In vitro effectiveness of acidic and alkline solutions on scolices of hydatid cyst

The most confident way for treatment of hydatid cyst is surgical operation. Spillage of the cyst contents during the operation is the major cause of recurrence after hydatid cyst surgery. Instillation of scolicidal agent into hydatid cyst is the most commonly employed measure to prevent this complication. In the present study, the scolicidal effect of highly acidic and alkaline solutions is investigated. Protoscoleces were collected aseptically from sheep livers containing hydatid cyst. Acidic solutions with pH 1, 2, 3, and 4 and alkaline solutions with pH 11, 12, 13, and 14 were used for 5,10, and 15 min in the experiments. Viability of protoscoleces was assessed by 0.1% eosin staining. Scolicidal effect of acidic solution with pH 1 after 5 min and with pH 2 and 3 after 10 min was 100%. Scolicidal effect of acidic solution with pH 2 and 3 after 5 min was 99.6% and 98.7%, respectively. Acidic solution with pH 4 after 5, 10, and 15 min killed 15.5%, 21.5%, and 22.6% of protoscoleces, respectively. Alkaline solution with pH 14 after 5 min and with pH 13 after 15 min killed all protoscoleces. The scolicidal effect of alkaline solution with pH 13 after 5 and 10 min was also 97.5% and 99.7%, respectively. These values for alkaline solution with pH 12 were 29.33%, 33.44% and 37.09%, respectively. The scolicidal effect of solution with pH 11 was 24.5%, 30.5%, and 31.3%, respectively. Although the in vitro scolicidal effect of highly acidic or alkaline solutions was satisfactory in our study, in vivo efficacy of these solutions and also possible side effects, remain to be more investigated.     

Low scolicidal effect of Ocimum bacilicum and Allium cepa on protoccoleces of hydatid cyst: an in vitro study

Hydatidosis in humans and animals is an economic and public health treat in many parts of the world, and surgery is still the main treatment for hydatid disease. One of the most important endpoints of hydatid cyst surgery is recurrence. The main cause of recurrence is dissemination of protoscolices during the surgical operation. Preoperative destruction of the cyst’s contents through instillation of a scolicidal agent into the hydatid cyst is the most commonly used approach to prevent this complication. Various scolicidal agents have been used for inactivation of the cyst content, but most are accompanied by adverse side effects. In the present study, the scolicidal effect of methanolic extract of Ocimum bacilicum and Allium cepa is investigated. Protoscoleces were aseptically collected from sheep livers containing hydatid cyst and were used in the experiments. O. bacilicum and A. cepa extracts were used in 2.5, 5, and 10 % concentration for 10, 20, 30, 40, 50, and 60 min. Viability of protoscoleces was confirmed by 0.1 % eosin staining. The scolicidal effect of O. bacilicum and A. cepa extracts was not satisfactory and a 10 % concentration of these extracts after 60 min of application, killed only 24.1 and 16.8 % of protoscolices, respectively. Result of this study showed that, as opposed to the fact that these extracts have a strong antibacterial effect, they have low scolicidal activity and cannot be used as scolicidal agent; on the contrary of in vitro inefficacy, the in vivo efficacy of these extracts and also their possible side effects remained to be examined.     

High scolicidal effect of Zataria multiflora on protoccoleces of hydatid cyst: an in vitro study

Hydatidosis in humans and animals is an economic and public health problem in many parts of the world, and surgery is still the main treatment for hydatid disease. One of the most important endpoints of hydatid cyst surgery is recurrence. The main cause of recurrence is dissemination of protoscolices during the surgical operation. Preoperative destruction of the cyst’s contents through instillation of a scolicidal agent into the hydatid cyst is the most commonly used approach to prevent this complication. Various scolicidal agents have been used for inactivation of the cyst content, but most are accompanied by adverse side effects. In the present study, the scolicidal effect of methanolic extract of Zataria multiflora is investigated. Protoscoleces were aseptically collected from sheep livers containing hydatid cyst and were used in the experiments. Z. multiflora extract was used at a concentration of 10 and 25 mg/ml for 1, 2, and 3 min. Viability of protoscoleces was confirmed by 0.1% eosin staining. Z. multiflora extract at a concentration of 10 mg/ml killed 68.9%, 93.7%, and 100% of protoccoleces after 1, 2, and 3 min respectively. The scolicidal effect of this extract at a concentration of 25 mg/ml was 100% after 1 min. The results of present study showed that methanolic extract of Z. multiflora has high scolicidal activity and might be used as an effective scolicidal agent. This is the first report on the scolicidal activity of Z. multiflora.     

