The Effect of Detopping the Field Bean Plants (Vicia faba L. minor) and Lack of the Pod Setting on the Symbiosis with Root Nodule Bacteria

The plants of field bean var. Nadwiślaνski were detopped after about 1 month since the beginning of flowering and after that all flower buds were removed from some plants. The effect of a change in the trophic balance on the setting and subsequent growth of root nodules was studied.
The removal of pods had the greater effect on the increase in the weight of the remaining organs than the detopping of plants. Detopping retarded the ageing of the plants and delayed the phase of the greatest reduction of the number of pods. The detopping of plants revealed their considerable compensative possibilities in terms of production of the pod weight and the potential of leaves for biomass production. The data obtained indicate that plant detopping may disturb the root–bacteria exchange of signals, necessary to initiate the nodule development, while the further growth of nodules depends, above all, on the amount of available photosynthates.
A very high correlation between the dry weight of the whole plant to the dry weight of leaves ratio and the weight of nodules was observed. It proves the importance of the trophic balance of the plant for the root nodule weight production.     

The Content of β-endorphin-like Immunoreactivity in Porcine Corpus Luteum and the Potential Roles of Progesterone, Oxytocin and Prolactin in the Regulation of β-endorphin Release from Luteal Cells in vitro

The amount of β‐endorphin‐like immunoreactivity (β‐END‐LI) in porcine corpora lutea from several stages of the oestrous cycle and the effects of progesterone, oxytocin, and prolactin on β‐END‐LI secretion in vitro by luteal cells were studied. Porcine corpora lutea obtained on days 1–5, 6–10, 11–13, 14–18, and 19–21 of the cycle were used to prepare extracts for β‐END‐LI determination. Additionally, corpora lutea from days 11–13 and 14–18 were enzymatically dissociated and isolated luteal cells were used for further study of β‐endorphin secretion in vitro. Cells were cultured in serum‐free defined M 199 medium (10⁶ cells/ml) at 37°C under 5% CO₂ in air, for 12 h. The influences of the following factors on β‐END‐LI secretion by luteal cells were tested: progesterone (10^–9, 10^–7 and 10^–5M ), oxytocin (0.01, 0.1, 1 and 10 ng/ml), and prolactin (0.1, 1, 10 and 100 ng/ml). The β‐END‐LI contents in extracts and media were measured by radioimmunoassay. The tissue concentration of β‐END‐LI was lowest on days 1–5 of the cycle (0.35 ± 0.03 ng/g wet tissue). Subsequently, it constantly increased to the highest value on days 14–18 (16.58 ± 0.52 ng/g wet tissue) and on days 19–21 it declined (11.10 ± 0.52 ng/g wet tissue). Progesterone at a low dose (10^–9 M ) resulted in significant (p < 0.05) increases and decreases in β‐END‐LI secretion by luteal cells from days 11–13 and 14–18, respectively. Higher doses of progesterone (10^–7 and 10^–5 M ) had no effect on β‐END‐LI release, compared with the control group. All dose‐levels of oxytocin used decreased β‐END‐LI secretion by luteal cells on days 11–13 and 14–18 of the cycle. Prolactin at doses of 0.1 and 1 ng/ml on days 11–13, and all doses tested on days 14–18 resulted in decreases in β‐END‐LI release from luteal cells. These results document evident changes in β‐END‐LI content in the pig corpus luteum during its development and indicate the potential roles of progesterone, oxytocin, and prolactin in luteal cell secretion of β‐END‐LI.     

Expression of proopiomelanocortin, proenkephalin and prodynorphin genes in porcine luteal cells

The objective of the study was to examine the expression of the genes coding for proopiomelanocortin (POMC), proenkephalin (PENK) and prodynorphin (PDYN) in porcine luteal cells isolated from corpora lutea (CL) collected on days 3-6, 8-10 and 13-16 of the oestrous cycle. Total RNA was purified from non-incubated cells and from cells incubated for 48 h in the absence or presence of luteinising hormone (LH). The semi-quantitative RT-PCR technique, involving coamplification of the target and control cDNA (beta-actin or 18S rRNA), was used to examine gene expression. It was found that the genes coding for opioid precursors are expressed in both non-incubated and incubated porcine luteal cells representing the early, mid- and late luteal phase. In non-incubated cells, only POMC mRNA content changed during CL development, whereas the expression of PENK and PDYN genes remained relatively constant. Additionally, the treatment of cells with LH markedly affected the expression of POMC and PENK, but no influence on PDYN expression was observed. The present study indicates that porcine luteal cells may produce opioid peptides and that gene expression of their precursors (except for PDYN) may be modulated in these cells by LH. Moreover, the present results support the involvement of opioid peptides in local regulation within the CL of the pig.     

