Textual research on Bungarus Parvus / 中国中药杂志

Bungarus Parvus, a precious animal Chinese medicinal material used in clinical practice, is believed to be first recorded in Ying Pian Xin Can published in 1936. This study was carried out to analyze the names, geographical distribution, morphological characteristics, ecological habits, poisonousness, and medicinal parts by consulting ancient Chinese medical books and local chronicles, Chinese Pharmacopeia, different processing standards of trditional Chinese medicine(TCM) decoction pieces, and modern literatures. The results showed that the earliest medicinal record of Bungarus Parvus was traced to 1894. In 1930, this medicinal material was used in the formulation of Annao Pills. The original animal, Bungarus multicinctus, was recorded by the name of "Bojijia" in 1521. The morphological characteristics, ecological habits, and poisonousness of the original animal are the same in ancient and modern records. The geographical distribution is similar between the ancient records and modern documents such as China Medicinal Animal Fauna. The dried body of young B. multicinctus is used as Bungarus Parvus, which lack detailed references. As a matter of fact, it is still inconclusive whether there are differences between young snakes and adult snakes in terms of active ingredients, pharmacological effects, and clinical applications. This study clarified the medicinal history and present situation of Bungarus Parvus. On the basis of the results, it is suggested that systematic comparison on young and adult B. multicinctus should be carried out to provide references for revising the medicinal parts of B. multicinctus.

Necrosis tisular local y trombocitopenia después de una intoxicación por mordedura de Bungarus multicinctusen una niña / Local tissue necrosis and thrombocytopenia following Bungarus multicinctus envenomation in a child

La intoxicación por mordedura de serpiente es un problema de salud pública global. En la población pediátrica, la intoxicación por mordedura de serpiente presenta características diferentes que en los pacientes adultos. La Bungarus multicinctus es una especie de elápido sumamente venenoso. Las presentaciones clínicas documentadas después de la intoxicación por mordedura de Bungarus multicinctus son reacciones locales mínimas, insuficiencia respiratoria, dolor generalizado e hiponatremia potencialmente mortal. Presentamos el caso de una intoxicación por mordedura de Bungarus multicinctus en una niña con manifestaciones clínicas atípicas, incluidas necrosis tisular grave y trombocitopenia con coagulopatía.

Snakebite envenoming is a global public health problem. The pediatric population poisoned by snakebite envenoming has different features than adult patients. Bungarus multicinctus is a highly venomous species of the elapid snake. The documented clinical presentations following Bungarus multicinctus envenoming are minimal local reactions, respiratory failure, general pain, and life-threatening hyponatremia. We present an uncommon case of Bungarus multicinctus envenomation in a girl with unusual clinical findings, including severe tissue necrosis and thrombocytopenia with coagulopathy.

Kinetic and toxicological effects of synthesized palladium(II) complex on snake venom (Bungarus sindanus) acetylcholinesterase

The venom of the krait (Bungarus sindanus), an Elapidae snake, is highly toxic to humans and contains a great amount of acetylcholinesterase (AChE). The enzyme AChE provokes the hydrolysis of substrate acetylcholine (ACh) in the nervous system and terminates nerve impulse. Different inhibitors inactivate AChE and lead to ACh accumulation and disrupted neurotransmission. Methods: The present study was designed to evaluate the effect of palladium(II) complex as antivenom against krait venom AChE using kinetics methods. Results: Statistical analysis showed that krait venom AChE inhibition decreases with the increase of Pd(II) complex (0.025-0.05 µM) and exerted 61% inhibition against the AChE at a fixed concentration (0.5 mM) of ACh. Kinetic analysis using the Lineweaver Burk plot showed that Pd(II) caused a competitive inhibition. The compound Pd(II) complex binds at the active site of the enzyme. It was observed that K m (Michaelis-Menten constant of AChE-ACh into AChE and product) increased from 0.108 to 0.310 mM (45.74 to 318.35%) and V max remained constant with an increase of Pd(II) complex concentrations. In AChE K Iapp was found to increase from 0.0912 to 0.025 µM (29.82-72.58%) and did not affect the V maxapp with an increase of ACh from (0.05-1 mM). K i (inhibitory constant) was estimated to be 0.029µM for snake venom; while the K m was estimated to be 0.4 mM. The calculated IC50 for Pd(II) complex was found to be 0.043 µM at constant ACh concentration (0.5 mM). Conclusions: The results show that the Pd(II) complex can be deliberated as an inhibitor of AChE.(AU)

