Snakebite Clinics and Pathogenesis: From Preclinical to Resource Mapping Studies

A special issue of Toxins (ISSN 2072-6651). This special issue belongs to the section "Animal Venoms".

Deadline for manuscript submissions: closed (31 December 2022) | Viewed by 28750

Special Issue Editors


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Guest Editor
Medical School, Universidade Federal de Roraima, Boa Vista, Brazil
Interests: immunotoxicology, especially regarding the discovery of human recombinant antivenoms and bioprospection of venom-derived peptides, such as scorpion neurotoxins and snake enzymes; clinical projects in the main hospital of Roraima aiming to understand the severity and pathological effects of snakebite envenomings—Snakebite Roraima
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Guest Editor
Fundação de Medicina Tropical Doutor Heitor Vieira Dourado and the Escola Superior de Ciências da Saúde, Universidade do Estado do Amazonas, Manaus, Brazil
Interests: G6PD deficiency; diagnostics; P. vivax epidemiology
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Guest Editor
Instituto Butantan, Sao Paulo, Brazil
Interests: snakebite envenomation; risk factors; antivenom treatment; snake venom; toxin inhibitors; novel molecules

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Guest Editor
Laboratório de Imunopatologia, Instituto Butantan, São Paulo 05503-900, Brazil
Interests: snake venoms; metalloproteinases; hemostasis; venom composition; venom variability; antivenoms; immunoassays
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear colleagues,

The issue of snakebite envenoming has been overlooked for many years, despite 2.7 million incidences of envenoming and about 81,000-138,000 deaths each year. In 2017, the World Health Organization (WHO) recognized the enormous health and economic impact associated with snakebite envenoming and consequently reintroduced it into its list of Category A Neglected Tropical Diseases (NTDs). Moreover, in 2019, WHO developed a strategy to reduce mortality and disability from snakebite enve­noming by 50% before 2030. This initiative has fostered growing interest from research agendas and public health authorities, and a diverse set of actions are currently taking place to better understand and reduce the impact of this disease. Clinical toxicology research groups, more specifically Toxinologists, are focusing their efforts on improving the understanding of various aspects of snakebite envenomings, ranging from basic biochemical and pharmacological studies to clinical aspects of envenoming and its therapies.

 

This Special Issue has one main goal: to disseminate knowledge of snakebite envenoming at a clinical level. We welcome expert reviews of clinical aspects of snakebites, as well as research papers on the following sub-topics:

 

- Clinical studies exploring snakebite envenomations;

- Immune response to snakebite envenomation and diagnostic tests;

- Burden and risk factors for different local and systemic manifestations of snakebite envenomations;

- Mapping resources for snakebite envenomation management;

- First aid/ prehospital care to reduce severity of local and systemic snakebite envenomations;

- One Health approach for snakebite envenomations;

- Relevant snakebite envenomation care and education packages;

- Innovative strategies to improve antivenom production and quality;

- Novel molecules able to neutralize venoms and toxins;

- Preclinical and clinical studies on the efficacy of immunoglobulins, toxin inhibitors and other therapies.

We look forward to receiving your contributions.

Dr. Manu Pucca
Dr. Wuelton M. Monteiro
Dr. Hui Wen Fan
Dr. Ana Maria Moura-Da-Silva
Guest Editors

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Keywords

  • snakebite envenomation
  • risk factors
  • health policies
  • prehospital care
  • antivenom treatment
  • snake venom
  • toxin inhibitors
  • novel molecules

Published Papers (10 papers)

