Background Despite global tuberculosis (TB) interventions, the disease remains one of the major public health concerns. Kenya is ranked 15th among 22 high burden TB countries globally. Methods A cross-sectional study was conducted in Western Kenya, which comprises 10 counties. A multistage sampling method was used where a single sub-county was randomly selected followed by sampling two high volume health facility from each sub-county. Identification of spoligotype profiles and their family distribution and lineage level were achieved by comparison with SITVIT database. Results Lineage distribution pattern revealed that the most predominant lineage was CAS 220 (39.8%) followed by Beijing 128 (23.1%). The other lineages identified were T, LAM, H, X, S and MANU which were quantified as 87 (15.7%), 67 (12.1%), 16 (2.8%), 10 (1.8%), 8 (1.4%) and 5 (0.9%) respectively. CAS and Beijing strains were the most predominant lineage in both HIV negative and positive TB patients. The Beijing lineage was also the most predominant in resistant M. tuberculosis strains as compared to wild type. A total of 12 (2.0%) were orphaned M. tuberculosis strains which were spread across all the 10 counties of the study site. In multivariate logistic regression adjusting for potential cofounders three potential risk factors were significant. HIV status (OR = 1.52, CI = 0.29–3.68 and P value of 0.001), Alcohol use (OR = 0.59, CI = 0.43–3.12 and P -value =0.001) and cross border travel (OR = 0.61, CI = 0.49–3.87 and P value = 0.026). Most M. tuberculosis clinical isolates showed genetic clustering with multivariate logistic regression indicating three potential risk factors to clustering. HIV status (OR = 1.52, CI = 0.29–3.68 and P value of 0.001), Alcohol use (OR = 0.59, CI = 0.43–3.12 and P-value =0.001) and cross border travel (OR = 0.61, CI = 0.49–3.87 and P value = 0.026). Conclusion There exist diverse strains of M. tuberculosis across the 10 counties of Western Kenya. Predominant distribution of clustered genotype points to the fact that most TB cases in this region are as a result of resent transmission other than activation of latent TB.

Abstract Inflammation is the immune system’s response to infection and injury. Although it is a natural and beneficial process, it has been implicated in the pathogeneses of arthritis, cancer, neurodegenerative, and cardiovascular disease. In this study, we evaluated the in vitro anti-inflammatory activity of the alcohol and aqueous extracts of marine gastropods Pachymelania aurita and Tympanotonos fuscatus (syn. Tympanotonus fuscatus) var. radula in peritoneal macrophage cells treated with lipopolysaccharide. Peripheral macrophages were harvested from mice following the injection of foetal calf serum into the peritoneal space. The cytotoxicity of the extracts on the macrophages and the maximum non-toxic dose was determined using the MTT assay. Cells were challenged with lipopolysaccharide and treated with crude extracts of T. fuscatus and P. aurita 24 h later. The antioxidant activity was also evaluated, using the DPPH assay. Both extracts of P. aurita and T. fuscatus exhibited significant anti-inflammatory activities in terms of their ability to inhibit the production of NO, TNF-α, IL-1 and PGE2, which are key mediators of the inflammation response. Also, the alcohol extracts proved to be as effective as vitamin C in the free radical scavenging of DPPH. Thus, the extracts from P. aurita and T. fuscatus are useful candidates for further drug studies, especially as drug candidates for the treatment of inflammatory conditions.

The majority of the viruses that currently cause diseases in humans and animals are RNA viruses, and more specifically arthropod-transmitted viruses. They cause diseases such as dengue, West Nile infection, bluetongue disease, Schmallenberg disease, and yellow fever, among others.

