Current status of cystic echinococcosis control in the Falkland Islands: has elimination been achieved?

Attempts to control cystic echinococcosis (CE) caused by Echinococcus granulosus in the Falkland Islands have been ongoing for over 50 years. No human cases have been recorded since the 1980s but there is a need to establish if the parasite has been completely eliminated from domestic animals. A study was carried out in 2018/2019 to identify dogs infected with E. granulosus using copro-antigen and copro-polymerase chain reaction (PCR) analysis. In addition, annual slaughter data were analysed to establish infection levels of E. granulosus and 2 other taeniid parasites. Results showed that 4 out of 589 dogs (0.7%) tested positive by copro-antigen analysis. Results from similar surveys carried out in 2010, 2012 and 2014 showed 17 (3%), 0 and 6 (1%) copro-antigen-positive dogs, respectively, with 8 dogs being confirmed by PCR in 2010. Annual abattoir data showed that from 2006 to 2020, 36 sheep were identified with E. granulosus (mean 0.0055%), 14 186 sheep with Taenia hydatigena (mean 2.2%) and 465 with Taenia ovis (mean 0.072%). Prevalences of T. hydatigena and T. ovis showed spontaneous rises in certain years where the infections could also be detected in lambs indicating that viable taeniid eggs were present. Observations of farm management procedures indicated that there were occasions when dogs could get access to infective taeniid material. In conclusion, E. granulosus is still present in sheep and dogs but at low prevalences. The increasing presence of T. hydatigena however, indicates that control measures are defective in some areas and there is potential for a re-emergence of CE.

Echinococcus granulosus cyst fluid enhances epithelial-mesenchymal transition.

Cystic echinococcosis is characterized by fluid-filled hydatid cysts in the liver and lungs. The cysts are surrounded by a host fibrous layer (the pericyst) which acts to isolate the parasite from surrounding tissues. Previous studies in liver cysts have indicated that the parasite may be a stimulating fibrosis. The aim of this study was to investigate whether hydatid cyst fluid (HCF) could influence the potential for fibrosis to occur in lung tissue by stimulating epithelial to mesenchymal transition (EMT) in a human lung epithelial cell line. An adenocarcinoma-derived alveolar basal epithelial cell line (A549) was used as a model for human alveolar epithelial cells (AEC II). These were cultured in vitro with HCF (UK sheep origin). Assays to investigate cell proliferation, cell migration and expression of cytoskeletal markers showed that HCF could stimulate changes indicative of EMT, including enhanced cell proliferation and migration; increased expression of mesenchymal cytoskeletal markers (fibronectin and vimentin) accompanied by a down-regulation of an epithelial marker (E-cadherin). Molecules within hydatid cyst fluid are capable of inducing phenotypic changes in A549 cells indicating that the parasite has the potential to modify lung epithelial cells which could contribute to fibrotic reactions.

Rodentolepis straminea (Cestoda: Hymenolepididae) in an urban population of Apodemus sylvaticus in the UK.

The presence of the cyclophyllidean cestode Rodentolepis straminea (Cestoda: Hymenolepididae), was confirmed by molecular DNA analysis from a wood mouse (Apodemus sylvaticus) population inhabiting urban woodland in Salford, Greater Manchester (UK) with a prevalence of 27.8%. It would appear that the only previously published record of this species in A. sylvaticus in the British Isles is that from south-west Ireland, where 24% of the wood mice examined were infected with R. straminea. This species has been recorded in studies on A. sylvaticus in continental Europe. The current report represents a new record for R. straminea on mainland Britain and a first study of helminth parasites in an urban wood mouse population.

Risk factors for Echinococcus coproantigen positivity in dogs from the Alay valley, Kyrgyzstan.

