Abstract in English:Scorpionism in the Americas occurs mainly in Mexico, northern South America and southeast Brazil. This article reviews the local scorpion fauna, available health statistics, and the literature to assess scorpionism in Central America. Notwithstanding its high toxicity in Mexico, most scorpion sting cases in Guatemala, Belize, El Salvador, Nicaragua, and Costa Rica are produced by species in the genus Centruroides that are only mildly toxic to humans despite the existence of ion channel-active toxins in their venoms. Regional morbidity is low with the exception of Panama, where an incidence of 52 cases per 100,000 inhabitants was recorded for 2007, with 28 deaths from 1998 to 2006. Taxa belonging to the genus Tityus (also present in the Atlantic coast of Costa Rica) are responsible for fatalities in Panama, with Tityus pachyurus being the most important species medically. Most Tityus species inhabiting Panama are also found in northern South America from which they probably migrated upon closure of the Panamanian isthmus in the Miocene era. Incorporation of Panama as part of the northern South American endemic area of scorpionism is thereby suggested based on the incidence of these accidents and the geographical distribution of Panamanian Tityus species.
Abstract in English:The genus Fusarium is known to produce mycotoxins that cause fusariosis in plants, animals and humans. Mycotoxins are among the virulence factors of this genus. Metabolic extracts of Fusarium oxysporum, isolated from a patient with onychomycosis and sterilized by filtration or autoclave, were inoculated intradermally into Wistar rats at concentrations of 0.25, 0.5 and 1 µg/µL, and the effects on their tegument were observed at 24 and 72 hours. After histological procedures and staining by hematoxylineosin, the sections were studied for their inflammatory-reaction intensity and for evidence of injury and tissue distortion. Inflammatory reactions in the dermis and the subcutaneous tissue were observed at all concentrations of the inoculated extract tested. There was a significant influx of neutrophils, mastocytes and lymphocytes, as well as a large quantity of macrophages. Apoptotic bodies and hyperemic blood vessels were observed. This reaction was directly related to the extract concentration, and was most intense in animals that received the 1 mg/µL dose. The maximum peak was observed at 24 hours. The autoclaved metabolic extract produced the same effects as the untreated one, indicating the presence of heat-resistant metabolites. In conclusion, the metabolic extracts obtained from sterilized culture filtrates of F. oxysporum are capable of inducing an inflammatory response within 24 hours in the dermis and subcutaneous tissue of rats.
Abstract in English:Cytotoxicity of venoms from eleven medically important snakes found in Southeast Asia (Naja kaouthia, Naja siamensis, Naja sumatrana, Ophiophagus hannah, Bungarus candidus, Bungarus fasciatus, Enhydrina schistosa, Calloselasma rhodostoma, Trimeresurus purpureomaculatus and Tropidolaemus sumatranus) was determined, based on the MTS cytotoxicity assay, which determines the survival of viable cells in monolayer MDCK and Vero cell cultures upon exposure to the snake venoms. Snake venom toxicity was expressed as the venom dose that killed 50% of the cells (CTC50) under the assay conditions. Venoms of C. rhodostoma (2.6 µg/mL, 1.4 µg/mL) and O. hannah were the most cytotoxic (3.8 µg/mL, 1.7 µg/mL) whereas N. siamensis venom showed the least cytotoxicity (51.9 µg/mL, 45.7 µg/mL) against Vero and MDCK cells, respectively. All the viper venoms showed higher cytotoxic potency towards both Vero and MDCK cell lines, in comparison to krait and cobra venoms. E. schistosa did not cause cytotoxicity towards MDCK or Vero cells at the tested concentrations. The cytotoxicity correlates well with the known differences in the composition of venoms from cobras, kraits, vipers and sea snakes.
Abstract in English:Although sea anemones are well known for being rich sources of toxins, including cytolysins and neurotoxins, their venoms and toxins have been poorly studied. In the present study, the venoms from five sea anemones (Heteractis crispa, Heteractis magnifica, Heteractis malu, Cryptodendrum adhaesivum and Entacmaea quadricolor) were obtained by the milking technique, and the potential of these venoms to kill cancer cells was tested on three cell lines (A549 lung cancer, T47D breast cancer and A431 skin cancer). The total protein level in the crude extract was determined by the bicinchoninic acid (BCA) protein assay. The cytotoxicity on different cell lines was assayed using the 3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyltetrazolium bromide (MTT) assay which measures survival based on the detection of mitochondrial activity and by the crystal violet assay, which measures survival based on the ability of cells to remain adherent to microplates. The results indicate that the sea anemone venom is cytotoxic to human cancer cells. The A549 cell line was the most sensitive of the cell lines tested with a significant reduction in viability observed at 40 µg/mL. H. malu, C. adhaesivum and E. quadricolor had a significant inhibitory effect on A431 cells. Furthermore, H. malu and C. adhaesivum had a significant inhibitory effect on T47D cell line at 40 µg/mL. In conclusion, the sea anemone venoms tested have the potential to be developed as anticancer agents.
