Leaf area development of ABA-deficient and wild-type peas at two levels of nitrogen supply

The ABA-deficient wilty pea (Pisum sativum L.) and its wild-type (WT) were grown at two levels of nitrogen supply (0.5 and 5.0 mM) for 5-6 weeks from sowing, to determine whether leaf ABA status altered the leaf growth response to N deprivation. Plants were grown at high relative humidity to prevent wilting of the wilty peas. Irrespective of N supply, expanding wilty leaflets had ca 50% less ABA than WT leaflets but similar ethylene evolution rates. Fully expanded wilty leaflets had lower relative water contents (RWC) and were 10-60% smaller in area (according to the node of measurement) than WT leaflets. However, there were no genotypic differences in plant relative leaf expansion rate (RLER). Growth of both genotypes at 0.5 mM N increased the RWC of fully expanded leaflets, but did not alter ethylene evolution or ABA concentration of expanding leaflets. Plants grown at 0.5 mM N showed a 20-30% reduction in RLER, which was similar in magnitude in both wilty and WT peas. Thus, leaf ABA status did not alter the leaf growth response to N deprivation.

Polymerase chain reaction tests for the identification of Ross River, Kunjin and Murray Valley encephalitis virus infections in horses

Objective To develop and validate specific, sensitive and rapid diagnostic tests using RT-PCR for the detection of Ross River virus (RRV), Kunjin virus (KV) and Murray Valley encephalitis virus (MVEV) infections in horses. Methods Primer sets based on nucleotide sequence encoding the envelope glycoprotein E2 of RRV and on the nonstructural protein 5 (NS5) of KV and MVEV were designed and used in single round PCRs to test for the respective viruses in infected cell cultures and, in the case of RRV, in samples of horse blood and synovial fluid. Results The primer pairs designed for each of the three viruses amplified a product of expected size from prototype viruses that were grown in cell culture. The identity of each of the products was confirmed by nucleotide sequencing indicating that in the context used the RT-PCRs were specific. RRV was detected in serums from 8 horses for which there were clinical signs consistent with RRV infection such that an acute-phase serum sample was taken and submitted for RRV serology testing. The RRV RT-PCR was analytically sensitive in that it was estimated to detect as little as 50 TCID50 of RRV per mL of serum and was specific in that the primer pairs did not amplify other products from the 8 serum samples. The RRV primers also detected virus in three independent mosquito pools known to contain RRV by virus isolation in cell culture. Samples from horses suspected to be infected with KV and MVEV were not available. Conclusion Despite much anecdotal and serological evidence for infection of horses with RRV actual infection and associated clinical disease are infrequently confirmed. The availability of a specific and analytically sensitive RT-PCR for the detection of RRV provides additional opportunities to confirm the presence of this virus in clinical samples. The RTPCR primers for the diagnosis of KV and MVEV infections were shown to be specific for cell culture grown viruses but the further validation of these tests requires the availability of appropriate clinical samples from infected horses.

The Australian paralysis may be the missing link tick in the transmission of Hendra virus from bats to horses to humans

Hendra virus is a new virus of the family Paramyxoviridae. This virus was first detected in Queensland, Australia, in 1994; although, it seems that the virus has infected fruit-eating bats (flying-foxes) for a very long time. At least 2 humans and 15 horses have been killed by this virus since it first emerged as a virus that may infect mammals other than flying-foxes. Hendra virus is thought to have moved from flying-foxes to horses, and then from horses to people. There is a reasonably strong hypothesis for horse-to-human transmission: transmission of virus via nasal discharge, saliva and/or urine. In contrast, there is no strong hypothesis for flying-fox-to-human transmission. I present evidence that the Australian paralysis tick, Ixodes holocyclus, which has apparently only recently become a parasite of flying-foxes, may transmit Hendra virus and perhaps related viruses from flying-foxes to horses and other mammals. (C) 2003 Elsevier Science Ltd. All rights reserved.