Echinococcus granulosus calreticulin: molecular characterization and hydatid cyst localization

Hydatidosis, whose etiological agent is the larval stage of the platyhelminth parasite Echinococcus granulosus, is a major zoonotic public health problem and causes great economic losses in many countries, affecting humans and livestock species. Calreticulin (CRT) is a multifunctional and conserved chaperone calcium-binding protein, present in every cell of higher organisms, except erythrocytes. In other parasites species, CRT emerges as a key modulator of several immunological aspects of their relationships with their vertebrate hosts. We report herein the cloning of two DNA segments coding for sequences of E. granulosus CRT (EgCRT). Together with another partial sequence available at the NCBI database, a complete EgCRT cDNA sequence is now proposed. EgCRT mRNA is equally expressed in fertile and infertile hydatid cysts germinal layers, as well as in protoscoleces. In hydatid cysts, EgCRT is a 50 kDa protein, expressed in germinal layer and protoscoleces, mainly in perinuclear and cytoplasmic zones.     

Skin disinfection with octenidine dihydrochloride for central venous catheter site care: a double-blind,randomized, controlled trial

To compare the efficacy of two commercially available, alcohol-based antiseptic solutions for preparation and care of central venous catheter (CVC) insertion sites, with and without octenidine dihydrochloride, a double-blind, randomized, controlled trial was undertaken in the haematology units and in one surgical unit of two university hospitals. Adult patients with a non-tunnelled CVC were randomly assigned to two different skin disinfection regimens at the insertion site: 0.1% octenidine with 30% 1-propanol and 45% 2-propanol, and as control 74% ethanol with 10% 2-propanol. Endpoints were (i) skin colonization at the insertion site; (ii) -positive culture from the catheter tip (≥15 CFU); and (iii) occurrence of CVC-associated bloodstream infection (defined according to criteria set by the CDC). Four hundred patients with inserted CVC were enrolled from May 2002 through April 2005. Both groups were similar in respect of patient characteristics and co-morbidities. Skin colonization at the CVC insertion site during the first 10 days was significantly reduced by octenidine treatment (relative difference octenidine vs. control: 0.21; 95%CI: 0.11–0.39, p <0.0001). -positive culture of the catheter tip was significantly less frequent in the octenidine group (7.9%) than in the control group (17.8%): OR = 0.39 (95%CI: 0.20–0.80, p 0.009). Patients treated with octenidine had a non-significant reduction in catheter-associated bloodstream infections (4.1% vs. 8.3%; OR = 0.44; 95%CI: 0.18–1.08, p 0.081). Side effects were similar in both groups. This randomized controlled trial supports the results of two observational studies demonstrating octenidine in alcoholic solution to be a better option than alcohol alone for the prevention of CVC-associated infections.     

Étude des aspects lésionnels de l’échinococcose hydatique chez l’Homme en Mauritanie: fertilité, histologie des kystes hydatiques et viabilité des protoscolex

A lesional study on human hydatidosis (localization, fertility of cysts, protoscolex viability, and histological structure) focused on 63 human hydatid cysts from patients operated at CHN of Nouakchott was conducted during the period 1997-2007. The authors report the following results: the annual surgical incidence rate was in the order of 1.2% per 100,000 inhabitants. The lung was the primary localization of cyst (52%) followed by the liver (33%), spleen (4%), brain (3%), heart (2%), breasts (2%), kidney (2%), diaphragm (1%), and peritoneum (1%). The pulmonary cysts were larger and more fertile (76%) with protoscoleces also more viable (70%) compared with liver cysts (67-62%). The histology of hydatid cysts revealed in most cases a histological structure consistent with that described in the literature: thick cuticle, membrane proligeous, and capsules visible.     

