Rotifers of Bahia State, Brazil: news records and limitations to studies / Rotíferos do Estado da Bahia, Brasil: novos registros e limitações dos estudos

A first checklist of Rotifera species in freshwater environments in Bahia State, in northeastern Brazil, is provided. The list includes sampling data from 26 aquatic environments (lotic and lentic) undertaken from 2010 to 2016. One hundred and fifty-five species were recorded, with 68 new records for the state. The family Brachionidae and Lecanidae were the most representative (54.8%). The greatest richness was recorded in the Colônia River (57 species). Those results reflect the low numbers of studies previously undertaken in the region, indicating more research needs to be focused on Rotifera biodiversity in Bahia, the fifth largest state in Brazil (567,295 km2) with large numbers of freshwater bodies.

É fornecida uma primeira lista de verificação das espécies de Rotifera em ambientes de água doce no Estado da Bahia, nordeste do Brasil. A lista de espécies inclui dados de amostragem de 26 ambientes aquáticos (lóticos e lênticos) de 2010 a 2016. Cento e cinquenta e cinco espécies foram registradas, com 68 novos registros para o estado. As famílias Brachionidae e Lecanidae foram as mais representativas (54,8%). A maior riqueza foi registrada no rio Colônia (57 espécies). Esses resultados provavelmente refletem o número de estudos na região, sugerindo mais pesquisas sobre a biodiversidade de Rotifera na Bahia, o quinto maior estado do Brasil (567.295 km2) com grande número de corpos aquáticos de água doce.

Rotifers of Bahia State, Brazil: News records and limitations to studies

Abstract A first checklist of Rotifera species in freshwater environments in Bahia State, in northeastern Brazil, is provided. The list includes sampling data from 26 aquatic environments (lotic and lentic) undertaken from 2010 to 2016. One hundred and fifty-five species were recorded, with 68 new records for the state. The family Brachionidae and Lecanidae were the most representative (54.8%). The greatest richness was recorded in the Colônia River (57 species). Those results reflect the low numbers of studies previously undertaken in the region, indicating more research needs to be focused on Rotifera biodiversity in Bahia, the fifth largest state in Brazil (567,295 km2) with large numbers of freshwater bodies.

Resumo É fornecida uma primeira lista de verificação das espécies de Rotifera em ambientes de água doce no Estado da Bahia, nordeste do Brasil. A lista de espécies inclui dados de amostragem de 26 ambientes aquáticos (lóticos e lênticos) de 2010 a 2016. Cento e cinquenta e cinco espécies foram registradas, com 68 novos registros para o estado. As famílias Brachionidae e Lecanidae foram as mais representativas (54,8%). A maior riqueza foi registrada no rio Colônia (57 espécies). Esses resultados provavelmente refletem o número de estudos na região, sugerindo mais pesquisas sobre a biodiversidade de Rotifera na Bahia, o quinto maior estado do Brasil (567.295 km2) com grande número de corpos aquáticos de água doce.

Rotifers of Bahia State, Brazil: News records and limitations to studies / Rotíferos do Estado da Bahia, Brasil: Novos registros e limitações dos estudos

A first checklist of Rotifera species in freshwater environments in Bahia State, in northeastern Brazil, is provided. The list includes sampling data from 26 aquatic environments (lotic and lentic) undertaken from 2010 to 2016. One hundred and fifty-five species were recorded, with 68 new records for the state. The family Brachionidae and Lecanidae were the most representative (54.8%). The greatest richness was recorded in the Colônia River (57 species). Those results reflect the low numbers of studies previously undertaken in the region, indicating more research needs to be focused on Rotifera biodiversity in Bahia, the fifth largest state in Brazil (567,295 km2) with large numbers of freshwater bodies.

É fornecida uma primeira lista de verificação das espécies de Rotifera em ambientes de água doce no Estado da Bahia, nordeste do Brasil. A lista de espécies inclui dados de amostragem de 26 ambientes aquáticos (lóticos e lênticos) de 2010 a 2016. Cento e cinquenta e cinco espécies foram registradas, com 68 novos registros para o estado. As famílias Brachionidae e Lecanidae foram as mais representativas (54,8%). A maior riqueza foi registrada no rio Colônia (57 espécies). Esses resultados provavelmente refletem o número de estudos na região, sugerindo mais pesquisas sobre a biodiversidade de Rotifera na Bahia, o quinto maior estado do Brasil (567.295 km2) com grande número de corpos aquáticos de água doce.

