Sympatric occurrence of Taenia saginata and Sarcocystis spp. in cattle from Narok County, Kenya: meat inspection findings with molecular validation.

The epidemiological picture of Taenia saginata infections in Kenya is fragmented with limited available data. Although Sarcocystis species are significant meat-borne parasites, few studies have explored their occurrence in Kenya. This study aimed to estimate the occurrence of bovine cysticercosis and screen for the presence of Sarcocystis spp. A meat inspection-based survey was conducted in ten abattoirs in Narok County, Kenya, and inspection for T. saginata cysticerci was limited to the Triceps brachii muscle. The apparent occurrence of the parasite was 5.4% (95% CI, 3.8, 7.6, n=573). Molecular confirmation of T. saginata was done via nested polymerase chain reaction targeting the mitochondrial 12S ribosomal RNA gene and restricted fragment length polymorphism. Sarcocystis species were identified using a multiplex polymerase chain reaction method targeting the 18S ribosomal RNA gene sequences and the mitochondrial cytochrome c oxidase subunit I gene. Of the 31 cystic lesions tested, 26/31 (83.9%) were confirmed to be T. saginata.Sarcocystis cruzi and S. hominis were detected in 8/31 (25.8%) and 1/31 (3.2%) of the cystic lesions, respectively. Co-infections of S. cruzi and T. saginata were found in 6/31 lesions (19.4%). The confirmation of bovine cysticercosis and S. hominis is suggestive of the presence of risky culinary and sanitation practices that facilitate transmission. This is the first report and molecular confirmation of Sarcocystis spp. in cattle in the country. The presence of both zoonotic S. hominis and pathogenic S. cruzi highlights an underexplored concern of veterinary and human health significance, warranting further epidemiological investigation.

A survey of intestinal helminths in domestic dogs in a human-animal-environmental interface: the Oloisukut Conservancy, Narok County, Kenya.

Dogs living in a domestic-wildlife interface can serve as reservoirs and sentinels of parasites shared among humans, domestic animals and wildlife. In Kenya, the epidemiology of intestinal parasites of dogs and their role as reservoirs of zoonoses is poorly understood, especially in domestic-wildlife interfaces. This study aimed to determine the occurrence of intestinal helminths in domestic dogs in the Oloisukut Conservancy. One hundred dog faecal samples were collected per rectum and examined microscopically following zinc chloride flotation and formal-ether concentration techniques. Genotyping of helminths was achieved by nested polymerase chain reaction of NADH dehydrogenase subunit 1, cytochrome oxidase 1 and partial sequencing. Nine genera were detected by microscopy in 65 (65%) dog faecal samples from 54/76 (71.05%) households. The most frequent helminths were hookworm (39%), Spirometra spp. (17%), taeniids (13%), Toxocara spp. (10%), Trichuris spp. (10%), Spirocerca lupi (5%), Physaloptera spp. (2%), Dipylidium caninum (1%) and Strongyloides spp. (1%). Ancylostoma caninum was the only hookworm species detected in dogs, while Taenia serialis and Taenia madoquae were detected in four and one faecal samples, respectively. This study reports for the first time the molecular detection of the cestodes Spirometra theileri, D. caninum and Mesocestoides sp. in dogs in Kenya. The presence of zoonotic helminths in dogs indicates that the residents of this conservancy are exposed to public health risks. The helminths reported here confirm the interaction of domestic dogs with wildlife. An integrated control programme involving the medical, veterinary and wildlife conservation professionals is needed to avert transmission of infectious diseases to humans, domestic animals and wildlife.

Prevalence and genotyping of Echinococcus granulosus sensu lato from livestock in north-eastern Kenya.

