The effect of a consortium of Penicillium sp. and Bacillus spp. in suppressing banana fungal diseases caused by Fusarium sp. and Alternaria sp.

AIMS: This study sought to utilize indigenous soil micro-organisms to suppress wilt-causing fungal pathogens of the banana. METHODS AND RESULTS: Fungal pathogens were isolated from wilt-affected rhizospheric soil, and potential antagonistic bacterial strains were isolated from healthy rhizospheric soil in the same area from which fungal pathogens were isolated. The antifungal activity of isolated micro-organisms against fungal pathogens was studied both in vitro and in vivo against fungal pathogens. It was found that Fusarium oxysporum and Alternaria sp. were pathogenic, while Penicillium sp., Bacillus velezensis and Bacillus subtilis were antagonistic. Moreover, it was seen that B. velezensis, B. subtilis and Penicillium sp. inhibited the growth of the two fungal pathogens in both in vitro and in vivo experiments. Further investigation indicated that B. velezensis, B. subtilis and Penicillium sp. were able to produce enzymatic antifungal compounds (chitinase and ß-1,3-glucanase). The spray application around rhizome revealed that a combination of Bacillus spp. and Penicillium sp. in greenhouse conditions gave the highest reduction in disease severity by up to 60% to both fungal pathogens among the treatments. CONCLUSIONS: Banana disease is seen to be induced not only by F. oxysporum but also by Alternaria sp. The isolated indigenous micro-organisms can effectively control both the pathogens. The combination of isolated antagonistic micro-organisms has thus demonstrated substantial potential for suppressing banana disease. SIGNIFICANCE AND IMPACT OF THE STUDY: An antagonistic consortium isolated in this study has demonstrated remarkable potential for controlling fungal diseases caused by Fusarium sp. and Alternaria sp. Therefore, the use of indigenous microflora to improve disease suppression of banana plants against soil-borne pathogens is a preferable approach.

On the distribution and natural history of a rarely encountered species: Goliathus (Fornasinius) klingbeili Zöller, Fiebig, & Schulze, 1995 (Coleoptera: Scarabaeidae: Cetoniinae).

Mudge et al. (2012) surveyed the Cetoniinae (Coleoptera: Scarabaeidae) of Ghana and recorded Goliathus (Fornasinius) higginsi Westwood, 1874, but no other species from this subgenus. We add the rare species Goliathus (Fornasinius) klingbeili Zöller, Fiebig, & Schulze, 1995 as a new country record for Ghana from the dense forests of the Volta and Eastern Regions. The species was previously reported as very rare in Togo (Zöller et al. 1995) in the northern dry part of the Bafilo Region. We record that the larvae develop in hyrax (Dendrohyrax dorsalis (Fraser, 1855) (Mammalia: Procaviidae) middens and can be locally common in this microhabitat. The purpose of this note is to record the new distributional and natural history data, discuss the taxonomy and bionomy of the subgenus Goliathus (Fornasinius) Bertoloni, 1852, and provide a key to species.

Increased genetic diversity of Viola tricolor L. (Violaceae) in metal-polluted environments.

Changes in DNA sequences affecting cryptic intraspecific variability are very important mechanisms of plant microevolutionary processes, initiating species diversification. In polluted environments, intra- and interpopulation changes at the molecular level proceed rapidly and lead to the formation of new ecotypes in a relatively short time. We used ISSR PCR fingerprinting data to analyze the genetic diversity and genetic structure of seven populations of Viola tricolor: four growing on soil contaminated with heavy metals (Zn, Pb, Cd; waste heaps) and three from control soil. The populations from the polluted sites showed higher genetic polymorphism (%(poly)=84%) and gene diversity (H(T)=0.1709) than the control populations (%(poly)=75% and H(T)=0.1448). The number of private markers we detected within metallicolous (MET) populations was more than double that found within non-metallicolous (NON) populations (15 vs. 7). The STRUCTURE and UPGMA analyses showed clear genetic differences between the NON and MET populations. Based on broad analyses of the genetic parameters, we conclude that the effect of these polluted environments on the genetic diversity of the MET populations, separating them from the NON populations, is evidence of microevolutionary processes at species level, leading to species divergence and the emergence of local ecotypes better adapted to their different environments.

Zinc protects Ceratophyllum demersum L. (free-floating hydrophyte) against reactive oxygen species induced by cadmium.

