Necrotic Wound Caused by Jararaca (Bothrops jararaca) in a Dog - Hyperbaric Oxygen Therapy (HBTO)

Background: Snakebites are the main responsible for envenoming in dogs and the bothropic venom remains the most common in Brazil, which can induce a necrotic skin wound. Hyperbaric oxygen therapy (HBOT) use 100% oxygen under high pressure and used to treat different wounds in human patients. To the authors’ knowledge, no reports regarding to use the HBOT in skin wound caused by snakebite (Bothrops jararaca) are present in the literature. The present clinical case aimed to describe the use of HBOT for the treatment of an extensive necrotic wound caused by jararaca snakebite in a dog.Case: A neutered 8-year-old mixed-breed dog, weighing 12 kg, was admitted with a 7-day history of extensive necrotic wound was identified in the face and neck causing by a snakebite, and no sign of pain. The procedure of HBOT (single sessions of 1.5 ATM, 45 min, repeated every 48 h, up to 12 sessions) was decided, and the complete blood cells, alanine aminotransferase, creatinine, creatine kinase, prothrombin time, activated partial thromboplastin time, wound clinical evaluation were measured at the following time-points: 2nd, 5th, 10th, and 12th sessions. At the 5th session was identified leukopenia, neutropenia and lymphopenia. Wound re-epithelialization was initiated after the 5th session, and the complete epithelialization was identified at the 12th session of HBOT. During the HBOT no side effects were identified. Three months after the HBOT finished, the animal returned to the clinic and the clinical status evolved positively, and the wound was completed healed.Discussion: This report described the treatment of an extensive necrotic skin wound caused by snakebite (Bothrops jararaca) in an 8-year-old, neutered, mixed-breed dog using the HBOT. The wound healing was achieved after 12 sessions, similar to the literature, which reported a ranging from 1 to 12 sessions. The HBOT protocol used in this case was similar as reported for human patients with chronic wounds due to the lack of HBOT protocols for animals. No reports regarding the use of HBOT for treat necrotic wound caused by snakebite was described in the literature, and to the authors’ knowledge, this is the first report in Brazil describing the use of HBOT in dogs. On the other hand, dogs with surgically induced skin wounds and treated with daily session of HBOT using the treatment protocol of 1.7 ATM (30 min) and 2.0 ATM (40 min) up to 7th day of treatment did not show significant results on healing [9]. This fact was associated with the HBOT achievement in the proliferative and remodeling phases of the healing process. The high intensity of HBOT was between the 5th and 10th session since the wound showed a higher area decrease rate and consequently increase of wound contraction. This period was corresponding to the 10th and 20th day of the healing process, which can be identified angiogenic activity, re-epithelialization, and collagen maturation. The decrease in PVC has been associated with the anticoagulant and/or hemorrhagic activity caused by the venom, and leukopenia, neutropenia and lymphopenia was related with possible bone marrow exhaustion. Single sessions of HBOT (1.5 ATM, 45 min, and repeat each 48 h, up to 12 session) induces healing of necrotic wound caused by snakebite (Bothrops jararaca) in an 8-year-old, neutered, mixed-breed dog without any side effects.Keywords: dog, healing, hyperbaric chamber, skin wound, snake.

Therapeutic possibilities of Bothrops jararaca in high dilution

Introduction: The knowledge and use of the venom of Bothrops jararaca in high dilutions is still quite limited. One of the important properties is the use of one of its components, bradykinin, for the development of antihypertensive medication known as captopril. Other situations, such as clinical, local and systemic should receive more depth to the composition of Materia Medica related to various medical actions on the man and mammals in general. The systemic action of the bite of this snake, includes hemostasis disorders, culminating as bleeding gums, in addition to sweating, hypertension, and hypothermia. The action includes local pain and swelling with bruising, bleeding and often blistering and tissue necrosis. The action on the immune system, through action on the complement C3 and other complement components may show its possible use in cases of bacterial infections, including mycobacteria, as presented in the study of 1970 Vanessa Birdsey, "Interactions of poisons toxic with the addition, "the journal of Immunology 1971. Today, this poison has a toxicology published by Anibal Melgarejo, "Venomous Animals of Brazil", 2003, which subsidizes the development of study for its use in high dilutions, and a comprehensive study of the biology of the animal itself. Published studies on biomolecular analysis add more details about the relations of the poison and mammals. All these characteristics suggest the use of poison as a homeopathic remedy. Objective: To investigate the therapeutic possibilities in high dilutions of the venom of the snake Bothrops jararaca, expanding its clinical use. Methodology: Methodological description of this poison in contemporary bases including: Origin, physical description chemistry, toxicology, pharmacology and medicine in preparation of high dilution, general action, specific actions on systems or organs, sensations, modalities, concomitants, etiological indications relations main clinics. Results: Defining the therapeutic indications such as modulation of the complement system, action on the cardiovascular system, among other uses, by Bothrops jararaca in high dilution. Conclusion: This evaluation can be used for different sources of products and allows the rational use of Bothrops jararaca in high dilution. The results can and should be complemented by clinical studies and pathogenetic. Bacterial infectious diseases such as tuberculosis and leprosy, and autoimmune disease LES and may receive treatment studies with the drug based on Bothrops jararaca snake venom because they are indirectly associated with them via similarity of the failure of complement, an important marker for bacterial the defense of mammals. Action on clinical aspects like hypertension, sweating, hypothermia and necrosis shall be seen. Perhaps the search for the stimulation of complement show a new pathway for the harmonization, long-predicted by Hahnemann, Hering and searched for among the many that followed the creator of this therapy.

