Phosphorus budget in the low-income, peri-urban area of Kibera in Nairobi (Kenya).

Kibera, located in Nairobi, Kenya is one of the largest (235,000 inhabitants) low-income areas in East Africa. Surface waters in Kibera show high pollution levels with respect to SRP (soluble reactive phosphorus; range: 2-10 mg P/L), coming from the uncontrolled wastewater discharges in the area. The different P production and consumption values in Kibera were estimated using interviews (155 interviewed) as well as detailed P house-keeping for five representative families. The results show that highest P consumption comes from food, in particular cereals. Highest P production came from urine (55% of the total) and faeces (31%), with relatively lower contributions from grey water and solid wastes. The overall P budget in Kibera amounted to around 9 x 10(3) kg P/month. This is equivalent to 0.47 g P/person yr, both for P production and consumption, with a relative error of 20%. Comparing with the estimated P outflows via the Kibera surface waters, around 65% of the P produced in Kibera will leave the area. In future ECOSAN techniques such as urine separation could well be applied for efficient recycling of these waste sources.

Subpopulations of T lymphocytes in Kenyan patients with visceral leishmaniasis.

Populations of peripheral blood T lymphocytes from patients with Kenyan visceral leishmaniasis were studied using specifically defined antisera (monoclonal antibodies, Ortho-mune OKT3, OKT4, OKT6, and OKT8). The levels of total T lymphocytes and circulating thymocytes were within the same range as those of clinically normal individuals. However, the proportions of the helper/inducer T cells were lower in untreated patients than in the controls (18.9% vs. 39.7%) while the levels of suppressor/cytotoxic T cells were higher than in the controls (40.5% vs. 27.8%). After successful antileishmania treatment these levels showed a gradual return towards normal over a period of one year. It was concluded that immunosuppression observed is due to the levels of peripheral blood helper/inducer and suppressor/cytotoxic T lymphocytes.

Chloroquine resistance in Plasmodium falciparum from Kenyan infants.

Forty-two infants, aged 6 to 24 months, infected with Plasmodium falciparum were identified in Kisumu, Kenya. Because of their age, all were presumably not semi-immune to malaria. Each infant was treated with 25 mg/kg chloroquine base and followed for 7 days. Forty-one infections were sensitive to chloroquine in vivo; asexual parasites disappeared in all by day 4 and were not present on days 5, 6, or 7. One infection was resistant in vivo; parasites disappeared by day 3 but recrudesced on day 4. Rieckmann micro in vitro tests for chloroquine were done on the 42 isolates. Interpretable results were found in 25. In vitro resistance was demonstrated in 18 (72%) isolates, including the patient with in vivo resistance; greater than or equal to 99% inhibition of schizont development only occurred in wells containing greater than or equal to 8 pmol chloroquine base (compared with less than or equal to 5.7 pmol/well for known sensitive isolates). This is the first demonstration of in vivo and in vitro chloroquine-resistant P. falciparum in a Kenyan. Comparison of these results with results from other studies carried out in the same area on the same area on older individuals suggests that the immune response may be playing a role in modifying the expression of resistance.

Contribution of adherent cells and serum components to immune suppression in Kenyan visceral leishmaniasis.

Adherent cells and serum components from Kenyan patients with visceral leishmaniasis were examined with the view to evaluating their contribution to cell-mediated immune suppression. Mitogens (phytohemagglutinin and concanavalin A) and antigens (purified protein derivative, streptokinase-streptodornase, and leishmania) were used as stimulants. Compared to the controls, the contribution of serum components to suppression in presence of any of the mitogens and antigens was not significant. The same applied to adherent cells, except in the presence of leishmania antigen where adherent cells contributed significantly (P less than 0.001). Removal of adherent cells from peripheral blood mononuclear cells of patients and controls considerably increased in vitro lymphocyte responses to both mitogens and antigens (by about twice), suggesting that in this study, the inhibition of in vitro lymphocyte responses to antigens and mitogens by adherent cells was a general phenomenon independent of the presence of the disease.

In vitro susceptibility of Plasmodium falciparum isolates from Jilore, Kenya, to antimalarial drugs.

