First parasitological, histopathological and molecular characterization of Echinococcus vogeli Rausch and Bernstein, 1972 from Cuniculus paca Linnaeus, 1766 in the Cerrado biome (Mato Grosso do Sul, Brazil).

Polycystic echinococcosis (PE) is caused by Echinococcus vogeli metacestodes (larval stage) in Neotropical countries. E. vogeli is trophically-transmitted between predators bush dogs (Speothos venaticus) and prey pacas (Cuniculus paca). In Brazil, reported PE cases are restricted to the Amazon biome. In this study, metacestodes from a paca hunted in Mato Grosso do Sul state (Cerrado biome) were identified morphological and histopathological techniques and further confirmed by molecular testing (sequencing of cytochrome C oxidase subunit I (cox1) gene) for the first time. Images of the whole liver showed superficial bubble-like hepatic masses. The parasitological analysis revealed large hooks (41.3 ±â€¯1.2 µm length/12.8 ±â€¯0.8 µm width) and small hooks (33.0 ±â€¯1.5 µm length/11.1 ±â€¯1.2 µm width), consistent with E. vogeli. Microscopically, the liver showed protoscoleces, a thick laminated layer, fibrosis, and inflammatory infiltrate in the adventitial layer. The DNA sequencing confirmed E. vogeli with 99% homology with sequences deposited in the GenBank. In addition, this finding greatly extends the geographic range of animal polycystic echinococcosis into the Cerrado. It is likely to occur in new biomes, where bush dogs and pacas share a given area in a trophic relationship.

A study of morphological aspects of cystic echinococcosis in sheep in Sardinia.

We have investigated the morphological and structural aspects of hydatid cysts recovered from sheep raised in southern Sardinia during two surveys conducted over the 20 years since the last control attempt at the end of the 1980s. In the first study carried out during 1995-1997, a few years after the last control attempt, a total of 10,334 cysts were examined, recovered from 1029 infected sheep out of a total of 1375 inspected (prevalence 74.84%). In the second survey, conducted ten years later during the period 2005-2010 in the absence of specific control measures, a total of 6249 cysts were recovered from 916 parasitized sheep out of the 1414 examined (prevalence 64.78%). Cysts were grouped into 5 different types: "Unilocular", "Multisepted", "Calcified", "Caseous", "Hyperlaminated". Unilocular: fertile, consist of a single fluid-filled cavity; multisepted: generally sterile, fluid filled, with cavity divided into spheroidal chambers (3-dozen); calcified: sterile, small, with internal chambers almost virtual due to the thickening of internal septa; caseous: sterile, the cavity filled with a thick yellowish matrix; hyperlaminated: sterile, with a virtual cavity filled with extensively folded and overlapping sheets of hyperproduced laminated tissue. A thorough knowledge and the precise characterization of each type of lesion, making it possible to establish detailed parameters for cyst classification, appears very useful for the harmonization of data collection. From a surveillance and control perspective it is proposed that, in addition to other relevant epidemiological information, data from meat inspection be further improved, including morpho-functional indications about cyst type.

Morphometric characteristics of the metacestode Echinococcus vogeli Rausch & Bernstein, 1972 in human infections from the northern region of Brazil.

Polycystic echinococcosis, caused by the larval stage (metacestode) of the small-sized tapeworm, Echinococcus vogeli, is an emerging parasitic zoonosis of great public health concern in the humid tropical rainforests of South and Central America. Because morphological and morphometric characteristics of the metacestode are not well known, hydatid cysts from the liver and the mesentery were examined from patients following surgical procedures. Whole mounts of protoscoleces with rostellar hooks were examined under light and confocal laser scanning microscopy. Measurements were made of both large and small hooks, including the total area, total length, total width, blade area, blade length, blade width, handle area, handle length and handle width. The results confirmed the 1:1 arrangement of hooks in the rostellar pad and indicated, for the first time, that the morphometry of large and small rostellar hooks varies depending upon the site of infection. Light and confocal microscopy images displayed clusters of calcareous corpuscles in the protoscoleces. In conclusion, morphological features of large and small rostellar hooks of E. vogeli are adapted to a varied environment within the vertebrate host and such morphological changes in calcareous corpuscles occur at different stages in the maturation of metacestodes.

Understanding the laminated layer of larval Echinococcus I: structure.