Abstract Introduction Globally anti-tuberculosis drug resistance is one of the major challenges affecting control and prevention of tuberculosis. Kenya is ranked among 30 high burden TB countries globally. However, there is scanty information on second line antituberculosis drug resistance among tuberculosis patients. Therefore, this study aimed at determining Mycobacterium tuberculosis drug resistant strain distribution pattern in 10 counties of Western Kenya among HIV positive and negative patients. Method A cross-sectional study was conducted in Western Kenya, which comprises 10 counties. A multistage sampling method was used where a single sub-county was randomly selected followed by sampling one high volume health facility from each sub-county. Consenting study subjects with at least two smear positive sputum at the time of enrolment were randomly selected. The collected sputum was decontaminated with N-acetyl-l-cysteine-sodium hydroxide (NALC-NaOH) and then stained with Ziehl Neelsen Stain before visualizing the presence of bacilli under microscope at ×100 magnification with oil immersion. Further, the identified bacilli were cultured and susceptibility test carried out using known first and second line antimycobacterial tuberculosis. HIV testing was carried out using Determine® HIV-1/2 rapid test (Abbot Diagnostics, Maidenhead, United Kingdom). Those who had smear converted were dropped from the study. Finally, drug susceptibility pattern across the 10 counties of Western Kenya was evaluated. Results Our study showed that Mycobacterium tuberculosis drug resistance among HIV negative and positive cases in Western Kenya was prevalent in all the 10 counties surveyed. Based on the drug susceptibility tests, 53.2% and 42.7% of the study samples were resistant to at least one antituberculosis drug among HIV negative and HIV positive patients respectively. The data analysis revealed that among the HIV-positive and HIV-negative patients, resistance to INH was predominant (28.5%, and 23.6%, respectively), followed by RIF (16.4% and 14.6% respectively). Second-line drug resistant strains identified among HIV negative patients included Ethionamide (0.3%), Gatifloxacin (0.3%), Amikacin (0.3%) and Capreomycin (0.3%). There was no second line drug monoresistance among HIV positive TB patients. Multi/poly drug resistance were noted among HIV-negative patients in, INH + AMK (0.7%), INH + PZA (1%), INH + GFX (0.7%, INH + ETO (0.7%, STY + ETO (1%), ETH + ETO (1.0%), INH + KAN (0.7%) and INH + CAP (0.7%) strains/cases at 95% confidence interval. Among HIV positive patients INH + GFX (1.1%), INH + ETO (0.4%) and INH + KAN (0.4%) strains of M. tuberculosis were identified with a confidence interval of 95%. Geographical distribution patterns analysis of M. tuberculosis drug polyresistant strains across the 10 counties were recorded. Among HIV TB patients, resistant strains were identified in Nyamira (INH + GFX, INH + KAN), Bungoma ((ETO + STY), Busia (ETH + ETO and STY + ETO) Homabay (RIF + AMK. ETO + ETH and ETO + STY), Kisumu (ETH + ETO and PZA + ETO) and in Kakamega, Kisii and Vihiga (INH + KAN and RIF + AMK). There was no M. tuberculosis polyresistant strain identified in Migori and Siaya counties. Among HIV positive TB patients, M. tuberculosis resistant strains were identified in three counties, Nyamira (INH + KAN) Homabay (INH + GFX and INH + AMK) and Kakamega (INH + GFX). There was no polyresistant M. tuberculosis strain identified in Migori, Bungoma, Kisii, Vihiga, Busia, Siaya and Kisumu Counties. Discussion The distribution patterns of M. tuberculosis drug resistance among HIV negative and positive TB patients could be as a result of reported high prevalence of HIV in Western Kenya counties especially the area under study. Tuberculosis is one of the opportunistic diseases that have been shown to be the major cause of AIDS among HIV infected patients. Resent reports by National AIDS Control Council shows that Kisumu, Siaya, Homabay, Migori, Busia have the overall leading in HIV prevalence in Kenya. The low prevalence of drug resistant strains among HIV tuberculosis patients could be as a result of drug adherence attitude adopted by HIV patients, availability of continuous counselling and close follow up and notification by healthcare workers and community health volunteers. Conclusion Drug resistant M. tuberculosis strains prevalence is still high among HIV negative and positive patients in Western Kenya with the most affected being HIV negative TB patients. It is therefore probable that the existing control measures are not adequate to control transmission of drug resistant strains. Further, miss diagnosis or delayed diagnosis of TB patients could be contributing to the emergence of M. tuberculosis drug polyresistant strains. Recommendation Based on the result of this study, regular TB drug resistance surveillance should be conducted to ensure targeted interventions aimed at controlling increased transmission of the tuberculosis drug resistant strains among HIV/AIDS and HIV negative patients. There is also need for improved drug resistant infection control measures, timely and rapid diagnosis and enhanced and active screening strategies of tuberculosis among suspected TB patients need to be put in place. Further, studies using a larger patient cohort and from counties across the country would shed much needed insights on the true national prevalence of different variants of M. tuberculosis drug resistance.