Twenty-six Plasmodium falciparum isolates obtained during a prophylaxis study at Jilore primary school, Malindi, Kenya, were adapted to in vitro culture and their susceptibility to 13 antimalarial drugs was tested by a modified radioisotopic method. Pyrimethamine, chloroquine, amodiaquine, cycloguanil, chlorcycloguanil, quinine, quinidine and sulfadoxine, and the experimental compounds MB 35769, mefloquine, WR 184806, parvoquone, and menoctone were used. The isolates could be divided into two groups with significantly different susceptibility to pyrimethamine, shown by a 755-fold difference in the mean ID50 values (2.77 +/- 1.98 x 10(-10) mol/l and 2.09 +/- 1.64 x 10(-7) mol/l). The mean susceptibility of the two groups differed 7.7-fold for chlorcycloguanil and 14.6-fold for cycloguanil, but were not significantly different for the other drugs. All isolates were more sensitive to amodiaquine than to chloroquine in vitro. The ratio of the geometric mean ID50 values of chloroquine to amodiaquine was 3.13. The ratio for the chemically related compounds parvoquone to menoctone was 5.63, quinine to quinidine was 5.58, and mefloquine to WR 184806 was 12.16.

Control of schistosome-transmitting snails in Kenya by the North American crayfish Procambarus clarkii.

Snail-transmitted trematode parasites such as schistosomes and liver flukes assume considerable medical and veterinary significance in tropical Africa. We have observed a strong negative association between the presence of medically important pulmonate snails and the crayfish Procambarus clarkii in freshwater habitats in Kenya. This crayfish, introduced into Kenya around 1970, readily consumes these snails in the laboratory. Field enclosure experiments indicate that crayfish exert a significant negative impact on the abundance of Biomphalaria pfeifferi, the intermediate host of the human blood fluke Schistosoma mansoni. It is likely that P. clarkii will continue to spread naturally in Kenya and that schistosome-transmitting snails will be excluded or reduced in numbers where crayfish are present. Procambarus clarkii may represent an alternative, biological means of snail control in East Africa.

Identification of snails infected with schistosomes by ELISA employing monoclonal antibodies: Schistosoma mansoni in laboratory snails (Biomphalaria glabrata) and in field snails (Biomphalaria pfeifferi) from Kenya.

An enzyme-linked immunosorbent assay (ELISA) employing monoclonal antibodies was used for detecting Schistosoma mansoni antigens in hemolymph of laboratory snails (Biomphalaria glabrata) in Kenya. Infected laboratory snails shedding cercariae were differentially identified by ELISA from uninfected snails with 100% sensitivity and specificity. Prepatent infections were detected by ELISA from 2 weeks after exposure to miracidia. Thus, ELISA revealed infection 3 weeks before maximal patency was reached (5-6 weeks post-exposure). Infected field snails (B. pfeifferi) shedding cercariae were differentially identified by ELISA, with 100% sensitivity and specificity, from uninfected field snails and from snails naturally infected with other trematodes (echinostomes and strigeids). Prepatent infections with S. mansoni were readily identified by ELISA in field snails. A case is demonstrated where infection rate, as determined by shedding test alone, was 9.8%, whereas the combined figure of prepatent and patent infection rates was 22.9%

Development of Leishmania major in Phlebotomus duboscqi and Sergentomyia schwetzi (Diptera: Psychodidae).

The extrinsic development of Leishmania major was observed in 2 man-biting sand flies, Phlebotomus duboscqi, a known vector, and Sergentomyia schwetzi, an assumed non-vector. Flies fed on a leishmanial lesion on the nose of a hamster were examined for infection at 0, 6, 12, 18, 24, 36, 48, and 60 hr and at approximately 24 hr intervals from day 3 to day 14 post-feeding. Infection rates, determined by light microscopy, were 47% (n = 258) in P. duboscqi and 5% (n = 162) in S. schwetzi. Transformation from amastigotes to "procyclic" promastigotes occurred in both species at 6-18 hr post-feeding. In P. duboscqi, the parasites multiplied rapidly and developed through as many as 10 forms, including at least 3 dividing-promastigote forms. Metacyclic promastigotes, the "infective" form, appeared at 6 days post-feeding, first in the region of the stomodeal valve, then in the pharynx, cibarium, and proboscis. In a single attempt 14 days post-feeding, a P. duboscqi transmitted L. major to a mouse by bite. In contrast, the parasites multiplied slowly in S. schwetzi, and did not develop beyond "procyclic" promastigotes. The parasites did not migrate anteriorly nor survive beyond 90 hr post-feeding, indicating that S. schwetzi is not a vector of L. major. Classical strategies for vector incrimination may be confounded by the isolation of non-infective early developmental forms of Leishmania from wild-caught non-vectors.