Echinococcosis, caused by the zoonotic cestodes Echinococcus granulosus (sensu lato) and Echinococcus multilocularis, is highly endemic in the Central Asian Republic of Kyrgyzstan, and is being identified increasingly as a public health problem, especially amongst pastoral communities. As domestic dogs are considered to be the main source of human infection, the identification of potential transmission pathways is of relevance when considering implementing an echinococcosis control scheme. The current report describes the results of an analytical study of canine Echinococcus coproantigen enzyme-linked immunosorbent assay (ELISA) prevalence in the Alay valley of southern Kyrgyzstan prior to the commencement of regular praziquantel dosing of dogs. A logistic regression model using a form of Bayes modal estimation was used to identify possible risk factors for coproantigen positivity, and the output was interpreted in a Bayesian context (posterior distributions of the coefficients of interest). The study found that sheepdogs had lower odds of coproantigen positivity, as did dogs in households with donkeys, where owners had knowledge of echinococcosis, and households which engaged in home slaughtering. Surprisingly, there was no evidence of an association between free roaming or previous praziquantel dosing and coproantigen positivity, as has been found in previous studies. Possible reasons for these findings are discussed in the context of the epidemiology of echinococcosis and potential intervention approaches.

Prevalence of Toxoplasma gondii in localized populations of Apodemus sylvaticus is linked to population genotype not to population location.

Toxoplasma gondii is a globally distributed parasite infecting humans and warm-blooded animals. Although many surveys have been conducted for T. gondii infection in mammals, little is known about the detailed distribution in localized natural populations. In this study, host genotype and spatial location were investigated in relation to T. gondii infection. Wood mice (Apodemus sylvaticus) were collected from 4 sampling sites within a localized peri-aquatic woodland ecosystem. Mice were genotyped using standard A. sylvaticus microsatellite markers and T. gondii was detected using 4 specific PCR-based markers: SAG1, SAG2, SAG3 and GRA6 directly from infected tissue. Of 126 wood mice collected, 44 samples were positive giving an infection rate of 34.92% (95% CI: 27.14-43.59%). Juvenile, young adults and adults were infected at a similar prevalence, respectively, 7/17 (41.18%), 27/65 (41.54%) and 10/44 (22.72%) with no significant age-prevalence effect (P = 0.23). Results of genetic analysis of the mice showed that the collection consists of 4 genetically distinct populations. There was a significant difference in T. gondii prevalence in the different genotypically derived mouse populations (P = 0.035) but not between geographically defined populations (P = 0.29). These data point to either a host genetic/family influence on parasite infection or to parasite vertical transmission.

Post-encystment/established immunity in cystic echinococcosis: is it really that simple?

In 1982, Rickard & Williams stated that immunity to larval taeniid cestodes could be divided into two different phases: the establishment phase, operating against oncospheres, and the established phase, operating against established metacestodes in the tissues. These concepts were largely based on experimental and veterinary data. As a result of subsequent research combined with clinical data, we can consider that, for metacestodes of Echinococcus spp, a single established phase is probably too simple a concept. These parasites show developmental changes in different hosts over time, the most significant being growth, fertility and degeneration. Recent clinical cyst classifications (WHO/IWGE) have highlighted that all hydatid cysts are morphologically not the same. This is important for treatment but fails to completely address variation in the host/parasite response and the understanding of parasite survival. This review attempts to relate the immunological and developmental data obtained from experimental, veterinary and medical studies to update our understanding of established immunity against E. granulosus. It proposes that a single 'Established' phase for E. granulosus could be subdivided into four additional phases: maturing; stable; unstable and degenerative. Combining this type of information with existing cyst classification systems could significantly benefit our understanding of the parasites immunobiology.

A molecular and ecological analysis of the trematode Plagiorchis elegans in the wood mouse Apodemus sylvaticus from a periaquatic ecosystem in the UK.

The prevalence of the digenean Plagiorchis sp. was investigated in a natural wood mouse population (Apodemus sylvaticus) in a periaquatic environment. Classical identification was complemented with the use of molecular differentiation to determine prevalence and verify species identity. Use of the complete ITS1-5.8S rDNA-ITS2 and partial 28S rDNA gene sequences have confirmed that the species reported at this location was Plagiorchis elegans and not Plagiorchis muris as reported previously. This underlines the difficulties in identification of these morphologically similar parasites. Plagiorchis elegans is typically a gastrointestinal parasite of avian species but has also been reported from small mammal populations. Although the occurrence of this digenean in A. sylvaticus in the UK is rare, in the area immediately surrounding Malham Tarn, Yorkshire, it had a high prevalence (23%) and a mean worm burden of 26.6 ± 61.5. The distribution of P. elegans followed a typically overdispersed pattern and both mouse age-group and sex were determined to be two main factors associated with prevalence. Male mice harboured the majority of worms, carrying 688 of 717 recovered during the study, and had a higher prevalence of 32.4% in comparison to only 8.7% in the small intestine of female mice. A higher prevalence of 43% was also observed in adult mice compared to 14% for young adults. No infection was observed in juvenile mice. These significant differences are likely to be due to differences in the foraging behaviour between the sexes and age cohorts of wood mice.