PURPOSE AND METHODS:In this study, the analgesic activity of the crude alcohol (acetone-methanol) and aqueous (in PBS, pH 7.2) extracts of the marine molluscs, Pachymelania aurita and Tympanotonus fuscatus, has been evaluated using the formalin test (for chronic antinociceptive) and the tail-flick (acute antinociceptive) pain models in male swiss albino mice. RESULTS:The results show that the extracts of P. aurita and T. fuscatus demonstrated high safety margins as single doses of up to 2000 mg/kg bwt proved to be well tolerated and non-lethal, although the alcohol extract of P. aurita caused necrosis in the liver and kidney when administered at a dose level of 2000 mg/kg bwt. In the formalin test, treatment with the aqueous extracts of P. aurita and T. fuscatus as well as the alcohol extract of T. fuscatus 30 min before the subcutaneous injection of 5% formalin to the paw of the mice resulted in a significant time- and dose-dependent reduction in total and phase 2a pain-related behavior and thus nociception. The extracts had no analgesic effect in tail-flick test up to the highest dose tested. CONCLUSION:Hence, the results from both models indicate that the site of their analgesic action is probably peripheral.

This study aimed to investigate the antimitotic and antiproliferation activities of crude acetone-methanol and aqueous extracts of two marine molluscs commonly found in the Niger Delta region of Nigeria; T.fuscatus and P.aurita, against human cancerous cell lines (DU145, Hep-2, and HCC1395) cell lines in vitro. The antimitotic activity of the extracts was evaluated using Allium cepa root meristematic cells. Antiproliferative activity of the plant extracts against the cancerous cell lines was compared with normal cell line (VeroE6). Doxorubicin was used as a positive control. Gene expression studies using qPCR for the proapoptotic genes, CASP3, CASP8 and P53 were also carried out. The alcohol extract of T.fuscatus (TFAC) exhibited the most promising activity against all the cancer cell lines tested (DU145 IC50 = 96.48 ± 1.36 μg/ml, HCC 1395 IC50 = 61.44 ± 2.45 μg/ml, Hep2 IC50 = 0.52 ± 0.36 μg/ml) and also had the highest selectivity index of 4.94, 7.78 and 921.97 for DU145, HCC 1395 and Hep-2 cells respectively. Furthermore, TFAC was the only extract that significantly upregulated the expression of caspase 3, caspase 8 and P53. Thus, these findings suggest potential exploitation of TFAC as an anticancer agent.

Scientific and technological progress envisages the solving the most important problems in the main areas of mineral processing, leading to the perfection of technological processes, improving quality indicators and reducing the cost of production. Rational use of minerals at the stages of their extraction and enrichment is a single indivisible problem, the main task of which is the fullest use of basic and rare minerals. Geo-resources include mineral deposits and subsurface materials (solid, liquid, gaseous, mixtures), underground space (cavity), subsurface energy (static, geo-dynamic, geomagnetic). Geo-resources are provisionally recognized as geoinformation, characterized by various attributes and properties of subsurface material (minerals, overburden, etc.), and the other geo-resources, including streams migrating into the depths (fluids, magma, etc.). It is clear that mineral resources are part of the subsurface material. Therefore, the most important aspect of geo-resources development today is the initiation of recycling of their industrial processing.

