Abstract in English:Propolis, a beehive product widely used in folk medicine as an antiinflammatory agent, has been attracting researchers' attention to scientifically elucidate its biological properties and therapeutic activities. The aim of this paper was to study the possible effect of propolis on natural killer activity, since propolis immunomodulatory action has been suggested, especially on non-specific immunity. Propolis was produced by africanized honeybees (Apis mellifera L.), collected throughout a whole year, and pooled by season. Hydroalcoholic solutions of propolis were prepared with each pool and administered to rats by gavage over three days. Natural killer activity of non-adherent spleen cells was evaluated by the 51Cr-release cytotoxicity assay against Yac-1 target cells. Our results indicated that the natural killer activity was increased in spleen cells from propolis-treated animals. There were no significant differences related to the seasonal effect on the immunomodulatory action of propolis.
Abstract in English:Electrocardiographic (ECG) changes were induced in dogs by injection of scorpion venom from Mesobuthus tamulus concanesis, Pocock. Venom (3 mg/kg body weight) was given subcutaneously (SQ) while 10 ml of scorpion antivenom (SAV) was administered intravenously (IV) to experimental dogs. Group 1 received only the venom; Groups 2, 3, and 4 received SAV at 0, 30, and 60 min, respectively, following envenoming. Thick, ropy and profuse salivation; muscle fasciculation; clonus and tetany-like contractions; frequent urination; and bowel emptying sometimes stained with bile and occasionally blood and bile were observed 20-25 minutes after envenoming. Following envenoming, hyperglycemia, increase in free fatty acid (FFA) levels, and reduction in triglyceride levels were observed in Groups 1, 3, and 4. There was an initial rise in insulin levels at 30 min followed by a reduction at 60 min. SAV caused a subsequent rise in insulin levels but there was a reduction in blood sugar to euglycemia levels and lipogenesis (reduction in FFA and increase in triglycerides levels) in Groups 3 and 4. Abnormal ECG changes and arrhythmias were not observed after SAV. Normal sinus rhythm was restored in Group 4. Scorpion envenoming with multi-system-organ failure (MSOF), characterized by a massive release of counter-regulatory hormones (catecholamines, glucagon, cortisol), angiotensin-II, and changes in insulin secretion, is a condition of fuel-energy deficits and an inability to utilize the existing metabolic substrates. These disturbances are reversed by SAV possibly through insulin release. It is concluded that SAV, under the laboratory conditions, effectively neutralizes, prevents, and reverses scorpion venom toxicity.
Abstract in English:The objective of this study was to detect the presence of local edema and hemorrhage caused by Crotalus durissus terrificus envenoming using magnetic resonance imaging (MRI). Six patients bitten by Crotalus durissus terrificus snakes were treated at the Emergency Unit and Tropical Diseases Unit of Botucatu School of Medicine, UNESP, São Paulo State, Brazil. After receiving specific serotherapy, the patients were submitted to MR of the bite site. Post contrast T1 and T2 spin-echo MRI were obtained revealing the following lesions: edema associated with hemorrhage in subcutaneous and muscular tissue (n=3), edema in subcutaneous tissue (n=2), and perimuscular hemorrhage (n=1). In this study, MR demonstrated a local effect of Crotalus durissus terrificus envenoming in inducing edema and hemorrhage, mainly in muscular tissues and perimuscular areas. These results indicate that Crotalus durissus terrificus venom cause a local muscular tissue damage in human envenoming represented by edema and hemorrhage.
Abstract in English:The pharmacological substances adenosine deaminase (ADA), histamine, and IgE are endogenously present in animals. They are implicated in allergy and asthma and detectable in blood serum. This research reports the presence of ADA, histamine, and IgE at varying levels in almost all major organs of mice. This research further reports that intramuscular injection of sub-lethal dose of Naja kaouthia venom disrupted homeostasis and lowered the levels of ADA, histamine, and IgE in the organs of mice. Adult Balb/C male mice were injected with a half lethal dose of the venom. The mice were sacrificed at 2, 8, and 24 hours post-injection and different organs were collected. Organs were homogenized, centrifuged, and the supernatants were assayed for ADA, histamine, and IgE using respective antisera by immunological test enzyme-linked immunosorbent assay (ELISA). Organs from mice injected with PBS served as controls. No major decrease in the levels of ADA, histamine, and IgE was observed after 2 hours of venom injection. However, tremendous decreases in the levels of ADA, histamine, and IgE was observed in organs 24 h post-injection. The highest decrease for ADA was observed in the brain, liver, lung, muscle, and testis; for histamine, in the heart, muscle, lung, and testis; and for IgE, in the bone, heart, lung, muscle, and testis. This is a first-hand investigation showing the effect of envenomation on the pharmacokinetics of ADA, histamine, and IgE in organs.