Identification of a new species of digenean Notocotylus malhamensis n. sp. (Digenea: Notocotylidae) from the bank vole (Myodes glareolus) and the field vole (Microtus agrestis).

Notocotylus malhamensis n. sp. is described from the caecum of the bank vole (Myodes glareolus) and the field vole (Microtus agrestis) from Malham Tarn Nature Reserve in North Yorkshire, UK. In total, 581 specimens were collected from rodents trapped at a wetland site (Tarn Fen) between July 2010 and October 2011 with a prevalence of 66·7% and mean intensity of 94·6 in the bank vole and 50% prevalence and a mean intensity of 4·3 in the field vole. This species appears to be most closely related to other previously described Notocotylus species infecting rodents in Europe but differs principally by the metraterm to cirrus sac ratio (1:1·5-1:1·2) in combination with a densely spinulated cirrus, simple caeca and a greater number of ventral glands in the lateral rows (14-17). The use of molecular differentiation was of limited use in this study due to a paucity of relevant information in the DNA sequence databases. However, the complete ITS1-5.8S rDNA-ITS2 and partial 28S gene sequences have been generated to provide a definitive tool for identification of this species in future studies. As far as we know this is the first report of a notocotylid infection in M. glareolus in the UK.

Prevalence and co-infection of Toxoplasma gondii and Neospora caninum in Apodemus sylvaticus in an area relatively free of cats.

The protozoan parasite Toxoplasma gondii is prevalent worldwide and can infect a remarkably wide range of hosts despite felids being the only definitive host. As cats play a major role in transmission to secondary mammalian hosts, the interaction between cats and these hosts should be a major factor determining final prevalence in the secondary host. This study investigates the prevalence of T. gondii in a natural population of Apodemus sylvaticus collected from an area with low cat density (<2·5 cats/km2). A surprisingly high prevalence of 40·78% (95% CI: 34·07%-47·79%) was observed despite this. A comparable level of prevalence was observed in a previously published study using the same approaches where a prevalence of 59% (95% CI: 50·13%-67·87%) was observed in a natural population of Mus domesticus from an area with high cat density (>500 cats/km2). Detection of infected foetuses from pregnant dams in both populations suggests that congenital transmission may enable persistence of infection in the absence of cats. The prevalences of the related parasite, Neospora caninum were found to be low in both populations (A. sylvaticus: 3·39% (95% CI: 0·12%-6·66%); M. domesticus: 3·08% (95% CI: 0·11%-6·05%)). These results suggest that cat density may have a lower than expected effect on final prevalence in these ecosystems.

Antibody responses in human cystic hydatid disease to the laminated layer of Echinococcus granulosus.

The laminated layer of hydatid cysts of Echinococcus granulosus represents a considerable amount of parasite material. Its antigenic role, however, is unclear. Extracts of laminated layer taken from sheep cysts were analysed in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS/PAGE) and were found to contain bands at 55 and 25-29 kDa, which reacted with an anti-sheep IgG antibody probe, indicating that these were likely to be host-contaminating components within the layer. However, the same bands were also recognised by a significant proportion of human hydatid patients, particularly by IgG4 antibodies, and not by negative control individuals. These individuals did not recognise immunoglobulin heavy and light chains in a sheep serum extract in the same manner. It seems likely that there are either host or parasite antigenic components at similar molecular weights or that certain parasite antigens may share epitopes with sheep immunoglobulins. The antigens at 25-29 kDa were found to be glycoproteins by lectin blot analysis and may be important markers of disease status.

The occurrence of the trematode Plagiorchis muris in the wood mouse Apodemus sylvaticus in North Yorkshire, UK.