Non-neurotoxic activity of Malayan krait (Bungarus candidus) venom from Thailand

Background: Envenoming by kraits (genus Bungarus) is a medically significant issue in South Asia and Southeast Asia. Malayan krait (Bungarus candidus) venom is known to contain highly potent neurotoxins. In recent years, there have been reports on the non-neurotoxic activities of krait venom that include myotoxicity and nephrotoxicity. However, research on such non-neurotoxicity activities of Malayan krait venom is extremely limited. Thus, the aim of the present study was to determine the myotoxic, cytotoxic and nephrotoxic activities of B. candidus venoms from northeastern (BC-NE) and southern (BC-S) Thailand in experimentally envenomed rats. Methods: Rats were administered Malayan krait (BC-NE or BC-S) venom (50 g/kg, i.m.) or 0.9% NaCl solution (50 L, i.m.) into the right hind limb. The animals were sacrificed 3, 6 and 24 h after venom administration. The right gastrocnemius muscle and both kidneys were collected for histopathological analysis. Blood samples were also taken for determination of creatine kinase (CK) and lactate dehydrogenase (LDH) levels. The human embryonic kidney cell line (HEK-293) was used in a cell proliferation assay to determine cytotoxic activity. Results: Administration of BC-NE or BC-S venom (50 g/kg, i.m.) caused time-dependent myotoxicity, characterized by an elevation of CK and LDH levels. Histopathological examination of skeletal muscle displayed marked muscle necrosis and myofiber disintegration 24 h following venom administration. Both Malayan krait venoms also induced extensive renal tubular injury with glomerular and interstitial congestion in rats. BC-NE and BC-S venoms (1000.2 g/ mL) caused concentration-dependent cytotoxicity on the HEK-293 cell line. However, BC-NE venom (IC50 =8 ± 1 g/mL; at 24 h incubation; n = 4) was found to be significantly more cytotoxic than BC-S venom (IC50 =15 ± 2 g/mL; at 24 h incubation; n = 4). In addition, the PLA2 activity of BC-NE venom was significantly higher than that of BC-S venom...

Non-neurotoxic activity of Malayan krait (Bungarus candidus) venom from Thailand

Envenoming by kraits (genus Bungarus) is a medically significant issue in South Asia and Southeast Asia. Malayan krait (Bungarus candidus) venom is known to contain highly potent neurotoxins. In recent years, there have been reports on the non-neurotoxic activities of krait venom that include myotoxicity and nephrotoxicity. However, research on such non-neurotoxicity activities of Malayan krait venom is extremely limited. Thus, the aim of the present study was to determine the myotoxic, cytotoxic and nephrotoxic activities of B. candidus venoms from northeastern (BC-NE) and southern (BC-S) Thailand in experimentally envenomed rats. Methods: Rats were administered Malayan krait (BC-NE or BC-S) venom (50 µg/kg, i.m.) or 0.9% NaCl solution (50 µL, i.m.) into the right hind limb. The animals were sacrificed 3, 6 and 24 h after venom administration. The right gastrocnemius muscle and both kidneys were collected for histopathological analysis. Blood samples were also taken for determination of creatine kinase (CK) and lactate dehydrogenase (LDH) levels. The human embryonic kidney cell line (HEK-293) was used in a cell proliferation assay to determine cytotoxic activity. Results: Administration of BC-NE or BC-S venom (50 µg/kg, i.m.) caused time-dependent myotoxicity, characterized by an elevation of CK and LDH levels. Histopathological examination of skeletal muscle displayed marked muscle necrosis and myofiber disintegration 24 h following venom administration. Both Malayan krait venoms also induced extensive renal tubular injury with glomerular and interstitial congestion in rats. BC-NE and BC-S venoms (100­0.2 µg/ mL) caused concentration-dependent cytotoxicity on the HEK-293 cell line. However, BC-NE venom (IC50 =8 ± 1 µg/mL; at 24 h incubation; n = 4) was found to be significantly more cytotoxic than BC-S venom (IC50 =15 ± 2 µg/mL; at 24 h incubation; n = 4). In addition, the PLA2 activity of BC-NE venom was significantly higher than that of BC-S venom. Conclusions: This study found that Malayan krait venoms from both populations possess myotoxic, cytotoxic and nephrotoxic activities. These findings may aid in clinical diagnosis and treatment of envenomed patients in the future.(AU)