Abstract in English:Scorpion venom neurotoxins are responsible for toxicity and pharmacological effects. They are active in sodium and potassium channels leading to an increase in the release of neurotransmitters, such as glutamate. Glutamate is found in large quantities in the hippocampus (HPC) and is involved in the long-term potentiation (LTP) induction. The HPC is known to be related to certain kinds of memory. The aim of this study is to evaluate the effects of Tityus serrulatus TS-8F toxin on rat behavior with emphasis on learning and memory. We analyzed the effects of different doses of TS-8F on rat behavior in home cages, open-field (habituation), inhibitory avoidance, T-maze, and hippocampus morphology. In the first two experiments, 0.05µg/animal dose of TS-8F did not cause convulsion but led to a decrease in locomotion (LO) frequency in the open-field first session. During the second session, rats receiving 0.03µg/animal TS-8F showed a decrease in LO and rearing frequency (RE); controls only showed decreased LO; and those receiving 0.05µg/animal showed no significant changes. In inhibitory avoidance, T-maze, and HPC morphology experiments no significant differences were observed. It is concluded that TS-8F may exert some influence in rat learning and memory and seems to be useful as a pharmacological tool. Further research is required to elucidate all possible uses of this toxin.
Abstract in English:Snake venoms frequently vary in composition. In this work, we compared the neurotoxic and myotoxic activities of 16 lots of Bothrops neuwiedii venoms from different regions of Brazil, using chick biventer cervicis preparations. The neuromuscular blockade varied from 2% to 100% after 120 min incubation with venoms (50 mug/ml). In all cases, this blockade was irreversible and concentration-dependent; at low concentrations (10-20 mug/ml), 15 of the 16 venom lots failed to abolish responses to acetylcholine (110 muM), but blocked responses to KCl (13.4 muM), and induced contracture. At 5-20 mug/ml, the most active venom totally blocked twitch-tension without affecting responses to acetylcholine and KCl. Polyacrylamide gel electrophoresis for basic proteins showed that the most active samples contained a band that was absent in the less active venoms. These results indicate that there may be considerable intraspecific variation in the neurotoxic activity of B. neuwiedii venoms, whereas myotoxic activity is less variable.
Abstract in English:The objective of this clinical study was to evaluate local tissue damage caused by Bothrops sp envenoming in relation to lesion type and damaged tissues using magnetic resonance imaging (MRI). Fifteen patients bitten by Bothrops snakes were treated at the Emergency Unit of the Tropical Diseases Unit at the University Hospital, Botucatu School of Medicine, UNESP, São Paulo State, Brazil. After receiving specific serotherapy, the patients were submitted to MR of the bite site. T1 spin-echo MRI were obtained revealing the following lesions: edema (n=9), edema associated with hemorrhage (n=5), and hemorrhage (n=1). Perimuscular areas (n=6) and subcutaneous tissues (n=5) were the most affected, followed by muscular tissues (n=4). It is important to mention that MRI did not show myonecrosis of the bite site, a widely reported finding in anatomical and histopathological experimental studies.
Abstract in English:Clinical samples from 71 bovine from different Brazilian states were processed for the analysis of anaerobe organisms with emphasis on the isolation and characterization of Clostridium spp. From these, eighty-nine Clostridium perfringens strains were recovered: 32 from liver, 19 from intestinal contents, 14 from kidney, 6 from rumen, 5 from nervous system, 4 from bone marrow, 2 from udder tract, blood, spleen and lung, and one from muscle. Four reference Clostridium perfringens types A, B, C, and D were used as controls in this study. All isolates were cultivated in appropriate media, and after centrifugation the supernatant and sediment were separated. From pure supernatant post exopolysaccharide (EPS) extraction, mouse toxigenicity tests were performed, determining protein and protein plus carbohydrate, respectively. ELISA was performed from sediments. The results showed that 51 (57.3%) of the isolates were toxigenic to mice when inoculated by intraperitoneal route; bacteria from different organs had variable patterns of toxigenicity. Toxigenicity of EPS extracts was only expressed when protein concentration was 0.04 mg/mL and between 0.31 and 0.5 mg/mL for carbohydrate. Isolates were characterized as toxigenic when showing optimum protein and carbohydrate concentrations.
