Virulence and pCM1 plasmid carriage are related to BOX-PCR fingerprint type in strains of Clavibacter michiganensis subsp. michiganensis that cause bacterial wilt and canker of tomato in Argentina

Clavibacter michiganensis subsp. michiganensis (Cmm) causes bacterial wilt and canker in tomato, producing important economic losses worldwide. Its virulence has been related to several putative virulence factors present on a chromosomal pathogenicity island and on plasmids pCM1 and pCM2, in strain NCPPB382. We genotypically characterized a collection of Cmm isolates from the main greenhouse tomato-producing areas of Argentina by BOX-PCR fingerprinting and screened for the presence of genes and plasmids involved in pathogenicity by PCR. In addition, we evaluated in vitro cellulolytic activity and virulence in planta of selected strains. BOX-PCR fingerprinting clustered strains into four groups. Group II was dominant and included the most virulent strains, while Group III was the smallest and had the least virulent strains. All local strains exhibited similar cellulolytic activity. Most of the examined strains carry two plasmids of similar size to those of NCPPB382, although there were strains with one or three plasmids. By PCR amplification of repA, pCM1 was detected only in strains belonging to Group III, which includes local strains closely related to reference strain NCPPB382. All analysed pathogenicity genes were widespread among strains, and so in strains belonging to Groups I and II, celA found on pCM1 in NCPPB382 could be found in the chromosome or in plasmids other than pCM1. This study contributes to a better understanding of the diversity of Cmm genetic profiles and virulence of strains present in Argentina. Such information could be useful for the selection of strains for screening of host resistance and development of resistant tomato varieties.     

Detection of BHV-1 in a Naturally Infected Bovine Fetus by a Nested PCR assay

Bovine herpesvirus 1 (BHV-1) is frequently associated with abortion in naturally and experimentally infected cattle. Most of the virus isolation and immunofluorescent antibody protocols described in the literature for detecting BHV-1 in bovine foetuses are rather laborious, costly and time-consuming. The detection is described of BHV-1 in the tissues of a naturally aborted bovine foetus by a nested PCR assay with no further hybridization procedures.Optimal results were achieved by filtering the foetal tissues on a chromatography column before DNA extraction, by using two pairs of primers in a nested PCR and by evaluating the amplification products on silver-stained polyacrylamide gels.This nested PCR was faster and easier to perform than the virus isolation test. To our knowledge, this is the first time that BHV-1 has been detected in the tissues of a naturally infected bovine foetus by means of a nested PCR. The test seems to be a practical alternative for rapid detection of BHV-1 in bovine foetus.     

Effects of sex and season in haematological parameters and cellular composition of spleen and head kidney of pejerrey (Odontesthes bonariensis)

Phylogenetic diversity in fish determines high interspecific variability in morphology as well as in physiological parameters. Moreover, several haematological variables and the organ composition of haemolymphopoietic sites may vary according to sex or season. The aim of this study was to establish the haematological parameters and the cellular composition of haemolymphopoietic organs in Odontesthes bonariensis, a commercially valuable fish species in Argentina, and also to determine gender or seasonal variations. Haematocrit exhibited the highest value in summer, while haemoglobin concentration was greater in summer and autumn. Erythrocyte count was higher in spring than autumn and winter, but did not differ with summer. The increase in these variables in seasons with higher water temperatures might be a compensatory mechanism to compensate the lower level of oxygen in the environment. Leucocyte formula and blast haemolymphopoietic cells in spleen and head kidney also showed annual variations since cells related to specific immune response, i.e., lymphocytes and thrombocytes, decrease in winter, whereas cells of the non-specific immune pathways, such as granulocyte, rise. The elevation of a particular type of circulating leucocyte was preceded by an increase in values of its precursor in blood in the previous season. Both, spleen and head kidney were active in haemolymphopoiesis, although with some differences in their activity during different seasons. Males showed higher values of circulating lymphoblasts and granulocytes than females, whereas females exhibited higher values of thrombocytes. This study corroborates the high interspecific variations in haematological parameters in fish that underlines the needing of basic studies in order to assess fish health status in new promising species for aquaculture.     

CHROMIUM TOXICITY IN HYBRID EUCALYPTUS (EUCALYPTUS UROPHYLLA S. T. BLAKE X GRANDIS W. HILL EX. MAIDEN) CUTTINGS

A greenhouse experiment was carried out to evaluate chromium (Cr) toxicity in urograndis (Eucalyptus urophylla S. T. Blake x grandis W. Hill ex. Maiden). Chromium nitrate was amended to Clark's nutrient solution (0.00, 0.04, 0.08, 0.16, 0.32 and 0.64 mmol L^?1 Cr), which was used to breed one urograndis cutting per pot (four replications). Regression analysis revealed that Cr rates of 0.08 mmol L^?1 or higher significantly decreased (P < 0.01) dry matter yield of shoot and root. Chromium accumulation order in plant was: root > stem > leaves. Critical Cr toxicity level in leaves was 0.74 mg kg^?1. Uptake and translocation of nutrients were affected differently by Cr. Mostly have their uptake reduced due to root damage. Magnesium, iron and manganese translocation increases, supposedly to maintain normal photosynthetic activity. Wilting is the main visual symptom of Cr toxicity. Urograndis may be suitable for contaminated sites phytostabilization and receive industrial waste fertilization.     

