Scolicidal Agent Research Articles

In vitro Protoscolicidal Effects of Cinnamomum zeylanicum Essential Oil and Its Toxicity in Mice.

Background:This study investigates the scolicidal effects of Cinnamomum zeylanicum essential oil against the protoscoleces of hydatid cysts and its toxicity in the mice model.Materials and Methods:Gas chromatography/mass spectroscopy analyses were used to identify the constituents of essential oil. Protoscoleces were treated with different concentrations of the essential oil (6.25–100 µL/mL) in each test tube for 5–30 min. The viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Forty-eight male NMRI mice were also used to determine the toxicity of C. zeylanicum essential oil (0.5–4 mL/kg).Results:The main components were found to be cinnamaldehyde (91.8%), ρ metoxicinamate (1.57%), and α pinene (1.25%). Findings indicate that C. zeylanicum essential oil with the concentrations of 100 and 50 µL/mL killed 100% of protoscoleces after 5 min of exposure. Also, the lower concentrations of C. zeylanicum essential oil motivated a late protoscolicidal effect. The LD50 value of intraperitoneal injection of C. zeylanicum essential oil was 2.07 mL/kg body weight after 48 h, and the maximum nonfatal dose was 1.52 mL/kg body weight. The results also showed that there was no significant toxicity following oral administration of C. zeylanicum essential oil for 2 weeks.Conclusion:The results exhibited the favorable scolicidal activity of C. zeylanicum, which could be applied as a natural scolicidal agent in hydatid cyst surgery.SUMMARY We evaluated the efficacy of Cinnamomum zeylanicum essential oil against hydatid cyst protoscolecesThe viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining)Forty-eight male NMRI mice were also used to determine the toxicity of C. zeylanicum essential oilC. zeylanicum with potent scolicidal activity could be applied as a natural scolicidal agent in surgery. Abbreviations used: GC/MS: Gas chromatography/mass spectrometry analysis; CE: Cystic echinococcosis; LD50: Lethal dose 50%; I.p: Intraperitoneally.

English

Medicinal plants are now recognized as most important alternative for chemical drugs resource, especially for anti-microbial targets. The current study aimed to determine the scolicidal effect of Allium paradoxium and Tanacetum parthenium as medicinal plants in vitro. Protoscoleces were exposed to different concentrations of chloroformic and hydroalcholic extracts (1, 10, 50 and 100 mg/mL) for 5, 10, 30, 60 and 120 min. Viability of protoscolices was confirmed by 0.1% Eosin staining. The results of this study showed that all extracts used were not significantly effective in comparison with the control group. Therefore, chloroformic and hydroalcholic extracts of A. paradoxium and T. parthenium could not be used during hydatid cyst treatment. However, there remains the need to perform advanced comparative clinical studies on the efficacy of A. paradoxium and T. parthenium and other scolicidal agents. Chloroformic and hydroalcoholic extract of A. paradoxium and T. parthenium, did not show any scolicidal activity at the concentration of 1, 10, 50 and 100 mg/ml. Key words: Allium paradoxium, Tanacetum parthenium, Echinococcus granulosus, in vitro.

Efficacy and Safety of Bunium Persicum (Boiss) to Inactivate Protoscoleces during Hydatid Cyst Operations.

Current scolicidal agents, which have been used for inactivation of protoscoleces during surgical procedures, are associated with adverse side effects including sclerosing cholangitis. This investigation aimed to evaluate the scolicidal effects of Bunium persicum (Boiss) essential oil against protoscoleces of hydatid cysts and also its toxicity in a mice model. Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (3.125-50 mcL/mL) were used for 5-30 min. The viability of protoscoleces was confirmed using the eosin exclusion test (0.1% eosin staining). Further, 48 male NMRI mice were used to determine the acute and sub-acute toxicity of B. persicum essential oil. The obtained results revealed that the B. persicum essential oil at the concentrations of 25 and 50 mcL/mL after 5 min of exposure killed 100% protoscoleces. The mean mortality rate of protoscoleces after 10 min of exposure to the concentration of 12.5 mcL/mL was 100%. Lower concentrations (6.25 and 3.125 mcL/mL) of B. persicum essential oil, however, indicated a delayed protoscolicidal effects. The LD50 value of intra-peritoneal injection of the B. persicum essential oil was 1.96 mL/kg body wt. No significant difference (p > 0.05) was observed in the clinical chemistry and hematologic parameters after oral administrations of B. persicum essential oil at the doses 0.05, 0.1, 0.2, and 4 mL/kg for 14 d. Our findings demonstrated the potent scolicidal activity of B. persicum with no significant toxicity; it might be used as a natural scolicidal agent in hydatid cyst surgery.