First Successful Puncture, Aspiration, Injection, and Re-Aspiration of Hydatid Cyst in the Liver Presenting with Anaphylactic Shock in Korea

Hydatid disease is a parasitic infestation caused by the larval form of Echinocococcus. In human, the most commonly affected organs are liver and lung. Most cysts remain clinically silent and are diagnosed incidentally or when complications occur. In Korea, hydatid disease is rare and surgically treated cases have been reported in the Korean literature. However, it is expected to confront this disease sooner or later, because of recent increase in traveling to the endemic area and industrial workers originating from those areas. With this trend, we experienced a case of hydatid cyst of the liver in a male patient from Uzbekistan. This patient was presented with anaphylactic shock combined with hydatid cyst. We successfully treated using ultrasound-guided transhepatic percutaneous drainage [termed puncture, aspiration, injection, and re-aspiration (PAIR)] of the hydatid cyst and concomitant albendazole instead of surgery. In this clinical case report, we describe all the course of the patient and recommend the PAIR as a first choice method for treatment of hepatic hydatid cyst.     

The damages of high intensity focused ultrasound to transplanted hydatid cysts in abdominal cavities of rabbits with aids of ultrasound contrast agent and superabsorbent polymer

The present study investigates the damages of high intensity focused ultrasound (HIFU) to transplanted hydatid cysts in abdominal cavities of rabbits with aids of ultrasound contrast agent (UCA) and superabsorbent polymer (SAP) alone or in combination. A rabbit model with transplanted hydatid cyst was established by implanting hydatid cyst isolated from infected sheep liver, and HIFU was used to ablate the transplanted cysts with the aid of UCA and SAP alone or in combination. The hydatid cyst with thin wall, good elasticity, approximately spherical, and a diameter of approximately 30 mm was selected for the following experiments. According to our previous studies, a mixture of 0.1 g SAP and 0.5 ml anhydrous ethanol, and the solution of 0.1 ml UCA SonoVue, or both materials were injected into different cyst before HIFU ablation, respectively. The cyst inoculated with the SAP and UCA alone or in combination was immediately implanted into the abdominal cavity of rabbit for HIFU ablation at a dosage of 100 W acoustic powers. The ablation mode was spot scanning at the speed of 3 mm/s. Every target point was scanned three times; every ablating time lasted 3 s. The distance of each ablated layer was 5 mm. The total ablation time depended on the volume of cyst. The comparison of ultrasound image for each layer of hydatid cyst was made before and after HIFU ablation. The protoscolices in ablated cysts were stained by trypan blue exclusion assay, and their structures were observed by light microscopy. To estimate ablation effects of HIFU to the walls of hydatid cysts, the ultrastructure changes of cyst walls were examined by electron microscopy. The pathological changes of rabbits’ skins through which ultrasound penetrated were observed to investigate the side effects of HIFU ablation. The results demonstrated that HIFU had some lethal effects to hydatid cysts in vivo, namely, echo enhancements of ultrasound images of cysts, increases in mortality rate of protoscolices from 15.19 % (HIFU alone) to 48.66 % (HIFU?+?SAP), 38.67 % (HIFU?+?UCA), and 67.75 % (HIFU?+?SAP?+?UCA), respectively, serious structural damages of protoscolices, and destructions or even disappearance of laminated layers and germinal layers in the walls of hydatid cysts ablated by HIFU aided with UCA and SAP alone or in combination. This study demonstrated that destructive effects of HIFU to transplanted hydatid cyst could be enhanced by UCA and SAP alone, but the destruction of HIFU aided with a combination of UCA and SAP to hydatid cysts was more effective than those aided with UCA or SAP alone. The enhanced thermal and cavitation effects of HIFU induced by UCA and SAP might be involved in the enhanced destructive effects of HIFU on hydatid cysts. There were no evidences of pathological changes on rabbits’ skins overlying the hydatid cysts after HIFU ablation. The results suggested that the rabbit model with transplanted hydatid cyst may serve as an optional animal model for the experiments of HIFU ablation to hydatid cyst in vivo, and the materials of UCA and SAP were proved as enhancing agents of HIFU ablation to hydatid cysts, and HIFU at a dosage of 100 W acoustic powers was a safe and feasible parameter to ablate the hydatid cysts in this special animal model. These results laid a theoretical foundation for improving HIFU therapy for cystic echinococcosis by inoculation of UCA and SAP into hydatid cysts.     