Rotifers of Bahia State, Brazil: News records and limitations to studies.

A first checklist of Rotifera species in freshwater environments in Bahia State, in northeastern Brazil, is provided. The list includes sampling data from 26 aquatic environments (lotic and lentic) undertaken from 2010 to 2016. One hundred and fifty-five species were recorded, with 68 new records for the state. The family Brachionidae and Lecanidae were the most representative (54.8%). The greatest richness was recorded in the Colônia River (57 species). Those results reflect the low numbers of studies previously undertaken in the region, indicating more research needs to be focused on Rotifera biodiversity in Bahia, the fifth largest state in Brazil (567,295 km2) with large numbers of freshwater bodies.

Changes in the composition of ichthyoplankton assemblage and plastic debris in mangrove creeks relative to moon phases.

Lunar influence on the distribution of fish larvae, zooplankton and plastic debris in mangrove creeks of the Goiana Estuary, Brazil, was studied over a lunar cycle. Cetengraulis edentulus, Anchovia clupeoides and Rhinosardinia bahiensis were the most abundant fish larvae (56·6%), independent of the moon phase. The full moon had a positive influence on the abundance of Gobionellus oceanicus, Cynoscion acoupa and Atherinella brasiliensis, and the new moon on Ulaema lefroyi. The full and new moons also influenced the number of zoeae and megalopae of Ucides cordatus, protozoeae and larvae of caridean shrimps, and the number of hard and soft plastic debris, both <5 and >5 mm. Micro and macroplastics were present in samples from all 12 creeks studied, at densities similar to the third most abundant taxon, R. bahiensis. Cetengraulis edentulus and R. bahiensis showed a strong positive correlation with the last quarter moon, when there was less zooplankton available in the creeks and higher abundance of microplastic threads. Anchovia clupeoides, Diapterus rhombeus, U. lefroyi and hard microplastics were positively associated with different moon phases, when calanoid copepods, Caridean larvae and zoeae of U. cordatus were highly available in the creeks. Cynoscion acoupa, G. oceanicus and A. brasiliensis were strongly associated with the full moon, when protozoeae of caridean shrimps and megalopae of U. cordatus were also highly available, as were hard and soft macroplastics, paint chips (<5 mm) and soft microplastics. The results reinforce the role of mangrove creeks as nursery habitats. The moon phases influenced the distribution of fish larvae species, zooplankton and plastic debris by changing their compositions and abundances in the mangrove creeks of the Goiana Estuary when under the influence of different tidal current regimes.

Extracts of Caesalpinia ferrea and Trichoderma sp. on the control of Colletotrichum sp. transmission in Sideroxylon obtusifolium seeds / Extratos de Caesalpinia ferrea e Trichoderma spp. no controle da tramissibilidade de Colletotrichum sp. em sementes de Sideroxylon obtusifolium (Sapotaceae)

ABSTRACT Recent research reports the importance of preserving plants in Brazilian semiarid regions, in this context, the scientific literature has reported different pharmacological studies from plant extracts with an antifungal potential, coming from forest species that can contribute as a control and management strategy in the transmission of phytopathogens. This study aimed to evaluate the effect of biotech treatments in controlling the transmission of Colletotrichum sp. in seeds of S. obtusifolium. In this study, 100 seeds were subjected to the following preventive treatments: fungicide Captan®, extract of Caesalpinia ferrea Mart. Ex. Tul., and biological control with Trichoderma spp. The biological control with Trichoderma spp. and the alternative control using C. ferrea extract provided a greater protection to seeds and seedlings of S. obtusifolium facing the transmissibility of Colletotrichum sp.The treatment based on plant extract is more efficient for this purpose only in large seeds and does not interfere on the germination percentage and speed. Therefore it is necessary to perform other studies with Trichoderma spp. and C. ferrea extract to test different doses of these products.

