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.

Molecular Characterization of Giardia duodenalis in Children in Kenya.

BACKGROUND: Giardia duodenalis is an important intestinal protozoan in humans worldwide with high infection rates occurring in densely populated and low resource settings. The parasite has been recorded to cause diarrhea in children. This study was carried out to identify G. duodenalis assemblages and sub-assemblages in children presenting with diarrhea in Kenya. METHODS: A total of 2112 faecal samples were collected from children aged ≤ 5 years and screened for the presence of Giardia cysts using microscopy. A total of 96 (4.5%) samples were identified as Giardia positive samples and were genotyped using glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi) and ß-giardin loci. RESULTS: The three markers successfully genotyped 72 isolates and grouped 2 (1.4) isolates as Assemblage A, 64 (88.9) as Assemblage B and 7 (9.7%) consisted of mixed infections with assemblage A and B. A further analysis of 50 isolates using GDH Polymerase Chain Reaction and Restriction Fragment Length Polymorphism (PCR-RFLP) categorized 2 assemblage A isolates as sub-assemblage AII while 6 and 14 assemblage B isolates were categorized into sub-assemblage BIII and BIV respectively. A mixed infection with sub-assemblage BIII and BIV was recorded in 28 isolates. Over half (55.6%) of Giardia infections were recorded among the children between 13 to 48 months old. CONCLUSION: This paper reports the first data on the assemblages and sub-assemblages of Giardia duodenalis in children representing with diarrhea in Kenya.

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.

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.