Browsing by Author "Anjili, Christopher O."
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Item Characterization and comparison of leishmania-like isolates from rodents, lizards and sand flies caught at Masinga location in Machakos district, Kenya(African Journal of Health Sciences, 2011-06) Kinuthia, Geoffrey K.; Lugalia, Reuben; Mwanyumba, Panuel J.; Ngeiywa, Moses M.; Anjili, Christopher O.; Tonui, Willy K.A laboratory based study was designed to characterize 43 cryo-preserved Leishmania-like flagellates. These Leishmania-like flagellates were originally obtained from non-human hosts that included spiny mice (Acomys subspinosus), plated lizards (Gerrosaurus major) and sand flies of the Genus Sergentomyia caught at Masinga location, Machakos District in Kenya. Morphological features and isoenzyme banding patterns of the flagellates were studied. The isoenzyme markers which were used for isoenzyme electrophoresis included Malate dehydrogenase (MDH), Phosphoglucomutase (PGM), Glucose phosphate isomerase (GPI), Glucose 6-phosphate dehydrogenase (G6PD), Malic enzyme (ME), 6 phosphogluconate dehydrogenase (6PGD) and Mannose phosphate isomerase (MPI). The isoenzyme banding patterns of the flagellates’ lysates were compared with those of six WHO Leishmania reference strains and those of seven well characterized reference strains of Trypanosoma, Crithidia, Herpetomonas and Leptomonas species. The results showed that the morphological changes of the Leishmania-like flagellates in the growth medium were indistinguishable from those of Leishmania WHO reference strains used. The isoenzyme profiles of the flagellates were all distinguishable from the reference strains used except for isolate NLB-1236 from G. major which had an enzyme profile identical to that of L. tropica (NLB-305) in 6 enzymes (MDH, GPI, MPI, ME, PGM, and G6PD). The banding pattern of isolate NLB-1261 from A. subspinosus was indistinguishable from that of L. major (NLB-326) in 3 enzymes only (MDH, GPI and ME) while isolate NLB-1231 from A. subspinosus had an enzyme profile identical to those of L. tropica (NLB-305) and L. arabica (NLB-664) in six enzymes (MDH, GPI, ME, PGM, MPI, and 6PGD). More than 80% of the Leishmania-like flagellates had enzyme profiles indistinguishable from each other, in all the isoenzyme markers. The morphological traits of the flagellates suggested that they were Leishmania or strains closely related to Leishmania. Isoenzyme analysis suggested that Sergentomyia sand flies most likely feed on both lizards (reptiles) and rodents (mammals). There is need to carry further investigations on NLB-1236 (from plated lizards), NLB-1261 (from wild spiny mice) and NLB-1231 (from wild spiny mice).Item Effects of Metarhizium anisopliae on sand fly populations in their natural habitats in Marigat sub- County, Baringo County, Kenya(Centre for Biotechnology Research & Development,, 2010) Ngure, Peter Kamau; Ngumbi, Philip; Kasili, Sichangi; Anjili, Christopher O.; Karanja, Robert M.; Kaburiae, Josyline C.; Mwangi, Milkah; Kinuthia, Geoffrey K.; Kiarie, Martha; Nzau, Anastasia; Kepha, Stella; Maniania, Nguya K.; Irungu, LucyLeishmaniasis is one of the neglected tropical diseases that pose a threat to people in the endemic regions. In Africa, transmission is through the bite of an infected female phlebotomine sand fly. Disease control with chemotherapy is logistically difficult, toxic and expensive. Entomopathogenic fungi (EPF) hold great potential as alternative vector control for sandflies as demonstrated by their efficacy in recent laboratory experiments. The impressive results have merited testing of the effect of EPF on sand flies in their natural habitats. The goal of this study was to test the effect of EPF on sand fly populations on a small scale in a leishmania-endemic area in Kenya. Entomopathogenic fungi isolates were cultured on Sabourand dextrose agar (SDA) in Petri dishes and incubated at 22-280C. Conidia were produced on long white rice substrate and harvested by sifting the substrate through a 295 μm mesh size sieve and stored at 4- 2 6 0C before being used in the experiments. Metarhizium anisopliae isolates were selected for use in the field after showing 76.8 and 100% mortality in laboratory bred target insects. 