Cystic echinococcosis in donkeys in eastern Africa.

Cystic echinococcosis (CE) is endemic in humans and domestic animals in eastern Africa. All the species of the Echinococcus granulosus sensu lato complex have been reported in this region except for E. equinus, possibly due to the small number of studies involving equids. This study reports the frequency of different Echinococcus species in donkeys from eastern Africa. A total of 5961 donkeys were examined during meat inspection in 3 slaughterhouses in Kenya. Identification of Echinococcus spp. was achieved through polymerase chain reaction-restriction fragment-length polymorphism and sequencing of the mitochondrial nicotinamide adenine dinucleotide (NADH) dehydrogenase subunit 1 gene. The prevalence of CE was 5.7% (337/5961). The 263 genotyped cysts belonged to E. equinus (n = 163), E. granulosus sensu stricto (n = 70), E. canadensis (G6/7) (n = 26) and E. ortleppi (n = 4). One donkey harboured a metacestode of Spirometra theileri. All E. equinus cases, except 2, originated from southern Ethiopia, whereas the other species were more evenly distributed across the study area. Most of the cysts belonging to E. equinus were fertile (111/163), while those of the other species were non-fertile. This is the first report of Echinococcus spp. in donkeys from sub-Saharan Africa and the first confirmation of E. equinus in East Africa. The frequent fertility of E. equinus cysts in donkeys affirms their suitability as intermediate hosts of this species, while low frequency and cyst fertility suggest a marginal role of donkeys in the transmission of E. granulosus s. s., E. canadensis (G6/7) and E. ortleppi.

Findings of a community screening programme for human cystic echinococcosis in a non-endemic area.

Cystic Echinococcosis (CE) is a zoonosis caused by infection with the larval stages of the taeniid cestodes of the species complex Echinococcus granulosus sensu lato. It is prevalent among transhumant communities in East Africa, including those residing in northern Kenya. The movement of livestock from these regions of high incidence to areas of low incidence creates an indirect risk of disease spill-over to humans. To assess possible establishment of the CE life cycle outside known endemic regions, we used a portable ultrasound scanner to screen for the presence of human CE in Bungoma County of western Kenya, an area which imports substantial numbers of cattle for slaughter from neighbouring pastoralist regions. Eight sentinel sites were purposively selected based on their proximity to slaughterhouses handling animals introduced from pastoralist regions, and necessary permissions to conduct the study were sought. Regression analyses were conducted to identify risk factors associated with the presence of abdominal and cystic lesions (CL). In total, 1002 participants were screened; of these, 654 (65.3%) were female and the median age was 43. Farming (n = 403; 43.4%) was the most frequent occupation, followed by professional (i.e. on regular salary) (n = 215; 23.1%), and business (n = 207; 22.3%) categories. Sixty-seven participants (6.7%) had abnormal ultrasound findings, of these, 7 (1.1%) had simple liver cysts/CL, as per WHO classification. As such, their outcome was inconclusive and they were not put on treatment but advised to attend follow-up investigations in a referral health facility. Other abnormal findings included splenomegaly (n = 14), ovarian cysts (n = 14), uterine fibroids (n = 10), polycystic kidneys (n = 6), and benign prostatic hyperplasia (n = 6). Age was unconditionally associated with the presence of presumptive CL. These results contribute to CE baseline data while providing insights on the implementation of ultrasound diagnosis in the field, as recommended by the WHO for targeted control of echinococcosis by 2030.

Diversity of Taenia and Hydatigera (Cestoda: Taeniidae) in domestic dogs in Kenya.