The destructive effects of high-intensity focused ultrasound on hydatid cysts enhanced by ultrasound contrast agent and superabsorbent polymer alone or in combination

Cystic echinococcosis, caused by the metacestode stage of Echincoccus granulosus, remains endemic in many regions around the world. The present work evaluated whether or not a superabsorbent polymer (SAP) and ultrasound contrast agent (UCA) alone or in combination could enhance damage efficacy of high-intensity focused ultrasound (HIFU) on hydatid cysts in vitro. HIFU of 100 W acoustic power, with the aid of 0.1 ml UCA and 0.1 g SAP alone or in combination, was used to ablate hydatid cysts in vitro. The comparison of ultrasound image for each layer of hydatid cyst before and after HIFU ablation was made immediately, and the protoscolices of the cysts were stained by eosin exclusion assay, and the structures of protoscolices were observed by light microscopy. To understand the destructive effects of HIFU, the pathological changes in cyst walls of hydatid cyst ablated with HIFU were examined. The results demonstrated that HIFU had some lethal effect on hydatid cysts: echo enhancement of ultrasound image, increase of mortality rate of protoscolices, serious structural damage of protoscolices, and complete destruction or even disappearance of laminated layer and germinal layer was observed in the group of HIFU combined with UCA and SAP alone or in combination. It was found that the destructive effect of HIFU aided with a combination of UCA and SAP to hydatid cysts was more effective than that of HIFU just aided with UCA or SAP alone. These results suggested that UCA and SAP might be used as a HIFU enhancing agent to improve the efficacy of HIFU ablation to hydatid cysts, which could be a possible therapeutic option for cystic echinococcosis.     

细粒棘球绦虫重组Bb—Eg95-EgA31疫苗诱导小鼠脾细胞增殖变化的研究

目的探讨细粒棘球绦虫（Eg）重组双歧杆菌（Bb）-Eg95-EgA31疫苗免疫和Eg原头节攻击后小鼠的囊重抑制率及脾细胞增殖的变化。方法将细粒棘球绦虫重组Bb-Eg95-EgA31疫苗分别采用皮下注射、肌肉注射、鼻腔内接种和口服灌胃4种途径免疫BALB／c小鼠,免疫后8周每鼠用50个Eg原头节攻击感染,感染后25周剖杀小鼠,分离细粒棘球蚴包囊并称重,计算囊重抑制率;取脾,分离脾细胞,用Eg粗抗原（EgAg）或刀豆素A（ConA）刺激培养,四甲基偶氮唑盐比色法（MTT法）检测免疫小鼠脾细胞增殖反应,同时设有空载体、Bb和MRS对照。结果以上4种疫苗接种组小鼠的囊重抑制率分别为45．33％、41．33％、70．67％和62．67％;疫苗接种组的脾细胞明显增殖;鼻腔内接种和口服免疫组的脾细胞增殖显著高于皮下和肌肉注射组。结论细粒棘球绦虫重组Bb-Eg95-EgA31疫苗可诱导小鼠产生特异性的细胞免疫反应。     

Failure of treatment with alpha-difluoromethylornithine against secondary multilocular echinococcosis in mice

In AKR/J mice inoculated intraperitoneally with protoscoleces ofEchinococcus multilocularis, the oral administration of alpha-difluoromethylornithine (DFMO) in drinking water had no obvious effect on the development of hydatid cysts. The DFMO treatment also caused no apparent depletion of three major polyamines (putrescine, spermidine, and spermine) in the cysts, although a slight reduction in the putrescine level was detected. The failure of DFMO treatment to reduce the growth of cysts in mice may result from the absence of ornithine decarboxylase in the parasite.     

细粒棘球绦虫抗氧化蛋白TPx的免疫定位

目的：研究抗重组细粒棘球绦虫抗氧化蛋白TPx（rEgTPx）多克隆抗体对天然EgTPx的结合活性，探讨EgTPx在原头蚴内的分布。方法：采集自然感染细粒棘球蚴的绵羊肝脏，在无菌条件下收集包囊内的原头蚴，经消化处理后制备石蜡切片。利用rEgTPx多克隆抗体，以间接免疫荧光法确定抗氧化蛋白EgTPx在原头蚴内的分布。结果：rEgTPx多克隆抗体能够特异性地结合天然EgTPx抗原表位，EgTPx广泛分布在原头蚴的体表皮层、皮下层和钙颗粒细胞内。结论：确定了EgTPx蛋白在细粒棘球绦虫原头蚴内的分布，为研究EgTPx在原头蚴发育的生物功能奠定基础。     

