Transcriptome profile of spleen tissues from locally-adapted Kenyan pigs (Sus scrofa) experimentally infected with three varying doses of a highly virulent African swine fever virus genotype IX isolate: Ken12/busia.1 (ken-1033).

BACKGROUND: African swine fever (ASF) is a lethal hemorrhagic disease affecting domestic pigs resulting in up to 100% mortality rates caused by the ASF virus (ASFV). The locally-adapted pigs in South-western Kenya have been reported to be resilient to disease and harsh climatic conditions and tolerate ASF; however, the mechanisms by which this tolerance is sustained remain largely unknown. We evaluated the gene expression patterns in spleen tissues of these locally-adapted pigs in response to varying infective doses of ASFV to elucidate the virus-host interaction dynamics. METHODS: Locally adapted pigs (n = 14) were experimentally infected with a high dose (1x106HAD50), medium dose (1x104HAD50), and low dose (1x102HAD50) of the highly virulent genotype IX ASFV Ken12/busia.1 (Ken-1033) isolate diluted in PBS and followed through the course of infection for 29 days. The in vivo pig host and ASFV pathogen gene expression in spleen tissues from 10 pigs (including three from each infective group and one uninfected control) were analyzed in a dual-RNASeq fashion. We compared gene expression between three varying doses in the host and pathogen by contrasting experiment groups against the naïve control. RESULTS: A total of 4954 differentially expressed genes (DEGs) were detected after ASFV Ken12/1 infection, including 3055, 1771, and 128 DEGs in the high, medium, and low doses, respectively. Gene ontology and KEGG pathway analysis showed that the DEGs were enriched for genes involved in the innate immune response, inflammatory response, autophagy, and apoptosis in lethal dose groups. The surviving low dose group suppressed genes in pathways of physiopathological importance. We found a strong association between severe ASF pathogenesis in the high and medium dose groups with upregulation of proinflammatory cytokines and immunomodulation of cytokine expression possibly induced by overproduction of prostaglandin E synthase (4-fold; p < 0.05) or through downregulation of expression of M1-activating receptors, signal transductors, and transcription factors. The host-pathogen interaction resulted in induction of expression of immune-suppressive cytokines (IL-27), inactivation of autophagy and apoptosis through up-regulation of NUPR1 [5.7-fold (high dose) and 5.1-fold (medium dose) [p < 0.05] and IL7R expression. We detected repression of genes involved in MHC class II antigen processing and presentation, such as cathepsins, SLA-DQB1, SLA-DOB, SLA-DMB, SLA-DRA, and SLA-DQA in the medium and high dose groups. Additionally, the host-pathogen interaction activated the CD8+ cytotoxicity and neutrophil machinery by increasing the expression of neutrophils/CD8+ T effector cell-recruiting chemokines (CCL2, CXCL2, CXCL10, CCL23, CCL4, CXCL8, and CXCL13) in the lethal high and medium dose groups. The recovered pigs infected with ASFV at a low dose significantly repressed the expression of CXCL10, averting induction of T lymphocyte apoptosis and FUNDC1 that suppressed neutrophilia. CONCLUSIONS: We provide the first in vivo gene expression profile data from locally-adapted pigs from south-western Kenya following experimental infection with a highly virulent ASFV genotype IX isolate at varying doses that mimic acute and mild disease. Our study showed that the locally-adapted pigs induced the expression of genes associated with tolerance to infection and repression of genes involved in inflammation at varying levels depending upon the ASFV dose administered.

Multi-drug resistant Salmonella enterica serovar Typhi isolates with reduced susceptibility to ciprofloxacin in Kenya.

BACKGROUND: Typhoid fever remains a public health concern in developing countries especially among the poor who live in informal settlements devoid of proper sanitation and clean water supply. In addition antimicrobial resistance poses a major challenge in management of the disease. This study assessed the antimicrobial susceptibility patterns of Salmonella enterica serotype Typhi (S. Typhi) isolated from typhoid fever cases (2004-2007). METHODS: A cross sectional study was conducted on 144 archived S. Typhi isolates (2004-2007) tested against 11 antimicrobial agents by quality controlled disk diffusion technique. Isolates resistant to ampicillin, chloramphenicol, and cotrimoxazole were considered Multidrug resistant (MDR). Thirty MDR isolates were selected randomly and further tested using minimum inhibitory concentration (MIC) E-test. RESULTS: Sixteen percent (23/144) of the isolates were susceptible to all the antibiotics tested while 68% were resistant to three or more of the 11 antibiotics tested. The isolates showed a high susceptibility to ceftriaxone (94%) and gentamicin (97%). A high percentage of resistance was observed for the conventional first-line antibiotics; ampicillin (72%), chloramphenicol (72%), and cotrimoxazole (70%). Sixty-nine percent of the isolates (100/144) showed reduced susceptibility to ciprofloxacin. All the 30 (100%) isolates selected for MIC test were susceptible to amoxicillin-clavulanic acid. All except one of the 30 isolates were susceptible to ceftriaxone while majority 21 (70%) recorded an intermediate susceptibility to ciprofloxacin with MIC of 0.12-0.5 µg/mL. CONCLUSION: A large proportion of S. Typhi isolates were MDR and also showed reduced susceptibility to ciprofloxacin. Fluoroquinolone resistance is emerging and this may pose a challenge in treatment of typhoid in future. There is need for routine surveillance to monitor this phenotype in clinical settings.