Evidence for Zn protection against Cd-induced reactive oxygen species in the free-floating hydrophyte Ceratophyllum demersum L. is presented in this paper. Metal treatments of 10 micromol/L Cd, 10 Cd micromol/L supplemented with Zn (10, 50, 100 and 200 micromol/L) and Zn-alone treatments of the same concentrations were used. Using 5,5 dimethyl pyrroline-N-oxide as the spin-probe, electron spin resonance spectra indicated a drastic increase in hydroxyl radicals (OH()) in Cd-10 micromol/L treatments, which was closely correlating with the enhanced formation of hydrogen peroxide (H(2)O(2)) and generation of superoxide radical (O(2)(-)) triggered by the oxidation of NADPH. The supplementation of adding Zn (10-200 micromol/L) to the Cd-10 micromol/L treatments significantly decreased the production of free radicals especially by eliminating the precursors of OH() through inhibition of NADPH oxidation. Cd-enhanced ROS production which substantially increased the oxidative products of proteins measured as carbonyls was effectively inhibited by Zn supplementation.

Formaldehyde and methanol biodegradation with the methylotrophic yeast Hansenula polymorpha. An application to real wastewater treatment.

The application of methylotrophic yeast Hansenula polymorpha to the treatment of methanol and formaldehydecontaining wastewater was experimentally verified. A variety of real wastewater samples originating from chemical industry effluent were examined. The yeast cell culture could grow in the wastewater environment, revealing low trophic requirements and a very high adaptation potential to poor cultivation conditions. The proliferation of cells was accompanied by a concomitant xenobiotic biodegradation. Grown, preadapted cellular suspension at a density of about 1 x 10(7) cells/ml proved to be able to utilize formaldehyde present in wastewater at concentrations up to 1750 mg/l, levels toxic to most microorganisms. The biological waste treatment method presented shows the enhanced potential by means of specific enzymatic activities of monocarbonic compound oxidations through methylotrophic pathway reactions. The need to obtain mutants highly resistant to formaldehyde has also been rationalized.

Arabidopsis FUSCA5 encodes a novel phosphoprotein that is a component of the COP9 complex.

The COP9 complex is a regulator essential for repression of light-mediated development in Arabidopsis. Using partial amino acid sequence data generated from purified COP9 complexes, we cloned the Arabidopsis cDNA encoding the 27-kD subunit of the COP9 complex and showed that it is encoded by the previously identified FUSCA5 (FUS5) locus. fus5 mutants exhibit constitutive photomorphogenic phenotypes similar to those of cop9 and fus6. Point mutations in FUS5 that led to a loss of FUS5 protein were detected in four fus5 allelic strains. FUS5 contains the PCI/PINT and mitogen-activated protein kinase kinase activation loop motifs and is highly conserved with the mammalian COP9 complex subunit 7 and the Aspergillus nidulans AcoB proteins. FUS5 is present in both complex and monomeric forms. In the COP9 complex, FUS5 may interact directly with FUS6 and COP9. Mutations in FUS6 and COP9 result in a shift in the electrophoretic mobility of FUS5. This shift can be mimicked by in vitro phosphorylation of FUS5 by plant extracts. These findings further support the hypothesis that the COP9 complex is a central and common regulator that may interact with multiple signaling pathways.

Thymic involution in aging. Prospects for correction.

The thymus produces several putative thymic hormones: thymosin alpha 1, thymulin, and thymopoietin, which have been reported to circulate and to act on both prothymocytes and mature T cells in the periphery, thus maintaining their commitment to the T cell system. These endocrine influences decline with age and are associated with "thymic menopause" and cellular immune senescence, which contribute to the development of diseases in the aged. Thymus endocrinology is characterized by the action of many hormones and hormone-like substances on the cellular components of the thymus, including thymocytes, thymic epithelial cells, and thymic stromal cells. The intrathymic environment is characterized by a complex network of paracrine, autocrine, and endocrine signals involving both interleukins and thymic peptides, which can be envisioned to operate in a synergistic network to carry the evolving T cell through its stepwise development to a mature T cell. Extrathymic influences regulating the secretory function of thymic epithelial cells and the stepwise evolution of T cells can be ascribed to circulating interleukins, mainly IL1 and IL2, derived from activation and secretion of leukocytes in the periphery. These interleukins act in a synergistic fashion at all levels of T cell development by the induction of high-affinity IL2 receptors and the resultant IL2-dependent proliferative responses. To determine whether exogenous administration of interleukins would induce T lymphocyte development in aged mice, we chemically thymectomized aged mice with a steroid hormone and treated them with mixed interleukins or thymic hormones such as thymosin. We found that mixed interleukins, but not thymosin, restored thymic weight and cellularity and enhanced thymocyte responses to interleukins and mitogen. Thymosin potentiated the effect.

Mixed interleukins and thymosin fraction V synergistically induce T lymphocyte development in hydrocortisone-treated aged mice.

