Abstract in English:Venoms represent a huge and essentially unexplored reservoir of bioactive components that may cure diseases that do not respond to currently available therapies. This review select advances reported in the literature from 2000 to the present about the new scenario of Hymenoptera venom composition. On account of new technologies in the proteomic approach, which presents high resolution and sensitivity, the combination of developments in new instruments, fragmentation methods, strategic analysis, and mass spectrometry have become indispensable tools for interrogation of protein expression, molecule interaction, and post- translational modifications. Thus, the biochemical characterization of Hymenoptera venom has become a major subject of research in the area of allergy and immunology, in which proteomics has been an excellent alternative to assist the development of more specific extracts for diagnosis and treatment of hypersensitive patients to Hymenoptera venoms.
Abstract in English:Staphylococcus aureus and coagulase-negative Staphylococcus (CoNS) are frequently found in nosocomial environments as the main pathogen in several infections. In 1961, reports of nosocomial S. aureus resistant to methicillin, the drug of choice against penicillin-resistant strains, required new alternatives and vancomycin started being used to treat infections caused by methicillin-resistant S. aureus (MRSA). Community-acquired methicillin-resistant S. aureus (CA-MRSA) was first reported in 1990 affecting patients without risk factors for infection with MRSA of hospital origin. MRSA of community origin harbor the genes responsible for the synthesis of Panton-Valentine leukocidin (PVL), a toxin associated with skin and soft tissue infections and that carries the staphylococcal cassette chromosome mec (SCCmec) type IV. CA-MRSA emergence has caused great impact on the worldwide medical community since the presence of this pathogen in patients without risk factors represents a high risk to public health.
Abstract in English:Viruses depend on cell metabolism for their own propagation. The need to foster an intimate relationship with the host has resulted in the development of various strategies designed to help virus escape from the defense mechanisms present in the host. Over millions of years, the unremitting battle between pathogens and their hosts has led to changes in evolution of the immune system. Snake venoms are biological resources that have antiviral activity, hence substances of significant pharmacological value. The biodiversity in Brazil with respect to snakes is one of the richest on the planet; nevertheless, studies on the antiviral activity of venom from Brazilian snakes are scarce. The antiviral properties of snake venom appear as new promising therapeutic alternative against the defense mechanisms developed by viruses. In the current study, scientific papers published in recent years on the antiviral activity of venom from various species of snakes were reviewed. The objective of this review is to discuss the mechanisms of resistance developed by viruses and the components of snake venoms that present antiviral activity, particularly, enzymes, amino acids, peptides and proteins.
Abstract in English:Leptospirosis is a zoonosis distributed worldwide, endemic mainly in humid subtropical and tropical countries, with epidemic potential. It affects a range of both wild and domestic animals, including sheep, which transport leptospires in their urine and, therefore, can infect other animals and humans who deal with them. Therefore, leptospirosis is characterized as an occupational zoonosis. In individual herds leptospirosis can cause severe economic loss due to miscarriages and outbreaks of mastitis with a significant reduction of milk production. The disease is caused by Leptospira interrogans, which was reclassified into 13 pathogenic species, and distributed into more than 260 serovars classified into 23 serogroups. The clinical signs of infection may vary depending on the serovar and host. In maintenance hosts, antibody production is generally low; there are relatively mild signs of the disease, and a prolonged carrier state with organisms in the kidneys. In incidental hosts, the disease may be more severe, with high titers of circulating antibodies and a very short or nonexistent renal carrier state. In general, young animals with renal and hepatic failure have more serious infections than adults. Several diseases may produce symptoms similar to those of leptospirosis, so that laboratory confirmation, through microscopic agglutination test, for example, is required. The effectiveness of treatment depends on early diagnosis and appropriate therapy, depending on clinical features, since leptospirosis can develop into chronic liver disease and nephropathy, progressing towards death. Improvements in habitation and sanitary conditions, rodent control, vaccination, isolation and treatment of affected animals are the main measures for the control of leptospirosis.
Abstract in English:In recent years, the search for natural plant products to fight viral diseases has been increasing. In this work, two Spondias species, namely S. mombin and S. tuberosa, found in Ceará state (Brazil), and their main phenolic components were evaluated against dengue virus. In vitro antiviral tests were performed against type-2 dengue virus by the MTT method and standard cytopathic effect reduction assay in C6/36 cells. Cytotoxicity was also evaluated by MTT. The presence of phenolic compounds quercetin, rutin, and ellagic acid in plant extracts was characterized by HPLC analysis. Both Spondias species extracts and components were nontoxic to the cells whereas rutin and quercetin displayed relevant antiviral activity with IC50 of 362.68 µg/mL and 500 µg/mL, respectively.