Wood mice Apodemus sylvaticus were trapped each September over a 13-year period, from 1993 to 2005, in a wooded area adjacent to Malham Tarn, Yorkshire, UK. Plagiorchis muris was found to be the dominant intestinal trematode and occurred in every year of sampling, with an overall prevalence of 16.9%. This appears to be the first record of P. muris in A. sylvaticus within the UK. The mean worm burden was 2.03 and the distribution of the parasite within mice was typically overdispersed. No difference in prevalence relative to host sex was evident although there was a higher prevalence of 21.9% in larger older mice compared with 9.1% in juveniles, which probably relates to a greater foraging activity in older mice. Annual prevalence values of P. muris infections varied significantly over the study period with higher prevalences being associated with years with a high spring/summer rainfall. The second intermediate hosts for P. muris include a variety of aquatic insect larvae and it is likely that the higher rainfall may result in the occurrence of temporary water bodies suitable for the development of insect larvae.

Canine echinococcosis in Turkana (north-western Kenya): a coproantigen survey in the previous hydatid-control area and an analysis of risk factors.

A study of Echinococcus granulosus infection in dogs, with risk-factor analysis, was carried out in the endemic area of northern Turkana district, Kenya, using necropsy on 42 strays and a coproantigen-ELISA survey of 161 owned animals. During the post-mortem examinations, 14 (33%) of the necropsied dogs were found infected with E. granulosus, with a mean burden of 540 worms (range=two to 4080 worms). The 26 necropsied dogs that came from the north-western Lokichoggio division--an area where, from 1983 to 1997, there had been a continuous programme of hydatid control--showed a similar prevalence of infection to the other dogs (34.6%) but a significantly lower mean burden, of 53 worms (range=two to 300). Forty-two (26%) of the animals tested for Echinococcus coproantigen were found positive. Although the dogs from the Lokichoggio division were more likely to be coproantigen-positive (29%) than those from the central Kakuma division (20%) or the north-eastern division (18%), the differences were not statistically significant. In questionnaire-based interviews, the owners of the dogs tested for coproantigens were asked about possible risk factors for canine infection with E. granulosus. Women were found to have twice the level of contact with dogs as men. The results of a univariate analysis of the dog-owners' responses revealed six factors that appeared to be significantly associated with a coproantigen-positive dog: non-restraint of the dog (P<0.001); dog fed on raw offal (P<0.001); the improper disposal of slaughter offal (P<0.001); the dog-owner's lack of knowledge about the transmission of echinococcosis (P=0.001); the dog not receiving anthelmintic treatment (P=0.003); and dog age < or =5 years (P=0.01). The results of a multivariate analysis confirmed that lack of dog restraint, access to raw offal, and young age of the dog (< or =5 years) each significantly increased the risk of coproantigen positivity (P, 0.005). Dogs that scavenged from cooking pots, were used to clean babies, had access to the inside of houses, and/or slept indoors appeared, however, to be at no increased risk of coproantigen positivity. The present results are discussed in relation both to older information on the epidemiology and role of human behaviour in the transmission of E. granulosus in Turkana, and the effects of the hydatid-control programme that ran continuously in the north-western division of Turkana between 1983 and 1997.

Human cystic echinococcosis in two Mongolian communities in Hobukesar (China) and Bulgan (Mongolia).

In order to investigate the prevalence of human cystic echinococcosis (CE) in traditional Mongolian communities in western Mongolia and Xinjiang (northwest China), studies were carried out between 1995 and 2000, on two ethnically identical populations in Hobukesar (China) and Bulgan (Mongolia). The prevalence of human hepatic CE in the two communities was significantly different. In Hobukesar, human CE prevalence by ultrasound was 2.7% (49/1844), while in Bulgan it was 0.2% (4/1609) (P<0.001). Dog surveys showed that coproantigen-positive rates or dog necropsy positives were similar in both communities (35.0% in Hobukesar and 35.7% in Bulgan). Comparing possible risk factors, there appeared to be some significant differences between the two communities, which might contribute to the observed difference in CE prevalence. These included: the proportion of herdsman or farmers recorded; the proportions of dog ownership and livestock ownership; and the proportion of families practising home slaughter. The presence of a Russian dog-dosing programme up to the mid-1980s may explain the lower prevalence of human CE in the Bulgan population; no similar programme operated in the China-administered Hobukesar community.