Praziquantel in human hydatidosis. Evaluation by preoperative treatment

The efficacy of praziquantel has been studied in human hydatic disease due to E. granulosus, using parasitological and pharmacological criteria of improvement. This evaluation uses a prospective therapeutic trial in 15 patients with one or several hydatic cysts in different sites. Before surgery, nine of these received daily 75 mg/kg of praziquantel in 2 courses of 10 days each. The other six patients are considered as controls. The protoscolices vitality is determined by direct optic microscopy and by intraperitoneal mouse inoculation. According to the results of optic microscopy, praziquantel sterilizes hydatic cysts: 19 sterilized cysts out of 26 coming from treated patients, and 4 out of 11 in control group. This effect seems to be emphasized in hepatic localization. Nevertheless, according to the results of mouse inoculation, praziquantel does not significantly reduce the pathogenicity of inoculated proscolices and germinal layer: 11 mice out of 39 are healthy in the treated group, and 5 out of 15 in controls. Moreover, the determination with a fluorimetric method has not detected praziquantel in the hydatid liquid issued from treated patients. The drug does not pass through the cyst wall, and so, cannot have a scolicidal activity. Praziquantel cannot be considered as a medical treatment in human hydatidosis.     

Albendazole treatment in cystic echinococcosis: pharmacokinetics and clinical efficacy of two different aqueous formulations

The pharmacokinetic (PK) behaviour and clinical efficacy of albendazole (ABZ) against hydatid cysts in mice were assessed after treatment with two different ABZ pharmaceutical formulations. BalbC mice received ABZ (0.5 mg/kg) prepared either as solution or suspension (50 microg/ml) for oral administration (PK study). Blood samples were collected up to 16 h post-treatment and processed to measure ABZ/metabolites concentrations in plasma. The clinical efficacy assessment was performed in BalbC mice infected 8 months earlier with Echinococcus granulosus protoscoleces. Infected animals were allocated into three experimental treatment groups: (a) untreated control, (b) ABZ-solution treated, (c) ABZ-suspension treated. Both treated groups received ABZ (0.5 mg/kg) administered under two different therapeutic schemes: dosing every 48 h over 30 days (regimen I) or treated every 12 h during 15 days (regimen II). Experimental mice were sacrificed 12 h after treatment, and cysts were recovered, weighed and processed for transmission electron microscopy. Enhanced ABZ sulphoxide (the main ABZ metabolite) concentration profiles were measured in animals treated with the ABZ solution. Any positive clinical response was obtained after treatment every 48 h (30 days therapy). However, consistent with the observed PK results, both ABZ formulations were clinically effective in infected mice treated with a 12-h dosing interval (15 days therapy).     

Albendazolesulphoxide concentrations in plasma and hydatid cyst and prediction of parasitological and clinical outcomes in patients with liver hydatidosis caused by Echinococcus granulosus

Aim

To investigate the relationship between plasma and cyst concentrations of albendazolesulphoxide (ASO) and their effects on parasitological findings and disease recurrence in patients with liver hydatidosis.

Methods

The study was conducted at the University Hospital for Infectious Diseases “Dr. Fran Mihaljević,” Zagreb, Croatia, between August 2006 and January 2011. Consecutive patients (N = 48, age 6-77 years) were treated with albendazole (3 × 5 mg/kg/d) over 28 days before surgical cyst removal (n = 34) or percutaneous evacuation (PAIR) (n = 14). Plasma ASO was determined on days 10 and 28 of treatment and cyst concentrations at surgery/PAIR.

Results

Disease recurred in 3 surgically treated patients. Variability of ASO concentrations was substantial. Plasma concentrations on day 10 were higher than on day 28 (geometric means ratio [GMR] 2.00; 95%CI 1.38-2.91, P < 0.001) and higher than cyst concentrations at the time of treatment (GMR = 1.58, 1.01-2.34, P = 0.045). Higher cyst (but not plasma) concentrations were independently associated with lower odds of protoscolex motility (OR = 0.23, 0.01-0.70, P < 0.001) and higher odds of protoscolex destruction (OR = 1.17, 1.04-1.46, P < 0.001). With adjustment for age and protoscolex motility, higher day 10 plasma concentrations (but not cyst concentrations) were associated with lower odds of disease recurrence (OR = 0.49, 0.09-0.97, P = 0.035). Plasma concentrations did not predict cyst concentrations.

