Genetic diversity of Kenyan Prosopis populations based on random amplified polymorphic DNA markers

Several Prosopis species and provenances were introduced in Kenya, either as a single event or repeatedly. To date, naturally established Prosopis populations are described as pure species depending on site, despite the aforementioned introduction of several species within some sites. To determine whether naturally established stands consist of a single or mixture of species, six populations from Bamburi, Bura, Isiolo, Marigat, Taveta and Turkwel were compared for relatedness with reference to Prosopis chilensis, Prosopis juliflora and Prosopis pallida using random amplified polymorphic DNA markers. Cluster analysis based on Nei’s genetic distance clustered Kenyan populations as follows: Marigat, Bura and Isiolo with P. juliflora, Bamburi with P. pallida and Taveta with P. chilensis, whereas the Turkwel population is likely to be a hybrid between P. chileneis and P. juliflora. Four populations had private markers, revealing germplasm uniqueness. Expected heterozygosity tended to be larger for Kenyan populations (ranging from 0.091 to 0.191) than in the three reference (ranging from 0.065 to 0.144). For the six Kenyan populations and two P. juliflora provenances from the Middle East, molecular variation was larger within populations than between population. Higher molecular variance among populations is attributed to their geographical separation and the low variation within populations is due to gene flow between individuals within a population. Overall, this study shows that (1) the Kenyan Prosopis populations are genetically isolated, (2) multiple introductions enhanced genetic diversity within sites and (3) P. juliflora and its hybrid are the most aggressive invaders.Key words: Prosopis chilensis, Prosopis juliflora, Prosopis pallida, multiple introductions, genetic diversity

Segurança e reação de hipersensibilidade tardia na pele de macacos vervet imunizados com antígeno sonicado de Leishmania donovani junto com adjuvantes

In this study, we report on the safety and skin delayed-type hypersensitivity (DTH), responses of the Leishmania donovani whole cell sonicate antigen delivered in conjunction with alum-BCG (AlBCG), Montanide ISA 720 (MISA) or Monophosphoryl lipid A (MPLA) in groups of vervet monkeys. Following three intradermal injections of the inoculums on days 0, 28 and 42, safety and DTH responses were assessed. Preliminary tumor necrosis factor alpha (TNF-&#945;) and interferon gamma (IFN-&#947;) levels were also measured and these were compared with DTH. Only those animals immunized with alum-BCG reacted adversely to the inoculum by producing ulcerative erythematous skin indurations. Non-parametric analysis of variance followed by a post-test showed significantly higher DTH responses in the MISA+Ag group compared with other immunized groups (p < 0.001). The MPLA+Ag group indicated significantly lower DTH responses to the sonicate antigen compared with the AlBCG+Ag group. There was a significant correlation between the DTH and cytokine responses (p < 0.0001). Based on this study we conclude that Leishmania donovani sonicate antigen containing MISA 720 is safe and is associated with a strong DTH reaction following immunization.Neste estudo reportamos segurança e resposta de hipersensibilidade tardia (DTH) do antígeno sonicado de células totais de Leishmania donovani introduzidos juntamente com alume-BCG (AIBCG) Montanide ISA 720 (MISA) ou lípide A monofosforilado (MPLA) em grupos de macacos vervet. Depois de três injeções intradérmicas do inóculo nos dias 0, 28 e 42 segurança e resposta DTH foram avaliados. Preliminarmente níveis de fator de necrose tumoral alfa (TNF-&#945;) e interferon gama (IFN-&#947;) foram também medidos e comparados com o DTH. Somente os animais imunizados com alume-BCG reagiram de maneira diversa ao inóculo produzindo indurações ulceradas e eritematosas na pele. Análise não paramétrica de variação seguida por um teste posterior mostraram resposta significantemente mais alta do DTH no grupo MISA + Ag quando comparado com outros grupos imunizados (p < 0.001). O grupo MPLA + Ag demonstrou resposta DTH significantemente menor do antígeno sonicado comparado com o grupo AIBCG + Ag. Houve correlação significante entre o DTH e a resposta às citocinas (p < 0.0001). Baseados neste estudo concluímos que o antígeno sonicado de Leishmania donovani contendo MISA 720 é seguro e está associado com forte reação DTH após imunização