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Research

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19 pages, 1146 KiB  
Article
“Two Cultures in Favor of a Dying Patient”: Experiences of Health Care Professionals Providing Snakebite Care to Indigenous Peoples in the Brazilian Amazon
by Felipe Murta, Eleanor Strand, Altair Seabra de Farias, Felipe Rocha, Alícia Cacau Santos, Evellyn Antonieta Tomé Rondon, Ana Paula Silva de Oliveira, Hiran Satiro Souza da Gama, Yasmim Vieira Rocha, Gisele dos Santos Rocha, Mena Ferreira, Vinícius Azevedo Machado, Marcus Lacerda, Manuela Pucca, Felipe Cerni, João Ricardo Nickenig Vissoci, Anna Tupetz, Charles J. Gerardo, Ana Maria Moura-da-Silva, Fan Hui Wen, Jacqueline Sachett and Wuelton Monteiroadd Show full author list remove Hide full author list
Toxins 2023, 15(3), 194; https://doi.org/10.3390/toxins15030194 - 03 Mar 2023
Cited by 9 | Viewed by 2083
Abstract
In the Brazilian Amazon, deaths and disabilities from snakebite envenomations (SBEs) are a major and neglected problem for the indigenous population. However, minimal research has been conducted on how indigenous peoples access and utilize the health system for snakebite treatment. A qualitative study [...] Read more.
In the Brazilian Amazon, deaths and disabilities from snakebite envenomations (SBEs) are a major and neglected problem for the indigenous population. However, minimal research has been conducted on how indigenous peoples access and utilize the health system for snakebite treatment. A qualitative study was conducted to understand the experiences of health care professionals (HCPs) who provide biomedical care to indigenous peoples with SBEs in the Brazilian Amazon. Focus group discussions (FGDs) were carried out in the context of a three-day training session for HCPs who work for the Indigenous Health Care Subsystem. A total of 56 HCPs participated, 27 in Boa Vista and 29 in Manaus. Thematic analysis resulted in three key findings: Indigenous peoples are amenable to receiving antivenom but not to leaving their villages for hospitals; HCPs require antivenom and additional resources to improve patient care; and HCPs strongly recommend a joint, bicultural approach to SBE treatment. Decentralizing antivenom to local health units addresses the central barriers identified in this study (e.g., resistance to hospitals, transportation). The vast diversity of ethnicities in the Brazilian Amazon will be a challenge, and additional studies should be conducted regarding preparing HCPs to work in intercultural contexts. Full article
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15 pages, 1949 KiB  
Article
Comparison of Protein Variation in Protobothrops mucrosquamatus Venom between Northern and Southeast Taiwan and Association with Human Envenoming Effects
by Liao-Chun Chiang, Kun-Yi Chien, Hung-Yuan Su, Yen-Chia Chen, Yan-Chiao Mao and Wen-Guey Wu
Toxins 2022, 14(9), 643; https://doi.org/10.3390/toxins14090643 - 18 Sep 2022
Cited by 7 | Viewed by 2078
Abstract
Reports of bite from Protobothrops mucrosquamatus (Pmu) are frequent in Taiwan, and its wide-spread distribution and diverse habitats drove us to investigate its envenoming effects and relevant venom variations. We used reversed-phase high-performance liquid chromatography and mass spectrometry to analyze 163 [...] Read more.
Reports of bite from Protobothrops mucrosquamatus (Pmu) are frequent in Taiwan, and its wide-spread distribution and diverse habitats drove us to investigate its envenoming effects and relevant venom variations. We used reversed-phase high-performance liquid chromatography and mass spectrometry to analyze 163 Pmu venom samples collected from northern and southeastern Taiwan. Twenty-two major protein fractions were separated and analyzed, and their contents were determined semi-quantitatively. The results showed that despite the trivial differences in the protein family, there is an existing variation in acidic phospholipases A2s, serine proteinases, metalloproteinases, C-type lectin-like proteins, and other less abundant components in the Pmu venoms. Moreover, clinical manifestations of 209 Pmu envenomed patients hospitalized in northern or southeastern Taiwan revealed significant differences in local symptoms, such as ecchymosis and blistering. The mechanism of these local effects and possibly relevant venom components were examined. Further analysis showed that certain venom components with inter-population variation might work alone or synergistically with others to aggravate the local effects. Therefore, our findings of the venom variation may help one to improve antivenom production and better understand and manage Pmu bites. Full article
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18 pages, 2847 KiB  
Article
A First Look at the Inhibitory Potential of Urospatha sagittifolia (Araceae) Ethanolic Extract for Bothrops atrox Snakebite Envenomation
by Antonio L. Vera-Palacios, Juan D. Sacoto-Torres, Josselin A. Hernández-Altamirano, Andres Moreno, Maria C. Peñuela-Mora, David Salazar-Valenzuela, Noroska G. S. Mogollón and José R. Almeida
Toxins 2022, 14(7), 496; https://doi.org/10.3390/toxins14070496 - 17 Jul 2022
Cited by 5 | Viewed by 3083
Abstract
Bothrops atrox snakebites are a relevant problem in the Amazon basin. In this biodiverse region, the ethnomedicinal approach plays an important role as an alternative to antivenom therapy. Urospatha sagittifolia (Araceae) is a plant used for this purpose; however, its neutralizing properties have [...] Read more.
