. Patterns of overall panmixia are in stark contrast to previous reports of localized subdivision among the same set of populations detected using hypervariable microsatellite markers. Because grouse cocks are territorial and show extreme natal philopatry and females are the dispersing sex, such discordance could be explained by sex-biased dispersal, with extensive female-mediated gene flow preventing mitochondrial DNA divergence. However, it is difficult to reconcile how effective dispersal of females would not homogenize both mitochondrial and nuclear structure simultaneously. We use a model that examines the spatial and temporal dynamics of diparentally and uniparentally inherited genes to show that, under realistic ecological scenarios and with specific differences in the dispersal of males and females, the local effective size of the nuclear genome can be less than that of the mitochondrial and the patterns of structuring we observe are meaningful.

The temporal relationship between the secretion of luteinizing hormone and testosterone in man.

It has been suggested recently that testosterone secretion by the human testis may be controlled by factors other than luteinizing hormone (LH). In order to re-examine this hypothesis, plasma LH and testosterone concentrations were determined throughout the day in eight studies. A new method of data analysis revealed that the levels of the two hormones were closely related, but that the testicular response to LH was sluggish. These results explain some inconsistencies in the literature. It was demonstrated that average values for LH varied throughout the day, with a morning maximum and an evening minimum. It was also shown that injections of LH releasing hormone in man resulted in an increase in plasma testosterone above control levels. These results are consistent with the concept that LH controls the major changes in testosterone secretion in men. They do not exclude, however, the possible existence of other factors which might affect the peripheral concentration of testosterone, such as changes in testicular blood flow.

Effect of maternal overnutrition during pregnancy on pituitary gonadotrophin gene expression and gonadal morphology in female and male foetal sheep at day 103 of gestation.

The aim was to determine whether nutritionally mediated restriction of placental growth alters foetal body growth, pituitary gonadotrophin gene expression and gonadal development at Day 103 of gestation. Embryos recovered from adult ewes inseminated by a single sire were transferred, singly, into the uteri of adolescent recipients. After transfer, adolescent ewes were offered a high (H, n=16) or moderate (M, n=12) level of a complete diet. Ewes were slaughtered at 103+/-0.2 days of gestation and foetal blood, brain, pituitary and gonads were collected. Mean placental weight was lower (P< 0.01) in H than in M groups but foetal weight and reproductive organ weights were similar. Maternal nutrition did not influence LHbeta or FSHbeta mRNA expression in either sex but FSHbeta mRNA expression was higher (P< 0.001) in female (n=11) than in male (n=17) foetal pituitaries. Mean foetal plasma gonadotrophin concentrations were not influenced by dietary intake in either sex. Plasma progesterone concentrations were lower (P=0.001) in foetuses derived from H compared with M intake dams. Compared with M foetuses (n=5), ovaries from H foetuses (n=6) had fewer primordial follicles (P< 0.05) and fewer follicles in total (P< 0.005). In contrast, maternal nutritional status did not influence either seminiferous cord or Sertoli cell numbers in male foetuses (H, n=10; M, n=7). It is concluded that high maternal nutrient intakes restricted placental growth and altered foetal ovarian follicular development prior to the end of the second third of gestation. The latter effect was independent of gonadotrophin secretion.Crown

Measurement of CO2 production by the doubly labeled water technique.

Recent applications of the doubly labeled water technique to the study of human metabolism have employed multiple sampling of body water over protracted periods, rather than the more traditional method of taking only an initial and final sample for isotopic analysis. In addition fractional turnovers of the body pools have been estimated by fitting curves to the sequential log-converted isotope enrichment against time. By manipulation of data collected in the field in a study of metabolism of vespertilionid bats, it is shown the curve-fitting technique results in an accurate estimate of CO2 production only when the rate of CO2 production is constant. Biologically realistic nonsteady-state conditions result in errors in estimates of CO2 production of up to 30%. In conditions where CO2 production is known to be temporally variable, the more traditional two-sample method may provide a more accurate estimate of CO2 production.

Effect of undernutrition of ewes from the time of mating on fetal ovarian development in mid gestation.