Taenia species of domestic dogs can cause cysticercosis and coenurosis in a wide range of intermediate hosts including humans. Most taeniids of dogs are globally distributed, but some wildlife-transmitted species can be specific for certain regions. Generally, little information exists on the species composition and frequency in most regions of the world, which impairs risk assessment and control strategies. This study determined the range of taeniid species in dogs in four widely spaced areas of Kenya by genetic identification of eggs in faeces collected from the environment. Individual taeniid eggs were characterised by nested polymerase chain reaction of NADH dehydrogenase subunit 1 and cytochrome C oxidase 1 genes, restriction fragment length polymorphism and partial sequencing. Overall 79/1621 (4.9%) faecal samples contained eggs of Taenia or Hydatigera (8.0% in Turkana, 4.8% in Isiolo, 3.8% in Maasai Mara and 1.3% in Meru). Taenia hydatigena and T. multiceps were the most frequent, found in 36 and 15 samples, respectively. Other eggs found in the faeces belonged to T. serialis (sensu lato), T. madoquae (the first record in domestic dogs), T. ovis, T. saginata and Hydatigera taeniaeformis. Polymorphism of nad1 sequences revealed 22 and 8 haplotypes of T. hydatigena and T. multiceps, respectively. The results show the involvement of dogs in both domestic and sylvatic transmission cycles. In addition to the species range, this study provides data on the intraspecific diversity of T. hydatigena and T. multiceps in Kenya, which will serve as baseline information for further studies into cysticercosis and coenurosis in livestock and humans in the region.

Prevalence and Genotyping of Echinococcus Species from Livestock in Kajiado County, Kenya.

Cystic Echinococcosis (CE) is a widespread neglected zoonotic disease and is caused by the larval stage of the dog tapeworm Echinococcus granulosus sensu lato. CE is more frequent in livestock-rearing areas and where people live a nomadic or seminomadic lifestyle such as in Kajiado County, Kenya. There is limited data on CE disease situation in the county of Maasailand; the present study, therefore, reports on the prevalence of CE in cattle, sheep, and goats and their relative importance in CE transmission in Kajiado County. In total, 1,486 livestock (388 cattle, 625 sheep, and 473 goats) slaughtered in two abattoirs were examined for the presence of hydatid cysts in various organs. Cyst isolates were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the NADH dehydrogenase subunit 1 gene (nad1). The overall prevalence of CE was 14.8% (220/1486), while prevalence per livestock species was 15.2% (72/473) in goats, 14.9% (93/625) in sheep, and 14.2% (55/388) in cattle. Out of the 421 cysts isolated, 389 cysts were successfully characterized to be either E. granulosus sensu stricto (s. s.), 356/389 (91.5%), E. canadensis (G6/7), 26/389 (6.7%), or E. ortleppi, 7/389 (1.8%). This record confirms predominance of E. granulosus s. s. in Maasailand and other parts of Kenya, while the importance of E. ortleppi and E. canadensis (G6/7) to the general CE burden in Maasailand might be higher than previously thought. More so, a higher infection pressure for humans by E. granulosus s. s. based on its abundance could be speculated. The study sheds significant light on CE situation in livestock in the nomadic/seminomadic society of the Maasai in Kajiado County and provides good bases to investigate human CE in the area.

Molecular characterization of Echinococcus species in dogs from four regions of Kenya.

Cystic echinococcosis is endemic both in livestock and humans in many parts of Kenya. However, very little data exists on Echinococcus infections in dogs, and therefore their role in maintaining the transmission cycles and environmental contamination with eggs of Echinococcus species is unknown. The study aimed to establish the prevalence and distribution of Echinococcus granulosus sensu lato causing infection in dogs in Kenya. A total of 1621 dog faecal samples were collected from the environment in four different regions and examined microscopically for the presence of taeniid eggs. Up to 20 individual taeniid eggs per faecal sample were picked, lysed and genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing of the NADH dehydrogenase subunit 1 (nad1) gene. Eleven percent (178/1621) of faecal samples had taeniid eggs, while 4.4% (71/1621) contained Echinococcus spp. eggs. Area-wise, the faecal prevalence of Echinococcus spp. was 9.2% (48/524) in Turkana, 4.0% (20/500) in Maasai Mara, 0.7% (2/294) in Isiolo and 0.3% (1/303) in Meru. E. granulosus sensu stricto (s. s.) was the dominant Echinococcus taxon, followed by E. canadensis (G6/7) that was detected in 51 and 23 faecal samples, respectively. E. ortleppi was detected in only 5 faecal samples. We report for the first time the presence of E. felidis eggs in two dog faecal samples (from Maasai Mara region). Mixed infections of these taxa were also found in faecal samples, including: E. granulosus s. s. and E. canadensis (G6/7) (n = 7), E. granulosus s. s. and E. ortleppi (n = 1) and all three species (n = 1). The dog data presented here confirm the differences in diversity and abundance of Echinococcus spp. between regions of Kenya, correspond well with previously published data from livestock, and tentatively suggest a role of domestic dogs as a link between domestic and sylvatic cycles of Echinococcus spp.