Bothrops atrox snakebites are a relevant problem in the Amazon basin. In this biodiverse region, the ethnomedicinal approach plays an important role as an alternative to antivenom therapy. Urospatha sagittifolia (Araceae) is a plant used for this purpose; however, its neutralizing properties have not been scientifically accessed. To fill this gap, we investigated the ability of U. sagittifolia to modulate the catalytic activity of Bothrops atrox venom, and their toxic consequences, such as local damage and lethality. The venom profile of B. atrox was assessed by chromatography and electrophoresis. Inhibition of the three main enzymatic and medically important toxins from the venom was evaluated using synthetic substrates and quantified by chromogenic activity assays. Additionally, the neutralization of lethality, hemorrhage and edema were investigated by in vivo assays. The possible interactions between venom proteins and plant molecules were visualized by polyacrylamide gel electrophoresis. Finally, the phytochemical constituents present in the ethanolic extract were determined by qualitative and quantitative analyses. The ethanolic extract reduced the activity of the three main enzymes of venom target, achieving ranges from 19% to 81% of inhibition. Our in vivo venom neuralizations assays showed a significant inhibition of edema (38.72%) and hemorrhage (42.90%). Additionally, lethality was remarkably counteracted. The highest extract ratio evaluated had a 75% survival rate. Our data support the biomedical value of U. sagittifolia as a source of natural enzyme inhibitors able to neutralize catalytically active B. atrox venom toxins and their toxic effects. Full article
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13 pages, 2223 KiB  
Article
Evaluating Antivenom Efficacy against Echis carinatus Venoms—Screening for In Vitro Alternatives
by Siddharth Bhatia, Avni Blotra and Karthikeyan Vasudevan
Toxins 2022, 14(7), 481; https://doi.org/10.3390/toxins14070481 - 13 Jul 2022
Cited by 8 | Viewed by 2334
Abstract
In India, polyvalent antivenom is the mainstay treatment for snakebite envenoming. Due to batch-to-batch variation in antivenom production, manufacturers have to estimate its efficacy at each stage of IgG purification using the median effective dose which involves 100–120 mice for each batch. There [...] Read more.
In India, polyvalent antivenom is the mainstay treatment for snakebite envenoming. Due to batch-to-batch variation in antivenom production, manufacturers have to estimate its efficacy at each stage of IgG purification using the median effective dose which involves 100–120 mice for each batch. There is an urgent need to replace the excessive use of animals in snake antivenom production using in vitro alternatives. We tested the efficacy of a single batch of polyvalent antivenom from VINS bioproducts limited on Echis carinatus venom collected from three different locations—Tamil Nadu (ECVTN), Goa (ECVGO) and Rajasthan (ECVRAJ)—using different in vitro assays. Firstly, size-exclusion chromatography (SEC-HPLC) was used to quantify antivenom–venom complexes to assess the binding efficiency of the antivenom. Secondly, clotting, proteolytic and PLA2 activity assays were performed to quantify the ability of the antivenom to neutralize venom effects. The use of both binding and functional assays allowed us to measure the efficacy of the antivenom, as they represent multiple impacts of snake envenomation. The response from the assays was recorded for different antivenom–venom ratios and the dose–response curves were plotted. Based on the parameters that explained the curves, the efficacy scores (ES) of antivenom were computed. The binding assay revealed that ECVTN had more antivenom–venom complexes formed compared to the other venoms. The capacity of antivenom to neutralize proteolytic and PLA2 effects was lowest against ECVRAJ. The mean efficacy score of antivenom against ECVTN was the greatest, which was expected, as ECVTN is mainly used by antivenom manufacturers. These findings pave a way for the development of in vitro alternatives in antivenom efficacy assessment. Full article
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14 pages, 598 KiB  
Article
Validation of a Culturally Relevant Snakebite Envenomation Clinical Practice Guideline in Brazil
by Gisele dos Santos Rocha, Altair Seabra Farias, João Arthur Alcântara, Vinícius Azevedo Machado, Felipe Murta, Fernando Val, Joseir Saturnino Cristino, Alícia Cacau Santos, Mena Bianca Ferreira, Leonardo Marques, Yasmim Vieira Rocha, André Sachett, Mailma Costa Almeida, Aline Alencar, Lisele Brasileiro, Érica da Silva Carvalho, Pedro Ferreira Bisneto, Marcus Lacerda, Anna Tupetz, Catherine A. Staton, João R.N. Vissoci, Elizabeth Teixeira, Charles J. Gerardo, Fan Hui Wen, Jacqueline Sachett and Wuelton Monteiroadd Show full author list remove Hide full author list
Toxins 2022, 14(6), 376; https://doi.org/10.3390/toxins14060376 - 28 May 2022
Cited by 10 | Viewed by 3151
Abstract
Snakebite envenoming (SBE) is a neglected tropical disease with significant global morbidity and mortality. Even when antivenom is available in low-resource areas, health workers do not receive adequate training to manage SBEs. This study aims to develop and validate a clinical practice guideline [...] Read more.
Snakebite envenoming (SBE) is a neglected tropical disease with significant global morbidity and mortality. Even when antivenom is available in low-resource areas, health workers do not receive adequate training to manage SBEs. This study aims to develop and validate a clinical practice guideline (CPG) for SBE management across Brazil. A panel of expert judges with academic and/or technical expertise in SBE management performed content validation. The content validity index (CVI) score was 90% for CPG objectives, 89% for structure and presentation and 92% for relevance and classified the CPG as valid. A semantic validation was performed by analyzing focus group discussions with doctors and nurses from three municipalities of the Brazilian Amazon, after a 5-day meeting during which the CPG was presented. Two central themes emerged: knowledge acquired during the meeting and recommendations for improving the CPG. Based on these results, the CPG was revised into a final version. This study presents the successful development and validation process of a CPG for SBE management, which is targeted to a specific low-resource, high-burden setting. This development and validation process can be adapted to other settings and/or other neglected tropical diseases. Full article