This study aimed at isolating and identifying bacteria and fungi with the capacity to degrade low density polyethylene (LDPE). The level of biodegradation of LDPE sheets with bacterial and fungal inoculums from different sampling points of Dandora dumpsite was evaluated under laboratory conditions. Incubation of the LDPE sheets was done for sixteen weeks at 37°C and 28°C for bacteria and fungi respectively in a shaker incubator. Isolation of effective candidates for biodegradation was done based on the recorded biodegradation outcomes. The extent of biodegradation on the polyethylene sheets was assessed by various techniques including weight loss analysis, Fourier Transform Infrared Spectroscopy (FTIR) and GC-MS. Fourier Transform Infra-Red spectroscopy (FTIR) analysis revealed the appearance of new functional groups attributed to hydrocarbon degradation after incubation with the bacteria and fungi. Analysis of the 16S rDNA and 18S rDNA sequences for bacteria and fungi respectively showed that bacteria belonging to genera Pseudomonas, Bacillus, Brevibacillus, Cellulosimicrobium, Lysinibacillus and fungi of genus Aspergillus were implicated as polyethylene degraders. An overall analysis confirmed that fungi are generally better degraders of polyethylene than bacteria. The highest fungal degradation activity was a mean weight reduction of 36.4±5.53% attributed to Aspergillus oryzae strain A5, 1 (MG779508). The highest degradation activity for bacteria was a mean of 35.72± 4.01% and 20.28± 2.30% attributed to Bacillus cereus strain A5,a (MG645264) and Brevibacillus borstelensis strain B2,2 (MG645267) respectively. Genus Aspergillus, Bacillus and Brevibacillus were confirmed to be good candidates for Low Density Poly Ethene bio-degradation. This was further confirmed by the appearance of the aldehyde, ether and carboxyl functional groups after FTIR analysis of the polythene sheets and the appearance of a ketone which is also an intermediary product in the culture media. To improve this degrading capacity through assessment of optimum conditions for microbial activity and enzyme production will enable these findings to be applied commercially and on a larger scale.

This study aimed at molecular and biochemical characterization of low-density polyethene (LDPE) degrading fungi and bacteria from Dandora dumpsite, Nairobi. Twenty bacterial and 10 fungal isolates were identified using 16S rDNA and 18S rDNA sequences for bacteria and fungi, respectively. The highest fungal degradation was attributed to Aspergillus oryzae strain A5,1 while the highest bacterial degradation was attributed to Bacillus cereus strain A5,a and Brevibacillus borstelensis strain B2,2, respectively. Isolates were screened for their ability to produce extracellular laccase and esterase; Aspergillus fumigatus strain B2,2 exhibited the highest presence of laccase (15.67 mm) while Aspergillus oryzae strain A5,1 exhibited the highest presence of esterase (14.33 mm). Alkane hydroxylase-encoding genes were screened for using primer AlkB 1 which amplified the fragment of size 870 bp. Four bacterial samples were positive for the gene. Optimum growth temperature of the fungal isolates was 30°C. The possession of laccase, esterase, and alkane hydroxylase activities is suggested as key molecular basis for LDPE degrading capacity. Knowledge of optimum growth conditions will serve to better utilize microbes in the bioremediation of LDPE. The application of Aspergillus oryzae strain A5,1 and Bacillus cereus strain A5,a in polyethene degradation is a promising option in this kind of bioremediation as they exhibited significantly high levels of biodegradation. Further investigation of more alkane degrading genes in biodegrading microbes will inform the choice of the right microbial consortia for bioaugmentation strategies.

The properties of a bloodmeal-induced trypanolysin from the midgut of the tsetse, G. m. morsitans was studied in vitro. The semi-purified trypanolysin from twice-fed tsetse had the highest trypanolysin activity against bloodstream trypanosomes followed by those once-fed and the unfed flies. Serum found to display trypanolysin activity. The trypanolysin had no trypsin activity nor even affected by the enzyme. In addition, trypanolysin was not affected by protease inhibitors such as soy bean trypsin inhibitor (STI), N-a-p-Tosyl-L-lysine chromethyl ketone (TLCK), phenylmethyl sulphonyl fluoride (PMSF), diisopropyl fluoro-phosphate (DFP) and tosylamide-2-phenylethyl chloromethyl ketone (TPCK). However, the activity was completely inhibited by diethyl pyrocarbonate (DEPC) and partially by aprotinin. The induction of trypanolysin activity by bloodmeal increased gradually reaching a peak at 72-120 h after the bloodmeal, and then decreased rapidly, with only 25% of the peak activity remaining after 192 h. The trypanolysin was inactivated during storage at 27°C and 4°C after 15 and 32 days, respectively. Similarly, heating the midguts trypanolysin to 60 80°C led to loss of activity. On the other hand, 50°C was found to be the optimum temperature for trypanolysin activity. The activity was also unstable by freeze-thaw at 80°C, -70°C, -20°C and 0°C after 33, 41, 55 and 63 days, respectively. Trypanolysin caused lyses of bloodstream-form T. b. brucei, while the procyclic trypanosomes were unaffected. The highest trypanolysin activity in different tsetse species was found with Glossina longipennis, followed by Glossina pallidipes, Glossina morsitans centralis, Glossina fuscipes fuscipes and G. m. morsitans. When the midgut homogenate was separated by anionexchange chromatography, the trypanolysin activity was recovered in the bound fraction. These results suggest that the midgut trypanolysin plays an important role in the establishment of trypanosome infections in tsetse.