Echinococcus larvae are protected by a massive carbohydrate-rich acellular structure, called the laminated layer. In spite of being widely considered the crucial element of these host-parasite interfaces, the laminated layer has been historically poorly understood. In fact, it is still often called 'chitinous', 'hyaline' or 'cuticular' layer, or said to be composed of polysaccharides. However, over the past few years the laminated layer was found to be comprised of mucins bearing defined galactose-rich carbohydrates, and accompanied, in the case of Echinococcus granulosus, by calcium inositol hexakisphosphate deposits. In this review, the architecture and biosynthesis of this unusual structure is discussed at depth in terms of what is known and what needs to be discovered.

Molecular and morphological characterisation of Echinococcus from food producing animals in India.

In view of the medical, veterinary and economic importance of hydatid disease in India, our study aimed to determine the prevalence and genotypes of Echinococcus present in domestic livestock in India. Out of 21,861 animals examined, cattle were found with the highest prevalence of hydatid cysts (5.10%) followed by buffaloes (3.81%), pigs (0.87%) and sheep (0.075%). Phylogenetic analysis of the cytochrome oxidase-1 gene revealed that the buffalo strain or G3 genotype was the predominant genotype (29/46) in all species of livestock followed by the cattle strain or G5 genotype (9/46), the G1 genotype or the common sheep strain (6/46) and the G2 genotype or Tasmanian Sheep strain (2/46). The ability of the G3 (buffalo) and G5 (cattle) genotypes of Echinococcus granulosus to infect and produce fertile hydatid cysts in pigs was also demonstrated for the first time. Both morphological and molecular results support earlier studies suggesting that Echinococcus of buffalo origin is phenotypically and genetically similar to the sheep (G1) and Tasmanian Sheep (G2) strains of Echinococcus, which adds further evidence to support its recognition as one species, viz., E. granulosus sensu stricto. Our molecular, morphological and biological characteristics also support earlier studies suggesting that Echinococcus of cattle origin, designated the G5 genotype, should be recognised as a separate species, viz. Echinococcus ortleppi. Finally, the study reveals that the prevalence of hydatidosis in urban centres in India has been showing a consistently declining trend over the past few decades, possibly owing to economic development and improved government legislation of abattoirs.

Alveolar Echinococcus species from Vulpes corsac in Hulunbeier, Inner Mongolia, China, and differential development of the metacestodes in experimental rodents.

Adults of alveolar Echinococcus species with different uterine structures were collected from Vulpes corsac in the Hulunbeier Pasture of Northeastern China in 2001. They were Echinococcus multilocularis Leuckart, 1863 (type No. 3, similar to E. m. multilocularis), with vaselike uterus; Echinococcus cf. sibiricensis Rausch et Schiller, 1954 (type No. 1), with pyriform uterus; and Echinococcus sp. (type No. 2) with spherical uterus at segment top. The metacestode development in rodents also differed among those 3 parasites. In the case of E. multilocularis (type No. 3), many germinal cells grew on the inner surface of early cysts, most of which metastasized into host tissue to form brood vesicles or from the germinal cell layer on the inner surface of the vesicle wall. Cells also had an appearance of proliferating by means of alveolar buds from alveolar tissue that developed outward to form new alveolar foci. In Echinococcus cf. sibiricensis (type No. 1), the formation of alveolar vesicles was due to the metastasizing of germinal tissue into host tissue; protoscoleces grew in the center of alveolar vesicles. In type No. 2 (Echinococcus sp.), the formation of the alveolar vesicle was by multiplication of germinal cell layers on the inner surface of alveolar cysts; protoscoleces grew from the germinal cell layer and mesh in the vesicles. On the basis of uterine structure and on differences in development of metacestodes in experimental rodents, we propose that the 3 types of Echinococcus represent 3 independent species: E. multilocularis, Echinococcus sibiricensis, and Echinococcus sp. (type No. 2-as yet under study).

Molecular and morphological characterization of Echinococcus in cervids from North America.