Evaluation of the Therapeutic Potential of Warburgia ugandensis, Prunus africana, and Piliostigma thonningii against Leishmania donovani in vitro and in Balb/c Mice

Leishmaniasis is a zoonotic disease caused by protozoan parasites of the genus Leishmania. Conventional chemotherapy remains to be the most preferred measure against leishmaniasis despite being associated with high toxicity and relapse rates. They are also expensive and require hospitalization. Plant-based compounds provide a better treatment alternative because they are effective, cheap, and less associated with toxicity and resistance. This study examined the therapeutic potential of Warburgia ugandensis, Prunus africana, and Piliostigma thonningii against Leishmania donovani infection in BALB/c mice. Anti-promastigote and toxicity studies were evaluated by incubating the test compound with promastigotes and Vero cells, respectively. Serum was obtained from the mice for total immunoglobulin gamma (IgG) quantification. For in vivo studies, the mice were infected with virulent Leishmania donovani then treated with methanolic extracts of Warburgia ugandensis, Prunus africana, and Piliostigma thonningii and control drug, pentostam (sodium stibogluconate). Treatment with the plant extracts and standard drug resulted to significant reduction in parasite burden. Outcomes in the mice treated with plant extracts were comparable to those treated with pentostam (P≥0.05). In the promastigote assay, all the test compounds killed more than half of the promastigotes at the highest concentration (500 µg/mL). Warburgia ugandensis, P. thonningii, and P. africana reduced the number of promastigotes from 2.0 × 106 to 7.7 × 103 , 72.0 × 103 , and 5.0 × 103 , respectively. Pentostam had the lowest IC50 (210 µg/mL), followed by Warburgia ugandensis (IC50 of 270 µg/mL). Piliostigma thonningii and P. africana were less toxic with IC50 of 720 µg/mL and 500 µg/mL, respectively. There was low production of IgG antibodies following treatment with the plant extracts and high levels in the untreated control

Estudo in vitro e in vivo da eficácia anti leishmaniótica de terapêutica combinada de Diminazene e Artesunate contra Leishmania donovani em camundongos Balb/c

The in vitro and in vivo activity of diminazene (Dim), artesunate (Art) and combination of Dim and Art (Dim-Art) against Leishmania donovani was compared to reference drug; amphotericin B. IC50 of Dim-Art was found to be 2.28 ± 0.24 µg/mL while those of Dim and Art were 9.16 ± 0.3 µg/mL and 4.64 ± 0.48 µg/mL respectively. The IC50 for Amphot B was 0.16 ± 0.32 µg/mL against stationary-phase promastigotes. In vivo evaluation in the L. donovani BALB/c mice model indicated that treatments with the combined drug therapy at doses of 12.5 mg/kg for 28 consecutive days significantly (p < 0.001) reduced parasite burden in the spleen as compared to the single drug treatments given at the same dosages. Although parasite burden was slightly lower (p < 0.05) in the Amphot B group than in the Dim-Art treatment group, the present study demonstrates the positive advantage and the potential use of the combined therapy of Dim-Art over the constituent drugs, Dim or Art when used alone. Further evaluation is recommended to determine the most efficacious combination ratio of the two compounds.A atividade in vitro e in vivo de Diminazene (Dim), Artezunate (Art) e a combinação Dim e Art (Dim-Art) contra Leishmania donovani foi comparada com a droga de referência Anfotericina B. IC50 da Dim-Art foi 2,28 ± 0,24 µg/mL enquanto aquelas de Dim e Art foram 9,16 ± 0,3 µg/mL e 4,64 ± 0,48 µg/mL respectivamente. O IC50 da Anfotericina B foi 0,16 ± 0,32 µg/mL contra a fase estacionária de promastigotas. A avaliação in vivo do modelo de L. donovani em camundongos Balb/c indicou que os tratamentos com a terapêutica de drogas combinadas em doses de 12,5 mg/kg por 28 dias consecutivos significantemente (p < 0,001) reduziu a carga parasitária no baço quando comparada a tratamentos com uma única droga dada nas mesmas dosagens. Embora a carga parasitária tenha sido levemente mais baixa (p < 0.05) no grupo Anfotericina B quando comparada com o grupo tratado Dim-Art, o estudo presente demonstra a vantagem positiva do uso potencial da terapêutica combinada Dim-Art sobre drogas como Dim ou Art quando usadas isoladamente. Posterior avaliação é recomendada para determinar a média de combinação mais eficaz dos dois compostos