Genetically determined artemisinin resistance in Plasmodium falciparum has been described in Southeast Asia. The relevance of recently described Kelch 13-propeller mutations for artemisinin resistance in Sub-Saharan Africa parasites is still unknown. Southeast Asia parasites have low genetic diversity compared to Sub-Saharan Africa, where parasites are highly genetically diverse. This study attempted to elucidate whether genetics provides a basis for discovering molecular markers in response to artemisinin drug treatment in P. falciparum in Kenya. The genetic diversity of parasites collected pre- and post- introduction of artemisinin combination therapy (ACT) in western Kenya was determined. A panel of 12 microsatellites and 91 single nucleotide polymorphisms (SNPs) distributed across the P. falciparum genome were genotyped. Parasite clearance rates were obtained for the post-ACT parasites. The 12 microsatellites were highly polymorphic with post-ACT parasites being significantly more diverse compared to pre-ACT (p < 0.0001). The median clearance half-life was 2.55 hours for the post-ACT parasites. Based on SNP analysis, 15 of 90 post-ACT parasites were single-clone infections. Analysis revealed 3 SNPs that might have some causal association with parasite clearance rates. Further, genetic analysis using Bayesian tree revealed parasites with similar clearance phenotypes were more closely genetically related. With further studies, SNPs described here and genetically determined response to artemisinin treatment might be useful in tracking artemisinin resistance in Kenya.

In the present study, a trypanolysin induced by component of blood meal in the midgut of tsetse G. m. morsitans was isolated and purified in three steps. The first step was achieved by separation of the midgut homogenate on a conventional anion-exchange chromatography column. The highest trypanolysin activity was recovered in the bound fractions (95%, 05 M NaCl). In the second step, isolation was achieved on a Mono Q anionexchange column by elution at 70-80% 1 M NaCl. The third step of isolation-purification was achieved by using epoxy-activated Sepharose 6-B-affinity chromatography column. In this case, the trypanolysin was eluted using 20 mM Tris-HCl. The purified native G. m. morsitans trypanolysin was of Mw ≈ 669 kDa, while ≈ 14 kDa trypanolysin was shown in denaturing trypanolysin SDS-PAGE gel for the same tsetse species. It was noticed that the purified trypanolysin was lipidated and also found to be glycosylated.

In the present study, a trypanolysin induced by component of blood meal in the midgut of tsetse G. m. morsitans was isolated and purified in three steps. The first step was achieved by separation of the midgut homogenate on a conventional anion-exchange chromatography column. The highest trypanolysin activity was recovered in the bound fractions (95%, 05 M NaCl). In the second step, isolation was achieved on a Mono Q anionexchange column by elution at 70-80% 1 M NaCl. The third step of isolation-purification was achieved by using epoxy-activated Sepharose 6-B-affinity chromatography column. In this case, the trypanolysin was eluted using 20 mM Tris-HCl. The purified native G. m. morsitans trypanolysin was of Mw ≈ 669 kDa, while ≈ 14 kDa trypanolysin was shown in denaturing trypanolysin SDS-PAGE gel for the same tsetse species. It was noticed that the purified trypanolysin was lipidated and also found to be glycosylated.

ABSTRACT The prevalence of a genetic polymorphism(s) at codon 268 in the cytochrome b gene, which is associated with failure of atovaquone-proguanil treatment, was analyzed in 227 Plasmodium falciparum parasites from western Kenya. The prevalence of the wild-type allele was 63%, and that of the Y268S (denoting a Y-to-S change at position 268) mutant allele was 2%. There were no pure Y268C or Y268N mutant alleles, only mixtures of a mutant allele(s) with the wild type. There was a correlation between parasite 50% inhibitory concentration (IC 50 ) and parasite genetic polymorphism; mutant alleles had higher IC 50 s than the wild type.