Many issues concerning the taxonomy of Echinococcus have been resolved in recent years with the application of molecular tools. However, the status of Echinococcus maintained in transmission cycles involving cervid intermediate hosts remains to be determined. The recent characterization of the parasite from cervids in Finland has highlighted the paucity of data available, particularly that from North America. In this study, we have characterized a large number of Echinococcus isolates from cervids from Western Canada on the basis of morphology and molecular genetic techniques. Our results support earlier studies suggesting that Echinococcus of cervid origin is phenotypically and genetically distinct to Echinococcus maintained in domestic host assemblages, and also confirms that Echinococcus of cervid origin does not constitute a genetically homogeneous group. However, our data do not support the existence of 2 distinct genotypes (strains/subspecies) with separate geographical distributions. Our data appear to support the existence of only 1 species in cervids, but additional isolates from cervids and wolves in other endemic regions should be characterized before a final decision is made on the taxonomic status of Echinococcus in cervids.

Echinococcus shiquicus n. sp., a taeniid cestode from Tibetan fox and plateau pika in China.

The taeniid cestode Echinococcus shiquicus n. sp. was found from the Tibetan fox Vulpes ferrilata and the plateau pika Ochotona curzoniae in the Qinghai-Tibet plateau region of China. In the adult stage, E. shiquicus from the foxes is morphologically similar to Echinococcus multilocularis. However, the new species is differentiated by its smaller rostellar hooks, fewer segments, distinct position of genital pore in the mature segment and fewer eggs in the gravid segment. Hydatid cysts of E. shiquicus found in the livers from the pikas were essentially unilocular but an oligovesicular cyst was also found. The data of mitochondrial and nuclear DNA sequences proved E. shiquicus to be a valid taxon.

Using morphometry of the larval rostellar hooks to distinguish Iranian strains of Echinococcus granulosus.

Human cystic echinococcosis is widely distributed throughout Iran, where sheep, cattle, goats, camels and other animals act as intermediate hosts. It appears that morphometry of the rostellar hooks on the protoscoleces of the causative parasite, Echinococcus granulosus, can be used to separate the strains or variants of the parasite to be found in humans and livestock in Iran. Multivariate statistical procedures, including principal-component analysis and discriminant-function analysis (DFA), were used to explore the morphometric data obtained from the larval hooks of Iranian samples of E. granulosus. Although five physical variables were initially considered, the results of the PCA indicated that just two factors (based on the length of hooks and number of hooks) accounted for 91.6% of the variance observed in the hook measurements. The results of the DFA allowed the correct classification of all the samples. The presence in Iran of the sheep and camel strains of the parasite was confirmed. The sheep strain was the most common variant encountered, being recovered from sheep, cattle, goats, humans and occasionally camels. Most of the metacestode samples from camels and a few of those from cattle (five of 30), sheep (one of 27) and humans (two of 23) were found to be of the camel strain. These results indicate that larval-hook morphology is a valid method for identifying E. granulosus strains in Iran, and one that is potentially useful for epidemiological studies.

Erroneous reports of the neotropical Echinococcus oligarthrus as a cause of echinococcosis in India.

Two cases of submandibular echinococcosis have been reported in adult residents of India (Kini et al., 1997; Sahni et al., 2000). It was stated in both publications that the cestode was E. oligarthrus, "a rare variety of E. granulosus", based on morphogical characteristics. It was also mentioned that 3 cases of E. granulosus with similar submandibular metacestode had been previously diagnosed in India. The metacestode is readily distinguishable from that of E. oligarthrus. The final hosts of E. oligarthrus are several neotropical species of wild cats and metacestode occurs in hystricognath, all of them absent in India. The larval stage of all 4 species of Echinococcus can be differentiated on the basis of the form and dimensions of rostellar hooks of protoescoleces. With the information provided in the two papers we concluded that the 2 cases of echinococcosis described in India were due to E. granulosus.

Study on the ecological distribution of alveolar Echinococcus in Hulunbeier Pasture of Inner Mongolia, China.

A study on the ecological distribution of alveolar Echinococcus was carried out in the Hulunbeier Pasture of Inner Mongolia, China during 1998 and 1999. Animals examined included wolves (Canis lupus), red foxes (Vulpes vulpes), sand foxes (Vulpes corsac), domestic dogs (Canis familiaris), Microtus brandti, Meriones unguiculatus, Citellus dauricus, Allactaga sibirica, Phodopus sungorus and Ochotona daurica. Three wolves were found to be infected with E. granulosus. Two sand foxes were infected with E. multilocularis. The majority of infections of alveolar echinococcus was found in M. brandti. Based on the structure of metacestodes found in the livers of naturally infected M. brandti, 3 main variants were observed. Type I had small alveolar cysts with thin cyst walls. Type II had a larger cyst with a thick cyst wall. Infection of laboratory mice with the gravid segments isolated from the naturally infected sand foxes led to the formation of mature Type I alveolar metacestodes in the lungs and Type II metacestodes in the livers of infected animals, respectively.