Classification, follow-up and recurrence of hepatic cystic echinococcosis using ultrasound images.

Ultrasound image and morphological structure of hepatic cystic echinococcosis (CE) were analysed in 277 human cases (385 hepatic hydatid cysts). These included 65 CE cases from community surveys carried out between 1995 and 2000 in 3 countries (China, Mongolia and Jordan) and 212 cases from a clinical hospital survey from Xinjiang, China. A new simplified WHO ultrasound classification for human CE was assessed, and considered useful. It is proposed that type, size and number in particular need to be included in the ultrasound classification of hepatic CE. For comparative purposes 6 categories of type were classified in the study as Type 0 to Type 5 (T0-T5): T0, univesicular without pathognomonic signs; T1, univesicular with pathognomonic signs; T2, cysts with sagging or floating laminated membrane; T3, cysts containing daughter cysts; T4, solid mass or mixed cysts; and T5, cysts with partial or full calcifications. This differs from the WHO classification wherein Type T3 cysts (daughter cysts present) are considered a pathological stage to occur in general prior to the sagging or floating membrane (T2) stage. Recurrent hydatid cysts in the liver were also studied based on morphological structures observed directly from surgical intervention. Case follow-up over 1-5 years since endocystectomy in the community surveys indicated 10% (2/10) recurrence of cysts in the residual surgical cavity. Recurrent CE included 2 (2/4) cases after percutaneous treatment.

Detection of antibodies to Staphylococcus aureus Toxic Shock Syndrome Toxin-1 using a competitive agglutination inhibition assay.

AIMS: To develop a competitive agglutination inhibition assay (CAIA) for the detection of anti-Toxic Shock Syndrome Toxin-1 (TSST-1) antibody in serum samples using a commercially available reverse passive agglutination assay (RPLA) kit. METHODS AND RESULTS: TSST-1 toxin and sera were incubated together, so that anti-toxin IgG would complex with the toxin. Latex particles sensitized with rabbit IgG anti-TSST-1 were added to test for un-complexed toxin. The sensitivity and specificity of the CAIA assay was determined relative to positive and negative ELISA results. The sensitivity (proportion of positive ELISA sera which tested positive by CAIA) was 66% whilst the specificity (proportion of ELISA negative sera which tested negative by CAIA) was 75%. Seven sera (14%) were negative by ELISA but positive for CAIA and 12 (18.8%) were positive for ELISA but negative for CAIA, suggesting some interference with the assays. Statistical analysis showed no significant difference between the methods in terms of the numbers of individuals testing positive (chi2, P = 0.04). CONCLUSIONS: The CAIA assay allowed detection of anti-TSST-1 within 18 h and was simple to read visually. SIGNIFICANCE AND IMPACT OF THE STUDY: The method is a useful test for individual serum samples and a preliminary investigation for medical screening of suspected toxic shock syndrome and is applicable in situations where antibody assays are not routinely used for anti-TSST-1 and also where sophisticated equipment (e.g. microtitre plate reader) is not available.

Echinococcosis: disease, detection and transmission.

Echinococcosis is one of the world's most geographically widespread parasitic zoonoses, with transmission occurring in tropical, temperate and arctic biomes. Most human infections are due to Echinococcus granulosus transmitted between domestic dogs and livestock, but this cosmopolitan species also cycles between wild carnivores (principally canids) and wild ungulates. The other species with significant zoonotic potential is E. multilocularis that occurs naturally in fox definitive hosts and small mammal intermediate hosts. These two species cause human cystic or alveolar echinococcosis respectively, which may be considered serious public health problems in several regions including developed countries. This review provides an introductory overview to the Supplement and summarises the biology and epidemiology of these two related cestodes with an emphasis on applied aspects relating to detection, diagnosis and surveillance in animal and human populations, and includes aspects of transmission ecology, and also considers aspects of community epidemiology and potential for control.

Rapid dot-ELISA for the detection of specific antigens in the cyst fluid from human cases of cystic echinococcosis.