Fatal neurotoxic envenomation following the bite of a greater black krait (Bungarus niger) in Nepal: a case report

Background Neurotoxic envenomation following bites by kraits (Bungarus species) is a leading cause of snakebite mortality in South Asia. Over a long time, this had been attributed only to one species, the common krait (Bungarus caeruleus). However, recent research has provided increasing evidence of the involvement of several krait species. Here, we report a fatal case of neurotoxic envenomation following the bite of a greater black krait (Bungarus niger) in Nepal. Case presentation A 33-year-old man was bitten in the outdoor corridor of his home in the eastern hills of Ilam district while handling a snake he thought to be non-venomous. He subsequently developed severe abdominal pain, frequent vomiting, and signs of neurotoxic envenomation leading to respiratory paralysis. The patient did not respond to Indian polyvalent antivenom given 4 h after the bite and died under treatment 8 h after the bite. This is the second time that a B. niger was observed in Nepal, the first documented case of envenomation by this species in the country and the sixth reported case worldwide. Conclusions Previous distribution records - from eastern India and western Nepal, from western hills in Nepal, and from lowland localities in India and Bangladesh - indicate risk of envenomation by B. niger throughout the low and intermediate elevations of Nepal up to at least 1,500 m above sea level. As very few people in Nepal bring killed snakes to healthcare centers and because there is a general belief among local people that there are no kraits in the hills, bites by B. niger are likely to be misdiagnosed and underreported.(AU)

Enzymatic and biochemical characterization of Bungarus sindanus snake venom acetylcholinesterase

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.(AU)

The establishment of PCR system to identify Bungarus multicinctus rapidly / 药学学报

The purpose of the present study is to establish a rapid and effective PCR method for the identification of B. multicinctus. Based on sequence alignment of B. multicinctus and its adulterants, we found that Cyt b gene is a good molecular genetic marker for the authentication of B. multicinctus. On the basis of the sequence data, a pair of highly specialized primers was designed. The templates were extracted by the DNA purification system. Key factors such as annealing temperature, concentration of Taq enzyme and cycle numbers were analyzed and optimized. The modified PCR program consisted of an initial denaturation step at 95 degrees C for 5 min, followed by 30 cycles of 95 degrees C for 30 s and 55 degrees C for 45 s and a final extension at 72 degrees C for 5 min. Thirteen samples of B. multicinctus were identified accurately from their 20 adulterants in 4 hours. The results indicated it is a highly accurate, rapid and applicable method for the authentication of B. multicinctus.

Specific antivenom for Bungarus candidus.