Trans-generational transfer of gregarious-phase traits in the desert locust Schistocerca gregaria (Forskål, 1775) is mediated by primer gregarizing pheromonal signals produced by ovipositing females that experience crowding. We monitored time-course proteomic events in eggs from solitary-reared locusts that had been exposed for 1, 3, 5, 7, 10 and 12 days to different levels of the sand-associated gregarizing signal originating from 0, 3, 5 or 10 ovipositions by crowd-reared females. Evidence for the phase transition was sought by comparing the protein patterns of embryos thus exposed with those from crowd-reared (gregarious) controls; this comparison was continued until the stage of the first instars. Expressed proteins were analysed by two-dimensional protein gel electrophoresis, and patterns from the different treatments within stages were compared by profile matching and χ2 analyses. Eggs derived from crowd- and solitary-reared females showed essentially similar protein patterns at early stages of embryogenesis; however, mature stages (particularly, days 10 and 12) and hatchlings demonstrated significantly different patterns. Protein patterns of eggs from solitary-reared females that were incubated in sand contaminated with the pheromonal signal and of the hatchlings that emerged were similar to those derived from gregarious females and dependent on the level of the pheromone to which the embryos had been exposed. The results confirm the gregarizing effect of the signal and constitute a useful basis for unravelling the mechanism of the signalling cascades associated with gene expressions triggered by the pheromone.

Tissue culture techniques are commonly used in plants as an efficient way to propagate and store valuable genotypes. Often, some of the regenerants differ from the parental type, a phenomenon called somaclonal variability. Assessment of nutritional value variability in crops that may arise from somaclonal variability during tissue culture propagation may have a strong impact on plant breeding, conservation of genetic resources and nutrition in the areas of use. It is particularly useful in the characterization of individual cultivars, and in determining duplications in germplasm collections and for selecting parents. The Sorghum bicolor (L) Moench tissue culture (TC) regenerants (Seredo, Mtama 1 and El Gardam) were developed at the Jomo Kenyatta University of Agriculture and Technology towards improvement for water stress tolerance for improved food production in the ASALs in Kenya. The study was conducted to evaluate the nutritional value of the parents and TC regenerants of Sorghum bicolor (L) Moench local cultivars (Seredo, Mtama 1 and El Gardam) in Kenya. For proximate composition significant (p≤0.05) differences were observed in parents and regenerants of the El‐Gardam (moisture, proteins and crude fiber), Mtama 1 (proteins) and Seredo (fats and crude fiber). The mineral compositions of the parents and regenerants of the cultivars were not significantly different (p ≤ 0.05) except for Zinc in Mtama 1 cultivar and Iron in both El‐Gardam and Mtama 1 cultivars. B‐vitamins showed significant differences (p ≤ 0.05) for both thiamine and Pyridoxine in El‐Gardam and Seredo. Significant variability (p ≤ 0.05) was shown phytates content in each cultivar. The parents were observed to have significantly higher amounts of Phytates than the regenerants within all the cultivars. The study recommends Mtama 1 regenerants with low anti‐nutrient appropriate for ASALs with respect to nutrient availability since anti‐nutrients in sorghum have been shown to impair the bioavailability of the other nutrients to the body.

Post-harvest pests are known to cause severe losses of beans in particular the bean weevil, Acanthoscelides obtectus, necessitating immediate and long term control measures. Convectional pesticides have been used for decades to protect stored grain from pests, but their harmful environmental impact and pest resistance resulting from the need for increased pesticide use have triggered researchers to search for eco-friendly, biodegradable and potent biopesticides. The use of natural products derived from metabolic activity of plants may constitute a new avenue of pest control. This paper describes phytochemical and biological investigations of leaves, root, and stem bark crude extracts and isolated compounds of selected Kenyan plants used to control the bean weevil. Bioassays tests of the compounds were done against brine shrimp to test their insecticidal activity against the bean weevil. Antibacterial and antifungal activity of the crude extracts of S. didymobotrya was also done. The extracts of S. didymobotrya showed high toxicity as well as adulticidal activity against bean weevil.