Biocontrol-based strategies for improving soil health and managing plant-parasitic nematodes in coffee production.

Coffee is an important commodity for Kenya, where production is steadily declining, despite a global rise in demand. Of the various constraints affecting production, plant-parasitic nematodes are a significant, but often overlooked, threat. As a perennial crop, treating plantations once infected with nematodes becomes difficult. The current study evaluated the drenching application of two biocontrol agents, Trichoderma asperellum and Purpureocillium lilacinum, for their nematode control efficacy, as well as their impact on the soil nematode community structure on mature, established coffee trees in Kenya. Seven Arabica coffee field trials were conducted over two years on trees of various ages. All the fields were heavily infested with Meloidogyne hapla, the first report of the species on coffee in Kenya. Both fungal biocontrol agents were detected endophytically infecting roots and recovered from soil but not until six months after initial applications. The population densities of M. hapla had significantly declined in roots of treated trees 12 months after the initial application, although soil nematode density data were similar across treatments. Based upon the maturity index and the Shannon index, treatment with T. asperellum led to improved soil health conditions and enrichment of diversity in the microbial community. Application of P. lilacinum, in particular, led to an increased abundance of fungivorous nematodes, especially Aphelenchus spp., for which P. lilacinum would appear to be a preferred food source. The soils in the trials were all stressed and denuded, however, which likely delayed the impact of such treatments or detection of any differences between treatments using indices, such as the functional metabolic footprint, over the period of study. A longer period of study would therefore likely provide a better indication of treatment benefits. The current study positively demonstrates, however, the potential for using biologically based options for the environmentally and climate-smart management of nematode threats in a sustainable manner on established, mature coffee plantations.

Safety and Effectiveness of Sodium Stibogluconate and Paromomycin Combination for the Treatment of Visceral Leishmaniasis in Eastern Africa: Results from a Pharmacovigilance Programme.

INTRODUCTION: In 2010, WHO recommended a new first-line treatment for visceral leishmaniasis (VL) in Eastern Africa. The new treatment, a combination of intravenous (IV) or intramuscular (IM) sodium stibogluconate (SSG) and IM paromomycin (PM) was an improvement over SSG monotherapy, the previous first-line VL treatment in the region. To monitor the new treatment's safety and effectiveness in routine clinical practice a pharmacovigilance (PV) programme was developed. METHODS: A prospective PV cohort was developed. Regulatory approval was obtained in Sudan, Kenya, Uganda and Ethiopia. Twelve sentinel sites sponsored by the Ministries of Health, Médecins Sans Frontières (MSF) and Drugs for Neglected Diseases initiative (DNDi) participated. VL patients treated using the new treatment were consented and included in a common registry that collected demographics, baseline clinical characteristics, adverse events, serious adverse events and treatment outcomes. Six-monthly periodic safety update reports (PSUR) were prepared and reviewed by a PV steering committee. RESULTS: Overall 3126 patients were enrolled: 1962 (62.7%) from Sudan, 652 (20.9%) from Kenya, 322 (10.3%) from Ethiopia and 190 (6.1%) from Uganda. Patients were mostly male children (68.1%, median age 11 years) with primary VL (97.8%). SSG-PM initial cure rate was 95.1%; no geographical differences were noted. HIV/VL co-infected patients and patients older than 50 years had initial cure rates of 56 and 81.4%, respectively, while 1063 (34%) patients had at least one adverse event (AE) during treatment and 1.92% (n = 60) had a serious adverse event (SAE) with a mortality of 1.0% (n = 32). There were no serious unexpected adverse drug reactions. CONCLUSIONS: This first regional PV programme in VL supports SSG-PM combination as first-line treatment for primary VL in Eastern Africa. SSG-PM was effective and safe except in HIV/VL co-infected or older patients. Active PV surveillance of targeted safety, effectiveness and key VL outcomes such us VL relapse, PKDL and HIV/VL co-infection should continue and PV data integrated to national and WHO PV databases.

Volatile constituents of redblush grapefruit (Citrus paradisi) and pummelo (Citrus grandis) peel essential oils from Kenya.