RESUMO Recentes pesquisas relatam a importância da preservação de plantas do semiárido brasileiro. Neste contexto, a literatura científica tem relatado diferentes estudos farmacológicos com extratos vegetais com potencial antifúngico proveniente de espécies florestais que podem contribuir como estratégia de controle e gerenciamento na transmissão de fitopatógenos. No presente estudo o objetivo foi avaliar o efeito de tratamentos biotecnológicos no controle da transmissibilidade de Colletotrichum sp. em sementes de S. obtusifolium. Neste estudo foram utilizadas 100 sementes submetidas aos seguintes tratamentos preventivos: fungicida Captan®, extrato de Caesalpinia ferrea Mart. Ex. Tul. e controle biológico com Trichoderma spp. O controle biológico com Trichoderma spp. e o alternativo com extrato de C. ferrea proporcionam maior proteção às sementes e plântulas S. obtusifolium quanto a transmissibilidade do Colletotrichum sp. O tratamento à base de extrato vegetal foi o mais eficiente para este fim, apenas em sementes de maior tamanho, por não interferir na porcentagem e velocidade de germinação. Portanto, faz-se necessário à realização de outros trabalhos com Trichoderma spp. e extrato de C. ferrea para testar doses diferentes desses produtos.

Pattern of inflammatory response to Loxosceles intermedia venom in distinct mouse strains: a key element to understand skin lesions and dermonecrosis by poisoning.

Envenomation caused by spiders Loxosceles induce intense dermonecrosis at the bite site and systemic disease. In this work we described the hyaluronidase and collagenase activities in vitro of the Loxosceles intermedia venom, but no phospholipase A2 activity. In vivo, we evaluated the effect of L. intermedia venom used different strain of mice, C57BL/6, BALB/c and Swiss. All mice developed paw edema after venom injection, persistent for 24 h in BALB/c and C57BL/6 mice. Histopathological analysis of the skin after venom injection revealed vascular congestion in Swiss mice and an inflammatory reaction in BALB/c and C57BL/6 mice. The mobilization of inflammatory cells from bone marrow, spleen and blood was investigated. Typical innate immune response with mobilization of myeloid cells and cytotoxic CD8 T lymphocytes was observed in C57BL/6 mice. In contrast, typical acquired/humoral immune response was observed in BALB/c mice, with preferential involvement of conventional B lymphocytes and CD4 T helper cells. The skin inflammation associated to mobilization of inflammatory cells indicated that mice models are strongly recommended to investigate specific cell types involved with immune response to the envenomation and mechanisms to inhibit skin lesions.

A comparison of virulence patterns and in vivo fitness between hospital- and community-acquired methicillin-resistant Staphylococcus aureus related to the USA400 clone.

Methicillin-resistant Staphylococcus aureus (MRSA) isolates genetically related to the CA-MRSA clone MW2/USA400 (ST1-SCCmecIV lineage) from the United States have emerged in hospitals in Rio de Janeiro and are associated with nosocomial bloodstream infections. To understand the virulence mechanisms involved in the adaptability of ST1 isolates as a hospital pathogen in Rio de Janeiro, we compared the virulence traits and fitness properties of the Brazilian isolates with those displayed by the CA-MRSA isolates from the United States. Similar to the USA400 from the United States, all the Brazilian isolates tested carried the genes encoding SEH and LukDE. In contrast, none of the Brazilian isolates carried the lukSF PVL, sea, sec, and sek genes. Competition experiments in mice demonstrated a significant increase in the fitness for the CA-MRSA isolates MW2 and USA400-0051 from the United States compared to other isolates. In the foreign body animal model, 83 % more North-American bacterial cells were recovered compared to the Brazilian ST1 isolates. Differences in gene expression of important virulence factors were detected. Transcription of rnaIII and psmα3 was increased about two-fold in the isolates from the United States, and sasG about two-fold in the Brazilian isolates. Thus, it is possible that the virulence attenuation observed among the Brazilian hospital isolates, associated with the acquisition of multiple resistant determinants, are consequences of microevolutionary events that contributed to the necessary fitness adjustment of this lineage, allowing a typically community-acquired MRSA (MW2/USA400) to emerge as a successful hospital pathogen (Brazilian ST1-SCCmecIV).