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12 pages, 3101 KiB  
Article
Development and Characterization of Anti-Naja ashei Three-Finger Toxins (3FTxs)-Specific Monoclonal Antibodies and Evaluation of Their In Vitro Inhibition Activity
by Ernest Z. Manson, Mutinda C. Kyama, Josephine Kimani, Aleksandra Bocian, Konrad K. Hus, Vladimír Petrilla, Jaroslav Legáth and James H. Kimotho
Toxins 2022, 14(4), 285; https://doi.org/10.3390/toxins14040285 - 16 Apr 2022
Cited by 4 | Viewed by 2688
Abstract
Antivenom immunotherapy is the mainstay of treatment for snakebite envenoming. Most parts of the world affected by snakebite envenoming depend on broad-spectrum polyspecific antivenoms that are known to contain a low content of case-specific efficacious immunoglobulins. Thus, advances in toxin-specific antibodies production hold [...] Read more.
Antivenom immunotherapy is the mainstay of treatment for snakebite envenoming. Most parts of the world affected by snakebite envenoming depend on broad-spectrum polyspecific antivenoms that are known to contain a low content of case-specific efficacious immunoglobulins. Thus, advances in toxin-specific antibodies production hold much promise in future therapeutic strategies of snakebite envenoming. We report anti-3FTxs monoclonal antibodies developed against N. ashei venom in mice. All the three test mAbs (P4G6a, P6D9a, and P6D9b) were found to be IgG antibodies, isotyped as IgG1. SDS-PAGE analysis of the test mAbs showed two major bands at approximately 55 and 29 kDa, suggestive of immunoglobulin heavy and light chain composition, respectively. The immunoaffinity-purified test mAbs demonstrated higher binding efficacy to the target antigen compared to negative control. Similarly, a cocktail of the test mAbs was found to induce a significantly higher inhibition (p-value < 0.0001) compared to two leading commercial brands of antivenoms on the Kenyan market, implying a higher specificity for the target antigen. Both the test mAbs and 3FTxs polyclonal antibodies induced comparable inhibition (p-value = 0.9029). The inhibition induced by the 3FTxs polyclonal antibodies was significantly different from the two antivenoms (p-value < 0.0001). Our results demonstrate the prospects of developing toxin-specific monoclonal-based antivenoms for snakebite immunotherapy. Full article
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24 pages, 2740 KiB  
Article
Profiling the Murine Acute Phase and Inflammatory Responses to African Snake Venom: An Approach to Inform Acute Snakebite Pathology
by Jaffer Alsolaiss, Chloe A. Evans, George O. Oluoch, Nicholas R. Casewell and Robert A. Harrison
Toxins 2022, 14(4), 229; https://doi.org/10.3390/toxins14040229 - 22 Mar 2022
Cited by 4 | Viewed by 3087
Abstract
Snake envenoming causes rapid systemic and local effects that often result in fatal or long-term disability outcomes. It seems likely that acute phase and inflammatory responses contribute to these haemorrhagic, coagulopathic, neurotoxic, nephrotoxic and local tissue destructive pathologies. However, the contributory role of [...] Read more.
Snake envenoming causes rapid systemic and local effects that often result in fatal or long-term disability outcomes. It seems likely that acute phase and inflammatory responses contribute to these haemorrhagic, coagulopathic, neurotoxic, nephrotoxic and local tissue destructive pathologies. However, the contributory role of acute phase/inflammatory responses to envenoming is under-researched and poorly understood—particularly for envenoming by sub-Saharan African venomous snakes. To provide data to help guide future studies of human patients, and to explore the rationale for adjunct anti-inflammatory medication, here we used an in vivo murine model to systematically assess acute phase and inflammatory responses of mice to ten African snake venoms. In addition to investigating snake species-specific effects of venom on the cardiovascular system and other key organs and tissues, we examined the response to intravascular envenoming by acute phase reactants, including serum amyloid A, P-selectin and haptoglobin, and several cytokines. Venoms of the spitting (Naja nigricollis) and forest (N. melanoleuca) cobras resulted in higher acute phase and inflammatory responses than venoms from the other cobras, mambas and vipers tested. Naja nigricollis venom also stimulated a 100-fold increase in systemic interleukin 6. Thin blood films from venom-treated mice revealed species-specific changes in red blood cell morphology, indicative of membrane abnormalities and functional damage, lymphopenia and neutrophil leukocytosis. Our ex vivo assays with healthy human blood treated with these venoms identified that N. nigricollis venom induced marked levels of haemolysis and platelet aggregation. We conclude that African snake venoms stimulate very diverse responses in this mouse model of acute systemic envenoming, and that venoms of the African cobras N. nigricollis and N. melanoleuca, in particular, cause marked inflammatory and non-specific acute phase responses. We also report that several African snake venoms cause haemolytic changes. These findings emphasise the importance of understanding acute responses to envenoming, and that further research in this area may facilitate new diagnostic and treatment approaches, which in turn may lead to better clinical outcomes for snakebite patients. Full article
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Review