Oral Tolerance Induction by Bothrops jararaca Venom in a Murine Model and Cross-Reactivity with Toxins of Other Snake Venoms

Oral tolerance is defined as a specific suppression of cellular and humoral immune responses to a particular antigen through prior oral administration of an antigen. It has unique immunological importance since it is a natural and continuous event driven by external antigens. It is characterized by low levels of IgG in the serum of animals after immunization with the antigen. There is no report of induction of oral tolerance to Bothrops jararaca venom. Here, we induced oral tolerance to B. jararaca venom in BALB/c mice and evaluated the specific tolerance and cross-reactivity with the toxins of other Bothrops species after immunization with the snake venoms adsorbed to/encapsulated in nanostructured SBA-15 silica. Animals that received a high dose of B. jararaca venom (1.8 mg) orally responded by showing antibody titers similar to those of immunized animals. On the other hand, mice tolerized orally with three doses of 1 µg of B. jararaca venom showed low antibody titers. In animals that received a low dose of B. jararaca venom and were immunized with B. atrox or B. jararacussu venom, tolerance was null or only partial. Immunoblot analysis against the venom of different Bothrops species provided details about the main tolerogenic epitopes and clearly showed a difference compared to antiserum of immunized animals.

Involvement of von Willebrand factor and botrocetin in the thrombocytopenia induced by Bothrops jararaca snake venom

Patients bitten by snakes consistently manifest a bleeding tendency, in which thrombocytopenia, consumption coagulopathy, mucous bleeding, and, more rarely, thrombotic microangiopathy, are observed. Von Willebrand factor (VWF) is required for primary hemostasis, and some venom proteins, such as botrocetin (a C-type lectin-like protein) and snake venom metalloproteinases (SVMP), disturb the normal interaction between platelets and VWF, possibly contributing to snakebite-induced bleedings. To understand the relationship among plasma VWF, platelets, botrocetin and SVMP from Bothrops jararaca snake venom (BjV) in the development of thrombocytopenia, we used (a) Wistar rats injected s.c. with BjV preincubated with anti-botrocetin antibodies (ABA) and/or Na2-EDTA (a SVMP inhibitor), and (b) VWF knockout mice (Vwf-/-) injected with BjV. Under all conditions, BjV induced a rapid and intense thrombocytopenia. In rats, BjV alone reduced the levels of VWF:Ag, VWF:CB, high molecular weight multimers of VWF, ADAMTS13 activity, and factor VIII. Moreover, VWF:Ag levels in rats that received BjV preincubated with Na2-EDTA and/or ABA tended to recover faster. In mice, BjV caused thrombocytopenia in both Vwf-/- and C57BL/6 (background control) strains, and VWF:Ag levels tended to decrease in C57BL/6, demonstrating that thrombocytopenia was independent of the presence of plasma VWF. These findings showed that botrocetin present in BjV failed to affect the extent or the time course of thrombocytopenia induced by envenomation, but it contributed to decrease the levels and function of plasma VWF. Thus, VWF alterations during B. jararaca envenomation are an ancillary event, and not the main mechanism leading to decreased platelet counts.

Bothrops Jararaca Snake Venom Modulates Key Cancer-Related Proteins in Breast Tumor Cell Lines

Cancer is characterized by the development of abnormal cells that divide in an uncontrolled way and may spread into other tissues where they may infiltrate and destroy normal body tissue. Several previous reports have described biochemical anti-tumorigenic properties of crude snake venom or its components, including their capability of inhibiting cell proliferation and promoting cell death. However, to the best of our knowledge, there is no work describing cancer cell proteomic changes following treatment with snake venoms. In this work we describe the quantitative changes in proteomics of MCF7 and MDA-MB-231 breast tumor cell lines following treatment with Bothrops jararaca snake venom, as well as the functional implications of the proteomic changes. Cell lines were treated with sub-toxic doses at either 0.63 μg/mL (low) or 2.5 μg/mL (high) of B. jararaca venom for 24 h, conditions that cause no cell death per se. Proteomics analysis was conducted on a nano-scale liquid chromatography coupled on-line with mass spectrometry (nLC-MS/MS). More than 1000 proteins were identified and evaluated from each cell line treated with either the low or high dose of the snake venom. Protein profiling upon venom treatment showed differential expression of several proteins related to cancer cell metabolism, immune response, and inflammation. Among the identified proteins we highlight histone H3, SNX3, HEL-S-156an, MTCH2, RPS, MCC2, IGF2BP1, and GSTM3. These data suggest that sub-toxic doses of B. jararaca venom have potential to modulate cancer-development related protein targets in cancer cells. This work illustrates a novel biochemical strategy to identify therapeutic targets against cancer cell growth and survival.