Double gene therapy with granulocyte colony-stimulating factor and vascular endothelial growth factor acts synergistically to improve nerve regeneration and functional outcome after sciatic nerve injury in mice.

Peripheral-nerve injuries are a common clinical problem and often result in long-term functional deficits. Reconstruction of peripheral-nerve defects is currently undertaken with nerve autografts. However, there is a limited availability of nerves that can be sacrificed and the functional recovery is never 100% satisfactory. We have previously shown that gene therapy with vascular endothelial growth factor (VEGF) significantly improved nerve regeneration, neuronal survival, and muscle activity. Our hypothesis is that granulocyte colony-stimulating factor (G-CSF) synergizes with VEGF to improve the functional outcome after sciatic nerve transection. The left sciatic nerves and the adjacent muscle groups of adult mice were exposed, and 50 or 100 µg (in 50 µl PBS) of VEGF and/or G-CSF genes was injected locally, just below the sciatic nerve, and transferred by electroporation. The sciatic nerves were transected and placed in an empty polycaprolactone (PCL) nerve guide, leaving a 3-mm gap to challenge nerve regeneration. After 6 weeks, the mice were perfused and the sciatic nerve, the dorsal root ganglion (DRG), the spinal cord and the gastrocnemius muscle were processed for light and transmission electron microscopy. Treated animals showed significant improvement in functional and histological analyses compared with the control group. However, the best results were obtained with the G-CSF+VEGF-treated animals: quantitative analysis of regenerated nerves showed a significant increase in the number of myelinated fibers and blood vessels, and the number of neurons in the DRG and motoneurons in the spinal cord was significantly higher. Motor function also showed that functional recovery occurred earlier in animals receiving G-CSF+VEGF-treatment. The gastrocnemius muscle showed an increase in weight and in the levels of creatine phosphokinase, suggesting an improvement of reinnervation and muscle activity. These results suggest that these two factors acted synergistically and optimized the nerve repair potential, improving regeneration after a transection lesion.

Pulsed ultrasound therapy accelerates the recovery of skeletal muscle damage induced by Bothrops jararacussu venom

We studied the effect of pulsed ultrasound therapy (UST) and antibothropic polyvalent antivenom (PAV) on the regeneration of mouse extensor digitorum longus muscle following damage by Bothrops jararacussu venom. Animals (Swiss male and female mice weighing 25.0 ± 5.0 g; 5 animals per group) received a perimuscular injection of venom (1 mg/kg) and treatment with UST was started 1 h later (1 min/day, 3 MHz, 0.3 W/cm², pulsed mode). Three and 28 days after injection, muscles were dissected and processed for light microscopy. The venom caused complete degeneration of muscle fibers. UST alone and combined with PAV (1.0 mL/kg) partially protected these fibers, whereas muscles receiving no treatment showed disorganized fascicules and fibers with reduced diameter. Treatment with UST and PAV decreased the effects of the venom on creatine kinase content and motor activity (approximately 75 and 48%, respectively). Sonication of the venom solution immediately before application decreased the in vivo and ex vivo myotoxic activities (approximately 60 and 50%, respectively). The present data show that UST counteracts some effects of B. jararacussu venom, causing structural and functional improvement of the regenerated muscle after venom injury.

Pulsed ultrasound therapy accelerates the recovery of skeletal muscle damage induced by Bothrops jararacussu venom.

We studied the effect of pulsed ultrasound therapy (UST) and antibothropic polyvalent antivenom (PAV) on the regeneration of mouse extensor digitorum longus muscle following damage by Bothrops jararacussu venom. Animals (Swiss male and female mice weighing 25.0 ± 5.0 g; 5 animals per group) received a perimuscular injection of venom (1 mg/kg) and treatment with UST was started 1 h later (1 min/day, 3 MHz, 0.3 W/cm(2), pulsed mode). Three and 28 days after injection, muscles were dissected and processed for light microscopy. The venom caused complete degeneration of muscle fibers. UST alone and combined with PAV (1.0 mL/kg) partially protected these fibers, whereas muscles receiving no treatment showed disorganized fascicules and fibers with reduced diameter. Treatment with UST and PAV decreased the effects of the venom on creatine kinase content and motor activity (approximately 75 and 48%, respectively). Sonication of the venom solution immediately before application decreased the in vivo and ex vivo myotoxic activities (approximately 60 and 50%, respectively). The present data show that UST counteracts some effects of B. jararacussu venom, causing structural and functional improvement of the regenerated muscle after venom injury.