2.0 x 1014 of dry conidia of the fungus were introduced into ten termite mounds through the shafts using a modified foot pump in Rabai, Marigat area of Baringo County, Kenya. The site harbours both Phlebotomus martini and P. duboscqi sandfly vectors. Sand fly densities, mycoses and survival after capture from the mounds were observed from week one post-treatment to week thirteen. Results indicate that the fungi in termite mounds caused three to ten-fold mortalities in the population of sand flies nine weeks after application. Longevity of sand flies collected from treated termite mounds was reduced to less than nine days after collection, whereas 90% of those from control mounds lived well over the nine days. Sand flies collected from the treated termite mounds during week nine had 100% mortality compared to 10.7% of those from the control mounds. Metarhizium anisopliae infection was observed in 42% of the sand fly cadavers collected from the treated termite mounds and 0% in the control group. The high susceptibility of sand flies to M. anisopliae suggests that fungi can be developed as potential vector control alternatives to synthetic chemical insecticides or preferentially be applied in integrated control strategies in order to gain maximum impact on adult sand fly populations. Further studies to determine the best methods for delivery and application in the diverse ecological settings of various leishmaniasis vectors are recommended.Item Efficacy of crude methanolic extracts of Allium sativum L. and Moringa stenopetala (Baker f.) Cufod. against Leishmania major(Int. J. Med. Arom. Plants, 2014-03) Gikonyo, Nicholas K.; Kinuthia, Geoffrey K.; Kabiru, Ephantus Wanjohi ; Anjili, Christopher O.; Kigondu, Elizabeth M.; Ngure, Veronica N.; Ingonga, Johnstone M.Leishmania major is a protozoan parasite responsible for cutaneous leishmaniasis (CL) in humans. CL is transmitted via a bite by infected female phlebotomine sand fly. Research on herbal therapy for leishmaniases is increasing globally because conventional drugs are costly, toxic and require a prolonged administration. In vitro and in vivo antileishmanial activities of dried Allium sativum (garlic) and Moringa stenopetala methanolic extracts against L. major were studied. Minimum inhibitory concentrations (MICs) of methanolic extracts of A. sativum (A) and M. stenopetala(M) against L. major were 3 and 5 mg/ml and IC50 of 863.12 and 1752.92 μg/ml respectively. The blend AM (1:1) hadIC50 of 372.1μg/ml and promastigotes’ viability of 71.03% compared to IC50 of 0.26 and 0.82μg/ml and promastigotes’viability of 18.41% and 12.22% for Pentostam and Liposomal amphotericin B respectively. Multiplication indices (MIs) of L. major amastigotes ranged from 43.67% to 45.93% after treatment with extracts A or M or blend AM at 125μg/ml and were significantly different (P < 0.05) from Liposomal amphotericin B at 12.5μg/ml. Oral extract A reduced significantly (P > 0.05) L. major caused foot pad lesions in BALB/c mice while oral extract M did not. Blend AM (ip) reduced the lesion sizes and its efficacy was close to Pentostam and Liposomal amphotericin B. Oral extract A had a high parasite reduction rate of 60.70% and average LDU of 0.22±0.15 compared to Pentostam at 66.40% and LDU of 0.18±0.08. In conclusion, methanolic extract of A. sativum showed anti-leishmanial activity both in vitro and in vivo and it decreased L. major caused foot pad lesions in BALB/c mice. Methanolic extracts of M. stenopetala (ip) reduced the amastigotes burden in spleens of BALB/c mice. A blend of garlic and moringa methanolic extracts (AM at 1:1) were active against L. major. The active ingredients in crude methanolic extracts of garlic and moringa plants should be established and tested against L. major when blended.Item In vitro activity of aqueous and methanol extracts of Callistemon citrinus (Family Myrtaceae) against Leishmania