Intoxicação natural por Clostridium botulinum tipo C: em grupo de aves domésticas

Clostridium botulinum causative of toxic infections due to toxin ingestion previously formed, occur in several ingestion species, mainly birds. In a poultry farm, located at São Paulo State, more than 3,000 birds have been attacked by this toxic infections, when the birds showed motionless, loss of weight, accelerated, breathing, resulting in death of the birds. The blood harvest for attainment of the serum and later necropsy was carried out throwgh the following samples: liver, gizzard, crop, feed, water and litter. After that, the toxin was extracted by gelatin-phosphate buffer and inoculation in mice, isolation of the agent in Blood. Ágar and Reinforced Clostridium Ágar and neutralization of toxin determine its type. The inoculation in mice showed positive results in samples of liver, gizzad, crop and symptoms like was waist, and death of the birds by limp paralysis. The colonies that have been isolated, suspected of Clostridium botulinum showed expected results and further analysis revealed positive results to botulinical toxin type C. The farms have to pay attention in the routine jobs, choicer, and elimination of carcass because they are essential to keep this problem away.

Interaction of Bacillus thuringiensis Cry1 and Vip3A proteins with Spodoptera frugiperda midgut binding sites

Vip3Aa, Vip3Af, Cry1Ab, and Cry1Fa were tested for their toxicities and binding interactions. Vip3A proteins were more toxic than Cry1 proteins. Binding assays showed independent specific binding sites for Cry1 and Vip3A proteins. Cry1Ab and Cry1Fa competed for the same binding sites, whereas Vip3Aa competed for those of Vip3Af. Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Is it all death feigning? Case in anurans

Anurans are known to feign death as a way to avoid or minimize the risk of predation. However, information on this defensive strategy is scattered and we believe that there is more than one behaviour type referred to as thanatosis. Here we review the literature, add original data, and propose definitions and new names that complement the present knowledge on the subject. We collected information on 334 individuals of 99 species in 16 families and grouped the recorded displays into two categories of tonic immobility: (1) thanatosis, death-feigning, or playing possum, and (2) shrinking or contracting. These two categories are treated as different behaviour types because of the display pattern (position of fore- and hindlimbs, eye opening), presence of skin toxins (shrinking is mostly displayed by toxic species, whereas thanatosis is mostly displayed by non-toxic species), social context (interaction with predators), and their putative or actual functions. © 2010 Taylor & Francis.

The acylpolyamines from spider venoms

Acylpolyamines are low molecular mass toxins occurring exclusively in the venoms from solitary wasps and some groups of spiders. Their chemical structures have been elucidated using hyphenated techniques of mass spectrometry, such as LC-MS and MS/MS, or through direct analysis with different types of NMR analyses. The chemical structures of the acylpolyamine toxins from the venoms of Nephilinae orb-web spiders appear to be organized into four parts based on the combinatorial way that the chemical building blocks are bound to each other. An aromatic moiety (part I) is connected through a linker amino acid (part II) to a polyamine chain (part III), which in turn may be connected to an optional tail (part IV). The polyamine chains were classified into seven subtypes according to the different combinations of chemical building blocks. These polyamine chains, in turn, are connected to one of three chromophore moieties: a 2,4-dihydroxyphenyl acetyl group, a 4-hydroxyindolyl acetyl group, or an indolyl acetyl group. They may be connected through an asparagine residue or sometimes through the dipeptide ornithyl asparagine. Also, nine different types of backbone tails may be attached to the polyamine chains. These toxins are noncompetitive blockers of ionotropic glutamate receptors with neuroprotective action against the neuronal death and antiepileptic effect. Thus, compounds of this class of spider venom toxin seem to represent interesting molecular models for the development of novel neuropharmaceutical drugs. © 2012 Elsevier B.V. All rights reserved.

Detection of enterotoxin A in coagulase-negative staphylococci isolated from nutrition students

Background: Staphylococcus is a clinically important genus because of its capacity to produce enterotoxins and to cause food poisoning. Staphylococci are the most frequent microorganisms of the skin and mucosal microbiota, with an estimated 20 to 40% of individuals carrying these bacteria on their hands or nose. Since nutrition professionals are involved in the handling and preparation of foods and are possible carriers of these bacteria, the objective of this study was to investigate the presence of Staphylococcus on the hands and in the nasal fossae of undergraduate nutrition students and to determine the enterotoxigenic capacity of these microorganisms. Methods and Findings: A total of 201 strains were isolated from the hands and nose of 61 nutrition students. Of these, 180 (89.5%) were identified as coagulasenegative staphylococci and 21 (10.5%) as S. aureus. Thirty-seven (18.4%) Staphylococcus isolates were producers of enterotoxin A. Toxin production was detected in 5 (19%) of the S. aureus isolates and in 31 (17.2%) of the coagulase-negative staphylococci. Conclusions: This study demonstrated a large number of enterotoxin-producing staphylococci on the hands and nose of nutrition students and professionals involved in the handling and preparations of foods. These findings indicate the need for adequate hygiene measures to prevent food poisoning. © iMedPub.