Ewes were fed either 150% (High, H) or 50% (Low, L) of their energy requirements for maintenance of liveweight during early gestation. Effects of maternal nutrition on fetal ovarian size, histological structure and steroidogenic capacity were studied at Day 47 and on ovarian size and structure at Day 62 of gestation. At Day 47 of gestation, there were significantly higher concentrations of oogonia in the ovaries of L fetuses than H fetuses (105.9 v. 76.9 germ cells mm(-2); s.e. 4.94; P < 0.001). The capacity of the ovaries to secrete oestradiol (pg/ovary/24 h) at Day 47 was not affected by treatment when they were incubated either with (H, 773; L, 740; s.e. 179; not significant, n.s.) or without (H, 260; L, 290; s.e. 92.7; n.s.) ovine luteinizing hormone (oLH). At Day 62 of gestation, the process of germ cell degeneration was less advanced in L than H fetal ovaries, as indicated by higher oocyte concentrations in the former (68.4 v. 48.6 germ cells mm(-2); s.e. 3.85; P < 0 01). There was a greater percentage of meiotic cells in L ovaries (76.5 v. 18.6; s.e. 5.82; P < 0.001). It is concluded that undernutrition of the ewe from the time of mating significantly retards ovarian development in fetal ovaries.

The effect of prenatal photoperiodic history on the postnatal endocrine status of female lambs.

Postnatal photoperiodic experience plays a pivotal role in determining the timing of ovarian activity in female lambs. This study examines whether a photoperiodic history gained while in utero is able to influence this timing. Pregnant Soay ewes were maintained in either long days (n = 7, 18 h light: 6 h dark; group PLD) or short days (n = 12, 6 h light: 18 h dark; group PSD) from 25 days of gestation. At birth, female lambs (n = 8 per group) were transferred to long days for 10 weeks, and then placed under short days until the end of the experiment at 38 weeks of age. Blood samples were collected from lambs on the day of birth and three times weekly for the duration of the study and the resulting plasma assayed for progesterone and prolactin. Although both gestational photoperiods produced, at best, abbreviated periods of ovarian activity, lambs born to ewes which experienced long days during gestation (group PLD) exhibited elevated plasma progesterone concentrations significantly earlier (P < 0.05) than lambs born to ewes exposed to short days during gestation (group PSD) (mean +/- SEM, 193 +/- 17 versus 244 +/- 14 days for PLD and PSD groups, respectively. Plasma prolactin concentrations in newborn lambs born between late December and early April were not affected by the ambient photoperiod, but reflected the artificial daylength experienced by their mothers during gestation. Lambs born to ewes maintained under long days during gestation (group PLD) had significantly higher prolactin concentrations on the day of birth than lambs born to ewes maintained under short days during gestation (group PSD) (45 +/- 5.4 ng/ml versus 7 +/- 3.7 ng/ml respectively, P < 0.001). The mean birth weight, rate of live weight gain and live body weight of lambs at the end of the experiment did not vary significantly between treatment groups. These results suggest that the ovine foetus is sensitive to photoperiodic information prior to birth, and develops a photoperiodic history which, under the present experimental conditions, modulates the subsequent endocrine status of the neonatal lamb.

Undernutrition of ewe lambs in utero and in early post-natal life does not affect hypothalamic-pituitary function in adulthood.

The effect of undernutrition in utero, during late gestation (from day 100), and early neonatal life on hypothalamic-pituitary function was investigated in female lambs born to ewes fed rations calculated to provide either 100% (high; H) or 70% (low; L) of the energy requirements to sustain a twin pregnancy. Following parturition in early spring, ewes and lambs were maintained on pasture with sward heights of 6 cm (H) or 4 cm (L) until week 8 of lactation and then sward heights of 5 cm (H) or 3 cm (L) until weaning at week 14. Mean lamb birth weights were 18% lower in L than H animals (P<0.05) and mean liveweights were 23% lower in the L animals (P<0.001) at weaning at 14 weeks of age. Liveweight differences were not significant at, or after, 26 weeks of age. There were no significant differences between pre-pubertal H and L animals, either before (26 weeks) or after ovariectomy (31 weeks), with respect to hypothalamic or pituitary activity, as measured by LH pulse frequency, pulse amplitude or mean plasma LH and FSH concentrations and the responses to GnRH injection as measured by LH peak amplitude, respectively. Similarly there were no differences in any of these variables in pubertal animals at 18 months of age. At 31 weeks of age, H animals had significantly lower pituitary GnRH receptor binding (P<0.01) and lower ERalpha mRNA content (P<0.05) than L lambs. There were no differences with treatment in the abundance of mRNA for LHbeta, FSHbeta or GnRH-receptor at 31 weeks of age or in pubertal animals aged 18 months, when there were no significant differences with treatment in GnRH receptor binding or ERalpha mRNA expression. It is concluded that effects on lifetime reproductive function of female sheep of undernutrition during late gestation and early neonatal life are unlikely to be expressed through permanent changes in hypothalamic-pituitary function and are therefore attributable to effects exerted directly on the ovary.