major(African Journal of Health Sciences, 2014-04) Anjili, Christopher O.; Kinuthia, Geoffrey K.; Kabiru, Ephantus Wanjohi; Gikonyo, Nicholas K.; Ingonga, Johnny M.; Kigondu, Elizabeth M.Leishmania major is a protozoan parasite that causes cutaneous leishmaniasis and the standard drugs are expensive and toxic. Cheaper and safer natural drugs are therefore needed. In this study, the in vitro efficacy of crude extracts of Callistemon citrinus were tested against L. major. Controls were anti leishmanial drugs pentostam and liposomal amphotericin B. The minimum inhibitory concentrations of C. citrinus crude aqueous and methanolic extracts were 5mg/ml and 1mg/ml respectively compared to 12.5μg/ml and 6.25μg/ml for pentostam and liposomal amphotericin B respectively. The IC50 for C. citrinus extracts against promastigotes ranged from 297.75 to 572.69μg/ml compared to 0.26 and 0.82μg/ml for pentostam and liposomal amphotericin B. The IC50 for C. citrinus extracts against vero cells ranged from 467μg/ml to 1314.65μg/ml. The promastigotes’ viability after treatment with aqueous and methanolic extracts was 69.58% and 75.74% respectively. At 125μg/ml, the aqueous and methanolic C. citrinus extracts had in vitro amastigotes’ infection rates (IRs) of 77.0±2.50 % and 77.5±3.50% respectively. The multiplication indices (MIs) and IRs of amastigotes treated with C. citrinus crude aqueous extracts and those treated with crude methanolic extracts differed insignificantly (P > 0.05). C. citrinus methanolic extracts stimulated production of about 20μM nitric oxide in BALB/c mice peritoneal macrophages suggesting immuno-modulatory role of the extracts. The crude aqueous and methanolic extracts of C. citrinus were therefore concluded to be relatively less toxic and possessed in vitro anti-leishmanial activity against L. major promastigotes and amastigotes.Item In vitro and in vivo activities of blends of crude aqueous extracts from Allium sativum L, Callistemon citrinus (Curtis) Skeels and Moringa stenopetala (Baker F) Cufodontis against Leishmania major(Int. J. Med. Arom. Plants, 2013-06) Kabiru, Ephantus Wanjohi; Kinuthia, Geoffrey K.; Anjili, Christopher O.; Gikonyo, Nicholas K.; Kigondu, Elizabeth M.; Ingonga, Johnstone M.Leishmania major caused cutaneous leishmaniasis leads to painful skin sores in humans and usual drugs are expensive, toxic, and require prolonged use. The in vitro and in vivo efficacy of aqueous crude extracts from Callistemon citrinus flowers (B), Allium sativum bulbs (C) and Moringa stenopetala leaves (A) against L. major was studied. Controls were pentostam, liposomal amphotericin B, and phosphate buffered saline (PBS). Dried and ground plant materials were soaked in distilled water at 70oC for 1.5 hours, filtered and freeze dried to obtain aqueous extracts. L. major infected BALB/c mice were treated orally or intra peritoneally (ip) with blends of the extracts. Minimum inhibitory concentrations (MICs) of single extracts ranged from 3 to 5mg/ml while IC50 from 297 to 575μg/ml compared to MICs of 12.50 and 6.25μg/ml and IC50 of 0.26 and 0.82μg/ml for pentostam and liposomal amphotericin B respectively. Blends of M. stenopetala and C. citrinus (AB), M. stenopetala and A. sativum (AC), and C. citrinus and A. sativum (BC) at concentrations based on MICs of individual extracts were active at ratios 1:1, 1:9 and 1:1 with promastigotes’ viabilities of 33.82%, 17.41% and 60.74 % respectively. IC50 for blends AB, AC, and BC ranged from 174μg/ml to 1314μg/ml against promastigotes. The individual extracts comprising blends AB, AC and BC interacted additively and synergistically in several combination ratios. Blend AC (1:1) at 125μg/ml had in vitro infection rate (IR) of 71% and multiplication index (MI) of 48.20% for L. major amastigotes compared to IR of 67% and MI of 47.51% for pentostam at 12.50μg/ml. Oral blend BC (1:1) reduced the mice footpad lesion size significantly (P < 0.05). Both oral blends BC and AC reduced mice spleen amastigotes by 48.33% and 60.94% with total LDUs of 6.35 ± 0.66 and 4.80 ± 0.95 respectively. Oral