Conclusion

Viability of protoscolices progressively decreased with increasing ASO concentrations in the cyst. Data strongly suggested that higher plasma concentrations reduced the risk of disease recurrence.Echinococcosis or hydatid cyst disease is an anthropozoonosis caused by the larvae of E. granulosus, E. multilocularis, E. vogeli, and E. oligarthus (1). In transient hosts (eg, sheep, cattle, pigs, humans), the parasite develops in the form of hydatid cyst(s) of different sizes. The cyst consists of the outer layer (ectocyst), made of dense fibrous tissue; the middle layer (endocyst), which is elastic and lamellar in structure; and the inner (germinative) layer, which gives rise to buds that develop into scolices. The cyst content is hydatid fluid, produced by the germinative layer. In humans, the most commonly invaded organs are the liver and the lungs, but practically all organs can be affected (1-3). In Croatia, the disease is caused exclusively by E. granulosus. According to the European Hospital Morbidity Database (4) for the period 2010-2012, age-adjusted annual hospital admission rates due to echinococcosis (ICD-B67) in Croatia varied between 0.011 and 0.0167/1000 population (ie, between 51 and 86 cases/y), indicating that the diseases is relatively rare but still stably present.Treatments for hydatid cyst disease include surgical removal of the cyst(s) (still most commonly used method); percutaneous aspiration of the cyst with instillation of a scolicidal agent (95% ethanol or hypertonic saline [15%-20%] or albendazole), ie, the PAIR (puncture, aspiration, instillation, and re-aspiration) procedure, which seems to have greater clinical efficacy and lower rate of complications than the surgical procedure; and treatment with benzimidazole anthelmintic drugs. The latter, pharmacological option might be used as a monotreatment in the case of smaller cysts or when invasive approaches are not feasible, but it is typically adjunctive treatment to surgery or PAIR used to prevent dissemination of scolices from ruptured cysts. In this setting, benzoimidazoles are used over at least 4 weeks before the definitive treatment (5-8). Among benzoimidazoles, albendazole is considered a cornerstone pharmacological treatment of echinococcosis. Although some controversies still exist regarding optimal dosing, the most widely accepted regimen implies administration of 10-15 mg/kg/d (5-8). Upon oral ingestion (with a fatty meal to increase bioavailability), albendazole is rapidly metabolized (first-pass metabolism in the liver) into the active form albendazolesulphoxide (ASO), which inhibits tubuline polymerization in the parasite microtubules and inactivates cell division (9). Greater systemic bioavailability is considered an important advantage of albendazole over mebendazole, the other member of the group (9). Although ASO has been shown to penetrate both the hepatic and non-hepatic cysts (9,10), the prognostic value of plasma and/or cyst concentrations has not been elucidated. Therefore, we aimed to relate ASO concentrations in the plasma and in the cysts, and to investigate their relationship with the parasitological and clinical outcomes in patients with liver hydatidosis treated with albendazole over one month prior to surgical treatment or PAIR.     

Pulmonary complications of cystic echinococcosis in children in Uruguay

Cystic echinococcal lesions surgically removed from 99 patients aged 1-15 years in Uruguay were histopathologically studied. The lesions were collected from the lungs, liver, peritoneum, omentum, pleura, ovary, appendix and perinephrium. All intact cysts were well-developed and unilocular. Pulmonary and hepatic fertile cysts in which protoscoleces were produced were in 75% and 47%, respectively, of all patients. Fertile cysts having many protoscoleces were in the lungs of a 1-year-old infant. Sixteen ruptured pulmonary cysts were found in the lungs of 13 patients and four ruptured hepatic cysts were found in the liver of three patients. Pulmonary complications were seen in patients having pulmonary echinococcal cysts. In lung tissue surrounding echinococcal cysts, pulmonary complications included chronic congestion, hemorrhage, bronchopneumonia, as well as interstitial pneumonia that included histological changes caused by bronchiolitis, organizing pneumonia and obstructive pneumonia. Liver tissue adjacent to echinococcal cysts showed atrophy of hepatic cells, proliferation of interstices, small focal hemorrhages, and infiltration of mononuclear cells. The present study shows that a close relationship exists between pulmonary echinococcosis and pneumonia. The term 'echinococcal pneumonia' is proposed for pneumonia accompanying pulmonary echinococcosis.     