The response of phospholipase C/protein kinase C and adenylyl cyclase/protein kinase A pathways in porcine theca interna cells to opioid agonist FK 33-824

Opioids were found as factors affecting porcine ovarian steroidogenesis. The mechanism of opioid action, however, on porcine theca interna cells is completely unknown. Therefore, the present study was designed to investigate the possible involvement of two intracellular pathways, phospholipase C/protein kinase C and adenylyl cyclase/protein kinase A, in opioid signal transduction in porcine theca cells treated with mu opioid receptor agonist, FK 33-824. Incubation of the cells for 4 h with FK 33-824 at the dose 1 nM resulted in decreases in inositol phosphate accumulation as well as androstenedione (A(4)), testosterone (T), and estradiol (E(2)) secretions. Protein kinase C (PKC) inhibitors, staurosporine (1-100 nM), D-sphingosine (10-500 nM), and PKCi (100-2000 nM), both added alone and together with the opioid agonist, depressed release of the steroid hormones. PKC activator, phorbol ester (PMA, 1-100 nM), used alone was without effect on theca cell steroidogenesis, but added in combination with FK 33-824 abolished inhibitory influence of the opioid on A(4), T, and E(2) output. The steroid hormone secretion by PKC-deficient theca cells was inhibited by the opioid agonist. FK 33-824 also suppressed PKC activity reducing [(3)H]PDBu specific binding to theca cells, whereas ionomycin (a positive control) increased labeled phorbol ester binding to the cells. In the next experiment, cAMP release from theca cells during 2 and 4 h incubations with FK 33-824 (1-100 nM), naloxone (10 microM; opioid receptor antagonist), and LH (100 ng/mL; a positive control) was examined. FK 33-824 at the dose 1 nM inhibited cAMP secretion during 2 h incubation, but had no effect during longer incubation. LH in a manner independent on incubation time multiplied cAMP release. Protein kinase A inhibitor, PKAi (100-2000 nM), alone and in combination with FK 33-824 (1 nM), inhibited A(4), T, and E(2) secretions by theca cells. PKA activator, 8BrcAMP (10-1000 microM), stimulated the steroid hormone release, but this stimulatory effect was diminished in the presence of FK 33-824. The results allow to suggest that opioid peptides affect porcine theca cell steroidogenesis and their acute action on the cells is connected with the inhibition of phospholipase C/protein kinase C and adenylyl cyclase/protein kinase A signal transduction systems.     

The Influence of Opioid Peptides on Steroidogenesis in Porcine Granulosa Cells

The present studies were undertaken to examine the influence of mu (beta-endorphin, DAMGO, FK 33-824), delta (met-enkephalin, leu-enkephalin, DPLPE) and kappa opioid receptor agonists (dynorphin A, dynorphin B, U 50488) used at different doses (1-1000 nM) alone and in combination with LH (100 ng/ml) on steroidogenesis in porcine granulosa cells derived from large follicles. The effects of mu, delta and kappa receptor agonists on both basal and LH-induced progesterone (P4) secretion were negligible. Agonists of mu opioid receptors reduced basal androstenedione (A4), testosterone (T) and oestradiol (E2) release. Co-treatment with LH entirely abolished the inhibitory effect of these agonists on A4 and E2 secretion and resulted in an increase in T release. The addition of delta receptor agonists was followed by a decrease in basal A4, T and E2 secretion. The cells incubated in the presence of LH increased the androgen production and abrogated the inhibitory effect of delta agonists on E2 output. Basal A4, T and E2 release was also suppressed by kappa receptor agonists. The presence of LH in culture media extended the inhibitory effect of these opioids on E2 output and caused either abolition of the inhibitory influence of kappa agonists or even augmentation of both androgen release in response to the opioids. In conclusion, these data support the involvement of three major types of opioid receptors in the regulation of porcine granulosa cell steroidogenesis.     