blend AB (1:1) lowered spleen amastigotes by 6.5% with total LDU of 11.49 ± 6.84. In conclusion, aqueous blends of C. citrinus, A. sativum and M. stenopetala extracts that interacted additively or synergistically were less toxic but active against L. major.Item Selected practices among rural residents versus the prevalence of Amoebiasis and Giardiasis in Njoro Distrct, Kenya(African Journal of Health Sciences, 2012) Anjili, Christopher O.; Geoffrey, Kinuthia K.; Afolayan, Funmilayo I. D.; Ngure, VeronicaThe study was designed to investigate on selected practices among rural population, and their likely contribution to the spread of amoebiasis (E. histolytica) and giardiasis (G. lamblia). A cross sectional study was carried out in three villages, namely Kikapu, Piave and Belbar in Njoro District, Kenya. Questionnaire, interviews and personal observations were used to obtain data from 336 randomly selected consenting individuals in homesteads in the three villages. A retrospective study was further carried out to establish the prevalence of E. histolytica and G. lamblia among outpatients attending two randomly selected health centers in the study area. Majority of the respondents’ highest education level was basic primary school education (61%), unemployment (98%) and with a monthly expenditure of less than 2200 Kenya Shillings or 28 US Dollars (81%). Eighty percent of the respondents were classified under low economic status. Eighty two percent reported lack of piped water and boiling of drinking water was less likely to occur among the low economic status respondents (Odds ratio (OR) = 0.423, 2 = 9.88; 95% CI of -5.74 to 6.58). However, washing of hands with soap after using a latrine seemingly was not influenced by economic status of the respondents (OR = 1; 2 = 0; 95% CI = 0). The level of education seemed to influence on the adoption of risky practices, such as, failure to boil drinking water was more likely to occur among respondents who had a low academic level (OR = 0.84, 2 = 0.04, 95% CI of -2.27 to 3.95). The stool tests records at Njoro PCEA health center showed that the prevalence of E. histolytica (20.83%) and G. lamblia (20.32%) were higher than the corresponding prevalence at Njoro County Council health center which were 1.34% and 0.00% respectively. Concurrent infections of E. histolytica and G. lamblia were absent in the two health centers. The trend of E. histolytica and G. lamblia followed an alternating pattern, in which an increase in one directly corresponded to a decrease in the other and vice versa in the two health centers. It was concluded that poverty and low education levels were significant factors that influenced on the adoption of risky lifestyles that were likely to enhance parasitic infections. There is a need for reliable diagnostic methods other than direct microscopy for E. histolytica and G. lamblia stool tests in order to minimize the wide variation of the results in the two health centers. Public health education should also be enhanced to discourage the adoption of risky practices.Item Selected practices among rural residents versus the prevalence of Amoebiasis and Giardiasis in Njoro Distrct, Kenya.(African Journal of Health Sciences, 2012) Ngure V.; Kinuthia, Geoffrey K.; Afolayan, Funmilayo I. D. ; Anjili, Christopher O.The study was designed to investigate on selected practices among rural population, and their likely contribution to the spread of amoebiasis (E. histolytica) and giardiasis (G. lamblia). A cross sectional study was carried out in three villages, namely Kikapu, Piave and Belbar in Njoro District, Kenya. Questionnaire, interviews and personal observations were used to obtain data from 336 randomly selected consenting individuals in homesteads in the three villages. A retrospective study was further carried out to establish the prevalence of E. histolytica and G. lamblia among outpatients attending two randomly selected health centers in the study area. Majority of the respondents’ highest education level was basic primary school education (61%), unemployment (98%) and