Analysis of total glucosinolates and chromatographically purified benzylglucosinolate in organic and conventional vegetables

The limited availability of foods that are free of pesticides has led Brazil to search for alternative production methods to meet the desires of consumers. Currently, organic cultivation represents a production system that complies with general expectations of producers and consumers. Organic cultivation is particularly interesting mainly because of its effect on plant secondary metabolite content, which may help plants to naturally combat pests; in humans, these substances can also contribute to the prevention of chronic diseases. We report on the extraction of glucosinolates (both as total glucosinolates and as benzylglucosinolate) with trifluoroacetic acid addition in a 70:30 MeOH:water (v/v). Total glucosinolates, determined by a thioglucosidase coupled assay, were measured in different Brassicaceae species and were similar to values reported in the literature. For broccoli, analyses were carried out separately on inflorescences, leaves and stalks; analyses were also conducted on thermally processed samples to simulate cooking. Furthermore, when the analysis was conducted on conventional and organic products, the highest concentrations of these substances were most often found in organically cultivated Brassicaceae. The benzylglucosinolate concentrations were evaluated on the same samples using HPLC. The concentration of benzylglucosinolate was significantly higher in organically cultivated vegetables, as well. © 2012 Elsevier Ltd.

Bothropstoxin-I reduces evoked acetylcholine release from rat motor nerve terminals: Radiochemical and real-time video-microscopy studies

Understanding the biological activity profile of the snake venom components is fundamental for improving the treatment of snakebite envenomings and may also contribute for the development of new potential therapeutic agents. In this work, we tested the effects of BthTX-I, a Lys49 PLA2 homologue from the Bothrops jararacussu snake venom. While this toxin induces conspicuous myonecrosis by a catalytically independent mechanism, a series of in vitro studies support the hypothesis that BthTX-I might also exert a neuromuscular blocking activity due to its ability to alter the integrity of muscle cell membranes. To gain insight into the mechanisms of this inhibitory neuromuscular effect, for the first time, the influence of BthTX-I on nerve-evoked ACh release was directly quantified by radiochemical and real-time video-microscopy methods. Our results show that the neuromuscular blockade produced by in vitro exposure to BthTX-I (1 μM) results from the summation of both pre- and postsynaptic effects. Modifications affecting the presynaptic apparatus were revealed by the significant reduction of nerve-evoked [3H]-ACh release; real-time measurements of transmitter exocytosis using the FM4-64 fluorescent dye fully supported radiochemical data. The postsynaptic effect of BthTX-I was characterized by typical histological alterations in the architecture of skeletal muscle fibers, increase in the outflow of the intracellular lactate dehydrogenase enzyme and progressive depolarization of the muscle resting membrane potential. In conclusion, these findings suggest that the neuromuscular blockade produced by BthTX-I results from transient depolarization of skeletal muscle fibers, consequent to its general membrane-destabilizing effect, and subsequent decrease of evoked ACh release from motor nerve terminals. © 2012 Elsevier Ltd.

Synthesis, characterization and antifungal activity of quaternary derivatives of chitosan on Aspergillus flavus

Two series of new chitosan derivatives were synthesized by reaction of deacetylated chitosan (CH) with propyl (CH-Propyl) and pentyl (CH-Pentyl) trimethylammonium bromides to obtain derivatives with increasing degrees of substitution (DS). The derivatives were characterized by 1H NMR and potentiometric titration techniques and their antifungal activities on the mycelial growth of Aspergillus flavus were investigated in vitro. The antifungal activities increase with DS and the more substituted derivatives of both series, CH-Propyl and CH-Pentyl, exhibited antifungal activities respectively three and six times higher than those obtained with commercial and deacetylated chitosan. The minimum inhibitory concentrations (MIC) were evaluated at 24, 48 and 72h by varying the polymer concentration from 0.5 to 16g/L and the results showed that the quaternary derivatives inhibited the fungus growth at polymer concentrations four times lower than that obtained with deacetylated chitosan (CH). The chitosans modified with pentyltrimethylammonium bromide exhibited higher activity and results are discussed taking into account the degree of substitution (DS). © 2012 Elsevier GmbH.