Feline leukaemia. ABCD guidelines on prevention and management

OverviewFeline leukaemia virus (FeLV) is a retrovirus that may induce depression of the immune system, anaemia and/or lymphoma. Over the past 25 years, the prevalence of FeLV infection has decreased considerably, thanks both to reliable tests for the identification of viraemic carriers and to effective vaccines.InfectionTransmission between cats occurs mainly through friendly contacts, but also through biting. In large groups of non-vaccinated cats, around 30–40% will develop persistent viraemia, 30–40% show transient viraemia and 20–30% seroconvert. Young kittens are especially susceptible to FeLV infection.Disease signsThe most common signs of persistent FeLV viraemia are immune suppression, anaemia and lymphoma. Less common signs are immune-mediated disease, chronic enteritis, reproductive disorders and peripheral neuropathies. Most persistently viraemic cats die within 2–3 years.DiagnosisIn low-prevalence areas there may be a risk of false-positive results; a doubtful positive test result in a healthy cat should therefore be confirmed, preferably by PCR for provirus. Asymptomatic FeLV-positive cats should be retested.Disease managementSupportive therapy and good nursing care are required. Secondary infections should be treated promptly. Cats infected with FeLV should remain indoors. Vaccination against common pathogens should be maintained. Inactivated vaccines are recommended. The virus does not survive for long outside the host.Vaccination recommendationsAll cats with an uncertain FeLV status should be tested prior to vaccination. All healthy cats at potential risk of exposure should be vaccinated against FeLV. Kittens should be vaccinated at 8–9 weeks of age, with a second vaccination at 12 weeks, followed by a booster 1 year later. The ABCD suggests that, in cats older than 3–4 years of age, a booster every 2–3 years suffices, in view of the significantly lower susceptibility of older cats.     

Feline rabies. ABCD guidelines on prevention and management

OverviewRabies virus belongs to the genus Lyssavirus, together with European bat lyssaviruses 1 and 2. In clinical practice, rabies virus is easily inactivated by detergent-based disinfectants.InfectionRabid animals are the only source of infection. Virus is shed in the saliva some days before the onset of clinical signs and transmitted through a bite or a scratch to the skin or mucous membranes. The average incubation period in cats is 2 months, but may vary from 2 weeks to several months, or even years.Disease signsAny unexplained aggressive behaviour or sudden behavioural change in cats must be considered suspicious. Two disease manifestations have been identified in cats: the furious and the dumb form. Death occurs after a clinical course of 1–10 days.DiagnosisA definitive rabies diagnosis is obtained by post-mortem laboratory investigation. However, serological tests are used for post-vaccinal control, especially in the context of international movements.Disease managementPost-exposure vaccination of cats depends on the national public health regulations, and is forbidden in many countries.Vaccination recommendationsA single rabies vaccination induces a long-lasting immunity. Kittens should be vaccinated at 12–16 weeks of age to avoid interference from maternally derived antibodies and revaccinated 1 year later. Although some vaccines protect against virulent rabies virus challenge for 3 years or more, national or local legislation may call for annual boosters.     

Environmental Influences on the Release of Ophiosphaerella agrostis Ascospores Under Controlled and Field Conditions

ABSTRACT Ophiosphaerella agrostis, the causal agent of dead spot of creeping bentgrass (Agrostis stolonifera), can produce prodigious numbers of pseudothecia and ascospores throughout the summer. The environmental conditions and seasonal timings associated with O. agrostis ascospore release are unknown. The objectives of this research were to (i) determine the influence of light and relative humidity on ascospore release in a controlled environment, (ii) document the seasonal and daily discharge patterns of ascospores in the field, and (iii) elucidate environmental conditions that promote ascospore release under field conditions. In a growth chamber, a sharp decrease (100 to approximately 50%; 25 degrees C) in relative humidity resulted in a rapid (1- to 3-h) discharge of ascospores, regardless of whether pseudothecia were incubated in constant light or dark. In the field, daily ascospore release increased between 1900 and 2300 h and again between 0700 and 1000 h local time. The release of ascospores occurred primarily during the early morning hours when relative humidity was decreasing and the canopy began to dry, or during evening hours when relative humidity was low and dew began to form. Few ascospores were released between 1100 and 1800 h when the bentgrass canopy was dry. The release of ascospores also was triggered by precipitation. Of the ascospores collected during precipitation events, 87% occurred within 10 h of the beginning of each event.     

Influence of Thermally Polluted Water on the Growth of Helophytes in the Vicinity of a Colliery Waste Tip

The impact of thermal pollution of leachate from a post-coal mine heap on three macrophyte species: Phragmites australis, Typha latifolia, and Scirpus sylvaticus was examined over the entire vegetation season. Hydrological measurements showed that the temperature of the leachate was ca 50?°C at the site of leachate inflow and decreased to ca 15?°C at the end of discharge canal. The annual temperature and conductivity of leachate from the two control sites, a polluted water stream in the vicinity of the waste tip and an unpolluted stream, differ significantly. However, only the temperature explained the differences in plant traits. In April, and in some cases in May, plants in the leachate were significantly higher than in those on the control sites in terms of biomass and plant height. Thermal pollution caused a phenological shift in all species and also caused Scirpus plants to die out more quickly. Temperature also affected the proportion flowering vs. vegetative individuals, e.g., none of Scirpus plants started to bloom.