with a monthly expenditure of less than 2200 Kenya Shillings or 28 US Dollars (81%). Eighty percent of the respondents were classified under low economic status. Eighty two percent reported lack of piped water and boiling of drinking water was less likely to occur among the low economic status respondents (Odds ratio (OR) = 0.423, 2 = 9.88; 95% CI of -5.74 to 6.58). However, washing of hands with soap after using a latrine seemingly was not influenced by economic status of the respondents (OR = 1; 2 = 0; 95% CI = 0). The level of education seemed to influence on the adoption of risky practices, such as, failure to boil drinking water was more likely to occur among respondents who had a low academic level (OR = 0.84, 2 = 0.04, 95% CI of -2.27 to 3.95). The stool tests records at Njoro PCEA health center showed that the prevalence of E. histolytica (20.83%) and G. lamblia (20.32%) were higher than the corresponding prevalence at Njoro County Council health center which were 1.34% and 0.00% respectively. Concurrent infections of E. histolytica and G. lamblia were absent in the two health centers. The trend of E. histolytica and G. lamblia followed an alternating pattern, in which an increase in one directly corresponded to a decrease in the other and vice versa in the two health centers. It was concluded that poverty and low education levels were significant factors that influenced on the adoption of risky lifestyles that were likely to enhance parasitic infections. There is a need for reliable diagnostic methods other than direct microscopy for E. histolytica and G. lamblia stool tests in order to minimize the wide variation of the results in the two health centers. Public health education should also be enhanced to discourage the adoption of risky practicesItem Toxicity and efficacy of aqueous crude extracts from Allium sativum, Callistemon citrinus and Moringa stenopetala against L. Major(Kabarak Journal of Research & Innovation, 2015) Gikonyo, Nicholas K.; Kinuthia, Geoffrey K.; Anjili, Christopher O.; Kabiru, Ephantus Wanjohi ; Kigondu, Elizabeth M.; Ingonga, Johnny M.Cutaneous leishmaniasis (CL) treatment involves pentavalent antimonials, amphotericin B, pentamidine, miltefosine among others. These drugs are toxic, costly, and require prolonged use. CL is a protozoan skin infection which may lead to disfiguring and stigmatization. In Kenya, CL is common in Baringo County where it is caused by Leishmania major and transmitted by infected female phlebotomine sand fly. Leishmaniases are common in poverty stricken areas where victims opt for local herbal therapies. Herbs used haven’t been tested scientifically to verify their toxicity and efficacy. The current study determines in vitro toxicity and in vivo efficacy of aqueous crude extracts of Moringa stenopetala, Callistemon citrinus, and Allium sativum against L. major. The IC50 of aqueous extracts against promastigotes ranged from 297μg/ml to 575μg/ml compared to Pentostam and liposomal amphotericin B with IC50 of 0.26μg/ml and 0.82μg/ml respectively. The viability of promastigotes upon exposure to extracts ranged from 52.55% to 60.57%. Similarly the IC50 of extracts against vero cells ranged between 467μg/ml to 2105μg/ml compared to 108μg/ml and 60μg/ml for pentostam and liposomal amphotericin B respectively. Orally administered A. sativum reduced L. major caused footpad lesions significantly (P < 0.05) when compared to control PBS. The efficacy of oral C. citrinus extracts (B) in reducing amastigotes in spleens of infected BALB/c mice was 82.99%, followed by oral M. stenopetala (A) at 66.96% and oral A. sativum (C) at 60.37% compared to pentostam and liposomal amphotericin B at 66.40% and 60.62% respectively. The difference between the mean total LDUs for aqueous oral C. citrinus extracts and control oral PBS was significant (P = 0.017). It was concluded that crude aqueous extracts of A. sativum, M. stenopetala, and C. citrinus show antileishmanial activity at low toxicity. Inclusion of garlic and moringa in the diets of people in leishmaniases foci should be emphasized.