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18 pages, 955 KiB  
Review
Cerebral Complications of Snakebite Envenoming: Case Studies
by Yu-Kai Huang, Yen-Chia Chen, Chia-Chun Liu, Hui-Chun Cheng, Anthony T. Tu and Kun-Che Chang
Toxins 2022, 14(7), 436; https://doi.org/10.3390/toxins14070436 - 27 Jun 2022
Cited by 4 | Viewed by 5445
Abstract
There are an estimated 5.4 million snakebite cases every year. People with snakebite envenoming suffer from severe complications, or even death. Although some review articles cover several topics of snakebite envenoming, a review of the cases regarding cerebral complications, especially rare syndromes, is [...] Read more.
There are an estimated 5.4 million snakebite cases every year. People with snakebite envenoming suffer from severe complications, or even death. Although some review articles cover several topics of snakebite envenoming, a review of the cases regarding cerebral complications, especially rare syndromes, is lacking. Here, we overview 35 cases of snakebite by front-fanged snakes, including Bothrops, Daboia, Cerastes, DeinagkistrodonTrimeresurus, and Crotalus in the Viperidae family; Bungarus and Naja in the Elapidae family, and Homoroselaps (rare cases) in the Lamprophiidae family. We also review three rare cases of snakebite by rear-fanged snakes, including Oxybelis and Leptodeira in the Colubridae family. In the cases of viper bites, most patients (17/24) were diagnosed with ischemic stroke and intracranial hemorrhage, leading to six deaths. We then discuss the potential underlying molecular mechanisms that cause these complications. In cases of elapid bites, neural, cardiac, and ophthalmic disorders are the main complications. Due to the small amount of venom injection and the inability to deep bite, all the rear-fanged snakebites did not develop any severe complications. To date, antivenom (AV) is the most effective therapy for snakebite envenoming. In the six cases of viper and elapid bites that did not receive AV, three cases (two by viper and one by elapid) resulted in death. This indicates that AV treatment is the key to survival after a venomous snakebite. Lastly, we also discuss several studies of therapeutic agents against snakebite-envenoming-induced complications, which could be potential adjuvants along with AV treatment. This article organizes the diagnosis of hemotoxic and neurotoxic envenoming, which may help ER doctors determine the treatment for unidentified snakebite. Full article
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Other