Genetic differentiation of the G6/7 cluster of Echinococcus canadensis based on mitochondrial marker genes.

Among the genotype/species causing cystic echinococcosis, the taxonomic status of Echinococcus canadensis is only partially resolved. Within E. canadensis, four genotypes (G6, G7, G8 and G10) have been described based on short mitochondrial sequences, of which G6 and G7 (the 'camel' and the 'pig' strain, respectively) are closely related and variously regarded as microvariants of a single strain G6/7. Globally, this G6/7 cluster is the second most important agent of human cystic echinococcosis and is the predominant Echinococcus taxon in large parts of sub-Saharan Africa. To add data on the internal structure and the geographical distribution of this cluster, we analysed diversity and population structure of 296 isolates of E. canadensis from sub-Saharan Africa, the Middle East and Europe using the complete mitochondrial cytochrome c oxidase subunit 1 (cox1) (1,608bp) and NADH dehydrogenase subunit 1 (nad1) (894bp) gene sequences. Polymorphism of the mtDNA loci gave 51 (cox1), 33 (nad1) and 73 (cox1-nad1 concatenated) haplotypes. African and Middle Eastern isolates mainly grouped in a star-like structure around a predominant haplotype, while the European isolates produced more diversified networks. Although the cox1 diagnostic sequence for G6 is frequent in the African/Middle Eastern sub-cluster, and that for G7 is common in the European isolates, numerous intermediate variants prevent a clear distinction into 'G6' or 'G7', and the entire taxon is best treated as a common haplotype cluster G6/7. Meanwhile, the G6/7 cluster is clearly distinct from sequences of wildlife isolates of G8 and G10 from the northern hemisphere, and sequences of the latter genotypes were remarkably distant from each other. It is clear from the present study that, based on mitochondrial data, G6/7 is a coherent genotypic entity within E. canadensis that retains substantial intraspecific variance, and sub-populations share common ancestral polymorphisms and haplotypes. This study provides the basis for wider biogeographic comparison and population genetics studies of this taxon.

Cystic Echinococcosis in Turkana, Kenya: The Role of Cross-Sectional Screening Surveys in Assessing the Prevalence of Human Infection.

Cystic echinococcosis (CE) is a neglected zoonotic disease caused by Echinococcus granulosus. Infection leads to formation of cysts within the viscera of the human host. In the 1980s, the transhumant population of northwest Turkana, Kenya, was found to have the highest prevalence of CE in the world. In 1983, AMREF Health Africa and the Kenya Medical and Research Institute launched a CE Control Program in northwest Turkana, screening and treating the local people. This epidemiological study of CE in Turkana analyses approximately 30 years of surveillance and surgical data. Cyst data were categorized using the World Health Organization CE ultrasound classification system before being analyzed for cyst, patient, and population characteristics, and surveillance data from 1985 are compared with more recent surveillance data to assess changes in prevalence in the control region since the commencement of control activities. In 1985, the prevalence of CE among the Turkana was 5.6%. In 2010-2011 and 2011-2012, calculated CE prevalence rates were 1.9% and 3.8%, respectively. Since the 1980s, the age distribution of people with CE in Turkana has shifted: initially, cases of CE appeared predominantly within younger age groups, but recent data reveal a higher prevalence within older age groups. The frequency of infection in females also significantly decreased. The reduction in CE prevalence from 5.6% in the 1980s to 1.9-3.8% in 2010-2012 and the shift in age distribution of CE-infected individuals over time indicate that the prevalence of CE in Turkana has decreased since the control program began.

A novel zoonotic genotype related to Echinococcus granulosus sensu stricto from southern Ethiopia.