The volatile constituents of cold-pressed peel essential oils of redblush grapefruit (Citrus paradisi Macfadyen forma Redblush) and pummelo (Citrus grandis Osbeck) from the same locality in Kenya were determined by GC and GC-MS. A total of 67 and 52 compounds, amounting to 97.9 and 98.8% of the two oils, respectively, were identified. Monoterpene hydrocarbons constituted 93.3 and 97.5% in the oils, respectively, with limonene (91.1 and 94.8%), alpha-terpinene (1.3 and 1.8%), and alpha-pinene (0.5%) as the main compounds. Sesquiterpene hydrocarbons constituted 0.4% in each oil. The notable compounds were beta-caryophyllene, alpha-cubebene, and (E,E)-alpha-farnesene. Oxygenated compounds constituted 4.2 and 2.0% of the redblush grapefruit and pummelo oils, respectively, out of which carbonyl compounds (2.0 and 1.3%), alcohols (1.4 and 0.3%), and esters (0.7 and 0.4%) were the major groups. Heptyl acetate, octanal, decanal, citronellal, and (Z)-carvone were the main constituents (0.1-0.5%). Perillene, (E)-carveol, and perillyl acetate occurred in the redblush grapefruit but were absent from the pummelo oil. Nootkatone, alpha- and beta-sinensal, methyl-N-methylanthranilate, and (Z,E)-farnesol were prominent in both oils.

Changes of the volatile profile and artifact formation in Daidai (Citrus aurantium) cold-pressed peel oil on storage.

The chemical changes and artifact formation in daidai (Citrus aurantium L. var. Cyathifera Y. Tanaka) cold-pressed peel oil upon storage at 20, 5, and -21 degrees C for 3, 6, and 12 months were investigated using capillary gas chromatography (GC) and GC-MS. Major changes occurred in the oil stored at 20 and 5 degrees C. No changes were found at -21 degrees C. Monoterpene hydrocarbons decreased from 98.0 to 66.4% upon 12 months at 20 degrees C, while sesquiterpene hydrocarbons and alcohols increased from 0.1 to 2.4% and from 0.3 to 7.9%, respectively. Notable decreases of germacrene D, myrcene, linalyl acetate, and limonene occurred. Prominent increases of cis-carveol, trans-beta-farnesene, trans-p-2,8-menthadien-1-ol, linalool, and beta-caryophyllene were found. Thirty-four artifact compounds constituting 17.0% of the total volatile compounds were formed upon 12 months at 20 degrees C. The artifacts consisted of 13 alcohols (6.0%), five carbonyl compounds (5.3%), seven esters (4.9%), three epoxides (0.4%), four hydrocarbons (0.3%), and two unidentified. The prominent artifact compounds were (+)-carvone, trans,trans-farnesyl acetate, sabinene hydrate, 1-octen-3-ol, cis,cis-farnesyl acetate, and dihydrocarveol acetate. The results could be applied for monitoring and control of the flavor quality of daidai essential oil and related products.

Effect of traditional fermentation and malting on phytic acid and mineral availability from sorghum (Sorghum bicolor) and finger millet (Eleusine coracana) grain varieties grown in Kenya.

Sorghum and finger millet grains are traditional staple foods in Kenya. However, they have naturally occurring anti-nutritional factors, such as phytic acid, that decrease their dietary availability. This work determined the effect of fermentation and malting on the phytic acid content of, and mineral availability in five varieties of sorghum and four varieties of finger millet grain grown in Kenya. Phytic acid ranged from 875.1 to 2,211.9 mg/100 g in sorghum. The levels in finger millet ranged from 851.6 to 1,419.4 mg/100 g grain. Fermentation resulted in a mean decrease of phytic acid in of 64.8% after 96 hours and 39.0% after 72 hours in sorghum grain. In finger millet, there was a mean decrease of 72.3% and 54.3% after 96 and 72 hours, respectively. Malting also resulted in a mean decrease of 23.9 and 45.3% after 72 and 96 hours, respectively. The extent of decrease of phytic acid differed among the grain varieties. Fermentation increased the rate of available iron, manganese, and calcium in both sorghum and finger millet. The available minerals were generally higher in finger millet than in sorghum after fermentation. Fermentation was also more effective than malting in reducing phytic acid in sorghum and finger millet. Simple traditional food processing methods can therefore be used to increase mineral availability.