Enhancement of sciatic nerve regeneration after vascular endothelial growth factor (VEGF) gene therapy.

AIMS: Recent studies have emphasized the beneficial effects of the vascular endothelial growth factor (VEGF) on neurone survival and Schwann cell proliferation. VEGF is a potent angiogenic factor, and angiogenesis has long been recognized as an important and necessary step during tissue repair. Here, we investigated the effects of VEGF on sciatic nerve regeneration. METHODS: Using light and electron microscopy, we evaluated sciatic nerve regeneration after transection and VEGF gene therapy. We examined the survival of the neurones in the dorsal root ganglia and in lumbar 4 segment of spinal cord. We also evaluated the functional recovery using the sciatic functional index and gastrocnemius muscle weight. In addition, we evaluated the VEGF expression by immunohistochemistry. RESULTS: Fluorescein isothiocyanate-dextran (FITC-dextran) fluorescence of nerves and muscles revealed intense staining in the VEGF-treated group. Quantitative analysis showed that the numbers of myelinated fibres and blood vessels were significantly higher in VEGF-treated animals. VEGF also increased the survival of neurone cell bodies in dorsal root ganglia and in spinal cord. The sciatic functional index and gastrocnemius muscle weight reached significantly higher values in VEGF-treated animals. CONCLUSION: We demonstrate a positive relationship between increased vascularization and enhanced nerve regeneration, indicating that VEGF administration can support and enhance the growth of regenerating nerve fibres, probably through a combination of angiogenic, neurotrophic and neuroprotective effects.

Mesenchymal stem cells in a polycaprolactone conduit enhance median-nerve regeneration, prevent decrease of creatine phosphokinase levels in muscle, and improve functional recovery in mice.

Although the majority of peripheral-nerve regeneration studies are carried out on the sciatic nerve, lesions of the upper extremities are more common in humans and usually lead to significant physical disabilities. The present study was driven by the hypothesis that a combination of strategies, namely grafts of mesenchymal stem cells (MSC) and resorbable polycaprolactone (PCL) conduits would improve median-nerve regeneration after transection. Mouse median nerves were transected and sutured to PCL tubes that were filled with either green fluorescent protein (GFP(+)) MSC in DMEM or with DMEM alone. During the post-operative period, animals were tested weekly for flexor digitorum muscle function by means of the grasping test. After 8 weeks, the proximal and middle portions of the PCL tube and the regenerating nerves were harvested and processed for light and electron microscopy. The flexor digitorum muscle was weighed and subjected to biochemical analysis for creatine phosphokinase (CK) levels. Scanning electron microscopy of the PCL tube 8 weeks after implantation showed clear signs of wall disintegration. MSC-treated animals showed significantly larger numbers of myelinated and unmyelinated nerve fibers and blood vessels compared with DMEM-treated animals. The flexor digitorum muscle CK levels were significantly higher in the MSC-treated animals, but muscle weight values did not differ between the groups. Compared with the DMEM-treated group, MSC-treated animals showed, by the grasping test, improved functional performance throughout the period analyzed. Immunofluorescence for S-100 and GFP showed, in a few cases, double-labeled cells, suggesting that transplanted cells may occasionally transdifferentiate into Schwann cells. Our data demonstrate that the polycaprolactone conduit filled with MSC is capable of significantly improving the median-nerve regeneration after a traumatic lesion.

Effect of perimuscular injection of Bothrops jararacussu venom on plasma creatine kinase levels in mice: influence of dose and volume.