Concurrent infection with Leishmania donovani and Leishmania major in a Kenyan patient: clinical description and parasite characterization.

Leishmania isolates aspirated a few months apart from the spleen of an indigenous adult male kala-azar patient from Baringo District, Kenya, were biochemically characterized and compared. The patient lived within a dual focus of L. donovani kalazar and L. major cutaneous leishmaniasis. A primary Leishmania isolate from splenic aspirates was cryopreserved (NLB-294). The patient was treated with sodium stibogluconate for kala-azar and discharged. Three months later, he had clinical relapse and returned for retreatment. During his second visit, the patient participated in a diagnostic study in which urine and nasopharyngeal samples were cultured for leishmaniasis. Urine, nasopharyngeal, and splenic samples were positive for Leishmania. Secondary isolates from splenic (NLB-294-I) and urine (NLB-318) cultures were cryopreserved and characterized by cellulose acetate electrophoresis (CAE) using 20 enzymes. Whereas the urine isolate was typed as L. donovani, the splenic aspirate culture revealed a mixed infection with L. donovani and L. major. The primary isolate (NLB-294) was then characterized and also showed a mixed infection. To exclude the possibility of protein post-translational modifications in electrophoretic assays, the primary and secondary isolates were grown and processed under identical cultural and lysis conditions, and compared using CAE. The results were identical to the first electrophoretic assays showing mixed promastigote banding patterns. Stationary-phase promastigotes of the secondary splenic isolate (NLB-294-I) inoculated subcutaneously, intraperitoneally, and intracardially into Syrian hamsters and BALB/c mice produced both kala-azar and cutaneous leishmaniasis within 6.5 months.(ABSTRACT TRUNCATED AT 250 WORDS)

Cellular and humoral immune responses in a population from the Baringo District, Kenya to Leishmania promastigote lipophosphoglycan.

In a cross-sectional house-to-house study in a leishmaniasis-endemic area in Kenya, the cellular and humoral immune response to Leishmania lipophosphoglycan (LPG) was determined. Clinical data, peripheral blood mononuclear cells, and plasma were obtained from 50 individuals over the age of eight years. Lymphoproliferation and interferon-gamma (IFN-gamma) production by these cells were examined. It was shown that cells from all six individuals in the population with a history of kala-azar responded to LPG in the lymphocyte proliferation assay, and four of these six responded in the IFN-gamma assay. In contrast, cells from 12 of 44 individuals from the study area with no history of kala-azar and none of the five Danish control samples responded to LPG. Antibodies against LPG were detected by enzyme-linked immunosorbent assay in 45 of 50 plasma samples. Our findings clearly show that mononuclear cells from kala-azar patients cured of infection were able to respond to the LPG preparation. The finding of a specific cellular immune response to LPG in 12 of 44 individuals with no history of kala-azar is consistent with previous epidemiologic studies, in which it has been shown that a proportion of L. donovani infections run a subclinical course. The high frequency of individuals with antibodies against LPG might indicate that a majority of the population had been exposed to the parasite.(ABSTRACT TRUNCATED AT 250 WORDS)

Impact of the crayfish Procambarus clarkii on Schistosoma haematobium transmission in Kenya.