Complete mitochondrial and two nuclear gene sequences of a novel genotype (GOmo) related to Echinococcus granulosus sensu stricto are described from a metacestode isolate retrieved from a human patient in southwestern Ethiopia. Phylogenetically, the genotype is positioned within the E. granulosus sensu stricto/Echinococcus felidis cluster, but cannot easily be allocated to either species. Based on different mitochondrial DNA markers, it is closest to the haplotype cluster that currently defines the species E. granulosus sensu stricto (which includes variants showing the widely cited G1, G2 and G3 sequences), but is clearly not part of this cluster. Pairwise distances between GOmo and E. granulosus sensu stricto are in the range of those between the most distant members of the Echinococcus canadensis complex (G6-10) that were recently proposed as separate species. At this stage, we prefer to list GOmo informally as a genotype rather than giving it any taxonomic rank because our knowledge rests on a single isolate from a dead-end host (human), and its lifecycle is unknown. According to data on molecularly characterised Echinococcus isolates from this region, GOmo has never been found in the usual livestock species that carry cystic echinococcosis and the possibility of a wildlife source of this newly recognised zoonotic agent cannot be excluded. The discovery of GOmo adds complexity to the already diverse array of cystic echinococcosis agents in sub-Saharan Africa and challenges hypotheses on the biogeographical origin of the E. granulosus sensu stricto clade.

Cystic echinococcosis in sub-Saharan Africa.

Cystic echinococcosis is regarded as endemic in sub-Saharan Africa; however, for most countries only scarce data, if any, exist. For most of the continent, information about burden of disease is not available; neither are data for the animal hosts involved in the lifecycle of the parasite, thus making introduction of preventive measures difficult. Available evidence suggests that several species or strains within the Echinococcus granulosus complex are prevalent in sub-Saharan Africa and that these strains might be associated with varying virulence and host preference. Treatment strategies (chemotherapy, percutaneous radiological techniques, but mainly surgery) predominantly target active disease. Prevention strategies encompass anthelmintic treatment of dogs, slaughter hygiene, surveillance, and health-educational measures. Existing data are suggestive of unusual clinical presentations of cystic echinococcosis in some parts of the continent, for which the causes are speculative.

Prevalence and diversity of cystic echinococcosis in livestock in Maasailand, Kenya.

Cystic echinococcosis (CE) is a zoonotic disease caused by several members of the Echinococcus granulosus species complex. In East Africa, several species/strains are known to occur in livestock and humans, but host preferences, relative frequencies and spatial distribution of these taxa are poorly known. Here, we contribute livestock data for Maasailand of southern Kenya. Total CE prevalence was 25.8 % in cattle (151/587), 16.5 % in sheep (71/430) and 10.8 % in goats (21/194), which is a significant increase compared to surveys done about three decades ago. The majority of cysts occurred in the liver (56 % in cattle, 70 % in sheep and 65 % in goats). Molecular characterization by PCR-RFLP and sequencing of parts of the mitochondrial nad-1 gene was done for a subsample of 285 cysts. E. granulosus G1 was dominant in all host species (200 of 201 cysts from cattle, 68 of 69 from sheep and 11 of 15 from goats); the remaining taxa were Echinococcus canadensis G6 (one cyst from sheep, four from goats) and Echinococcus ortleppi (one cyst from cattle). Considering cyst fertility, sheep appear to be the most important hosts for E. granulosus G1, while goats were found to be suitable hosts for E. canadensis G6 (three of four cysts were fertile). For the first time, E. ortleppi was found in cattle from southern Kenya. Our data show an intense and possibly increasing level of CE transmission in southern Kenya, and the predominance of E. granulosus G1, which appears to be particularly pathogenic to humans, calls for urgent control measures.

Molecular evidence of the camel strain (G6 genotype) of Echinococcus granulosus in humans from Turkana, Kenya.

Cystic echinococcosis (CE) is a zoonotic helminthic disease, which is widely distributed throughout the world. Although G1 is the Echinococcus granulosus genotype most commonly involved in CE in humans, the prevalence of infection with other genotypes, such as G6, may be higher than previously thought. We performed molecular analysis to identify which E. granulosus genotypes are the causative agents of CE in humans in Kenya's Turkana district. During a Hydatid Control Programme in 1993-1994, 71 cyst fluid isolates of E. granulosus were collected during PAIR (puncture, aspiration, injection, re-aspiration) sessions. DNA was amplified for two genes from 59 isolates. Of these, 49 isolates (83%) were identified as G1 and 10 (17%) as G6. This is the highest prevalence of G6 detected in humans of the Old World, and our results suggest that, in highly contaminated environments, G6 might be of greater public health significance than previously believed.