Chemical composition and scolicidal activity of Zataria multiflora Boiss essential oil

Zataria multiflora Boiss (Lamiaceae) commonly grows in Iran is a popular medicinal plant with various pharmacological activities mentioned in traditional Iranian medicine and modern phytotherapy. This study was designed to evaluate the chemical composition and scolicidal effects of Z. multiflora essential oil on the protoscoleces of hydatid cysts on an in vitro model. The components of the Z. multiflora essential oil were identified by GC/MS analysis. Protoscoleces were aseptically aspirated from the livers of naturally infected sheep. Various concentrations of essential oil, thymol and carvacrol were used for 5–30 minutes. Eosin exclusion test was used to determine the viability of protoscoleces. The main components were thymol (41.8%), carvacrol (28.8%), and p-cymene (8.4%). Findings showed that essential oil at the concentrations of 12.5 and 6.25 μL/mL killed 100% protoscoleces after 5 and 20 minutes of exposure, respectively. In addition, thymol and carvacrol at the concentrations of 100 μg/mL and 100 μL/mL killed 100% protoscoleces after 10 minutes incubation, respectively. Obtained results in this investigation for the first time demonstrated that Z. multiflora essential oil and its main components might be a natural source for the production of new scolicidal agents.

In vitro protoscolicidal effects of various concentrations of Ziziphora tenuior L. extract at different exposure times.

Hydatidosis is one of the most important zoonotic diseases and surgery is still the main treatment for this problem. One of the side effects of hydatid cyst surgery is recurrence, thus, searching and assessment of some new agents such as medicinal plant extracts are very important. In the present study, the scolicidal effect of ethanolic extract of Ziziphora tenuior (Z. tenuior) was investigated. Protoscolices were aseptically collected from sheep livers containing hydatid cyst and used in the experiments. Z. tenuior extract was used at concentration of 3-100 mg/ml for 10-60 min. Viability of protoscolices was determined by 0.1% eosin staining. Based on our results, Z. tenuior extract at concentration of 10 mg/ml killed all protoscolices after 20 min. However, this medicinal plant at concentration of 25 mg/ml destroyed all protoscolices in a shorter exposure time (10 min). Therefore, the scolicidal activity of the extract at 10 and 25 mg/ml concentrations was considerably effective in lower concentrations and shorter exposure times. The findings of this study showed that the ethanolic extract of Z. tenuior produces high scolicidal activity; it may be used as an appropriate and effective scolicidal agent in hydatidosis surgery. This is the first report on the protoscolicidal activity of Z. tenuior.

Chemical composition, efficacy and safety of Pistacia vera (var. Fandoghi) to inactivate protoscoleces during hydatid cyst surgery.

At present, various scolicidal agents have been used for inactivation of protoscoleces during hydatid cyst surgery, however, they are associated with serious adverse side effects including sclerosing colangititis (biliary tract fibrosis), liver necrosis and methaemoglobinaemia. This investigation was designed to evaluate the chemical composition and in vitro scolicidal effects of Pistacia vera (var. Fandoghi) essential oil against protoscoleces of hydatid cysts and also its toxicity in mice model. The components of the P. vera essential oil were identified by gas chromatography/mass spectroscopy (GC/MS) analysis. Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (25-200μl/mL) were used for 5-30min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). In addition, forty male NIH mice were used to determine the acute and sub-acute toxicity of P. vera essential oil for 2 and 14 days, respectively. The main components of P. vera essential oil were limonene (26.21%), α-pinene (18.07%), α-thujene (9.31%) and α-terpinolene (9.28%). Findings of the present study demonstrated that the P. vera essential oil at the concentrations of 100 and 200μl/mL killed 100% protoscoleces after 10 and 5min of exposure, respectively. The LD50 values of intraperitoneal injection of the P. vera essential oil was 2.69ml/kg body weight, and the maximum nonfatal doses were 1.94ml/kg body weight. No significant difference (P>0.05) was observed in the clinical chemistry and hematological parameters following oral administrations of P. vera essential oil at the doses 0.1, 0.2, and 0.4ml/kg for 14 days. The obtained findings demonstrated new chemical composition and promising scolicidal activity of the P. vera with no significant toxicity which might be used as a natural scolicidal agent in hydatid cyst surgery.