Corporal and testicular biometry in wild boar from birth to 12 months of age

The aim of this tudy was to evaluate corporal and testicular development in wild boars (Sus scrofa scrofa) from birth to 12 months of age, evaluating body weight, biometric testicular parameters, and gonadosomatic index. Thirty-nine male wild boars from a commercial farm licensed by IBAMA were used in the study. The animals were weighed and assigned to 13 experimental groups. The testes were recovered through unilateral orchiectomy, weighed on an analytical balance and measured for length, width and thickness. Body weight and testicular measures increased with the age, up to 12 months, and were more accelerated in the first and ninth months. Initially the testicular growth pattern, between zero and nine months, followed the body growth, and the gonadosomatic index varied from 0.07 to 0.09%. Between 9 and 11 months, the testicular growth was superior to the body growth, and the gonadosomatic index varied from 0.09 to 0.16%. Finally, after 11 months of age, testicular and body growth had a similar behavior. In conclusion, body weight, testicular biometry, and gonadosomatic index development accelerated in the ninth month.

Aloe vera-based formula as emollient on horses' hooves

The present study aimed at developing an Aloe vera-based formula for topical use on horse hoof and evaluating whether the treatment affects hooves growth and balance. Six healthy male horses between the ages of 3 and 17 years (12±5.25) were used, all semi-confined animals for breeding purposes. Before beginning A. vera treatment, animals underwent two trimming procedures with a 45 days-interval. After the second trimming, one of the forelimbs and one of the hindlimbs of 4 horses was weekly treated by topical application of the glycolic extract of A. vera at 20%. The contralateral limb, randomly chosen, received the extract at 50%. The hooves of the other animals were treated with propylene glycol. Treatment was done for 225 days and, during this time, animals underwent periodic trimming. Variables related to growth and balance of the hooves were measured before and after trimming. Data were analyzed using chi-square test and regression analysis at 5% significance. Growth rate of the hooves was not related to treatment. On the other hand, the 50% extract was related to the majority of the hooves in balance (p<0.05). Results suggest that a weekly topical treatment with A. vera glycolic extract does not improve the growth rate of the hooves; however, when applied at a high concentration, it improves their balance.

Trace elements in wild and orchard honeys

The present study aims the identification and quantification of trace elements in two types of honey samples: Orchard honey and Wild honey from mainland Portugal. Chemical elements content was assessed by Instrumental Neutron Activation Analysis (INAA). Concentrations were determinated for Ag, As, Br, Ca, Cl, Cs, Cu, Fe, K, La, Mg, Mn, Na, Rb, Sb, Sc, U, V and Zn. The nutritional values of both honey types were evaluated since this product contains some elements that are essential dietary nutrients for humans. Physical properties of the honey samples, such as electrical conductivy and pH, were assessed as well.

Chromatographic behaviour of monoclonal antibodies against wild-type amidase from Pseudomonasaeruginosa on immobilized metal chelates

The aim of this work was to devise a one-step purification procedure for monoclonal antibodies (MAbs) of IgG class by immobilized metal affinity chromatography (IMAC). Therefore, several stationary phases were prepared containing immobilized metal chelates in order to study the chromatographic behaviour of MAbs against wild-type amidase from Pseudomonas aeruginosa. Such MAbs adsorbed to Cu(II), Ni(II), Zn(II) and Co(II)-IDA agarose columns. The increase in ligand concentration and the use of longer spacer arms and higher pH values resulted in higher adsorption of MAbs into immobilized metal chelates. The dynamic binding capacity and the maximum binding capacity were 1.33 +/- 0.015 and 3.214 +/- 0.021 mg IgG/mL of sedimented commercial matrix, respectively. A K(D) of 4.53 x 10(-7) M was obtained from batch isotherm measurements. The combination of tailor-made stationary phases of IMAC and the correct selection of adsorption conditions permitted a one-step purification procedure to be devised for MAbs of IgG class. Culture supernatants containing MAbs were purified by IMAC on commercial-Zn(II) and EPI-30-IDA-Zn(II) Sepharose 6B columns and by affinity chromatography on Protein A-Sepharose CL-4B. This MAb preparation revealed on SDS-PAGE two protein bands with M(r) of 50 and 22 kDa corresponding to the heavy and light chains, respectively. Copyright (C) 2011 John Wiley & Sons, Ltd.