The effect of dose and volume of a perimuscular injection of Bothrops jararacussu venom on myonecrosis of skeletal muscle was studied in mice. An increase of the venom dose (0.25 to 2.0 micro g/g) at a given volume (50 micro l) resulted in an increase in plasma creatine kinase (CK) levels 2 h after injection. Plasma CK activity increased from the basal level of 129.27 +/- 11.83 (N = 20) to 2392.80 +/- 709.43 IU/l (N = 4) for the 1.0 micro g/g dose. Histological analysis of extensor digitorum longus muscle 4 h after injection showed lesion of peripheral muscle fibers, disorganization of the bundles or the complete degeneration of muscle fibers. These lesions were more extensive when higher doses were injected. Furthermore, an increase in volume (12.5 to 100 micro l) by dilution of a given dose (0.5 micro g/g) also increased plasma CK levels from 482.31 +/- 122.79 to 919.07 +/- 133.33 IU/l (N = 4), respectively. These results indicate that care should be taken to standardize volumes and sites of venom injections.

Effect of perimuscular injection of Bothrops jararacussu venom on plasma creatine kinase levels in mice: influence of dose and volume

The effect of dose and volume of a perimuscular injection of Bothrops jararacussu venom on myonecrosis of skeletal muscle was studied in mice. An increase of the venom dose (0.25 to 2.0 æg/g) at a given volume (50 æl) resulted in an increase in plasma creatine kinase (CK) levels 2 h after injection. Plasma CK activity increased from the basal level of 129.27 ± 11.83 (N = 20) to 2392.80 ± 709.43 IU/l (N = 4) for the 1.0 æg/g dose. Histological analysis of extensor digitorum longus muscle 4 h after injection showed lesion of peripheral muscle fibers, disorganization of the bundles or the complete degeneration of muscle fibers. These lesions were more extensive when higher doses were injected. Furthermore, an increase in volume (12.5 to 100 æl) by dilution of a given dose (0.5 æg/g) also increased plasma CK levels from 482.31 ± 122.79 to 919.07 ± 133.33 IU/l (N = 4), respectively. These results indicate that care should be taken to standardize volumes and sites of venom injections

Effect of suramin on myotoxicity of some crotalid snake venoms.

We investigated the protective effect of suramin, an enzyme inhibitor and an uncoupler of G protein from receptors, on the myotoxic activity in mice of different crotalid snake venoms (A.c. laticinctus, C.v. viridis, C.d. terrificus, B. jararacussu, B. moojeni, B. alternatus, B. jararaca, L. muta). Myotoxicity was evaluated in vivo by injecting im the venoms (0.5 or 1.0 mg/kg) dissolved in physiological saline solution (0.1 ml) and measuring plasma creatine kinase (CK) activity. Two experimental approaches were used in mice (N = 5 for each group). In protocol A, 1 mg of each venom was incubated with 1.0 mg suramin (15 min, 37 degrees C, in vitro), and then injected im into the mice at a dose of 1.0 mg/kg (in vivo). In protocol B, venoms, 1.0 mg/kg, were injected im 15 min prior to suramin (1.0 mg/kg, iv). Before and 2 h after the im injection blood was collected by orbital puncture. Plasma was separated and stored at 4 degrees C for determination of CK activity using a diagnostic kit from Sigma. Preincubation of some venoms (C.v. viridis, A.c. laticinctus, C.d. terrificus and B. jararacussu) with suramin reduced (37-76%) the increase in plasma CK, except for B. alternatus, B. jararaca or L. muta venoms. Injection of suramin after the venom partially protected (34-51%) against the myotoxicity of B. jararacussu, A.c. laticinctus and C.d. terrificus venom, and did not protect against C.v. viridis, L. muta, B. moojeni, B. alternatus or B. jararaca venoms. These results show that suramin has an antimyotoxic effect against some, but not all the North and South American crotalid snake venoms studied here.