In vitro effect of sodium arsenite on Echinococcus granulosus protoscoleces

Cystic echinococcosis (CE) caused by the metacestodes of Echinococcus granulosus is an important cosmopolitan zoonosis. Surgery is the main treatment option for CE. Meanwhile, chemotherapy is used as an significant adjunct to surgery. However, the benzimidazole carbamate group and the existing scolicidal agents may not be as effective as hoped. In this study, we aimed to explore the in vitro effect of sodium arsenite (NaAsO2) on Echinococcus granulosus protoscoleces, the causative agents of CE. Protoscoleces of E. granulosus were incubated in vitro with 4, 8, 12, 16, and 20μM NaAsO2. Viability and changes in morphology were investigated by 0.1% eosin staining. The ultrastructural alterations were observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Additionally, caspase-3 activity was measured by colorimetric assay. Obvious protoscolicidal effect was seen with NaAsO2 at concentrations of 16μM and 20μM. Protoscolex mortality was 83.24% (16μM) and 100% (20μM) after 6 days post-incubation. SEM showed that the primary site of drug damage was the tegument of the protoscoleces. TEM analysis demonstrated that the internal tissues were severely affected and revealed an increase in the number of lipid droplets and vacuoles after treatment with 16μM NaAsO2. Meanwhile, the caspase-3 activity significantly increased in protoscoleces after 24h of NaAsO2 incubation compared to the untreated controls. Our study demonstrated the clear in vitro scolicidal effect of NaAsO2 against E. granulosus protoscoleces. However, the in vivo efficacy, specific mechanism, and any possible side effects of NaAsO2 remain to be investigated.

Protoscolicidal effects of chenodeoxycholic acid on protoscoleces of Echinococcus granulosus

Dissemination of protoscoleces-rich fluid during surgical operation for cystic echinococcosis is a major cause of its recurrence. Instillation of a scolicidal agent into hydatid cysts to reduce the risk of spillage of viable protoscoleces is an integral part of the surgical technique employed by many surgeons. In this study, the protoscolicidal effect of chenodeoxycholic acid (CDCA) was investigated. Freshly isolated protoscoleces were subjected to CDCA treatment (500, 1000, 2000, and 3000 μmol/L), and the effects on protoscoleces were investigated with the help of 0.1% eosin staining, electron microscopy, and colorimetric assay of caspase-3 like activity. Dose-dependent mortality of Echinococcus granulosus protoscoleces was observed within a few days of CDCA treatment. The treated protoscoleces showed loss of viability, and morphological changes such as contraction of the soma region, formation of blebs, rostellar disorganization, loss of hooks, destruction of microtriches, and formation of vesicles, lipid droplets, and lamellar bodies. Apoptosis was evident in the treated protoscoleces, as compared to the control group, which were cultivated for nearly 3 months. Our study indicates a therapeutic potential for CDCA as a protoscolicidal agent against E. granulosus. However, further studies are needed to test the long-term effects of CDCA in animal models.

In vitro effectiveness of Curcuma longa and Zingiber officinale extracts on Echinococcus protoscoleces.

Hydatid disease is an important economic and human public health problem with a wide geographical distribution. Surgical excision remains the primary treatment and the only hope for complete cure of hydatosis. The most important complications arising from surgical excision, however, is recurrence, which is due to dissemination of protoscolices during the surgery. Pre-surgical inactivation of the contents of the hydatid cyst by injection of scolicidal agent into the cyst has been used as adjunct to surgery in order to overcome the risk of recurrence. In the present study, ethanolic extracts of turmeric (Curcuma longa) and ginger (Zingiber officinale) were tested as scolicidal agent for Echinococcus protoscoleces. Protoscoleces were collected aseptically from sheep livers containing hydatid cysts. Three concentrations (10, 30 and 50 mg/ml) of each extract were investigated and viability of the protoscoleces was tested by 0.1% eosin staining. Ginger extract showed the strongest scolicidal effect (100%) after 20 min at a concentration of 30 mg/ml and 10 min at 50 mg/ml. The maximum scolicidal effect of turmeric was 93.2% after 30 min at a concentration of 50 mg/ml. It is concluded that turmeric and ginger extracts have high scolicidal activity and could be used as effective scolicidal agents against Echinococcus protoscoleces.