In vitro and in vivo antileishmanial efficacy of a combination therapy of diminazene and artesunate against Leishmania donovani in BALB /c mice

The in vitro and in vivo activity of diminazene (Dim), artesunate (Art) and combination of Dim and Art (Dim-Art) against Leishmania donovani was compared to reference drug; amphotericin B. IC50 of Dim-Art was found to be $2.28 \pm 0.24 \mu$ g/mL while those of Dim and Art were $9.16 \pm 0.3 \mu$ g/mL and $4.64 \pm 0.48 \mu$ g/mL respectively. The IC50 for Amphot B was $0.16 \pm 0.32 \mu$ g/mL against stationary-phase promastigotes. In vivo evaluation in the L. donovani BALB/c mice model indicated that treatments with the combined drug therapy at doses of 12.5 mg/kg for 28 consecutive days significantly ($p < 0.001$) reduced parasite burden in the spleen as compared to the single drug treatments given at the same dosages. Although parasite burden was slightly lower ($p < 0.05$) in the Amphot B group than in the Dim-Art treatment group, the present study demonstrates the positive advantage and the potential use of the combined therapy of Dim-Art over the constituent drugs, Dim or Art when used alone. Further evaluation is recommended to determine the most efficacious combination ratio of the two compounds.Comment: 4 Pages, 3 Figure

Rotavirus group : a genotype circulation patterns across Kenya before and after nationwide vaccine introduction, 2010-2018

Background Kenya introduced the monovalent G1P [8] Rotarix® vaccine into the infant immunization schedule in July 2014. We examined trends in rotavirus group A (RVA) genotype distribution pre- (January 2010–June 2014) and post- (July 2014–December 2018) RVA vaccine introduction. Methods Stool samples were collected from children aged < 13 years from four surveillance sites across Kenya: Kilifi County Hospital, Tabitha Clinic Nairobi, Lwak Mission Hospital, and Siaya County Referral Hospital (children aged < 5 years only). Samples were screened for RVA using enzyme linked immunosorbent assay (ELISA) and VP7 and VP4 genes sequenced to infer genotypes. Results We genotyped 614 samples in pre-vaccine and 261 in post-vaccine introduction periods. During the pre-vaccine introduction period, the most frequent RVA genotypes were G1P [8] (45.8%), G8P [4] (15.8%), G9P [8] (13.2%), G2P [4] (7.0%) and G3P [6] (3.1%). In the post-vaccine introduction period, the most frequent genotypes were G1P [8] (52.1%), G2P [4] (20.7%) and G3P [8] (16.1%). Predominant genotypes varied by year and site in both pre and post-vaccine periods. Temporal genotype patterns showed an increase in prevalence of vaccine heterotypic genotypes, such as the commonly DS-1-like G2P [4] (7.0 to 20.7%, P < .001) and G3P [8] (1.3 to 16.1%, P < .001) genotypes in the post-vaccine introduction period. Additionally, we observed a decline in prevalence of genotypes G8P [4] (15.8 to 0.4%, P < .001) and G9P [8] (13.2 to 5.4%, P < .001) in the post-vaccine introduction period. Phylogenetic analysis of genotype G1P [8], revealed circulation of strains of lineages G1-I, G1-II and P [8]-1, P [8]-III and P [8]-IV. Considerable genetic diversity was observed between the pre and post-vaccine strains, evidenced by distinct clusters. Conclusion Genotype prevalence varied from before to after vaccine introduction. Such observations emphasize the need for long-term surveillance to monitor vaccine impact. These changes may represent natural secular variation or possible immuno-epidemiological changes arising from the introduction of the vaccine. Full genome sequencing could provide insights into post-vaccine evolutionary pressures and antigenic diversity