Abstract in English:This study evaluates the mortality and average survival rates of captive female Philodryas olfersii and Philodryas patagoniensis snakes maintained for venom production. Also, two factors likely to reduce captive survival were studied - body condition at admission and seasonality. Mortality peaks occurred during the second month in captivity. More than half the individuals were dead at the end of the third month. This suggests that the first three months in captivity are the most critical in terms of survival and adaptation. Females collected and admitted during spring and summer lived less time than those collected in autumn and winter. As gravidity and egg-laying occur during spring and summer, we suggest that the lower survival rates in these seasons may be due to high costs and stress involved in these reproductive events. Unexpectedly, body mass and body condition were poor predictors of survival in captivity. Our results have important implications in maintaining snakes for venom production. We propose some prophylactic measures to minimize the deleterious impacts of captivity during the adaptation period.
Abstract in English:The snake genus Lachesis provokes 2 to 3% of snakebites in Colombia every year. Two Lachesis species, L. acrochorda and L. muta, share habitats with snakes from another genus, namely Bothrops asper and B. atrox. Lachesis venom causes systemic and local effects such as swelling, hemorrhaging, myonecrosis, hemostatic disorders and nephrotoxic symptoms similar to those induced by Bothrops, Portidium and Bothriechis bites. Bothrops antivenoms neutralize a variety of Lachesis venom toxins. However, these products are unable to avoid coagulation problems provoked by Lachesis snakebites. Thus, it is important to ascertain whether the envenomation was caused by a Bothrops or Lachesis snake. The present study found enzyme linked immunosorbent assay (ELISA) efficient for detecting Lachesis acrochorda venom in a concentration range of 3.9 to 1000 ng/mL, which did not show a cross-reaction with Bothrops, Portidium, Botriechis and Crotalus venoms. Furthermore, one fraction of L. acrochorda venom that did not show crossreactivity with B. asper venom was isolated using the same ELISA antibodies; some of its proteins were identified including one Gal-specific lectin and one metalloproteinase. This test may be useful to physicians, since it could be applicable for tracking the kinetic distribution of antigens in patients or experimentally envenomed animals.
Abstract in English:Helicobacter pylori, a gram-negative bacterium, possesses two important virulence factors: the vacuolating toxin (vacA), and the cytotoxin-associated gene product (cagA). The aim of the present study was to evaluate the presence of H. pylori in the stomach and oral cavity of humans and compare the cagA and vacA genotypes of H. pylori found in different samples (stomach, saliva and dental plaque) from the same patient. Gastric biopsies, saliva and dental plaques were obtained from 62 dyspeptic adults. DNA was extracted and evaluated for the presence of H. pylori and the alleles cagA and vacA. Persons with gastritis had a higher frequency of H. pylori -positive samples in the stomach while positive samples from gastric biopsies were significantly correlated with those from the oral cavity. There was a high H. pylori frequency in patients while the cagA gene was associated with vacA s1 alleles in gastric biopsies. Our results suggest a reservoir of the species in the oral cavity and that, in one patient, more than one H. pylori strain may exist in the saliva, dental plaque and stomach. We found a relationship between gastric infection and the bacterium in the oral cavity, with the cytotoxin genotype varying between saliva and dental plaque.