BACKGROUND: Bungarus candidus (Malayan krait) snake is a neurotoxin snake. Previous treatment after snakebite was mainly respiratory support until the patient had spontaneous breathing. Recently specific antivenom for the Bungarus candidus snake was produced by the Queen Saovabha Memorial Institute and distributed in June 2004. The present article is the first report on the clinical response to the specific antivenom for Bungarus candidus. OBJECTIVE: To analyze the signs and symptoms of patients after snakebite and the response of the patients after receiving specific antivenom for Bungarus candidus snake. STUDY DESIGN: Retrospective chart review. MATERIAL AND METHOD: Four cases of Bungarus candidus snakebite were identified and divided into two groups. Group I (Case 1, 2, and 3) had received specific antivenom for Bungarus candidus while group 2 (case 4) had not. Onset, signs and symptoms after snakebite, antivenom dosage, and response time after receiving antivenom were analyzed. RESULTS: The first three patients received specific antivenom for Bungarus candidus and the fourth patient did not receive any. All four patients developed neurological signs and symptoms from this neurotoxic venom. In case 1, 2, and 4, the first signs and symptoms were dyspnea, difficulty with speech, and opening the eyelids at 50 minutes (30-60 minutes). The onset ofother signs and symptoms included respiratory paralysis with intubation 3 hours (2-4 hours), full ptosis 3.66 hours (3-4 hours), mydriasis and fixedpupils 4.33 hours (4-5 hours), no response to stimuli 5.66 hours (4-10 hours), tachycardia 5.5 hours (47 hours), and hypertension 14 hours (4-24 hours). The first two patients received specific antivenom for Bungarus candidus after being bitten at 10 and 12 hours, respectively. The first clinical response in case 1, were 12 hours after receiving 16 vials, and in case 2, were 20 hours after receiving 16 vials. These were slight movement of feet phalanxes. At 40 hours after receiving specific antivenom 30 vials in case 1 and 32 vials in case 2, they were able to respond to commands, motor power changed from grade 0 to grade 1 and there was 50% elevated eyebrows. The motor power changedfrom grade I to grade 4 with 100% elevation of eyebrows from full ptosis was 65 hours after receiving specific antivenom 60 vials in case 1 and 70 hours after receiving specific antivenom 87 vials in case 2. The patients had spontaneous opening ofeyelids at 90 hours after receiving 80 vials for case I and 88 hours after receiving 87 vials for case 2. Case 2 was extubated on day 4 after the snakebite while case 1 was extubated later on day 10 because of superimposing pneumonia. The third case had delayed onset of signs and symptoms of neurotoxicity compared to the other three patients. Dyspnea, difficulty with speech, and opening eyelids occurred at 5 hours after the snakebite. No response to stimuli and respiratory paralysis occurred at 20 hours after the snakebite. His consciousness improved 10 hours after receiving 3 vials of specific antivenom. This was noted by being able to respond to commands and the motor power changed to grade 2 however, full ptosis was still present up to 24 hours. After receiving 23 vials ofspecific antivenom, he accidentally extubated himself however, he could breathe adequately using a mask with a bag. His motor power changed to grade 4 with 100% elevated eyebrows but full ptosis 34 hours after receiving 38 vials of specific antivenom. He could spontaneously open his eyelids 40 hours after receiving 38 vials specific antivenom. Cases 1, 2, and 3 had persistent mydriasis andfixed pupils until discharge. Case 4 did not receive specific antivenom for Bungarus candidus. He did not respond to stimuli 10 hours after snakebite and he was treated with respirator and symptomatic treatment. On day 2, his blood pressure dropped, he was on dopamine to raise his BP On day 3, he developed ventricular fibrillation. Defibrillation was administered and ECG returned to normal. He was given further supportive care. On day 7, he was discharged at the request of his relatives without any improvement. CONCLUSION: The patients who received specific antivenom had more rapid improvement ofsigns and symptoms comparing to the patient who did not receive the antivenon.

Preliminary studies with a neurotoxin obtained from Bungarus caeruleus venom

The neurotoxin purified from the venom of Bungarus caeruleus causes a neuromuscular blockade on acetylcholine-induced muscle twitch response in isolated frog rectus abdominis muscle preparation. Neuromuscular blockade produced by d-tubocurarine on acetylcholine-induced muscle twitch response in an isolated frog rectus abdominis muscle preparation was reversed to normal muscle twitch response in presence of neostigmine. Whereas the purified neurotoxin produced an irreversible neuromuscular blockade in presence of the same strength of neostigmine. As it is already known, botulinum toxin, which also brings about neuromuscular blockade, is effectively used as a drug in the treatment of painful movement disorders. Since the purified toxin also causes paralysis of the muscle, we propose its possible efficacy in the treatment of neuromuscular disorders.(AU)

High specific PCR identification of Bungarus multicinctus and its adulterants / 中国中药杂志