Abstract in English:Currently, medically significant scorpion species belong to the Buthidae family and are represented by the genera Androctonus, Buthus, Mesobuthus, Hottentotta, Parabuthus, Tityus, Centruroides, Leiurus. Although Leiurus was originally considered a monotypic genus, four additional species have since been described. Leiurus abdullahbayrami (previously identified as L. quinquestriatus in Turkey) was classified as a new Leiurus species. This is the first report conducted on the lethality and biologic effects of L. abdullahbayrami scorpion venom in mice. In this study, the electrophoretic protein pattern of its venom was also determined. Two protein bands with molecular masses of 4 and 6 kDa were more strongly detected than other protein bands in the venom sample. Electrophoresis showed that L. abdullahbayrami scorpion venom possesses both short- and long-chain neurotoxins. The median lethal dose of this venom was found to be 0.19 mg/kg by subcutaneous (SC) injection in mice. Animals experimentally envenomed with L. abdullahbayrami venom exhibited hyperexcitability, agitation, aggressive behavior, squeaking and fighting, tachypnea, weakness, convulsions, and death due to cardiac and respiratory failure. In further studies, the potency of antivenom should be investigated in relation to the scorpion venom. Molecular and pharmacological studies are also required to identify and characterize L. abdullahbayrami scorpion venom.
Abstract in English:Wasp venom is mixture of complex proteins that have several physical and pharmacological properties. The photochemical detoxification of Vespa orientalis venom is expected to generate photooxidized venom sac extract (PVSE). Antigenically active PVSE is obtained by exposing the venom sac extract (VSE) of Vespa orientalis to ultraviolet radiation in the presence of methylene blue. The aim of the present work was to evaluate the effect of PVSE on learning and memory of rats. Detoxification of PVSE was evident since treated mice had longer survival time than the group of mice treated with VSE. Photooxidized VSE of V. orientalis revealed enhancement on learning and memory by shortening the time to reach food (TRF) in T-maze. In a 28-day study with rats, we observed that PVSE significantly decreased transfer latency (TL) in elevated plus maze (EPM), significantly increased step down latency (SDL), diminished step down errors (SDE) and time spent in shock zone (TSS) in step down avoidance test. Thus, we concluded that although there is a possibility of employing PVSE in the treatment of Alzheimer, dementia or neurodegenerative illness as a non-herbal and non-synthetic alternative for patients who do not respond to available therapy, further investigation is still required.
Abstract in English:Envenomations caused by different species of Bothrops snakes result in severe local tissue damage, hemorrhage, pain, myonecrosis, and inflammation with a significant leukocyte accumulation at the bite site. However, the activation state of leukocytes is still unclear. According to clinical cases and experimental work, the local effects observed in envenenomation by Bothrops alternatus are mainly the appearance of edema, hemorrhage, and necrosis. In this study we investigated the ability of Bothrops alternatus crude venom to induce macrophage activation. At 6 to 100 ¼g/mL, BaV is not toxic to thioglycollate-elicited macrophages; at 3 and 6 ¼g/mL, it did not interfere in macrophage adhesion or detachment. Moreover, at concentrations of 1.5, 3, and 6 ¼g/mL the venom induced an increase in phagocytosis via complement receptor one hour after incubation. Pharmacological treatment of thioglycollate-elicited macrophages with staurosporine, a protein kinase (PKC) inhibitor, abolished phagocytosis, suggesting that PKC may be involved in the increase of serum-opsonized zymosan phagocytosis induced by BaV. Moreover, BaV also induced the production of anion superoxide (O2_) by thioglycollate-elicited macrophages. This BaV stimulated superoxide production was abolished after treating the cells with staurosporine, indicating that PKC is an important signaling pathway for the production of this radical. Based on these results, we suggest that phagocytosis and reactive oxygen species are involved in the pathogenesis of local tissue damage characteristic of Bothrops spp. envenomations.