The Louisiana red swamp crayfish, Procambarus clarkii, which was introduced into east Africa in the 1950s or 1960s, has since widely dispersed. Previous work by our group has shown that P. clarkii can reduce populations of the molluscan intermediate hosts of human schistosomes through predatory and competitive interactions. Here, we investigate whether crayfish can reduce populations of Bulinus africanus and consequently, Schistosoma haematobium prevalence in school children. Children from 6 primary schools in the Machakos and Kitui Districts of Kenya were selected for study. Schools were divided into 3 experimental-control pairs. At experimental schools, crayfish were introduced into nearby aquatic habitats harboring Bulinus africanus snails and serving as S. haematobium transmission sites. Snail habitats near control schools did not receive crayfish. Six months after crayfish introduction, all infected children were treated with praziquantel. Children were then monitored quarterly for 2 years, at which time infection and reinfection rates were compared statistically between the paired schools. In one such pair, crayfish failed to establish, resulting in neither snail control nor a reduction in transmission. At the second pair of schools, the numbers of snails were decreased by the presence of crayfish, but a clear difference in infection rates in children could not be detected, primarily because drought conditions kept overall transmission rates low. At the third school pair, crayfish established well in experimental habitats, snail numbers decreased precipitously, and children at the experimental school were significantly less likely to acquire S. haematobium infections than children at the control school. Our results indicate that under certain environmental circumstances, P. clarkii exerts a significant impact on the transmission of human schistosomiasis in Kenya. Important questions remain regarding the impact of P. clarkii on Kenyan freshwater ecosystems, not the least of which is its potential to significantly influence the epidemiology of schistosomiasis in east Africa.

The Kenyan Saradidi community malaria project: I. Response of Plasmodium falciparum isolates to chloroquine in 1981 and 1982.

In 1981, 41 schoolchildren from Saradidi, Kenya, infected with Plasmodium falciparum were treated with chloroquine. All 41 infections were sensitive in vivo: parasitaemia cleared by day 3 and remained absent through day 7. All 17 (of 27) isolates successfully tested for chloroquine sensitivity in vitro were sensitive in the Rieckmann macro test: greater than 1% schizont development did not occur at chloroquine concentrations of greater than 1.25 nmol per ml of blood. In 1982 in the same area 20 P. falciparum infections were sensitive in vivo: parasitaemia cleared by day 5 and did not recur through day 7. Two of the 20 isolates were resistant in vitro with persistent schizont development at greater than 1% of control values at chloroquine concentrations of 1.5 and 3.0 nmol/ml in the macro test and 16 and 32 pmol/well in the Rieckmann micro test (compared with inhibition at less than or equal to 1.25 nmol/ml and less than or equal to 5.7 pmol/well, respectively, for sensitive isolates). In a modified 48-hour test, growth of two additional isolates was not inhibited until chloroquine concentration of 0.06 nmol per ml of medium, a pattern intermediate between that observed with known chloroquine-sensitive (less than or equal to 0.03 nmol/ml) and resistant (greater than or equal to 0.1 nmol/ml) P. falciparum isolates. The results demonstrate a changing pattern of the in vitro response of P. falciparum isolates in Saradidi to chloroquine.

High-dose sodium stibogluconate treatment of cutaneous leishmaniasis in Kenya.

Cutaneous leishmaniasis caused by Leishmania aethiopica usually responds poorly to conventional doses of pentavalent antimonial drugs. We treated three patients with cutaneous leishmaniasis acquired in Kenya, presumed or documented to be caused by L. aethiopica, with intravenous sodium stibogluconate, 18 to 20 mg Sb/kg body-weight twice daily for 30 days. All patients had a good response to treatment, with disappearance of parasites from skin smears and cultures after 14 to 27 days, clinical healing of the lesions, and no recurrence during a three to 18-month follow-up. Side effects of treatment were minor. We conclude that this high dose sodium stibogluconate regimen is safe and effective for treating cutaneous leishmaniasis caused by L. aethiopica in Kenya.

Response of Plasmodium falciparum to dihydrofolate reductase inhibitors in Malindi, Kenya.