Abstract in English:Leishmaniasis is a neglected disease endemic in five continents. It is a severe disease that may lead to death, and its early detection is important to avoid severe damage to affected individuals. Molecular methods to detect Leishmania are considered alternatives to overcome the limitations presented by conventional methods. The aim of this study was to develop multiplex PCR systems able to detect small amounts of target DNA of Leishmania infantum and Leishmania braziliensis, and the gene coding for glyceraldehyde-3-phosphate dehydrogenase (G3PD) in mammals, enabling quality evaluation of the sample simultaneously with detection of the specific target. The systems created for G3PD recognition were combined with detection systems for L. infantum and L. braziliensis to compose multiplex PCR systems for visceral (mVL) and cutaneous (mACL) leishmaniasis diagnosis. The multiplex PCR systems developed were assessed in blood samples from five different species of mammal reservoirs involved in the disease cycle in Brazil, and 96 and 52 human samples from patients with suspected visceral leishmaniasis (VL) and cutaneous leishmaniasis (ACL), respectively. Three G3PD detection systems were created (G3PD1, G3PD2 and G3PD3) with different product sizes, G3PD2 was chosen for the formation of multiplex PCR systems. The two multiplex PCR systems (mVL and mACL) were reproducible in all species evaluated. Results of test samples (sensitivity, specificity and efficiency) suggest its use in routine diagnosis, research activities in medicine and veterinary medicine. Additionally, the systems designed to detect the G3PD gene are capable of combining with other targets used for molecular diagnosis of infectious diseases. Concerning leishmaniasis, the multiplex PCR systems can be used in epidemiological studies for the detection of new and classic reservoirs, which may contribute to the reliability of results and development of actions to control the disease.
Abstract in English:Some constituents of snake venom have been found to display a variety of biological activities. The antibacterial property of snake venom, in particular, has gathered increasing scientific interest due to antibiotic resistance. In the present study, king cobra venom was screened against three strains of Staphylococcus aureus [including methicillin-resistant Staphylococcus aureus (MRSA)], three other species of gram-positive bacteria and six gram-negative bacteria. King cobra venom was active against all the 12 bacteria tested, and was most effective against Staphylococcus spp. (S. aureus and S. epidermidis). Subsequently, an antibacterial protein from king cobra venom was purified by gel filtration, anion exchange and heparin chromatography. Mass spectrometry analysis confirmed that the protein was king cobra L-amino acid oxidase (Oh-LAAO). SDS-PAGE showed that the protein has an estimated molecular weight of 68 kDa and 70 kDa under reducing and non-reducing conditions, respectively. The minimum inhibitory concentrations (MIC) of Oh-LAAO for all the 12 bacteria were obtained using radial diffusion assay method. Oh-LAAO had the lowest MIC value of 7.5 µg/mL against S. aureus ATCC 25923 and ATCC 29213, MRSA ATCC 43300, and S. epidermidis ATCC 12228. Therefore, the LAAO enzyme from king cobra venom may be useful as an antimicrobial agent.
Abstract in English:The venomous Levantine viper, Macrovipera lebetina lebetina is endemic to Cyprus. The objective of this study was to investigate in vitro cytotoxicity, in vivo lethality, and antivenom production followed by a re-immunization schedule in mice against Macrovipera lebetina lebetina venom. The LD50 value was estimated as 7.58 mg/kg within 24 hours by different venom doses administrated intraperitoneally in mice. Freund's complete and incomplete adjuvants were used for first and second immunization of mice in antivenom production. A cell-based assay was performed to determine the effects of Macrovipera lebetina lebetina venom and antivenom neutralizing potency on L929 cell viability. The snake venom toxicity and cytotoxicity were examined and comparison of results showed good correlation, the LD50 value was tenfold higher than the IC50 value. The IC50 value was 0.62 ± 0.18 mL after 48 hours treatment while the calculated value was 1.62 ± 0.25 mL for the culture media totally refreshed after two hours treatment with venom. The in vitro efficacy of antivenom against Macrovipera lebetina lebetina venom was found to be low. This is the first report that describes the in vivo and in vitro toxic effects of Macrovipera lebetina lebetina venom and antivenom production against this species.
Abstract in English:All snakebites registered in Rio de Janeiro state, Brazil, between 2001 and 2006 were analyzed. Of these, the snake species involved was identified in 2,431 cases, most of which were caused by Bothrops (2,347). Most victims were male (78.4%) and the age group most frequently attacked was between 20 and 39 years (38.4%). As for severity, 1,118 (45.9%) were classified as minor, 748 (30.7%) as moderate, and 209 (8.6%) as severe. Progression towards complete cure occurred in 1,567 patients; 39 had sequelae; three died; and clinical outcomes were not monitored in 822 cases. The time between bite and first medical attention was less than three hours in 74.6% of cases. Most occurred in municipalities with vast areas of preserved Atlantic Forest: Angra dos Reis (208), Rio de Janeiro (197), Parati (186), Teresópolis (134), and Petrópolis (110). Accident frequency was highest between November and April, in which the daily average is practically twice that observed in other months. Although results show that Rio de Janeiro state has a good level of medical care, it could be improved by creating multidisciplinary teams that include doctors, biologists, and nurses. The Vital Network for Brazil helps to promote a stimulating environment for this type of training, with both traditional courses and distance learning. Training hours must be increased to improve the skills of professionals responsible for victim care.