Effect of suramin on myotoxicity of some crotalid snake venoms

We investigated the protective effect of suramin, an enzyme inhibitor and an uncoupler of G protein from receptors, on the myotoxic activity in mice of different crotalid snake venoms (A.c. laticinctus, C.v. viridis, C.d. terrificus, B. jararacussu, B. moojeni, B. alternatus, B. jararaca, L. muta). Myotoxicity was evaluated in vivo by injecting im the venoms (0.5 or 1.0 mg/kg) dissolved in physiological saline solution (0.1 ml) and measuring plasma creatine kinase (CK) activity. Two experimental approaches were used in mice (N = 5 for each group). In protocol A, 1 mg of each venom was incubated with 1.0 mg suramin (15 min, 37ºC, in vitro), and then injected im into the mice at a dose of 1.0 mg/kg (in vivo). In protocol B, venoms, 1.0 mg/kg, were injected im 15 min prior to suramin (1.0 mg/kg, iv). Before and 2 h after the im injection blood was collected by orbital puncture. Plasma was separated and stored at 4ºC for determination of CK activity using a diagnostic kit from Sigma. Preincubation of some venoms (C.v. viridis, A.c. laticinctus, C.d. terrificus and B. jararacussu) with suramin reduced (37-76 percent) the increase in plasma CK, except for B. alternatus, B. jararaca or L. muta venoms. Injection of suramin after the venom partially protected (34-51 percent) against the myotoxicity of B. jararacussu, A.c. laticinctus and C.d. terrificus venom, and did not protect against C.v. viridis, L. muta, B. moojeni, B. alternatus or B. jararaca venoms. These results show that suramin has an antimyotoxic effect against some, but not all the North and South American crotalid snake venoms studied here

Effect of heparin and antivenom on skeletal muscle damage produced by Bothrops jararacussu venom.

We examined the effect of treatment with heparin and polyvalent antivenom on mice muscle Extensor digitorum longus (EDL) regeneration, after damage induced by injection of Bothrops jararacussu crude venom over the muscle of the right posterior limb. The mice were separated into groups and each group received treatment, by intravenous route with either high molecular weight heparin (H), low molecular weight heparin (LMWH), polyvalent antivenom (PAV) or with the combination of PAV plus H or PAV plus LMWH at 15 minutes and 4 hours after the injection of the venom. Myotoxicity was measured by the increase in plasma creatine kinase (CK) activity at two hours after the injection of the venom. The histological changes in EDL at 1, 3, 7 and 21 days after the injection of the venom were analyzed by light microscopy. In each group the normal and regenerated muscle fibers were quantified using Scion Image computer program. We also evaluated in vitro, the influence of these substances in the proteolytic and phospholipase activities of the venom. Heparins decreased the proteolytic activity of the venom but did not affect its phospholipase activity. However the PAV antagonized both activities. PAV and its combinations showed antimyotoxic activity, according to the magnitude of CK plasma levels. At 21 days the regeneration was observed in all animals, also in those that received only the venom. All treatments, except LMWH, promote a significant increase in the number of muscle fibers.

A novel thienylhydrazone, (2-thienylidene)3,4-methylenedioxybenzoylhydrazine, increases inotropism and decreases fatigue of skeletal muscle.

This study was designed to investigate the effects on single skeletal muscle fibers of a novel thienylhydrazone, referred to as LASSBio-294, which is a bioisoster of pyridazinone compounds that inhibit the cyclic AMP-specific phosphodiesterase (PDE) 4. Twitch and fatigue were analyzed in single skeletal muscle fibers isolated from either the semitendinous or the tibialis anterior muscles dissected from the frog Rana pipiens. LASSBio-294 (12.5-100 microM) increased twitch tension, accelerated the maximal rate of tension decay during relaxation, and had very little effect in the maximal rate of tension development of muscle fibers directly stimulated at < or =30 Hz. The positive inotropic effect of LASSBio-294 developed slowly, reaching its maximum at 40 min and was inversely proportional to the frequency of stimulation, becoming negligible at 60 and 90 Hz. The concentration-response relationship for LASSBio-294-induced potentiation of twitch tension was bell-shaped, with maximal effect occurring at 25 microM. In addition, LASSBio-294 reduced development of fatigue induced by tetanic stimulation of the muscle fibers and reduced the time needed for 80% prefatigue tension recovery after fatigue had developed to 50% of the maximal pretetanic force. These effects of LASSBio-294 can be fully explained by stimulation of the sarcoplasmic reticulum Ca2+ pump and could be ascribed to an increase in cellular levels of cyclic AMP due to PDE inhibition. The novel thienylhydrazone LASSBio-294 may be useful for treatment of patients suffering from conditions in which muscle fatigue is a debilitating symptom (e.g., chronic heart failure).