Sodium stibogluconate (SSG) & paromomycin combination compared to SSG for visceral leishmaniasis in East Africa: a randomised controlled trial.

BACKGROUND: Alternative treatments for visceral leishmaniasis (VL) are required in East Africa. Paromomycin sulphate (PM) has been shown to be efficacious for VL treatment in India. METHODS: A multi-centre randomized-controlled trial (RCT) to compare efficacy and safety of PM (20 mg/kg/day for 21 days) and PM plus sodium stibogluconate (SSG) combination (PM, 15 mg/kg/day and SSG, 20 mg/kg/day for 17 days) with SSG (20 mg/kg/day for 30 days) for treatment of VL in East Africa. Patients aged 4-60 years with parasitologically confirmed VL were enrolled, excluding patients with contraindications. Primary and secondary efficacy outcomes were parasite clearance at 6-months follow-up and end of treatment, respectively. Safety was assessed mainly using adverse event (AE) data. FINDINGS: The PM versus SSG comparison enrolled 205 patients per arm with primary efficacy data available for 198 and 200 patients respectively. The SSG & PM versus SSG comparison enrolled 381 and 386 patients per arm respectively, with primary efficacy data available for 359 patients per arm. In Intention-to-Treat complete-case analyses, the efficacy of PM was significantly lower than SSG (84.3% versus 94.1%, difference = 9.7%, 95% confidence interval, CI: 3.6 to 15.7%, p = 0.002). The efficacy of SSG & PM was comparable to SSG (91.4% versus 93.9%, difference = 2.5%, 95% CI: -1.3 to 6.3%, p = 0.198). End of treatment efficacy results were very similar. There were no apparent differences in the safety profile of the three treatment regimens. CONCLUSION: The 17 day SSG & PM combination treatment had a good safety profile and was similar in efficacy to the standard 30 day SSG treatment, suggesting suitability for VL treatment in East Africa. CLINICAL TRIALS REGISTRATION: www.clinicaltrials.govNCT00255567.

Geographical variation in the response of visceral leishmaniasis to paromomycin in East Africa: a multicentre, open-label, randomized trial.

BACKGROUND: Visceral leishmaniasis (VL) is a major health problem in developing countries. The untreated disease is fatal, available treatment is expensive and often toxic, and drug resistance is increasing. Improved treatment options are needed. Paromomycin was shown to be an efficacious first-line treatment with low toxicity in India. METHODS: This was a 3-arm multicentre, open-label, randomized, controlled clinical trial to compare three treatment regimens for VL in East Africa: paromomycin sulphate (PM) at 15 mg/kg/day for 21 days versus sodium stibogluconate (SSG) at 20 mg/kg/day for 30 days; and the combination of both dose regimens for 17 days. The primary efficacy endpoint was cure based on parasite-free tissue aspirates taken 6 months after treatment. FINDINGS: Overall, 135 patients per arm were enrolled at five centres in Sudan (2 sites), Kenya (1) and Ethiopia (2), when the PM arm had to be discontinued due to poor efficacy. The trial has continued with the higher dose of PM as well as the combination of PM and SSG arms. These results will be reported later. Baseline patient characteristics were similar among treatment arms. The overall cure with PM was significantly inferior to that with SSG (63.8% versus 92.2%; difference 28.5%, 95%CI 18.8% to 38.8%, p<0.001). The efficacy of PM varied among centres and was significantly lower in Sudan (14.3% and 46.7%) than in Kenya (80.0%) and Ethiopia (75.0% and 96.6%). No major safety issues with PM were identified. CONCLUSION: The efficacy of PM at 15 mg/kg/day for 21 days was inadequate, particularly in Sudan. The efficacy of higher doses and the combination treatment warrant further studies.