Abstract in English:Two-dimensional gel electrophoresis (2DE) is an important tool for investigating the complexity of snake venom proteomes. Apart from applications based on whole proteome analysis, we suggest that 2DE can be used as an assay to guide the progress of protein purification. The aim of this study was to prove the feasibility of this concept by using it to purify rhodocetin from Calloselasma rhodostoma venom. Rhodocetin (α subunit) spot on the 2DE profile of C. rhodostoma venom was first identified and confirmed by mass spectrometry, with a molecular mass of 16 kDa and calculated pI of 5.16. Rhodocetin was subsequently purified by successive anion-exchange and gel filtration chromatography. Every peak from both chromatography profiles was collected and tested on 2DE. The presence of rhodocetin (α subunit) spot in the 2DE profile of the peak DP2 indicated the presence of the protein. The purified compound was used to spike the crude venom. A spiked spot with a 1.6-fold increase in intensity was observed and its position matched to that of rhodocetin (α subunit) on the 2DE profile. Together, these spots confirmed the identity of the purified compound as rhodocetin. Hence, our results have demonstrated the effectiveness of the concept we now term 2DE-guided purification.
Abstract in English:The lethal and enzymatic activities of venom from Naja sumatrana (Equatorial spitting cobra) were determined and compared to venoms from three other Southeast Asian cobras (Naja sputatrix, Naja siamensis and Naja kaouthia). All four venoms exhibited the common characteristic enzymatic activities of Asiatic cobra venoms: low protease, phosphodiesterase, alkaline phosphomonoesterase and L-amino acid oxidase activities, moderately high acetylcholinesterase and hyaluronidase activities and high phospholipase A2. Fractionation of N. sumatrana venom by Resource® S cation exchange chromatography (GE Healthcare, USA) yielded nine major protein peaks, with all except the acidic protein peak being lethal to mice. Most of the protein peaks exhibit enzymatic activities, and L-amino acid oxidase, alkaline phosphomonoesterase, acetylcholinesterase, 5'-nucleotidase and hyaluronidase exist in multiple forms. Comparison of the Resource® S chromatograms of the four cobra venoms clearly indicates that the protein composition of N. sumatrana venom is distinct from venoms of the other two spitting cobras, N. sputatrix (Javan spitting cobra) and N. siamensis (Indochinese spitting cobra). The results support the revised systematics of the Asiatic cobra based on multivariate analysis of morphological characters. The three spitting cobra venoms exhibit two common features: the presence of basic, potentially pharmacologically active phospholipases A2 and a high content of polypeptide cardiotoxin, suggesting that the pathophysiological actions of the three spitting cobra venoms may be similar.
Abstract in English:In order to evaluate the pathogenicity of yeasts isolated from vaginal secretion of pregnant and non-pregnant women - stored in mineral oil at the URM Mycology Collection, Department of Mycology, Federal University of Pernambuco - 30 samples belonging to the genera Candida, Rhodotorula, Trichosporon, and Kloeckera, were studied regarding their pathogenic characteristics, ability to grow at room temperature (28°C ± 1°C), 37°C, and 42°C for 72 hours, and production of both phospholipase and proteinase. Results showed that all 30 isolates (100%) were able to grow at room temperature and 37°C, and that 17 samples (57%) were able to grow at 42°C. Evaluation of enzymatic activity showed protease activity in only two isolates (7%), namely C. maritima and C. obtusa. Phospholipase activity was detected in 20 isolates (67%) using soy lecithin as substrate at different temperatures. The characterization of yeasts isolated from vaginal secretion and determination of their enzymatic activity may contribute to understanding the epidemiology of vulvovaginitis and assist in the treatment of patients.
Abstract in English:Several antibodies, including anticardiolipin antibodies (ACA), have been detected among chronically infected hepatitis C virus (HCV) patients. The present work aimed at ascertaining the clinical significance of ACA levels among HCV infection associated with two commonly encountered diseases, thrombocytopenia and arteriovenous-shunt malfunction. Six groups were studied, 11 HCV-positive thrombocytopenic patients (group I), 14 HCV-positive non-thrombocytopenic patients (group II) and 15 healthy controls (group III), 11 anti-HCV-positive hemodialysis patients with non-functioning shunt (group IV), 14 anti-HCV-positive hemodialysis patients with patent shunt (group V) (Bain Medical Equipment Co., China) and 15 healthy controls (group VI). Anticardiolipin antibody (ACA) assay was performed on all patients and controls whereas tumor necrosis factor α (TNF-α) assay was carried out on thrombocytopenic patients and controls. Thrombocytopenic groups presented an inverse correlation between IgG ACA levels and both thrombocytopenia and TNF-α levels. During the follow-up period, no other clinical manifestations related to ACA were developed. Hemodialysis groups showed a significant elevation in IgG ACA levels in groups IV and V compared to the controls, with statistically higher levels in group IV than group V. Three group IV patients were hypercholesterolemic. We can conclude that induction of proinflammatory cytokines such as TNF-α by persistent HCV infection may promote the generation of ACA. Complications of HCV, including thrombocytopenia and thrombosis in arteriovenous shunt, are more strongly correlated with IgG ACA than with IgM ACA.