Triacontyl p-coumarate: An inhibitor of snake venom metalloproteinases

Snake venom metalloproteinases (SVMPs) participate in a number of important biological, physiological and pathophysiological processes and are primarily responsible for the local tissue damage characteristic of viperid snake envenomations. The use of medicinal plant extracts as antidotes against animal venoms is an old practice, especially against snake envenomations. Such plants are sources of many pharmacologically active compounds and have been shown to antagonize the effects of some venoms and toxins. The present study explores the activity of triacontyl p-coumarate (PCT), an active compound isolated from root bark of Bombacopsis glabra vegetal extract (Bg), against harmful effects of Bothropoides pauloensis snake venom and isolated toxins (SVMPs or phospholipase A2). Before inhibition assays, Bg or PCT was incubated with venom or toxins at ratios of 1:1 and 1:5 (w/w; venom or isolated toxins/PCT) for 30 min at 37 °C. Treatment conditions were also assayed to simulate snakebite with PCT inoculated at either the same venom or toxin site. PCT neutralized fibrinogenolytic activity and plasmatic fibrinogen depletion induced by B. pauloensis venom or isolated toxin. PCT also efficiently inhibited the hemorrhagic (3MDH-minimum hemorrhagic dose injected i.d into mice) and myotoxic activities induced by Jararhagin, a metalloproteinase from B. jararaca at 1:5 ratio (toxin: inhibitor, w/w) when it was previously incubated with PCT and injected into mice or when PCT was administered after toxin injection. Docking simulations using data on a metalloproteinase (Neuwiedase) structure suggest that the binding between the protein and the inhibitor occurs mainly in the active site region causing blockade of the enzymatic reaction by displacement of catalytic water. Steric hindrance may also play a role in the mechanism since the PCT hydrophobic tail was found to interact with the loop associated with substrate anchorage. Thus, PCT may provide a alternative to complement ophidian envenomation treatments. © 2012 Elsevier Ltd. All rights reserved.

Hepatoencephalopathy syndrome due to Cassia occidentalis (Leguminosae, Caesalpinioideae) seed ingestion in horses

Cassia occidentalis is a bush from the Leguminosae family, subfamily Caesalpinoideae, and is a toxic plant of veterinary interest due to the occasional contamination of animal rations. This report describes the clinical and histopathological findings of an outbreak of C.occidentalis poisoning in horses. Twenty mares were poisoned after consuming ground corn contaminated with 8% of C.occidentalis seeds. Of the 20 animals affected, 12 died: 8 mares were found dead, 2 died 6h after the onset of clinical signs compatible with hepatic encephalopathy and the 2 other animals were subjected to euthanasia 12h after the onset of the clinical signs. The remaining 8 mares presented with mild depression and decreased appetite, but improved with treatment and no clinical sequelae were observed. In 6 animals that underwent a necropsy, an enhanced hepatic lobular pattern was noted and within the large intestine, a large number of seeds were consistently observed. Hepatocellular pericentrolobular necrosis and cerebral oedema were the main histological findings. In one mare, there was mild multifocal semimembranosus rhabdomyocytic necrosis and haemorrhage. Seeds collected from intestinal contents and sifted from the culpable feedstuff were planted. Examination of the leaves, flowers, fruits and seeds of the resultant plants identified C.occidentalis. Horses poisoned by C.occidentalis seeds demonstrate clinical signs associated with hepatoencephalopathy and frequently die suddenly. Lesions primarily involve the liver and secondarily, the central nervous system. Cassia occidentalis poisoning should be considered a differential diagnosis in horses with hepatoencephalopathy and special caution should be taken with horse rations to avoid contamination with seeds of this toxic plant. © 2012 EVJ Ltd.

A simple, rapid method for the extraction of whole fire ant venom (Insecta: Formicidae: Solenopsis)

The invasive fire ant Solenopsis invicta is medically important because its venom is highly potent. However, almost nothing is known about fire ant venom proteins because obtaining even milligram-amounts of these proteins has been prohibitively challenging. We present a simple and fast method of obtaining whole venom compounds from large quantities of fire ants. For this, we separate the ants are from the nest soil, immerse them in dual-phase mixture of apolar organic solvent and water, and evaporate each solvent phase in separate. The remaining extract from the aqueous phase is largely made up of ant venom proteins. We confirmed this by using 2D gel electrophoresis while also demonstrating that our new approach yields the same proteins obtained by other authors using less efficient traditional methods. © 2013 Elsevier Ltd.

Insect/Bt-cotton interactions: Are immunological variables and hemocyte ultrastructure in Podisus nigrispinus (Hemiptera: Pentatomidae) affected?