Cystic echinococcosis (CE) is a zoonotic disease of cosmopolitan distribution and caused by the larval stage of the dog tapeworm, Echinococcus granulosus sensu lato (s.l.). CE occurs in the wider African continent and in Kenya, notably in the Maasailand and Turkana regions; however, recent studies demonstrate its presence in other parts of Kenya. This study determined the occurrence of CE in livestock (camels, goats, sheep and cattle) in Isiolo, Garissa and Wajir counties, and characterized the species of E. granulosus s.l. present. An abattoir survey was used to determine the presence of CE in various organs in livestock. Polymerase chain reaction-restriction fragment length polymorphism and sequencing of the mitochondrial NADH dehydrogenase subunit 1 gene was used for genotyping. A total of 1368 carcasses from 687 goats, 234 camels, 329 sheep and 118 cattle were inspected for the presence of hydatid cysts. The overall proportion of infections was 29.1% in camels, 14.4% in cattle, 9.9% in goats and 8.2% in sheep. The liver was the most infected organ, while only the lung of camels harboured fertile cysts. Of the 139 cysts genotyped, 111 (79.9%) belonged to Echinococcus canadensis (G6/7) and 20 (14.4%) to E. granulosus sensu stricto. One and two cysts were identified as Taenia saginata and unknown Taenia species, respectively. There was a significant association between county of origin and species of the animal with occurrence of CE. This study reports, for the first time, the characterization of Echinococcus species in livestock from Garissa and Wajir counties, and the current situation in Isiolo county. The fertility of cysts in camels and frequency of E. canadensis (G6/7) in all livestock species indicate that camels play an important role in the maintenance of CE in the north-eastern counties of Kenya.

Molecular identification of zoonotic hookworms in dogs from four counties of Kenya.

All canine hookworms are known to be zoonotic, causing infections ranging from transient skin irritations to prolonged 'creeping eruptions', eosinophilic enteritis and even patent intestinal infections. There is little information on canine hookworm species and their public health significance in sub-Saharan Africa. This study determined the prevalence and species of hookworms in dogs from different climatic zones of Kenya. Dog faecal samples were collected from the environment, and hookworm eggs were isolated by zinc chloride flotation and subjected to DNA extraction. Polymerase chain reaction (PCR) assays targeting the internal transcribed spacer (ITS) 1 and 2, 5.8S and 28S ribosomal RNA of Ancylostoma spp. and Uncinaria stenocephala were performed, and hookworm species were identified by PCR-restriction fragment length polymorphism (RFLP) or DNA sequencing. Hookworm eggs were detected by microscopy in 490/1621 (30.23%, 95% CI 28.01-32.54) faecal samples. Estimates of faecal prevalence were high in counties receiving higher rainfall (Narok 46.80%, Meru 44.88%) and low in those with a more arid climate (Isiolo 19.73%, Turkana 11.83%). In a subset of 70 faecal samples, Ancylostoma caninum (n = 59) was the most common species, followed by A. braziliense (n = 10) and A. cf. duodenale (n = 1). This study reports for the first time the detection of A. cf. duodenale in dog faeces and zoonotic hookworm species in Kenyan dogs. These findings emphasize the need for control measures such as enforcing laws for restraining stray dogs, regular deworming of dogs, and public health awareness programmes aimed at informing communities on outdoor use of footwear.

Prevalence and genotyping of Echinococcus granulosus in sheep in Narok County, Kenya.

Cystic echinococcosis (CE) is a zoonotic disease caused by the larval stage of Echinococcus granulosus species (sensu lato, s.l.). In East Africa, several species/strains occur in livestock, wildlife, and humans, but there is limited information on frequencies of infection by different genotypes in the various mammalian hosts. We have obtained data on E. granulosus infection prevalence in sheep sampled from abattoirs in Narok County, southern Kenya. We inspected carcasses for the presence of hydatid cysts in 180 sheep randomly selected in five sub-locations. The overall prevalence was 16.0% (144/900 animals), with the majority of cysts (50.7%) found in the liver, followed by the lungs (36.8%), while infections involving the liver and lungs were detected in 12.5% of the sheep. PCR-RFLP genotyping of the mitochondrial nad-1 gene in all the 343 cysts identified E. granulosus G1-G3 (sensu stricto, s.s.) as the only genotype. The majority of the cysts (62.1%) were fertile, and 35.2% were sterile, while 2.7% were calcified. Considering cyst fertility, 73.02% of lung cysts were fertile compared to 53.4% in liver cysts. Our data extends previous CE studies in livestock and indicates a high level of CE infection of sheep in Narok, with a predominance of E. granulosus s.s., which is highly pathogenic and commonly infects humans. Given the high fertility rates observed in the cysts, there is an urgent need to determine whether there is a significant incidence of human infection in Narok, and initiate "One Health" control measures.

Cystic echinococcosis in Turkana, Kenya: 30 years of imaging in an endemic region.