Abstract in English:Hypnale hypnale (hump-nosed pit viper) has been recently identified as one of the medically important venomous snakes in Sri Lanka and on the southwestern coast of India. The characterization of its venom is essential for understanding the pathophysiology of envenomation and for optimizing its management. In the present study, the biological properties of Hypnale hypnale venom and venom fractions obtained using Resource Q ion exchange chromatography were determined. The venom exhibited toxic activities typical of pit viper venom, comparable to that of its sister taxon, the Malayan pit viper (Calloselasma rhodostoma). Particularly noteworthy were its high activities of thrombin-like enzyme, proteases, phospholipase A2, L-amino acid oxidase and hyaluronidase. The thrombin-like enzyme was mainly acidic and distributed over several chromatography fractions, indicating its existence in multiple isoforms. The hemorrhagic and necrotic activities of the venom were likely associated with the proteolytic enzyme found mainly in the basic fraction. Phospholipase A2 and phosphomonoesterase exist in both acidic and basic isoforms, while L-amino acid oxidase and hyaluronidase are highly acidic. The venom clotting activity on fibrinogens showed distinct species specificity in the following increasing order for clotting time: bovine < rabbit < goat < human < horse < < dog, and was comparable to that of C. rhodostoma venom. Its clot formation on human fibrinogen is gradual and prolonged, a phenomenon suggestive of consumptive coagulopathy as a complication observed clinically. At an intramuscular sublethal dose, the venom did not cause acute kidney injury in a rodent model, contrary to the positive control group treated with Daboia russelii venom. Nephrotoxicity may result from higher venom doses in the context of coagulopathy, as a complication provoked by venom hematoxicity.
Abstract in English:Rhynocoris fuscipes is a potential predator of many economically important pests in India. In the present study, its venomous saliva (VS) was collected by milking and diluted with HPLC grade water to different concentrations (200, 400, 600, 800 and 1000 ppm). Microinjection of Rhynocoris fuscipes VS was more toxic than its oral administration in Helicoverpa armigera (cotton bollworm) and Spodoptera litura (tobacco cutworm). Thus, R. fuscipes VS was found to be toxic to third instar S. litura and H. armigera with respective LD50s of 846.35 and 861.60 ppm/larva at 96 hours after microinjection. The current results showed that VS of Rhynocoris fuscipes caused mortality of H. armigera and S. litura. Active peptides from VS may be isolated, identified and assessed for their impact in order to ascertain how they alter the physiology of these pests, information that could be applicable in pest management programs.
Abstract in English:The guidelines of the Brazilian National Health System (SUS) state that the municipal authorities are responsible for the vaccination of the population. The present study examined the types of immunobiologicals, amounts and reasons for their destruction and disposal by the Municipal Health Secretariat in the city of Bauru, São Paulo state, Brazil, between 2008 and 2009. This study comprises a descriptive, exploratory and retrospective work that employed search of bibliographic data, collection of secondary data from forms of immunobiologicals disposal and interview of the agent responsible for the disposal of these products in the municipality. It was observed that the total numbers of unused vaccines in 2008 and 2009 were similar (4523 and 4395, respectively), being the most discarded: the diphtheria, tetanus and pertussis (DTP) vaccine in 2008 and DTP, BCG and influenza in 2009. It was found that the amount of discarded vaccines could be reduced since the reasons for that were predictable. Moreover, the current study emphasized that although there is a municipal regulation for the disposal of immunobiologicals, the city still requires a better structure to handle such problem.
Abstract in English:The present study reports a snakebite in a horse in the state of Pará, Brazil. At initial evaluation the animal was reluctant to walk and had tachycardia, tachypnea, severe lameness, bleeding on the pastern and swelling around the left hind leg. Blood samples from the bleeding sites, took on the first day, showed leukocytosis and neutrophilia, whereas biochemical values of urea and creatinine were significantly increased. The chosen treatment was snake antivenom, fluid therapy, antibiotics, anti-inflammatory agents and diuretic drugs. On the fourth day of therapy, the hematological values were within normal parameters. There was improvement related to the clinical lameness and swelling of the limb. However, a decrease in water intake and oliguria were observed. On the seventh day the animal died. Necropsy revealed areas of hemorrhagic edema in the left hind limb and ventral abdomen; the kidneys presented equimosis in the capsule, and when cut they were wet. Moreover, the cortex was pale, slightly yellow and the medullary striae had the same aspect. Based on these data, we concluded that the snakebite in the present study was caused by Bothrops spp. and that renal failure contributed to death.