Analysis of the role of interleukins in T cell ontogeny in vitro indicates that the regulation of T cell development involves interleukins (ILs) as well as thymic hormones (THs). In order to assess their respective roles in T lymphocyte development in vivo, chemically thymectomized mice were treated with ILs and THs. After 2 days of hydrocortisone treatment, aged mice showed acute thymic involution (weight was less than 30% of control) and reduced spleen size (less than 80% of control) with progressive recovery to 8 days. After 2 days of hydrocortisone treatment, adult mice were injected for 5 days with mixed buffy coat interleukins (BC-IL; 50 units IL2 equivalence), purified IL2 (50 units), rIL1 beta (4 ng), and thymosin fraction V (TF5; 100 micrograms). The animals were sacrificed and spleens and thymuses were analyzed for weight, cellularity, T cell number, subsets, and function as determined by proliferative responses to concanavalin A and ILs. BC-IL treatment increased the recovery of spleen and thymus weights and cellularity with corresponding augmentation of number and function of T lymphocytes; neither IL1 or IL2 or their combination had this effect. TF5 had no effect alone but strongly potentiated the effect of BC-IL on T lymphocyte function. These data indicate that BC-IL in combination with thymic peptides potently promotes T lymphocyte development. The combination may be therapeutically relevant for immunorestoration.

Chronic beta-adrenergic antagonist treatment affects human T lymphocyte responsiveness "in vitro".

The aim of the present study was the evaluation of the effect of long-term propranolol treatment on some immune parameters in humans. The subjects were 12 duodenal ulcer patients, aged 19-23 years. They were given propranolol in a dose of 40 mg three times a day orally. Blood samples were drawn before and after one-month propranolol treatment. The drug did not significantly affect mitogen (PHA, Con A, PWM)- induced lymphoproliferative response, although a tendency of a slight inhibition of proliferation was observed. In all patients treated with propranolol absolute peripheral blood lymphocyte counts increased. Propranolol did not significantly influence one-way MLR, while it enhanced proliferation in the AMLR test in all patients. The drug increased spontaneous and PHA-induced IL-2R expression as well as IL-2 generation. The study showed that chronic beta-adrenoceptor blockade with propranolol may modify some immune functions in humans.

Some mechanisms of immunosuppressive action of epinephrine in humans.

The studies were performed on 12 men (9 duodenal ulcer patients and 3 healthy controls) aged 19-23 years. They were given epinephrine in a dose of 0.014 mg/kg subcutaneously. Blood samples were drawn before and 30 and 150 min. after drug injection. The results obtained in the duodenal ulcer patients and controls revealed the same direction of changes. Epinephrine induced a significant increase in the absolute numbers of all investigated peripheral blood mononuclear cell subpopulations (total lymphocytes, T, Th, Ts, NK cells and monocytes) observed at 30 min. following injection, while they nearly normalized at 150 min. A Th/Ts cell number ratio slightly, but significantly, decreased. Epinephrine did not affect spontaneous IL-2 receptor expression and it significantly inhibited PHA-induced IL-2 receptor expression and IL-2 generation at 30 min. following injection. NK cell activity also decreased 30 min. after epinephrine administration and it was still lowered at 150 min. The drug very distinctly inhibited proliferative response in the autologous mixed lymphocyte reaction test.

Effect of epinephrine, propranolol, and verapamil on the chemiluminescence of neutrophils in duodenal ulcer patients.

To answer the question whether beta-adrenergic receptor agonists or antagonists and calcium channel blocking agents affect activation of neutrophils in vivo, the chemiluminescence (CL) test was employed. The intensity of emitted photons was amplified by luminol. The effect of investigated agents was measured after stimulation of isolated peripheral blood polymorphonuclear leukocytes (PMNLs) with opsonized zymosan particles. The investigations were performed on 9 duodenal ulcer patients, aged 19-23 years, after informed consent. Single subcutaneous dose of epinephrine (0.014 mg/kg) induced a marked increase in PMNL number and a moderate, but significant, decrease in CL 30 min after injection. Verapamil (0.15 mg/kg intravenously) diminished CL, propranolol (0.1 mg/kg intravenously) enhanced CL, but 150 min after injections CL was approaching the initial values. The obtained results suggest that the investigated compounds may modify the PMNL function in vivo.

Short-term somatostatin infusion affects T lymphocyte responsiveness in humans.

Peripheral blood lymphocytes (PBL) function was analysed in 16 young men with duodenal ulcers after one-hour intravenous infusion of somatostatin (SMS) at a dose of 250 micrograms/h. Proliferative responses of PBL from SMS-treated patients were significantly diminished compared with pre-treatment values, after stimulation with PHA, PWM or Con A. Spontaneous IL-2R expression was moderately increased after SMS infusion but PHA-induced IL-2R expression was not affected by this drug. Alloantigen and autoantigen stimulation of PBL showed no significant changes in the proliferative response after SMS infusion. NK cell activity was similarly unaffected. These observations establish a link between SMS exposure and possible development of immune dysfunction.

Immunomodulatory effect of naloxone in duodenal ulcer patients.