Abstract in English:This study applied a socioeconomic questionnaire designed to evaluate the frequency of intestinal parasites and characterize epidemiological, nutritional, and immunological variables in 105 HIV/AIDS patients - with and without parasitic infections, attending the Day Hospital in Botucatu, UNESP, from 2007 to 2008. Body mass index was calculated and the following tests performed: parasitological stool examinations; eosinophil, IgE, CD4+ T and CD8+ T lymphocyte cell counts; albumin test; viral load measure; and TNF-α, IFN-γ, IL-2, IL-5 and IL-10 cytokine levels. Results were positive for parasitic intestinal infections in 12.4% of individuals. Most patients had good socioeconomic conditions with basic sanitation, urban dwellings, treated water supply and sewage, good nutritional and immunological status and were undergoing HAART. Parasites were found at the following frequencies: Entamoeba - five patients (38.5%), Giardia lamblia - four (30.7%), Blastocystis hominis - three (23.0%), Endolimax nana - two (15.4%), and Ascaris lumbricoides - one (7.7%). There were no significant differences between the two groups for eosinophils, albumin, IgE, CD4+ T and CD8+ T lymphocytes, INF-γ, IL-2, or IL-10. Most patients also showed undetectable viral load levels. Significant differences were found for TNF-α and IL-5. These results show the importance of new studies on immunodeficient individuals to increase understanding of such variables.
Abstract in English:This study analyses venom from the elapid krait snake Bungarus sindanus, which contains a high level of acetylcholinesterase (AChE) activity. The enzyme showed optimum activity at alkaline pH (8.5) and 45ºC. Krait venom AChE was inhibited by substrate. Inhibition was significantly reduced by using a high ionic strength buffer; low ionic strength buffer (10 mM PO4 pH 7.5) inhibited the enzyme by 1. 5mM AcSCh, while high ionic strength buffer (62 mM PO4 pH 7.5) inhibited it by 1 mM AcSCh. Venom acetylcholinesterase was also found to be thermally stable at 45ºC; it only lost 5% of its activity after incubation at 45ºC for 40 minutes. The Michaelis-Menten constant (Km) for acetylthiocholine iodide hydrolysis was found to be 0.068 mM. Krait venom acetylcholinesterase was also inhibited by ZnCl2, CdCl2, and HgCl2 in a concentrationdependent manner. Due to the elevated levels of AChE with high catalytic activity and because it is more stable than any other sources, Bungarus sindanus venom is highly valuable for biochemical studies of this enzyme.
Abstract in English:Yeasts are becoming a common cause of nosocomial fungal infections in immunocompromised patients. Such infections often develop into sepsis with high mortality rates. The aim of this study was to evaluate some of the numerous factors associated with the development of candidemia. Medical records were retrospectively analyzed of 98 Candida spp. patients. Results showed that the most prevalent risk factors for developing candidemia were: antibiotics and antifungal agents (93.9% and 79.6%, respectively); the use of central venous catheter (93.9%); mechanical ventilation (73.5%); and parenteral nutrition (60.2%). The main species of Candida found were: C. parapsilosis (37.76%), C. albicans (33.67%); and others (28.57%). C. glabrata showed the highest mortality rate (75%), followed by C. tropicalis (57.1%) and C. albicans (54.5%). The elevated mortality rate found in this study indicates that preventive measures against candidemia must be emphasized in hospitals.
Abstract in English:Nearly half of Hymenoptera stings affect the head and neck region of victims, but reports on oropharyngeal bee stings are very few. We describe the case of a patient with odynophagia and suffocation in mass envenomation. He had a retained bee stinger whose removal was delayed for more than 24 hours following the sting, due to persisting angioedema. Odynophagia receded after removal of the stinger and treatment with paracetamol, steroids and metronidazole. The patient also developed rhabdomyolysis, renal failure and hepatitis that were treated with conservative therapy. Oropharyngeal stings can simulate symptoms of persisting angioedema in victims of mass envenomation.