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8 pages, 781 KiB  
Case Report
Neurotoxicity and Other Clinical Manifestations of a Common European Adder (Vipera berus) Bite in Romania
by Gabriela Viorela Nițescu, Coriolan Emil Ulmeanu, Maria-Dorina Crăciun, Alina Maria Ciucă, Alexandru Ulici, Ioan Ghira and Davide Lonati
Toxins 2022, 14(7), 500; https://doi.org/10.3390/toxins14070500 - 18 Jul 2022
Cited by 4 | Viewed by 1573
Abstract
Most cases of envenomation by common European vipers (Vipera berus) have not been reported to have neurotoxic manifestations. However, these manifestations have been demonstrated in some cases of envenomation by subspecies of V. berus, found in the Carpathian Basin region [...] Read more.
Most cases of envenomation by common European vipers (Vipera berus) have not been reported to have neurotoxic manifestations. However, these manifestations have been demonstrated in some cases of envenomation by subspecies of V. berus, found in the Carpathian Basin region of south-eastern Europe. Here, we report the case of a 5-year-old girl from the south of Romania who presented symptoms of neurotoxicity, as well as other systemic and local symptoms, after being bitten by an adder of the V. berus subspecies. Treatment consisted of monovalent antivenom, a corticosteroid, and prophylactic enoxaparin. Neurotoxic manifestations of envenomation as well as other local and systemic symptoms improved within 5 days of treatment. The presented case shows that venom from V. berus subspecies found in the Carpathian Basin can have neurotoxic effects. This case also confirmed the efficacy of monospecific antivenom treatment in bringing about rapid and complete remission, following envenomation. Full article
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9 pages, 1699 KiB  
Case Report
Medical Management after Lancehead Snakebite in North Amazon: A Case Report of Long-Term Disability
by Isadora S. Oliveira, Carla B. Ananias, Jilvando M. Medeiros, Michelle V. S. Franco, Isabela G. Ferreira, Felipe A. Cerni, Eliseu A. Sandri, Wuelton M. Monteiro and Manuela B. Pucca
Toxins 2022, 14(7), 494; https://doi.org/10.3390/toxins14070494 - 16 Jul 2022
Cited by 6 | Viewed by 1840
Abstract
Snakebites are a major public health problem in indigenous communities in Brazil, leading to acute local and systemic damage with resulting deficiencies. Long-term musculoskeletal disabilities related to snakebites have been a neglected area of research. Bothrops (lancehead) snakes are responsible for most of [...] Read more.
Snakebites are a major public health problem in indigenous communities in Brazil, leading to acute local and systemic damage with resulting deficiencies. Long-term musculoskeletal disabilities related to snakebites have been a neglected area of research. Bothrops (lancehead) snakes are responsible for most of the permanent sequelae related to snakebites in Latin America. Here, we present a case report of a 32-year-old male indigenous patient who was envenomed by a Bothrops species. The patient was clinically followed for a period of approximately 2 years and 6 months, during which time he experienced a loss of musculoskeletal tissue and required several medical procedures such as debridement, tissue reconstruction, and physical therapy, which resulted in a recovery of mobility, though with a permanent sequelae in gait. This case report shows how snakebites have a significant impact on health systems, as victims require physiotherapy, plastic surgery, and orthopedics services, as well as social support for reintegration into their local communities. Full article
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