The response of Plasmodium falciparum isolates to dihydrofolate reductase inhibitors (DHFRI) was examined in Malindi, Kenya. All 20 infected children treated with pyrimethamine/sulphadoxine responded. In contrast, after treatment with pyrimethamine, parasitaemia in 9 of 14 infections failed to clear or recrudesced during the seven-day follow-up. In a 48-hour in vitro test, five of six isolates resistant to pyrimethamine in vivo had a minimal inhibitory concentration (MIC) to pyrimethamine greater than or equal to 300 nmoles/1 compared with less than or equal to 100 nmoles/1 for the four sensitive isolates; four isolates did not grow. MIC to M-B 35769, an experimental DHFRI structurally similar to pyrimethamine were the same (six isolates) or 10-fold lower (three isolates). In the laboratory four of five isolates adapted to in vitro culture had the same MICs as in the field while one isolate became less responsive to both drugs. Cycloguanil (the active metabolite of proguanil) was more active in vitro in the laboratory than pyrimethamine or M-B 35769.

Chlorproguanil/dapsone for the treatment of non-severe Plasmodium falciparum malaria in Kenya: a pilot study.

Chlorocycloguanil, the active metabolite of chlorproguanil, was synergistic in vitro with dapsone against 2 culture-adapted Plasmodium falciparum isolates from Kenya; maximal synergy occurred at lower concentrations that it did with pyrimethamine and sulfadoxine. 48 children with asymptomatic P. falciparum infections were treated with chlorproguanil (at a target dose of 1.2 mg/kg) and dapsone (target dose of 1.2 or 2.4 mg/kg); all were free of parasitaemia by day 7. The following numbers had recurrences on days 14, 21, and 28, respectively: 1 of 48, 7 of 47, and 7 of 40. All 39 children treated with pyrimethamine (target dose 1.2 mg/kg) and sulfadoxine (target dose 24 mg/kg) were cleared of infection, while the following had recurrences on days 14, 21, and 28: 1 of 39, 2 of 38, and 2 of 36. The rate of decrease in parasitaemia was the same in the 2 groups, and there was no change in haematocrit or haemoglobin during the follow-up. The rate of recurrence in the children receiving chlorporguanil/dapsone was higher, probably because these drugs have a much shorter clearance time than pyrimethamine/sulfadoxine. Chlorproguanil/dapsone is an effective combination for treating P. falciparum malaria and deserves further study.

Experimental transmission of Leishmania major to vervet monkeys (Cercopithecus aethiops) by bites of Phlebotomus duboscqi (Diptera: Psychodidae).

Experimental transmission of Leishmania major to vervet monkeys (Cercopithecus aethiops) was accomplished by bites of Phlebotomus duboscqi sandflies. Three-day-old, laboratory-reared P. duboscqi were fed on leishmanial lesions on hamsters infected with L. major. The flies were re-fed on monkeys 10 d after infection. Five adult male vervet monkeys were used in concurrent transmission trials. Two of the monkeys received subcutaneous inoculations with stationary-phase promastigotes (2 x 10(6) promastigotes in 0.1 ml of medium) on the base of the tail. Putatively infected P. duboscqi were allowed to feed on the remaining 3 monkeys at sites on the base of the tail and on the right eyebrow. Challenges by sandfly bites resulted in multiple leishmanial lesions at all bite sites and, consequently, more lesion area than was produced by needle challenges. Post-feeding dissection of sandflies indicated that multiple lesions could be caused by bites of a single fly, and that probing alone, without imbibing blood, was sufficient for transmission. These first experimental transmissions of L. major to vervets by bites of P. duboscqi demonstrate that sandfly challenge is an efficient alternative to needle challenge, making available a unique Leishmania-sandfly-non-human primate model for use in vaccine development.

Variability in the metabolism of proguanil to the active metabolite cycloguanil in healthy Kenyan adults.

Extensive metabolizers (EM) and poor metabolizers (PM) of the malaria chemoprophylactic drug proguanil have been identified by measuring the proguanil/cycloguanil ratio in urine following a single dose of the pro-drug. The pharmacokinetic characteristics of proguanil were similar in 8 EM and 8 PM subjects, but there were significant differences between the 2 groups with respect to cycloguanil pharmacokinetics. In none of the PM subjects could cycloguanil be detected in whole blood samples at any time after proguanil dosage. Plasma cycloguanil was measureable in only 2 of 8 PM subjects, despite an analytical sensitivity in the high-performance liquid chromatographic assay of 1 ng/ml cycloguanil. A comparatively high proportion of Black Kenyan adults appear to metabolize proguanil poorly, possibly because they lack the specific mixed function oxidase which will accept proguanil as substrate.