Cystic echinococcosis (CE), a widespread, complex zoonosis, causes chronic disease associated with high morbidity. The pastoral Turkana people of Kenya have one of the highest prevalence rates of CE in the world. Between 1983 and 2015, a CE control program in the Turkana region used ultrasound (US) screening surveys and surgical outreach visits to evaluate CE prevalence and treat those with the disease. As the gold standard modality for diagnosing CE, US reveals a great deal of information about the disease in affected populations. The aim of this study is to discuss the characteristics of untreated CE in the Turkana people as revealed by US data collected during the CE control program and evaluate disease presentation, factors influencing the risk of transmission, and the timeline of disease progression. Data were obtained from written patient notes from US screenings and images; cysts were classified using the World Health Organization (WHO) standardized US classification of CE. Findings include greater prevalence of cysts, later stages of cysts, and multiple cysts in older age groups, with no multiple cysts occurring in patients under six years of age, which are consistent with the assertion that rates of exposure, transmission, and infection increase with age in endemic regions. Findings also raise questions regarding the timeline of disease progression, and factors potentially influencing disease transmission within this and other endemic populations. A comprehensive survey focusing on cultural and community observations (e.g., changing behaviors, hygienic practices, etc.) may provide more detailed information regarding factors that facilitate transmission.

The natural history of cystic echinococcosis in untreated and albendazole-treated patients.

The World Health Organization (WHO) treatment protocols for cystic echinococcosis (CE) are based on the standardized ultrasound (US) classification. This study examined whether the classification reflected the natural history of CE in untreated and albendazole-treated patients. Data were collected during mass US screenings in CE endemic regions among transhumant populations, the Turkana and Berber peoples of Kenya and Morocco. Cysts were classified using the WHO classification. Patient records occurring prior to treatment, and after albendazole administration, were selected. 852 paired before/after observations of 360 cysts from 257 patients were analyzed. A McNemar-Bowker χ2 test for symmetry was significant (p<0.0001). 744 observations (87.3%) maintained the same class, and 101 (11.9%) progressed, consistent with the classification. Regression to CE3B occurred in seven of 116 CE4 cyst observations (6.0%). A McNemar-Bowker χ2 test of 1414 paired before/after observations of 288 cysts from 157 albendazole-treated patients was significant (p<0.0001). 1236 observations (87.4%) maintained the same class, and 149 (10.5%) progressed, consistent with the classification. Regression to CE3B occurred in 29 of 206 CE4 observations (14.1%). Significant asymmetry confirms the WHO classification's applicability to the natural history of CE and albendazole-induced changes. Regressions may reflect the stability of CE3B cysts.

Echinococcus spp. in central Kenya: a different story.

Research on cystic echinococcosis (CE) has a long history in Kenya, but has mainly concentrated on two discrete areas, Turkana and Maasailand, which are known to be foci of human CE in Africa. Here, we report on a survey for CE in livestock from central to northeastern Kenya, from where no previous data are available. A total of 7,831 livestock carcasses were surveyed. CE prevalence was 1.92% in cattle (n = 4,595), 6.94% in camels (n = 216), 0.37% in goats (n = 2,955) and 4.62% in sheep (n = 65). Identification of the parasite was done using an RFLP-PCR of the mitochondrial nad1 gene, which had been validated before against the various Echinococcus taxa currently recognized as distinct species. From a total of 284 recovered cysts, 258 could be identified as Echinococcus granulosus sensu stricto (n = 160), E. ortleppi (n = 51) and E. canadensis (n = 47) by RFLP-PCR of nad1. In cattle, fertile cysts occurred mostly in the lungs and belonged to E. ortleppi (31 of 54), while the vast majority were sterile or calcified cysts of E. granulosus s.s.. Most fertile cysts in camels belonged to E. canadensis (33 of 37); sterile or calcified cysts were rare. Goats harboured fertile cysts of E. ortleppi (n = 3)--which is the first record in that host species--and E. canadensis (n = 1), while all cysts of E. granulosus were sterile. Only sterile cysts were found in the three examined sheep. Typically, all cysts in animals with multiple infections belonged to the same species, while mixed infections were rare. Our data indicate that the epidemiological situation in central to northeastern Kenya is clearly different from the well-studied pastoral regions of Turkana and Maasailand, and the apparently low number of human CE cases correlates with the infrequent occurrence of E. granulosus s.s.

A survey for Echinococcus spp. of carnivores in six wildlife conservation areas in Kenya.