Effect of Pistacia atlantica Fruit and Leaf Extracts on Hydatid Cyst Protoscolices.

Treatment of human hydatidosis is mainly surgical, with chemical treatment being reserved as a coadjuvant treatment. Use of effective protoscolicidal agents during surgery of hydatid disease is essential to reduce the recurrence rate. The aim of this study was to investigate the scolicidal effects of Pistacia atlantica leaf and fruit hydroalcoholic extracts on protoscolices of hydatid cyst. Echinococcus granulosus protoscolices were obtained from 50 sheep infected with hydatid cysts. Various concentrations of plant hydroalcoholic extracts were used in different exposure times for viability assay of protoscolices. The scolicidal effects of the hydroalcoholic extracts of the leaf, fresh and dried fruits were significant compared to the control groups (P < 0.05). Among the Pistacia atlantica extracts tested, 0.1% (mg/ml) concentrations of fresh fruit extract (99.09 ± 1.27) and leaf extract (89.25 ± 18.42) had strong scolicidal effects in 360 min, of exposure times and the mortality rate decreased with the lower concentration. Information from the current study has the strong scolicidal effect of fresh fruit hydroalcoholic extract of Pistacia atlantica on protoscoleces, which may be used as a scolicidal agent during the surgery techniques.

Efficacy of Myrtus communis L. to Inactivate the Hydatid Cyst Protoscoleces

ABSTRACTPurpose: The present study aims to investigate the scolicidal effects of Myrtus communis L. essential oil against protoscoleces of hydatid cysts and also its toxicity in mice model. Materials and Methods: Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (12.5–100 μl/ml) were used for 5–30 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Moreover, 48 male NMRI mice were used to determine the acute and sub-acute toxicity of M. communis essential oil. One-way ANOVA with Tukey's post-hoc test was used to assess differences between experimental groups. Results: Findings of the present study demonstrated that the M. communis essential oil at the concentration of 100 μl/ml after 5 min of exposure killed 100% protoscoleces. Similarly, the mean mortality rate of protoscoleces after 10 min of exposure to concentration of 50 μl/ml was 100%. However, lower concentrations (12.5 and 25 μl/ml) of M. communis essential oil provoked a delayed protoscolicidal effects. The LD50 values of intraperitoneal injection of the M. communis essential oil was 2.23 mL/kg body wt. No significant difference (p > .05) was observed in the clinical chemistry and hematological parameters following oral administrations of M. communis essential oil at the doses 0.05, 0.1, 0.2, and 0.4 mL/kg for 14 days. Conclusion: The results showed potent scolicidal activity of M. communis with no significant toxicity, which might be used as a natural scolicidal agent in hydatid cyst surgery.

Arsenic trioxide negatively affects Echinococcus granulosus.

Spillage of cyst contents during surgery is the major cause of recurrences of hydatidosis, also called cystic echinococcosis (CE). Currently, many scolicidal agents are used for inactivation of the cyst contents. However, due to complications in the use of those agents, new and more-effective treatment options are urgently needed. The aim of this study was to investigate the in vitro efficacy of arsenic trioxide (ATO) against Echinococcus granulosus protoscolices. Protoscolices of E. granulosus were incubated in vitro with 2, 4, 6, and 8 μmol/liter ATO; viability of protoscolices was assessed daily by microscopic observation of movements and 0.1% eosin staining. A small sample from each culture was processed for scanning and transmission electron microscopy. ATO demonstrated a potent ability to kill protoscolices, suggesting that ATO may represent a new strategy in treating hydatid cyst echinococcosis. However, the in vivo efficacy and possible side effects of ATO need to be explored.

Chemical composition, protoscolicidal effects and acute toxicity of Pistacia atlantica Desf. fruit extract

The present study was designed to evaluate the chemical composition and scolicidal effects of Pistacia atlantica Desf. extract against protoscoleces of hydatid cysts and its acute toxicity in mice model. Various concentrations of the methanolic extract (5–50 mg/mL) were used for 10–60 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1%). Acute toxicity was also determined in mice model. The main components were β-myrcene (41.4%), α-pinene (32.48%) and limonene (4.66%). Findings demonstrated that P. atlantica extract at the concentrations of 25 and 50 mg/mL after 20 and 10 min of exposure killed 100% protoscoleces. The LD50 of the intraperitoneal injection of the P. atlantica methanolic extract was 2.43 g/kg and the maximum non-fatal dose was 1.66 g/kg. Obtained results showed the potential of P. atlantica extract as a natural source with no significant toxicity for the production of new scolicidal agent to use in hydatid cyst surgery.