A new rural focus of cutaneous leishmaniasis caused by Leishmania tropica in Kenya.

We have identified a new rural focus of cutaneous leishmaniasis caused by Leishmania tropica in Muruku sublocation, Salama location, Laikipia district, Rift Valley province, Kenya. Based on a few available case histories, previous reports of L. tropica in Kenya indicated a tentative geographical distribution. Recently 6 indigenous Kenyans from the new focus, who had never travelled outside Kenya, developed cutaneous lesions on the face and/or extremities found to contain Leishmania by culture and smear. Most of the patients manifested the typical 'urban' dry sore which grew slowly into a nodule measuring 2 x 1 cm to 9.5 x 3 cm, and after some months formed a central crust surrounded by small satellite papules. After treatment with Pentostam (sodium stibogluconate), about 40% of the sores failed to heal completely, either scarring centrally with fulminating papules at the edges and spreading peripherally, or healing but then recrudescing at the edge of the scar. Stationary-phase promastigotes from culture isolates were analysed by cellulose acetate electrophoresis. Isoenzyme profiles of 6 isolates were compared with those of World Health Organization reference strains using 12 enzyme loci; they were indistinguishable from those of 2 L. tropica reference strains. All 6 case sites lay within a radius of 4 km. Several other suspected cases from the same area are being investigated.

Field application of an ELISA using redefined Leishmania antigens for the detection of visceral leishmaniasis.

Two soluble antigens from Leishmania donovani of 116 kDa and 70 kDa molecular mass, and a soluble mixture of crude antigens, were used in an enzyme-linked immunosorbent assay (ELISA) for the detection of visceral leishmaniasis (VL) in the field, and compared with the direct agglutination test (DAT). The tests were carried out on 8 VL patients, 34 normal individuals from an area endemic for the disease, and 68 former visceral leishmaniasis patients 1-5 years after treatment. The 70 kDa ELISA and the DAT had a sensitivity and specificity of 100% (95% confidence interval 63-100%), while the 116 kDa ELISA and the soluble crude antigen ELISA were 37.5% (9-76%) and 50% (16-84%) sensitive, respectively. When using ELISA (116 kDa or 70 kDa), 68-69% of sera tested 1-2 years, and 92-94% of sera tested 5 years, after treatment were negative. In contrast, when DAT or ELISA with crude antigen were used, the negativity rate was 31% 1-2 years, and 53% 5 years, after treatment. DAT was therefore not an accurate test for diagnosis in the field. The use of the 70 kDa antigen in ELISA was an accurate alternative to DAT in the detection of VL.

Antifolate drug resistance and point mutations in Plasmodium falciparum in Kenya.

Due to increased chloroquine resistance, the antifolate/sulpha drug combinations are becoming increasingly important in the chemotherapy of falciparum malaria. However, point mutations in the dihydrofolate reductase gene lead to resistance to the antifolate drugs. We therefore investigated the prevalence of the 6 reported point mutations in this gene among field isolates of Plasmodium falciparum from Kenya, to determine if the mutations correlated with resistance to pyrimethamine and the biguanides cycloguanil and chlorcycloguanil. We used a mutation-specific polymerase chain reaction technique to test for these reported mutations in 21 Kenyan isolates and 4 reference lines. We also amplified and directly sequenced the dihydrofolate reductase coding sequence from these parasites to confirm the results and test for other possible mutations. Of the reported mutations, we found S108N, which is the central mutation of pyrimethamine resistance, and mutations N51I and C59R, which modulate the levels of resistance and may confer decreases in response to cycloguanil that are folate and p-aminobenzoic acid dependent. No isolate possessed the paired point mutations S108T and A16V, or I164L and S108N, which have been associated with cycloguanil resistance in previous studies. These results provided supportive evidence for the combined use of a cycloguanil-class drug (e.g., chlorproguanil) and a sulpha drug (e.g., dapsone) against P.falciparum malaria in Kenya.