To investigate the presence of Echinococcus spp. in wild mammals of Kenya, 832 faecal samples from wild carnivores (lions, leopards, spotted hyenas, wild dogs and silver-backed jackals) were collected in six different conservation areas of Kenya (Meru, Nairobi, Tsavo West and Tsavo East National Parks, Samburu and Maasai Mara National Reserves). Taeniid eggs were found in 120 samples (14.4%). In total, 1160 eggs were isolated and further analysed using RFLP-PCR of the nad1 gene and sequencing. 38 of these samples contained eggs of Echinococcus spp., which were identified as either Echinococcus felidis (n=27) or Echinococcus granulosus sensu stricto (n=12); one sample contained eggs from both taxa. E. felidis was found in faeces from lions (n=20) and hyenas (n=5) while E. granulosus in faeces from lions (n=8), leopards (n=1) and hyenas (n=3). The host species for two samples containing E. felidis could not be identified with certainty. As the majority of isolated eggs could not be analysed with the methods used (no amplification), we do not attempt to give estimates of faecal prevalences. Both taxa of Echinococcus were found in all conservation areas except Meru (only E. felidis) and Tsavo West (only E. granulosus). Host species identification for environmental faecal samples, based on field signs, was found to be unreliable. All samples with taeniid eggs were subjected to a confirmatory host species RLFP-PCR of the cytochrome B gene. 60% had been correctly identified in the field. Frequently, hyena faeces were mistaken for lion and vice versa, and none of the samples from jackals and wild dogs could be confirmed in the tested sub-sample. This is the first molecular study on the distribution of Echinococcus spp. in Kenyan wildlife. The presence of E. felidis is confirmed for lions and newly reported for spotted hyenas. Lions and hyenas are newly recognized hosts for E. granulosus s.s., while the role of leopards remains uncertain. These data provide the basis for further studies on the lifecycles and the possible link between wild and domestic cycles of cystic echinococcosis in eastern Africa.

MOLECULAR CHARACTERISATION OF ECHINOCOCCUS GRANULOSUS SPECIES/STRAINS IN HUMAN INFECTIONS FROM TURKANA, KENYA.

BACKGROUND: Cystic echinococcosis (CE) or hydatid disease is a neglected, economically important zoonotic disease endemic in pastoralist communities, in particular the Turkana community of Kenya. It is caused by the larval stage of the highly diverse species complex of Echinococcusgranulosus sensu lato (s.l). The situation on the genetic diversity in humans in Kenya is not well established. OBJECTIVE: To characterise Echinococcus granulosus (s.1) species/strains isolated from humans undergoing surgery in Turkana, Kenya. DESIGN: A Cross sectional study. SETTING: The Kakuma Mission Hospital and Centre for Microbiology Research, Kenya Medical Research Institute. SUBJECTS: Eighty (80) parasite samples from 26 subjects were analysed by Polymerase chain reaction--Restriction fragment length polymorphism (PCR-RFLP) targeting the nad 1 gene for molecular characterization. RESULTS: Two different genotypes of E. granulosus were identified from the samples analysed: E. granulosus sensu stricto (G1-G3) 85% of the samples analysed and E. canadensis G6/7 (15%). Most of the hydatid cysts (35%) were isolated from the liver. Other sites where cysts were isolated from include: kidney, abdomen, omentum, retroperitonium and the submandibular. Majority of cysts presented as CE1 (50%) and CE3B (42%) images according to WHO ultrasound classification. Both males and females were infected with E. granulosus s.s but only the females showed infection with E. canadensis G6/7. Chi-square test revealed significant difference between age of individuals and cysts classification by ultrasound. In addition, there was an association between cyst presentation (single or multiple) and genotype whereby all the E. canadensis G6/7 cases presented as single cysts in the infected persons. CONCLUSION: This study corroborates previous reports that E. canadensis G6/7 strain is present in Turkana, a place where initially only E. granulosus s.s (G1-G3) was known to be present and that E. granulosis (G1-G3) remains the most widespread genotype infecting humans in the Turkana community.

Echinococcosis in sub-Saharan Africa: emerging complexity.

Cystic echinococcosis occurs in most regions of sub-Saharan Africa, but the frequency of this zoonosis differs considerably among and within countries. Especially human cases seem to be focally distributed. A number of environmental and behavioural factors partially explain this pattern, i.e. density of livestock, presence of dogs, uncontrolled slaughter, and hygiene. In addition, the various taxa of Echinococcus spp. are known to differ considerably in infectivity to different host species including humans. Genetic characterizations of isolates, which are necessary to evaluate the impact of this factor - so far done in only a few countries - indicate that the diversity of Echinococcus spp. in Sub-Saharan Africa is greater than on any other continent. The very incomplete data which are available show that sympatrical taxa may infect different hosts, others may be geographically restricted, some life cycles involve livestock, others wild animals. Possible implications of this complexity for public health, livestock economy and conservation are briefly discussed.