Recent studies have shown that ingestion by the army worm Spodoptera frugiperda of Cry1Ac toxin from Bt cotton promotes histochemical and ultrastructural changes in the digestive cells of the predatory pentatomid bug Podisus nigrispinus. Therefore, mindful of the changes in the midgut of the predator, which represents the first line of defence in this insect, our aim was to test the hypothesis that the consumption of Bt cotton-fed S. frugiperda by P. nigrispinus might lead to alterations in components of the immune system of P. nigrispinus. The Cry1Ac toxin level in the leaves of Bt cotton, nitric oxide, phenoloxidase activity, and total proteins were quantified by ELISA. Total and differential hemocyte counts were evaluated, and hemocyte ultrastructure analysis was undertaken. We found that ingestion of the prey fed daily with approximately 23 ± 0.70 ng g-1 Cry1Ac by wet weight of leaves, and expressed by the Bt cotton, induces small ultrastructural changes in the predator's granulocytes and plasmatocytes. However, these changes did not affect the total number and differential and humoral variables analyzed for the bug's hemocytes. © 2013 Copyright Taylor and Francis Group, LLC.

Persistent inflammation in the CNS during chronic EAE despite local absence of IL-17 production

Experimental autoimmune encephalomyelitis (EAE) is an artificially induced demyelination of the central nervous system (CNS) that resembles multiple sclerosis in its clinical, histopathological, and immunological features. Activated Th1 and Th17 cells are thought to be the main immunological players during EAE development. This study was designed to evaluate peripheral and local contribution of IL-17 to acute and chronic EAE stages. C57BL/6 mice were immunized with MOG plus complete Freund's adjuvant followed by pertussis toxin. Mice presented an initial acute phase characterized by accentuated weight loss and high clinical score, followed by a partial recovery when the animals reached normal body weight and smaller clinical scores. Spleen cells stimulated with MOG produced significantly higher levels of IFN-γ during the acute period whereas similar IL-17 levels were produced during both disease stages. CNS-infiltrating cells stimulated with MOG produced similar amounts of IFN-γ but, IL-17 was produced only at the acute phase of EAE. The percentage of Foxp3+ Treg cells, at the spleen and CNS, was elevated during both phases. The degree of inflammation was similar at both disease stages. Partial clinical recovery observed during chronic EAE was associated with no IL-17 production and presence of Foxp3+ Treg cells in the CNS. © 2013 Sofia Fernanda Gonçalves Zorzella-Pezavento et al.

Proteome and phosphoproteome of Africanized and European honeybee venoms

Honey bee venom toxins trigger immunological, physiological, and neurological responses within victims. The high occurrence of bee attacks involving potentially fatal toxic and allergic reactions in humans and the prospect of developing novel pharmaceuticals make honey bee venom an attractive target for proteomic studies. Using label-free quantification, we compared the proteome and phosphoproteome of the venom of Africanized honeybees with that of two European subspecies, namely Apis mellifera ligustica and A. m. carnica. From the total of 51 proteins, 42 were common to all three subspecies. Remarkably, the toxins melittin and icarapin were phosphorylated. In all venoms, icarapin was phosphorylated at the 205Ser residue, which is located in close proximity to its known antigenic site. Melittin, the major toxin of honeybee venoms, was phosphorylated in all venoms at the 10Thr and 18Ser residues. 18Ser phosphorylated melittin-the major of its two phosphorylated forms-was less toxic compared to the native peptide. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Molecular epidemiology of methicillin-resistant Staphylococcus aureus in a burn unit from Brazil

Methicillin-resistant Staphylococcus aureus (MRSA) poses a threat for patients in burn units. Studies that mix epidemiological designs with molecular typing may contribute to the development of strategies for MRSA control. We conducted a study including: molecular characterization of Staphylococcal Chromosome Cassette mecA (SCCmec), strain typing with pulsed field gel electrophoresis (PFGE) and detection of virulence genes, altogether with a case-case-control study that assessed risk factors for MRSA and for methicillin-susceptible S. aureus (MSSA), using S. aureus negative patients as controls. Strains were collected from clinical and surveillance cultures from October 2006 through March 2009. MRSA was isolated from 96 patients. Most isolates (94.8%) harbored SCCmec type III. SCCmec type IV was identified in isolates from four patients. In only one case it could be epidemiologically characterized as community-associated. PFGE typing identified 36 coexisting MRSA clones. When compared to MSSA (38 isolates), MRSA isolates were more likely to harbor two virulence genes: tst and lukPV. Previous stay in other hospital and admission to Intensive Care Unit were independent risk factors for both MRSA and MSSA, while the number of burn wound excisions was significantly related with the former (OR = 6.80, 95%CI = 3.54-13.07). In conclusion, our study found polyclonal endemicity of MRSA in a burn unit, possibly related to importing of strains from other hospitals. Also, it pointed out to a role of surgical procedures in the dissemination of MRSA strains. © 2013 Elsevier Ltd and ISBI. All rights reserved.