Open Conservative Surgical Management of Cystic Echinococcosis in a Tertiary Care Hospital, Nepal.

Cystic echinococcosis (CE) or hydatid disease caused by E. granulosus in Nepal is amenable to surgical treatment. Aim of the study is to evaluate the efficacy of surgical treatment of CE, by open partial pericystectomy with albendazole as adjuvant. Material of this prospective study were the consecutive series of 33 patients operated for CE, over a period of 8 years, at a single centre. Clinical examination, ultrasonography (USG) and computed tomography (CT) were used for establishing diagnosis. Patients were prescribed perioperative albendazole. Povidone iodine 10% (betadine)was used as contact scolicidal agent during operation. Cysts were evacuated from livers, lungs, retroperitoneum by partial pericystectomy. CE of mesentery was completely excised. Descriptive statistics was obtained using EPI- info windows version soft ware. A total of 33 patients were operated for CE; 24 were females and 9 males. Age ranged from 4 years to 80 years. Organs/ site involved were: liver - 24, lungs - 4, combined liver and lungs - 2, retroperitoneum - 2 and mesentery - 1. Complication - bile leak for 2 weeks in an operated CE of liver. There was no mortality. Hospital stay (in days) was - mean 14 (range 7to21). Follow up for 3 years (average 2years) showed no recurrence. Evacuation of CE by partial pericystectomy is an effective, safe and simple procedure, and gives excellent cure rate with perioperative albendazole therapy.

Acute rupture of a peritoneal hydatid cyst

Summary Echinococcus granulosus, the pathogen responsible for hydatid disease, mostly settles in the liver and lungs but affects the peritoneum less frequently. Rupture of a cyst into the peritoneal cavity is a potentially life-threatening incident. Although numerous studies on ruptured hepatic hydatid cysts have been published, few cases of peritoneal cyst rupture have been reported. We describe the case of a 19-year-old woman who presented with an acute abdomen and allergic reactions after a fall. Ultrasonography and computed tomography revealed a hydatid cyst of the liver and ruptured pelvic hydatid cyst. First, the patient received appropriate measures to prevent anaphylactic shock and later underwent emergency surgery. Partial cystectomy of the ruptured pelvic hydatid cyst, peritoneal washing, and unroofing of the large unruptured hepatic hydatid cyst were conducted. Albendazole was administered postoperatively for 3 months. No recurrence was noticed during 3 years of follow-up. Although rarely documented, acute rupture of a peritoneal hydatid cyst is the most severe complication of peritoneal echinococcosis. Typically after trauma, it must be considered in the presence of an acute abdomen with allergic reactions. Ultrasonography and computed tomography have high sensitivity in demonstrating rupture of a hydatid cyst. Emergency surgery is the only effective treatment and should aim at the complete removal of a cyst, if possible, and peritoneal washing with scolicidal agents. Additional studies should be conducted to evaluate the feasibility of laparoscopy. Albendazole should be prescribed postoperatively to prevent recurrence. Mortality is closely related to anaphylaxis; hence, early and accurate diagnosis and appropriate preventive measures are crucial.

Scolicidal activity of biosynthesized silver nanoparticles against Echinococcus granulosus protoscolices

Cystic hydatid disease (CHD), a helminth infection with various clinical complications caused by the larval stage of the dog tapeworm Echinococcus granulosus is considered as a public health problem in different regions of the world. To date, scolicidal agents have been broadly applied for inactivation of the fertile cysts and these scolicidal agents have several side effects on patients. Therefore, the objective of this study was to evaluate the scolicidal efficacies of synthesized silver nanoparticles (AgNPs) derived from the aqueous aerial extract of Penicillium aculeatum against protoscolices of CHD in-vitro. Protoscolices were aseptically aspirated from infected livers of sheep with CHD. Various concentrations (0.025, 0.05, 0.1 and 0.15 mg/mL) of green synthesis of Ag-NPs and different exposure times (10, 30, 60 and 120 min) were used against protoscolices of CHD. Viability of protoscolices was confirmed by 0.1% eosin staining. The findings showed that the Ag-NPs at all concentrations have high scolicidal effects. The concentrations 0.1 and 0.15 mg/mL after 120 min of exposure times showed 83% and 90% mortality rate, respectively. The least scolicidal activity of biosynthesized Ag-NPs was 40% (0.025 mg/mL and 10 min). The current investigation indicated that applying biogenic Ag-NPs may be considered as a potential scolicidal agent for CHD surgery due to being economical, safer and non-toxic compared to the used chemical materials. However, further studies are required to evaluate the efficacy of Ag-NPs in vivo.