Data from: Evidence of high genetic diversity and significant population structuring in Vachellia tortilis (Forsk.) Galasso & Bonfi population in Kenya

Context: Vachellia tortilis is an important dryland tree species valued for fuelwood and fodder production, however, no strategy has been put in place for sustainable management of the species genetic resources. Furthermore, there is inadequate information on the species population genetics to facilitate development of such strategies. Aim: We evaluated the amount and structure of neutral genetic diversity of V. tortilis population in Kenya and provided recommendations necessary for improvement and conservation of the species genetic resources. We hypothesised that the current genetic diversity of V. tortilis is high because of it is demographic history and that no population structuring was expected to occur due to the presumed long distance and effective gene flow within the species. Methods: Leaf tissues were collected from fifteen putative natural populations of V. tortilis covering the whole distribution range in Kenya. DNA was isolated from the leaf tissues and analysed using microsatellite markers. In total, 450 trees were genotyped using 10 polymorphic nuclear microsatellite loci, and genetic diversity and population structure parameters determined Results: We found high levels of genetic diversity within the populations with a mean gene diversity at 0.85. However, significant population differentiation was evident (FST = 0.026, P = 0.007; RST = 0.032, P = 0.004) despite large number of migrants per generation (Nm = 5.3). Population structure detected suggests presence of two clusters, although, some populations showed mixed ancestry. The groups reflect the influence of geographic patterns and historical population gene flow. Conclusion: There exist high genetic diversity in V. tortilis in Kenya with significant population structuring into two clusters. We recommend consideration of the two distinct groups in the development of the species improvement, breeding and conservation programmes. Such programmes should ensure maintenance of the majority of the extant genetic diversity

Immunopathological Features Developing in the Mosquito Midgut after Feeding on Anopheles gambiae Mucin-1 / Interleukin-12 cDNA Immunized Mice

The mosquito midgut is the organ into which the blood meal passes and in which, within a peritrophic membrane secreted by the epithelium, the blood is retained during digestion and absorption. The mosquito midgut is lined with an actin filled microvilli that are exposed to the harsh environment of the gut lumen such as food particle abrasion, digestive hydrolases and attack by pathogens and parasites that are taken in by the blood meal. These microvilli are protected them these effects by the peritrophic matrix, the glycocalyx and the mucin proteins that line their epithelial surfaces. Immunization of BALB/c mice with AgMUC1/IL-12 cDNA has been shown to kill mosquitoes when fed on these mice. Mucin is one of the proteins produced in the mosquito midgut after a blood meal. The fine structure of the mosquito midgut epithelium interacting with immune factors such as antibodies or immune cells is of special significance for interpreting early events in the interaction between the mosquito midgut lining and the specific immune components present in the blood of AgMUC1/IL-12 cDNA immunized BALB/c mice. Following bright light microscopy, scanning electron and transmission electron microscopic observations of the features seen in mosquito midgut sections from An. gambiae mosquitoes fed on BALB/c mice immunized with AgMUC1/IL-12 cDNA, the most likely immune mechanisms responsible for mosquito killing could be cell mediated, most likely antibody dependent cellular cytotoxicity. Both necrotic and apoptotic processes that could be the cause of mosquito death were seen to take place in the cells lining the midgut epithelium