Echinococcus granulosus: occurrence of ecdysteroids in protoscoleces and hydatid cyst fluid

Both free ecdysteroids and hydrolysable polar conjugated ecdysteroids were detected in protoscoleces of Echinococcus granulosus from the equine host, and in hydatid cyst fluid from the same source. Comparisons were made of hydatid cyst fluid from E. granulosus infections of three intermediate host species: horses, sheep and humans. Ecdysone and 20-hydroxyecdysone were identified in both protoscoleces and hydatid cyst fluids by high-performance liquid chromatography monitoring fractions by radioimmunoassay, and by capillary gas chromatography/mass spectrometry (selected ion monitoring). The free ecdysteroid fractions of hydatid cyst fluid from horses and sheep also contained several unidentified, chromatographically unique, immunoreactive compounds which were refractory to hydrolysis with a crude Helix pomatia aryl sulphatase enzyme preparation.     

Flubendazole and ivermectin in vitro combination therapy produces a marked effect on Echinococcus granulosus protoscoleces and metacestodes

The aim of the present work was to evaluate the in vitro efficacy of the flubendazole (FLBZ) and ivermectin (IVM) combination against Echinococcus granulosus protoscoleces and metacestodes. Protoscoleces and groups of ten peritoneal cysts obtained from BALB/c mice were incubated with the two drugs, either separately or in combination, at the following final concentrations: 10 μg/mL FLBZ, 1 μg/mL FLBZ, 1 μg/mL IVM, 10 μg/mL FLBZ + 1 μg/mL IVM, and 1 μg/mL FLBZ + 1 μg/mL IVM. The maximum protoscolicidal effect was found with the combination 10 μg/mL FLBZ + 1 μg/mL IMV. After 1 day of incubation, the presence of numerous blebs in the tegument of protoscoleces was observed. Ultrastructural studies revealed that the primary site of damage was the tegument of the parasite. The effect of the two drugs on hydatid cysts obtained from mice was more rapidly detected in cysts treated with the combination of FLBZ + IVM than when drugs were used separately. Ultrastructural studies revealed that the germinal layer of treated cysts lost the multicellular structure feature and underwent considerable degenerative changes after in vitro treatment. The outcomes obtained demonstrated the favorable effect of the combination of FLBZ and IVM against E. granulosus.     

Sepsis of vascular catheters. II: In vitro disinfection of colonized tubing

Preparatory to development of in situ disinfection of implanted catheters, silicone rubber tubing colonized by incubation with Staphylococcus aureus, Staphylococcus epidermidis, or Klebsiella pneumoniae was used to test the efficacy of various chemicals in vitro. Protocols sterilizing all segments colonized for 24 h (n = 30) were immersion into 50% povidone iodine for 5 and 60 min, 100% povidone iodine for 5, 15, and 60 min, 1.2 x 10(3) ppm chlorine dioxide for 15 and 60 min, and 1.2 x 10(3) ppm chlorine dioxide buffered to pH 5.1 for 60 min. Immersion in up to 2% chlorhexidine, 7.4% formaldehyde, or 6% sodium hypochlorite for up to 60 min failed to sterilize all segments. None of 117 control segments were sterilized. Segments colonized for seven days were sterilized by immersion into 100% povidone iodine for 15 or 60 min. Use of 1.2 x 10(3) ppm chlorine dioxide for 60 min sterilized 97% of segments tested. Lower concentrations and shorter exposure times failed to sterilize all segments. Eighteen silicone rubber catheters, colonized on the outer surface, were all sterilized within 24 h by a chlorine dioxide solution placed in the lumen and diffusing through the wall to kill the bacteria.     

Ultrastructural changes in hydatid cyst walls obtained from human cases,exposed to different therapeutic approaches

Recurrence of cystic echinococcosis as a result of treatment failure is frequently reported to cause a major problem in management of such serious parasitic infection. The deeply seated innermost germinal layer of hydatid cysts is a relatively delicate layer, yet responsible for viability maintenance of this parasitic stage. In this study, a trial was done to explore the ultrastructural changes in germinal and laminated layer of the hydatid cyst for the first time in human cases exposed to different therapeutic approaches which were done earlier to the final open surgical intervention. Four groups were included: group 1 did not receive any earlier form of treatment; group 2 was previously treated with only medical therapy; group 3 was treated with a single course of medical treatment, plus a single PAIR technique; group 4 was treated with multiple courses of medical treatment plus multiple PAIR techniques. Complete alteration of ultrastructural features of germinal and laminated layers were observed only with samples from group 4, indicating a kind of failure of the therapeutic approaches used in group, 1, 2, and 3, unless repeated in group 4 to achieve a real change regarding the fitness of the parasitic cystic lesions. Searching for more effective, safe, therapeutic method is highly recommended which may end the suffering of the affected patients.