Optimization of fluoride-containing bioactive glasses as a novel scolicidal agent adjunct to hydatid surgery

We have presented a novel scolicidal agent made from fluoride-containing bioactive glass (FBG). The samples were characterized by XRD and the ion release behavior of the samples was evaluated at 37°C. The scolicidal activity, cellular response and antibacterial activity and biocompatibility of the different concentrations of the FBG (ranging from 0.5 to 2mgml−1) were evaluated in vitro and in vivo. The results showed an easy diffusion of the fluoride through the glass matrix in an F concentration-dependent manner. The FBG showed an intensive scolicidal property, so that toxicity effect begun from 5min and the samples with 20, 10, 5 and 0% fluoride showed 98±2, 93±5.8, 76.2±6 and 5.8±1.7% scolicidal activity, respectively, after 8h exposing time. Our data revealed that scolicidal activity of the FBG increased with the increase of F ratio and concentrations of the bioactive glass. On the other hand, all the synthesized FBG samples found to be cyto-biocompatible when tested in vitro (MTT and LDH assays) and in vivo (no significant infiltration of inflammatory cells compared with control, p>0.05), and antibacterial against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. The results demonstrated that the synthesized FBG might have a potential application in prevention of post-surgery infections, especially hydatidosis.

Zataria multiflora Bioss: lethal effects of methanolic extract against protoscoleces of Echinococcus granulosus.

The present study was aimed to evaluate the lethal effects of Zataria multiflora Boiss (Lamiaceae) methanolic extract against Echinococcus granulosus protoscoleces. Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (2.5-20mg/mL) were used for 10-60min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Obtained results showed that Z. multiflora extract at the concentration of 20mg/mL after 10min of exposure killed 100% protoscoleces. The mean of mortality rate of protoscoleces after 20min of exposure to the concentration of 10mg/mL was also 100%. Lower concentrations of Z. multiflora extract provoked a delayed protoscolicidal activity. The findings indicated potential of Z. multiflora methanolic extract as a natural source for the producing of new scolicidal agent for use in hydatid cyst surgery.

Povidone-iodine toxicity in a child posted for laparoscopic removal of hepatic and renal hydatid cysts.