The effect of a single dose of the opiate receptor antagonist naloxone (NAL) on some immune parameters in chronic duodenal ulcer patients and healthy controls was investigated. Twelve hospitalized men, aged 19-23 years (nine duodenal ulcer patients and three controls) were given NAL intravenously at a dose of 0.03 mg/kg. Blood samples were drawn before and 30 and 150 min after NAL injection. Results obtained in duodenal ulcer patients and healthy subjects revealed the same direction of changes despite a rather wide scatter. Baseline NK cell activity was lower in duodenal ulcer patients than in healthy persons. In approximately one fourth of patients the changes in individual parameters observed following NAL injection were slight. Numbers of total lymphocytes, Th, Ts, NK cells, monocytes and Th/Ts ratio did not significantly change after NAL administration. T-lymphocyte counts moderately decreased at 30 min after NAL injection. NAL did not affect spontaneous IL-2 receptor expression and it moderately increased PHA-induced IL-2 receptor expression in most of the investigated persons. IL-2 generation and NK cell activity slightly, but significantly, increased at 30 min following NAL injection. NAL markedly inhibited lymphocyte proliferation in an AMLR test 30 min after its administration. Most of the investigated parameters returned to their initial levels after 150 min following NAL administration. The studies showed that not only endorphins and enkephalins may have an immunomodulatory action, but NAL, their antagonist, may also affect some functions of the immune system in humans, although its action is transient.

Modulation of autologous mixed lymphocyte reaction by pharmacological agents affecting adrenergic receptor.

The effect of epinephrine and propranolol administration in vivo on autologous mixed lymphocyte reaction (AMLR) was investigated. AMLR inhibition by epinephrine and potentiation of AMLR after propranolol injection was found. Presumable mechanisms of these changes in the light of beta-receptor agonist and antagonist are discussed.

Effect of a single intravenous dose of verapamil on some immune parameters in duodenal ulcer patients.

The effect of a single intravenous dose of calcium channel blocking agent verapamil (VER) on immune parameters in humans remains uncertain. In this study the effects of VER on lymphocyte subpopulations, interleukin 1 (IL-1) and 2 (IL-2) production in vitro, autologous mixed lymphocyte reaction (AMLR), IL-2 receptor expression (Tac positive cells) and natural killer (NK) cell activity were assessed. The investigations were undertaken on 12 hospitalized men, aged 19-23 years, with small abdominal complaints. None of them had active duodenal ulcer disease, while in the remaining no signs of organic disease were found. VER was given intravenously in a dose of 0.15 mg/kg and examinations were performed on peripheral blood mononuclear cells (PBL) drawn before and 30 and 150 min following VER injection. The single VER dose induced a significant, but transient increase in T, T helper, T suppressor lymphocytes and monocytes, and decrease in IL-1 and IL-2 generation as well as diminished Tac antigen expression. These data provide evidence that calcium channel blockers may transiently disturb immunoregulation.

Propranolol enhances in vitro interleukin 2 receptor expression on human lymphocytes.

Interleukin 2 receptor (IL-2R) expression was assessed, using anti-Tac monoclonal antibody, after incubation of human lymphocytes with the beta-adrenoceptor antagonist propranolol (PRO). One-hour incubation of mononuclear cells (MNC) with PRO resulted in an increase of Tac positive cells. The greatest rise was observed with a PRO concentration of 10(-6) M (29% ring-stained cells compared with 0% in the control group). Cells cultured for 3 days with PRO showed a lower proportion of Tac positive cells (11%). Fifty-one per cent of MNC cultured with PHA for 3 days expressed IL-2R. Addition of PRO to PHA cultures increased IL-2R expression even more (65%). Enhancement of IL-2R expression after only 1 h incubation with PRO may be due to exposure of formerly synthesized receptor, which can be uncovered by some agents such as PRO.

The stimulatory effect of a single intravenous dose of propranolol of some immune parameters in humans.

Propranolol (PRO) is a beta-adrenergic receptor antagonist. The studies were aimed to evaluate the effect of a single intravenous PRO injection at a dose of 0.1 mg/kg on some immune parameters, and were performed on 12 men, including 9 duodenal ulcer patients and 3 healthy subjects, aged 19-23 years (mean 20.5 +/- 1.2). Blood samples were drawn before, 30 and 150 min following the injection. Results obtained in duodenal ulcer patients and healthy subjects revealed the same type of changes. A single PRO injection did not significantly affect total peripheral blood lymphocyte, Th cell and monocyte counts; however, it slightly but significantly lowered numbers of T and Ts cells, which was accompanied by a rise in Th/Ts cell number ratio and NK cell numbers. PRO significantly augmented spontaneous as well as PHA-stimulated IL-2 receptor expression, IL-2 generation, NK cell activity and AMLR observed at 30 and 150 min after the drug injection. Possible mechanisms of these changes are discussed.