<p><b>OBJECTIVE</b>To develop a convenient and effective method for the identification of Bungarus multicinctus.</p><p><b>METHOD</b>Based on the sequence of Cyt b gene fragment of B. multicinctus and its adulterants, a pair of highly specific primer (HJL- and HJH-) were designed for distinguishing B. ulticinctus from other species of snake. To establish specific PCR reaction condition, the primers were employed to amplify the DNA templates extracted from B. multicinctus and 6 other species of snake, under different annealing temperature. Using this method, B. multicinctus was identified from 18 samples bought from many drugstores.</p><p><b>RESULT</b>A 230 bp DNA fragment was amplified from B. multicinctus in PCR with annealed temperature at 67 degrees C, whereas no DNA fragment was amplified from other snake samples under the same reaction condition, B. multicinctus could be clearly distinguished from others by PCR reaction with the highly specific primers. In the present study, 18 sample, bought from different drugstores, were also identified by the highly specific PCR with the primers. The results indicated that 14 samples were B. multicinctus and the other 4 were adulterant, which was consistent with the conclusion of authentication based on morphological.</p><p><b>CONCLUSION</b>The primers designed in the present study were highly specific for B. multicinctus.</p>

Management of snake-bite in rural Maharashtra: a 10-year experience.

BACKGROUND: A high incidence of snake-bite envenomation has been reported from Marathwada, Maharashtra. This study analysed the pattern of snake-bites and their management in a rural area of India over a 10-year period. METHODS: A total of 633 patients with snake-bite admitted to the Rural Community Centre and Punde Hospital in Mukhed taluka, Nanded district (Marathwada) of Maharashtra, between 1992 and 2001, were analysed retrospectively. The local and systemic manifestations of snake-bite, response to antisnake venom, atropine and neostigmine, the treatment of complications and the outcome were analysed. RESULTS: Of the 633 patients, 427 (67.5%) had been bitten by poisonous snakes and 206 (32.5%) by non-poisonous snakes. The majority of snake-bites (68.9%) occurred between May and November. Those affected were mainly farmers (228 [36%]), students (191 [30.2%]) and housewives (175 [27.6%]). Of the 427 envenomed by poisonous snakes, 274 (64.2%) were by Echis carinatus (saw-scaled viper), 71 (16.6%) by cobra, 42 (9.8%) by krait and 40 (9.4%) by Russell viper. The requirement of antisnake venom for treating neurotoxic envenomation was 40-320 ml and for Echiscarinatus and Russell viper bites it was 20-250 ml. Among those envenomed by poisonous snakes, the mortality was 4.7% (n=20). CONCLUSION: Snake-bite is a common life-threatening emergency in the study area. We observed an occupational risk and a seasonal incidence of snake-bite. Knowledge of the varied clinical manifestations of snake-bite are important for effective management. Ready availability and appropriate use of antisnake venom, close monitoring of patients, institution of ventilatory support and early referral to a larger hospital when required help in reducing the mortality. Most patients with snake-bites can be successfully managed even in small rural hospitals with limited facilities.

Neurotoxic envenoming following bites by the Malayan krait (Bungarus candidus).

The author reports three cases of patient bitten by the Malayan krait (Bungarus candidus). Within two to six hours after bites, patients developed ptosis, ophthalmoplegia, dysarthria, dysphagia and generalized paralysis requiring assisted ventilation. After ventilatory support and other supportive treatments, all patients gradually recovered to normal activity.

Krait bite requiring high dose antivenom: a case report.

Anti snake venom (ASV) is the most specific therapy available for treatment of snakebite envenomation. The ASV available in Nepal are polyvalent ASV produced in India and are effective against envenomation by cobra and krait, the two most common species found in Eastern Nepal. Neurotoxic signs respond slowly and unconvincingly and continuous absorption of venom may cause recurrent neurotoxicity. Therefore, close observation and continuous administration of ASV is essential to save the victim. We report a case of neurotoxic envenomation due to bite by common krait (Bangarus caeruleus). The victim required very high dose of polyvalent ASV for reversal of neurological manifestations.