Morphological and genetic characterization of Echinococcus granulosus in the Slovak Republic.

The present study was undertaken to characterize more thoroughly aetiological agent of cystic echinococcosis that still represents a health problem in the Slovak Republic. Sequencing of the mitochondrial ND1 gene, Random Amplified Polymorphic DNA (RAPD) analysis and isoenzyme analyses were used to characterize genetically protoscolices of 37 isolates from pigs and cattle and two isolates from humans collected in different regions of the Slovak Republic. Solely the G7 genotype (pig strain) was identified by ND1 sequences in all 14 pigs isolate examined by this method. This genotype was also found in two human patients and in two cattle. The exclusive finding of the G7 variant has implications for the implementation of control strategies given its unique developmental and biological properties. Despite striking morphological variability, a low degree of the intra-strain variability was found in the Slovak Echinococcus granulosus isolates with the genetic methods used. This was particularly associated with the shape and size of hooks, number of testes and shape of ovaries and uterus. GPI and MDH enzymes were found suitable for the discrimination of G7 and G1 (=cosmopolitan sheep strain) variant.

Granulomatous peritonitis in hydatid disease.

We report a rare case of persisting granulomatous inflammation within the peritoneal cavity due to previous leakage from a liver hydatid cyst, with the granulomata bearing hooklets originating from protosolices of Echinococcus granulosus. The patient was treated with a full three-month course of albendazole. We believe this represents a rare case of 'pseudotuberculous peritonitis'.

Molecular and morphological characterization of Echinococcus granulosus of human and animal origin in Iran.

Iran is an important endemic focus of cystic hydatid disease (CHD) where several species of intermediate host are commonly infected with Echinococcus granulosus. Isolates of E. granulosus were collected from humans and other animals from different geographical areas of Iran and characterized using both DNA (PCR-RFLP of ITS1) and morphological criteria (metacestode rostellar hook dimensions). The sheep and camel strains/genotypes were shown to occur in Iran. The sheep strain was shown to be the most common genotype of E. granulosus affecting sheep, cattle, goats and occasionally camels. The majority of camels were infected with the camel genotype as were 3 of 33 human cases. This is the first time that cases of CHD in humans have been identified in an area where a transmission cycle for the camel genotype exists. In addition, the camel genotype was found to cause infection in both sheep and cattle. Results also demonstrated that both sheep and camel strains can be readily differentiated on the basis of hook morphology alone.

Epidemiology and strain characteristics of Echinococcus granulosus in the Benghazi area of eastern Libya.

The incidence of surgically confirmed cystic echinococcosis in eastern Libya was estimated to be at least 4.2 cases/100,000, with significantly more female cases than male. The prevalences of infection with Echinococcus granulosus among 1087 sheep, 881 goats, 428 camels and 614 cattle from the same region, determined postmortem in abattoirs, were 20%, 3.4%, 13.6% and 11%, respectively. Infection in the livestock was age-dependent and, generally, the female animals were more often infected than the male. The measurements of rostellar hooks on protoscoleces collected from sheep and cattle were similar but significantly different from the corresponding measurements of parasites of human or camel origin. However, when a portion of the cytochrome c-oxidase subunit I (cox1) gene from each of 30 protoscolex samples from Libya (12 from cattle, three from humans, five from camels and 10 from sheep) was sequenced, the sequences were all found to be identical to that published for the common sheep strain of E. granulosus.

Could ivermectin have a synergic effect with albendazole in hydatidosis therapy?

The efficacy of a combination of ivermectin (IVM) and albendazole (ABZ) used as a chemoprophylactic treatment and as treatment of secondary hydatidosis is described for the first time. IVM treatment alone was not effective against Echinococcus granulosus, either when the protoscolices were recently inoculated or when they had developed to the metacestode stage. However, the efficacy of IVM and ABZ when used in combination as a prophylactic treatment was 95.72% and 87% with respect to the number and the wet weight of cysts, respectively. These results were higher than in the treatment of secondary hydatidosis, which were 44.8% and 45.26%, respectively. The ultrastructural changes in the germinal layer of the cysts after the treatments are also described.