Fertility of the early postpartum, lactating domestic rabbit.

Sixty-four crossbred primiparous lactating does each suckling six pups were allocated at random into four groups and were mated on either Day 1, 2, 3, or 4 post partum (where Day 0 = the day of parturition). They were subsequently killed on Day 10 post coitum (where Day 0 = the day of mating) to assess fertility. There were no significant differences between treatment groups in their mating response (97% overall), ovulation response (77% overall), implantation response (83% overall), implantation rate (8.7 overall), or preimplantation mortality rate (24% overall). Ovulation rate was significantly increased in does mated on Days 3 and 4 (13.3 and 13.1, respectively), compared with those mated on Day 1 (10.2, P<0.05) and Day 2 (9.6, P<0.01) post partum. From these results we conclude that fertility is high throughout the early postpartum period in the lactating rabbit.

Effects of naloxone, metaclopramide and domperidone on plasma levels of prolactin and LH following suckling in the female rabbit.

Saline, naloxone, domperidone or metaclopramide was injected into lactating rabbits immediately before suckling. Blood samples were taken prior to injection (0 minutes) and then at 15, 30, 45 and 60 minutes after the start of suckling, after which the samples were assayed for plasma prolactin and LH concentrations. In all the does there was a significant increase in prolactin concentration, which was highest 15 minutes after the start of suckling, and which declined exponentially thereafter to levels significantly higher than before suckling. The increase in prolactin concentration was similar in does given saline and naloxone, but it was significantly enhanced in does given metaclopramide; with domperidone the increase was intermediate and not significantly different from that following treatment with saline. In does given saline, domperidone, and metaclopramide plasma LH concentrations declined slowly during the hour after suckling but the concentration was increased significantly in does given naloxone. The inverse correlations between prolactin and LH were low weak and were not significant.

Bat rabies, public health and European bat conservation.

Most records of European bat lyssaviruses (EBLVs) are confined to three species - the serotine bat for EBLV1 (900 records) and Daubenton's bat and the pond bat for EBLV2 (25 records). High levels of seroprevalence, which may vary from year to year, are also recorded. All bat vectors of EBLVs are synanthropic, some exclusively so. Despite this, there have been only five cases of human rabies resulting from EBLV infection in the 590 million people of greater Europe during the last 35 years. These have triggered major programmes of surveillance in many European countries. The emphasis on active versus passive surveillance and the intensity with which they have been carried out has varied from country to country. Both involve cooperation between bat researchers, virologists and public health officials and the latter, in particular, engages amateur bat workers and members of the public. Bat NGOs throughout Europe have worked to persuade the public not to handle bats or to do so only with gloved hands and, in the case of bat workers, to receive pre-exposure immunization. They have also countered negative media coverage of bat rabies. Householders with bat roosts in their dwellings have in general been persuaded to retain their bats. Attempts have been made to persuade all European countries to establish comparable EBLV surveillance programmes. In the last 25 years, virologists, public health officials, bat biologists and conservationists, both amateur and professional have worked closely and collaboratively for the protection of the public and the conservation of bats, with little polarization of views.

Seasonal changes in testosterone levels and androgen-dependent organs in male moles (Talpa europaea).

Seasonal changes in testicular and plasma testosterone levels and in androgen-dependent organs were determined in moles breeding at 53 degrees N. Although the testes contain up to 30 times more testosterone during spermatogenesis than during sexual quiescence, appreciable quantities of this hormone are present during late summer and early autumn. Annual spermatogenesis lasts for only 2 months, but in some moles spermatozoa remain in the epididymis for up to 3 months after the testes have begun to involute, and may therefore be available to inseminate females coming into a second oestrus.

Light and electron microscopical observations on the male excurrent duct system of the common shrew (Sorex araneus).

In sexually active common shrews (Sorex araneus) the rete testis is a simple sac-like structure situated superficially between the equator and the anterior pole of the testis, and its epithelium glycogen-rich cells. In the ductuli efferentes, the tight junctions of the epithelial cells are poorly developed. The epididymis can be divided cytologically into initial, middle and terminal segments, and some variations are apparent in the fine structure of the principal cells throughout the duct. In the initial segment and in the most proximal part of the middle segment, mitochondrion-rich cells occur sporadically in the epithelium. The ductus deferens is divided into three anatomical regions, a tortuous pars epididymica, a straight middle portion, and an ampullary region. The distal half of the straight portion is distended, and appears to function, together with the terminal segment of the epididymis, as a sperm reservoir.