Sir, Povidone-iodine (PI) is an antiseptic solution consisting of polyvinylpyrrolidone with water and iodine (available iodine 1%). The 10% solution has been used effectively as a scolicidal agent for hydatid cysts. We hereby discuss the case of a child posted for laparoscopic removal of hepatic and renal hydatid cysts, who developed metabolic acidosis with acute renal failure. A 10-year-old child (weight 26 kg) presented with history of pain in epigastrium. Pain was dull, mild to moderate, and there was no radiation. Systemic examination revealed hepatomegaly. Computed tomography scan abdomen revealed well-defined cystic lesion of 12.5 × 8.5 × 11.4 cm in the left lobe of liver extending to lesser sac and perihepatic area along with small cysts in right kidney. Routine blood investigations and liver function tests were normal. Anti-Echinococcus IgG antibody was positive by Enzyme Linked Immunosorbent Assay. Diagnosis of hydatid cyst of liver and kidney was made, and the child was posted for elective laparoscopic removal of hepatic and renal cysts. Baseline haemodynamic parameters were normal for age. Inhalational induction performed with sevoflurane (8%) in oxygen. After administering fentanyl (60 μg) and vecuronium (3 mg), trachea was intubated. A total of 2.5 L of 10% PI (Microwin®10% w/v) was injected into hepatic hydatid cyst. An hour into the procedure, suddenly heart rate increased from 110 to 180/min along with hypercarbia (end-tidal CO255mmHg) and hyperthermia (from 35.1 to 37.4°C). Injection paracetamol (500 mg) and a bolus (0.5 L) of normal saline were infused. An arterial blood gas analysis showed pH 7.14, pCO236.5 mmHg, pO2135 mmHg, SO297.7%, K+7.4 mmol/L, Na+127 mmol/L, Ca+0.81 mmol/L, Cl − 249 mmol/L, lactate 4.3 mmol/L, base deficit - 15 mmol/L, HCO312.7 mmol/L with Hb of 16 gm%. Blood pressure and airway pressures were normal. Urine output decreased and did not improve after first fluid bolus so a further fluid bolus of 0.5 L of balanced salt solution (sterofundin®) and injection frusemide 15 mg intravenous (IV) were administered. Urine output improved and presumptive diagnosis of anaphylaxis due to systemic entry of hydatid cyst fluid was suspected. Injection hydrocortisone and pheniramine maleate were administered. Patient was shifted to intensive care unit (ICU). Investigations revealed haemoglobin 17 g/dL, leukocyte count 49.6 × 103/μL, serum creatinine 0.9 mg/dL, serum albumin 1.9 g/dL, and potassium 5.2 mEq/L in the immediate post-operative period. Liver, renal, and thyroid function tests were in normal range. Severe metabolic acidosis persisted and thus plan for haemodialysis was made. Tachycardia was fluid responsive and Sterofundin®100 ml/h, albumin (20%) 25 ml/h and hydrocortisone 6 mg/h were started. Bedside echocardiography revealed good cardiac contractility, under filled heart chambers and inferior vena cava diameter of 0.4 – 0.8 cm with collapsibility. After 3–4 h of initiation of haemodialysis, acidosis was corrected and heart rate settled. On post-operative day 1, the patient was fully awake and obeying commands. Trachea was extubated after completion of dialysis. Hydrocortisone infusion was tapered and stopped. Patient was kept for another 2 days in ICU for observation before shifting to ward. In its complexed form (i.e. PI), iodine vapour pressure is reduced essentially to zero, and it becomes soluble in water. The available iodine is released at very slow rate from the complex, prolonging germicidal action of free iodine. Povidone-iodine is highly nephrotoxic and due to its renal elimination, iodine intoxication can result in lethal vicious cycle and anuria.[1] Clinically, patients with PI intoxication can present with hypotension, nausea, vomiting, acute renal failure, hypothyroidism, confusion, metabolic acidosis, seizures, blindness, fever, rash, uterine infarction, elevated hepatic enzymes and iodine-induced haemolysis with decreased serum haptoglobin and lactate dehydrogenase levels.[1] Serum iodine levels could not be done as this analysis was not available in our hospital. Labbe et al. reported a case where abnormalities of cardiac conduction, lactic acidosis, acute renal failure, hypocalcaemia and thyroid dysfunction were features of intoxication.[2] In another case report of iodine toxicity in a child with mediastinitis, after median sternotomy, continuous PI irrigation was carried out and the child died due to its toxicity.[3] Iodinated contrast medium (IV) can cause contrast-induced acute kidney injury, but even with non-intravascular use of iodinated compounds, iodine absorption through mucosal, burned skin or interstitial tissues, can cause nephrotoxicity.[4] Early implementation of renal replacement therapy can be lifesaving.

English

Surgical removal of intact hydatid cyst is the most effective treatment for hydatid disease. Recurrence of hydatid cyst is mainly due to dissemination of protoscolices (PSCs) rich fluid during the surgical operation. Therefore, preoperative instillation of a scolicidal agent into the cyst is a common practice with adverse side effects of the used drugs. All-trans retinoic acid (atRA) is the physiological mediator of most of the functions of vitamin A, particularly as a cellular differentiation and apoptosis regulating factor. We hypothesized that instillation of atRA could provide an alternative safe scolicidal approach. We tested the scolicidal effects and ultrastructural changes imposed by atRA on human hydatid cyst PSCs in vitro. Freshly isolated hydatid cyst PSCs were subjected to atRA (at 16.7, 1.67, 0.167 &micro;M and 16.7 nM/L). Changes in protoscolices viability (0.1% eosin exclusion) and morphology (scanning and transmission electron microscopy; SEM and TEM) were investigated. Dose-dependent PSCs death within few minutes to 7 days of exposure to atRA was observed. SEM demonstrated ultrastructural damages including rosteller disorganization, loss of hooks and distortion of hooks morphology. TEM revealed loss of the integrity of the internal tissues of PSCs, an increased vacuolization, formation of large lipid droplets in the distal cytoplasm and aberrant, rounded abnormally large sized mitochondria. atRA is a promising alternative to the available synthetic and chemical scolicidal agents. However, in vivo scolicidal activities of atRA and the possible side effects necessitate further studies. Key words: Hydatid cyst, protoscolices, retinoic acid, ultrastructure, scolicidal activity. &nbsp