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.

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.

A study of cystic echinococcosis in slaughter animals in three selected areas of northern Turkana, Kenya.

In an attempt to establish the prevalence of cystic echinococcosis, a study was conducted in slaughter animals in three divisions of northern Turkana, Kenya. A total of 5752 goats, 588 sheep, 381 cattle and 70 camels were examined at slaughter. Echinococcus granulosus metacestodes were found in 19.4% of the cattle, 3.6% of sheep, 4.5% of goats and 61.4% of camels. The prevalence of cystic echinococcosis in cattle, sheep and goats was higher in Lokichogio than in either Kakuma or Central divisions. On the other hand, the prevalence of the disease in camels was higher in Central (84.6%) than either Lokichogio (70.6%) or Kakuma (50%). The differences in prevalence rates in different study areas are attributed to differences in environmental conditions, livestock stocking intensity and cross-border migration of livestock.

Application of ultrasonography in prevalence studies of hydatid cysts in goats in north-western Turkana, Kenya and Toposaland, southern Sudan.

A study was done to determine the prevalence of hydatid cysts in goats using ultrasonography. A total of 1,390 goats were examined, 43,6 % (606/1,390) of them from north-western Turkana, Kenya, and 56,4% (784/1,390) from Toposaland, southern Sudan. Hydatid cysts were visualized in 1,82 % (11/ 606) of the goats from north-western Turkana and 4,34% (34/784) of those from Toposaland. Unlike abattoir surveys, the prevalence data obtained in this study were unbiased because entire flocks were examined. The lower prevalence rate of the disease in goats from Turkana was attributed to the hydatid disease control programme in that area, which is absent in Toposaland.

Prevalence of intestinal parasites among children in southern Sudan.

A study was conducted in southern Sudan to determine the prevalence of intestinal parasites among school children. A total of 275 stool samples which were examined using formol-ether concentration techniques yielded 15 different species of parasites. Hook worm with a prevalence of 13.1% was the predominant nematode followed by Strongyloides stercoralis (3.3%), Trichostrongylus (2.5%), Schistosoma mansoni (2.2%) and Trichuris trichiura (1.8%). Ascaris lumbricoides and cestodes were not detected in this population. Intestinal protozoans were common. Entamoeba coli (37.8%), Entamoeba histolytica (28.4%) and Giardia lamblia (9.8%). Children in the age group 6-10 years old were the most affected followed by the 11-15 year-old age group. The infection rate was slightly higher in males than females.

Oxfendazole treatment for cystic hydatid disease in naturally infected animals.

Few chemotherapeutic agents are available for the medical management of hydatid disease caused by the parasite Echinococcus granulosus. In order to test the potential of oxfendazole for the treatment of infection with this parasite, nine infected goats and four sheep were given oxfendazole twice weekly at a dose of 30 mg/kg of body weight for 4 weeks and monitored by ultrasound for an additional 4 weeks. Efficacy was finally evaluated by postmortem examination, including determination of protoscolex viability and cyst wall histology. In treated animals, protoscolices were dead or absent in 97% of cysts from oxfendazole-treated animals compared to 28% of cysts from untreated control animals. On postmortem examination, 53% of cysts from treated animals were found to be grossly degenerate. A sample of those cysts that appeared potentially viable all demonstrated evidence of severe damage to the cyst wall. By light microscopy, cysts showed severe disorganization of the adventitial layer with invasion of inflammatory cells and in some cases frank necrosis with no apparent adventitial layer. The follow-up period for assessment of the drug's ability to cause complete degeneration and resorption of cysts was relatively short. This study, however, indicates that oxfendazole is at least as effective as and is easier to administer than albendazole for the treatment of hydatid disease.

Evaluation of diagnostic ultrasound as a mass screening technique for the detection of hydatid cysts in the liver and lung of sheep and goats.

Ultrasound examination of the liver and lung followed by post-mortem examination was performed in 16 sheep and 284 goats. Thirty-one (10.3%) were positive for hydatid cysts on ultrasound examination and 46 (15.3%) were positive on post-mortem examination. Twenty-one positive on post-mortem examination were falsely identified as negative on ultrasound examination. Of the 254 animals negative on post-mortem examination, six (2.4%) were falsely identified as positive on ultrasound examination. The sensitivity and specificity of ultrasound examination for detecting hydatid cysts in sheep and goats was 54.36% and 97.64%, respectively (positive predictive value: 80.64%; negative predictive value: 92.19%).