Potential role of the CD38/cADPR signaling pathway as an underlying mechanism of the effects of medetomidine on insulin and glucose homeostasis

ObjectiveTo investigate the CD38/cADPR signaling pathway as possible underlying mechanism of the effects of medetomidine on insulin and glucose homeostasis.AnimalsThirty–two C57BL/6 mice of both sexes.MethodsWild–type (WT) and CD38–knockout (CD38^?/?) mice received medetomidine (50 μg kg^?1) or a similar volume of 0.9% NaCl (control) by intraperitoneal (IP) injection (each group n = 8). The mice were euthanized 45 minutes later with sodium pentobarbital IP and blood was sampled via cardiac puncture. Insulin and glucose concentrations were measured by radioimmunoassay and by the oxygen rate method, respectively. Data were analyzed with anova and Bonferroni post hoc (5% significance) and are shown as mean ± SD.ResultsPlasma insulin and glucose concentrations were similar between WT and CD38^?/? mice under control conditions. As compared to controls, medetomidine administration produced a statistically significant decrease in plasma insulin concentrations in the WT mice whereas the decrease in the CD38^?/? mice was not statistically significant. Correspondingly, medetomidine caused a significantly greater increase in plasma glucose concentrations in the WT than in the CD38^?/? mice.ConclusionThe CD38/cADPR signaling pathway may be one underlying mechanism of the glucose and insulin effects of the alpha–2 adrenergic receptor agonist medetomidine and likely other drugs of its’ class.     

Descriptive Analyses of Deficiency Symptoms in Calla Lily Plants

The objective of this work was to describe the nutrition deficiencies symptoms in calla lily (Zantedeschia aethiopica). Seedlings were maintained in complete nutrition solution and with omission of nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), sulfur (S), iron (Fe), boron (B), and manganese (Mn), totalizing 10 treatments. The first deficiencies symptoms appeared in plants cultivated in N omission, followed by treatments with omission of S, B, K, Fe, P, and Ca. Individual omissions of N and B prevented blooming, while S and K deficiencies resulted in production of small and greenish inflorescences with deformities. Reddish spots in leaves were observed in plants growing in solution with omission of P. Root development was affected either by omission of B or S. Death of the apical bud and formation of a “black center” in calla lily rhizome was caused by B deficiency. Nitrogen, B, and S are the most required nutrients for calla lily plants. The mineral composition of calla lily leaves is affected by nutritional deficiencies     

The metamorphosis of supernova SN 2008D/XRF 080109: a link between supernovae and GRBs/hypernovae

The only supernovae (SNe) to show gamma-ray bursts (GRBs) or early x-ray emission thus far are overenergetic, broad-lined type Ic SNe (hypernovae, HNe). Recently, SN 2008D has shown several unusual features: (i) weak x-ray flash (XRF), (ii) an early, narrow optical peak, (iii) disappearance of the broad lines typical of SN Ic HNe, and (iv) development of helium lines as in SNe Ib. Detailed analysis shows that SN 2008D was not a normal supernova: Its explosion energy (E approximately 6x10(51) erg) and ejected mass [ approximately 7 times the mass of the Sun (M(middle dot in circle))] are intermediate between normal SNe Ibc and HNe. We conclude that SN 2008D was originally a approximately 30 M(middle dot in circle) star. When it collapsed, a black hole formed and a weak, mildly relativistic jet was produced, which caused the XRF. SN 2008D is probably among the weakest explosions that produce relativistic jets. Inner engine activity appears to be present whenever massive stars collapse to black holes.     

Compounds from Ageratum conyzoides: isolation, structural elucidation and insecticidal activity

This work aimed at identifying plant compounds with insecticidal activity against Diaphania hyalinata (L.) (Lepidoptera: Pyralidae), Musca domestica (L.) (Diptera: Muscidae), Periplaneta americana (L.) (Blattodea: Blattidae) and Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae). The plant species used were: basil (Ocimum selloi Benth.), rue (Ruta graveolens L.), lion's ear (Leonotis nepetaefolia L.), Jimson weed (Datura stramonium L.), 'baleeira' herb (Cordia verbenaceae L.), mint (Mentha piperita L.), wild balsam apple (Mormodica charantia L.) and billy goat weed (Ageratum conyzoides L.). Firstly, the insecticidal activities of hexane and ethanol plant extracts were evaluated against adults of R. dominica. Among them, only the hexane extract of A. conyzoides showed insecticidal activity. The hexane extract of this plant species was therefore fractionated by silica gel column chromatography to isolate and purify its bioactive chemical constituents. Three compounds were identified using IR spectra, (1)H NMR, (13)C NMR, HMBC and NOE after gel chromatography: 5,6,7,8,3', 4', 5'-heptamethoxyflavone, 5,6,7,8,3'-pentamethoxy-4', 5'-methylenedioxyflavone and coumarin. The complete assignment of (13)C NMR to 5,6,7,8,3'-pentamethoxy-4', 5'-methylenedioxyflavone was successfully made for the first time. 5,6,7,8,3'-Pentamethoxy-4', 5'-methylenedioxyflavone did not show any insecticidal activity against the four insect species tested. 5,6,7,8,3', 4', 5'-Heptamethoxyflavone showed low activity against D. hyalinata and R. dominica and was not toxic to M. domestica or P. americana. In contrast, coumarin showed insecticidal activity against all four insect pest species tested, with the following order of susceptibility: R. dominica < P. americana < D. hyalinata < M. domestica after 24 h exposure.