A rapid dot-ELISA was developed for the detection of specific antigens in samples of cyst fluid from human cases of Echinococcus granulosus infection, permitting the confirmation of cystic echinococcosis (CE). Peroxidase-conjugated antibodies against antigen B (derived from the fluid in the cysts of sheep infected with E. granulosus) were used to test cyst-fluid samples from 22, surgically confirmed cases of human CE and 21 domestic animals (horses, sheep, buffalo, a cow and a camel) with CE. All of these samples were found to be strongly positive in the dot-ELISA, by direct reading with the naked eye. In contrast, fluid samples from seven, non-parasitic liver cysts of human origin were all negative, and a sample taken from the residual cavity left in a patient after previous CE surgery showed a weakly positive reaction. The antigen-detection assay, which can be completed within 10 min, may be a useful 'bedside' test for the differential diagnosis of cysts during surgery or percutaneous treatment for suspected CE.

In-vitro susceptibility of hydatid cysts of Echinococcus granulosus to nitric oxide and the effect of the laminated layer on nitric oxide production.

Murine hydatid cysts of Echinococcus granulosus were incubated in vitro in the presence of nitric oxide produced from S-nitroso-N-acetylpenicillamine (SNAP) or interferon-gamma activated peritoneal macrophages. In both situations, evidence of cyst damage and death was observed by microscopy in over 77% of cysts after 3 days, indicating that intact hydatid cysts could be susceptible to a Th1 driven macrophage attack. A crude extract of the laminated layer from cysts was found to be able to reduce the production of nitric oxide from activated macrophages in vitro and in vivo and this may have been due to phagocytosis of laminated layer fragments by the macrophages. The results indicate that, although cysts may be susceptible to the effects of nitric oxide, the laminated layer may be involved in downregulating nitric oxide production.

Cystic echinococcosis in semi-nomadic pastoral communities in north-west China.

In order to determine the prevalence of human cystic echinococcosis (CE) in semi-nomadic traditional pastoralist groups in north-west China, 2 large community studies were undertaken in Altai and Tacheng Prefectures in 1990/91 and 1995/96, respectively. The Kekergash community (Altai) comprised mainly ethnic Kazakhs, whereas the Narenhebuke community (Tacheng) comprised mainly Mongolians. Populations were screened for CE by abdominal ultrasound scan (US) and serological tests. The total prevalence of confirmed human CE was higher in Narenhebuke (2.7%, 49/1844) than in Kekergash (0.9%, 17/1861; P < 0.01). Within each community there was no significant difference of CE prevalence between the Kazakh and Mongolian groups, although Han Chinese exhibited twice the rate of CE (4.9%) in Narenhebuke compared to the dominant Mongolian population. For each community, human CE prevalence increased with age and there was a greater risk associated with the practice of home slaughter of livestock. Dogs were screened for Echinococcus granulosus infection and re-infection levels using a highly specific coproantigen test. The proportion of dogs with positive coproantigen tests was significantly higher in Narenhebuke (36.0%, 50/139) compared to Kekergash (17.8%, 16/90). In Narenhebuke the re-infection levels of dogs, as determined by coproantigen positivity, were higher in the winter quarters (49.4%, 39/79) compared to the summer quarters (18.3%, 11/60; P < 0.01). Furthermore, coproantigen re-test positivity was 25% at 3 months and 29.2% at 7 months. Highest dog coproantigen positivity was obtained over the winter period.

Apoptosis in human Jurkat T cells after culture with live Taenia crassiceps cysticerci in vitro.

This study examines the effects of Taenia crassiceps cysticerci on the viability of a human T lymphocyte cell line (Jurkat). Both budding and non-budding T. crassiceps metacestodes were cultured over 24 and 48 h in the presence of Jurkat cells. Cell viability decreased with increasing numbers of cysticerci, particularly budding cysticerci. Single cell gel electrophoresis (comet) analysis, which grades DNA damage, showed a significant increase in apoptosis at 24 and 48 h. The morphology of treated cells was determined using acridine orange with the classical morphology of apoptotic bodies seen to increase with increasing cysticerci numbers over time. These results indicate that parasite-induced apoptosis occurs during murine cysticercosis. Such a mechanism may be important in survival of other metacestode infections of medical or veterinary importance.