Abstract in English:Goat spermatozoa were incubated in vitro in Tris-citrate buffer, pH 7.2, containing 0, 40, 80, or 160 mug of Naja haje venom/mL buffer for 4 hours. During incubation, the percentages of sperm motility were decreased, while percentages of dead spermatozoa were increased in a time and concentration-dependent manner. The effect of venom concentrations on the ultrastructure of incubated spermatozoa was examined hourly by scanning (SEM) and transmission (TEM) electron microscopy. SEM results showed plasma membrane wrinkling at heads of some spermatozoa after 2 h incubation with 40mug venom. Most spermatozoa suffered membrane wrinkling after 4 h incubation. However, incubation with 80 mug venom caused membrane fractures in most sperm heads after 1 h incubation. The extent and depth of these fractures were increased after 2-3 h incubation. After 4 h incubation, plasma membrane focal erosion of many spermatozoa heads was common. Incubation with 160 mg venom induced sperm head swollen plasma membranes after 1 h incubation. Ruptured and disintegrated membranes were seen after 2 h; lysis and removal of external surface of spermatozoa head plasma membranes were recorded after 3-4 h incubation. TEM indicated slightly swollen areas on the sperm head plasma membrane, but showed normal nuclei, acrosomes, and tail regions after 2 h incubation in 40 mug cobra venom. The swollen areas were accompanied by sperm head membrane disintegration as well as membrane irregularities and distortion of tail mitochondrial cristae after 3-4 h incubation. However, incubation with 80 mug venom showed focal areas of membrane lysis and discontinuity in the sperm heads and tails increasing with incubation time. Severe axoneme and tail longitudinal fiber degeneration and increased numbers of distorted mitochondrial cristae were also observed after 3-4 h incubation. Spermatozoa incubation with 160 mug venom increased severity of plasma membrane dissolution, disintegration, and rupture. Axoneme disorganization and tail longitudinal fiber fusion were seen after 1-2 h; complete deformation of the motility system was recorded after 3-4 h incubation. Nuclei were normal, but acrosomes were partially damaged, and distorted mitochondrial cristae were increased after 4 h incubation. The results indicated that cobra venom induced a concentration and time-dependent alterations on spermatozoa plasma membrane as well as obvious deformation on the motility system and tail mitochondria, which are responsible for sperm viability and fertilizing ability.
Abstract in English:Micrurus spixii venom was studied after fractionation by Sephadex G-100 SF gel filtration chromatography. Several enzymatic activities and biological effects were investigated in whole venom and fractions. The venom was resolved in four peaks in a range of about 73.2-10.7 kDa molecular weight. Alkaline phosphatase and acetylcholinesterase activities were found in peak I, and procoagulant activity was seen in peak II. Phospholipase A2, hemorrhagic, and proteolytic activities were detected in peak III. A second procoagulant factor and proteinase were present in peak IV. Thrombin-like enzyme and direct hemolytic activities were not found in any assayed samples.
Abstract in English:This study reports the isolation of an Ophidian Paramyxovirus (OPMV) in sputum of a captive rattlesnake (Crotalus durissus terrificus) kept in a serpentarium located in Botucatu, São Paulo State, Brazil. Polymerase chain reaction (PCR) and nested-PCR were performed for the identification of the isolated virus.
Abstract in English:This report refers to a female Pinscher treated at the Veterinary Hospital of the Botucatu School of Veterinary Medicine and Animal Husbandry (FMVZ)-São Paulo State University (UNESP), Botucatu, 14 hours after a suspected spider envenoming. The animal showed spastic limbs, dyspnea, salivation, hypothermia, muscular fasciculation, non-responsive mydriasis, and claudication of the right hindlimb. The spider was later identified by the Center for the Study of Venoms and Venomous Animals-CEVAP/UNESP, Botucatu as Phoneutria nigriventer. The treatment with potent analgesic was efficient.