Evaluation of three PCR assays for the identification of the sheep strain (genotype 1) of Echinococcus granulosus in canid feces and parasite tissues.

The performance of 3 PCR assays for the identification of the G1 sheep genotype of Echinococcus granulosus was evaluated using tissue and canid fecal samples. The "Dinkel" and "Stefanic" primers were the most sensitive in detecting E. granulosus DNA in feces of necropsied dogs (73.7% and 100%, respectively). The "Abbasi" primers detected 52.6% of E. granulosus infected dogs but were the most species-specific, cross-reacting only with Echinococcus shiquicus (tissue 90.9%; feces 75%). The Stefanic primers were the least specific (tissue, 27.3%; feces, 25%) for E. granulosus. The Dinkel primers also showed inter-species cross-reactivity (tissue, 63.6%; feces, 100%) but were found to be strain-specific for the E. granulosus G1 sheep genotype. Improvement of PCR tests for Echinococcus species and subspecific variants should rely on the use of less-conserved genes and development of protocols that improve the quality and quantity of DNA extracted from feces.

Hydatid cysts: does every picture tell a story?

Ultrasound imaging is an effective aid to the detection of abdominal cystic echinococcosis. Recently, the images produced have provided more information than just a diagnosis. Different cyst morphologies have been classified into types, and such classifications can be considered to represent a natural history of cyst development. Understanding how cyst structures alter with time is crucial to evaluating the effectiveness of different treatment regimes and to understanding natural degeneration of cysts that can occur. However, the developmental events seen in ultrasound need to be matched more closely to immunological and cellular events if we are to have a fuller appreciation of disease progression and regression. At a community level, the relative proportions of cyst types could also provide epidemiological information on disease transmission.

Epidemiology and control of echinococcosis in sub-Saharan Africa.

Cystic echinococcosis (CE) is highly endemic among the nomadic pastoral tribes of East Africa, but is rare amongst the agriculturally based communities. Echinococcus granulosus infections are common in dogs from all countries in sub-Saharan Africa where they have been examined. Transmission of E. granulosus to humans is affected by such factors as prevalence of the parasite in domestic dogs, behaviors of humans towards dogs, and heterogenicity of the parasite and susceptibility of humans to infection. Sheep and goats appear to be the most common domestic intermediate hosts, but recent studies suggest that camels are equally important intermediate host, especially in Sudan and Turkana. At least five of ten E. granulosus genotypes are infective to humans in sub-Saharan African. Most human cases of CE are caused by the sheep strain (GI) and camel strain (G6) of E. granulosus. Other strains occurring in the area may include a lion strain, the horse strain (G4 or Echinococcus equinus) and the cattle strain (G5 or Echinococcus ortleppi).

A PCR system for detection of species and genotypes of the Echinococcus granulosus-complex, with reference to the epidemiological situation in eastern Africa.

We describe the development of a specific and sensitive PCR/semi-nested PCR system for the rapid diagnosis of Echinococcus granulosus genotype G1, E. granulosus genotype G6/7, and Echinococcus ortleppi (G5). Diagnosis of G1 and the group G5/6/7 is performed by a simple PCR, while discrimination between E. ortleppi (G5) and G6/7 involves a subsequent semi-nested PCR step. The target sequence for amplification is part of the mitochondrial 12S rRNA gene. Specificity of the PCRs was 100% when evaluated with isolates of 16 species of cestodes, including Echinococcus multilocularis, Echinococcus equinus, E. ortleppi and three strains of E. granulosus (G1, G6 and G7). Sensitivity threshold was 0.25pg of DNA. This new approach was compared with published protocols of restriction fragment length polymorphism-PCR and sequencing of mitochondrial cytochrome c oxidase subunit 1 and NADH dehydrogenase 1 genes using Echinococcus isolates of human, sheep, goat, camel, cattle and pig origin from Kenya and Sudan. Additionally, two internal DNA probes were developed, one hybridising only with G1, the other with G5, G6 and G7 amplification products. Preliminary epidemiological results obtained with this PCR approach include the detection of a camel strain (G6) infection for the first time in a human patient from eastern Africa, and the first reports of E. ortleppi (G5) in livestock from Kenya and the Sudan.