Structural characterization and immunogenicity in wild-type and immune tolerant mice of degraded recombinant human interferon Alpha2b

Purpose: This study was conducted to study the influence of protein structure on the immunogenicity in wild-type and immune tolerant mice of well-characterized degradation products of recombinant human interferon alpha2b (rhIFNα2b). Methods: RhIFNα2b was degraded by metal-catalyzed oxidation (M), cross-linking with glutaraldehyde (G), oxidation with hydrogen peroxide (H), and incubation in a boiling water bath (B). The products were characterized with UV absorption, circular dichroism and fluorescence spectroscopy, gel permeation chromatography, reverse-phase high-pressure liquid chromatography, sodium dodecyl sulfate polyacrylamide gel electrophoresis, Western blotting, and mass spectrometry. The immunogenicity of the products was evaluated in wild-type mice and in transgenic mice immune tolerant for hIFNα2. Serum antibodies were detected by enzyme-linked immunosorbent assay or surface plasmon resonance. Results: M-rhIFNα2b contained covalently aggregated rhIFNα2b with three methionines partly oxidized to methionine sulfoxides. G-rhIFNα2b contained covalent aggregates and did not show changes in secondary structure. H-rhIFNα2b was only chemically changed with four partly oxidized methionines. B-rhIFNα2b was largely unfolded and heavily aggregated. Nontreated (N) rhIFNα2b was immunogenic in the wild-type mice but not in the transgenic mice, showing that the latter were immune tolerant for rhIFNα2b. The anti-rhIFNα2b antibody levels in the wild-type mice depended on the degradation product: M-rhIFNα2b > H-rhIFNα2b ∼ N-rhIFNα2b ≫ B-rhIFNα2b; G-rhIFNα2b did not induce anti-rhIFNα2b antibodies. In the transgenic mice, only M-rhIFNα2b could break the immune tolerance. Conclusions: RhIFNα2b immunogenicity is related to its structural integrity. Moreover, the immunogenicity of aggregated rhIFNα2b depends on the structure and orientation of the constituent protein molecules and/or on the aggregate size.

Cultural innovation and transmission of tool use in wild chimpanzees:evidence from field experiments

Animal Cognition, V.6, pp. 213-223

Wild species used in the traditional pharmacopoeia of Trás-os-Montes (Portugal)

XX Symposium of Brazilian Medicinal Plants & X International Congress of Ethnopharmacology. S. Paulo, Brasil.

Surveillance of arbovirus infections in the atlantic forest region, State of São Paulo, Brazil: I. detection of hemagglutination-inhibition antibodies in wild birds between 1978 and 1990

We report data related to arbovirus antibodies detected in wild birds periodically captured from January 1978 to December 1990 in the counties of Salesópolis (Casa Grande Station), Itapetininga and Ribeira Valley, considering the different capture environments. Plasmas were examined using hemagglutination-inhibition (HI) tests. Only monotypic reactions were considered, except for two heterotypic reactions in which a significant difference in titer was observed for a determined virus of the same antigenic group. Among a total of 39,911 birds, 269 birds (0.7%) belonging to 66 species and 22 families were found to have a monotypic reaction for Eastern equine encephalitis (EEE), Venezuelan equine encephalitis (VEE), Western equine encephalitis (WEE), Ilheus (ILH), Rocio (ROC), St. Louis encephalitis (SLE), SP An 71686, or Caraparu (CAR) viruses. Analysis of the data provided information of epidemiologic interest with respect to these agents. Birds with positive serology were distributed among different habitats, with a predominance of unforested habitats. The greatest diversity of positive reactions was observed among species which concentrate in culture fields.

New records of Histoplasma capsulatum from wild animals in the Brazilian Amazon

Twenty-eight isolates of Histoplasma capsulation were obtained from eight species of forest mammals from the States of Amazonas, Pará and Rondônia in the Amazon Region of Brazil. Primary isolates were obtained by inoculating triturated liver and spleen tissue intradermally and intraperito-neally in hamsters. Mycological diagnosis in hamsters presenting lesions was confirmed by histopathology and culture on Sabouraud dextrose-agar. Infected hamsters developed signs of disease within two to nine months; all had disseminated visceral lesions and most also had skin lesions at the sites of inoculation. None of the hamsters inoculated with skin macerates of the original hosts developed histoplasmosis, and histopathological examination of the viscera of the wild hosts failed to reveal H. capsulation. Prevalence of infection was considerably higher in females than in males both for the opossum Didelphis marsupialis and for total wild animals (479) examined. It is proposed that canopy-dwelling mammals may acquire the infection from conidia borne on convective currents in hollow trees with openings at ground-level.