Milk fatty acid variability and association with polymorphisms in SCD1 and DGAT1 genes in White Fulani and Borgou cattle breeds.

The stearoyl-CoA desaturase 1 (SCD1) A293V and acyl CoA: diacylglycerol acyltransferase 1 (DGAT1) K232A polymorphisms have been associated with significant variation in bovine milk fatty acid composition and unsaturation indices in western cattle breeds. This study aimed to estimate the milk fatty acid variability in indigenous Borgou and White Fulani cattle breeds of Benin, and the effects of the SCD1 A293V and DGAT1 K232A polymorphisms on milk and fatty acid composition and unsaturation indices. Thus, 85 Borgou and 96 White Fulani cows were genotyped for the SCD1 A293V and DGAT1 K232A polymorphisms and their milk and fatty acid composition and unsaturation indices were determined. Borgou presented milk with higher linoleic acid (P < 0.001), oleic acid (P < 0.05), C18 index (P < 0.001), total unsaturation index (P < 0.05), and lower total saturated fatty acid (SFA) compared to White Fulani. The SCD1 VV genotype was associated with higher protein and lactose contents in White Fulani (P < 0.05). In Borgou, the SCD1 AV genotype was associated with higher C14 and total unsaturation indices (P < 0.01), while the SCD1 V allele was associated with decrease in C14 index (P < 0.05). In White Fulani, the SCD1 VV genotype was associated with lower C18:1 cis-9 content (P < 0.05) while the DGAT1 K allele was associated with increased total SFA (P < 0.05), and decreased C18 index (P < 0.05), total unsaturation index (P < 0.01) and total monounsaturated fatty acid (P < 0.01). The SCD1 A293V and DGAT1 K232A may serve as genetic markers to improve milk fatty acid traits in Borgou and White Fulani breeds.

Assessing actors in rural markets of sheep and goats in the Nyando Basin of Western Kenya: a key to improving productivity from smallholder farms.

Livestock traders are a key conduit for incentives in livestock production systems. Their actions affect producer prices, investment decisions, and their livelihoods. However, smallholder farmers in rural communities of eastern Africa often have limited understanding of the marketing process and the interactions among market actors. This study was undertaken following the introduction of improved indigenous goat and sheep breeds in Climate Smart Villages of Nyando in western Kenya. We sought to understand the marketing of sheep and goats in five key rural markets used by the smallholder farmers of Nyando, to generate information on key market actors, their functions in the value chain, and their preferences for attributes in sheep and goats. Most traders (65%) in all the markets were men. Four different types of traders operated in the markets, producers, retailers, wholesalers, and brokers, each making a profit. Preference for selling sheep or goats and the animal attributes desired by the traders differed depending on the surrounding community. Markets in Kericho County of Nyando traded more goats than sheep while those in Kisumu County of Nyando traded more sheep than goats. There were no clear policies guiding pricing of animals. Prices offered to producers depended on previous days, and the number of animals available for sale at the marketing point. The collaborative group actions of the producers in the Climate Smart Villages did not extend beyond the level of production to the marketing of their livestock. A critical change is required for the smallholder farmers to derive better incomes from improved quality of animals following adoption of improved breeding and management practices. The results illustrate the need to engage rural market actors when implementing livestock improvement programs for smallholder farmers for better farm gate prices of animals, and thus achieve the improved incomes envisioned.

Partial HIV C2V3 envelope sequence analysis reveals association of coreceptor tropism, envelope glycosylation and viral genotypic variability among Kenyan patients on HAART.

BACKGROUND: HIV-1 is highly variable genetically and at protein level, a property it uses to subvert antiviral immunity and treatment. The aim of this study was to assess if HIV subtype differences were associated with variations in glycosylation patterns and co-receptor tropism among HAART patients experiencing different virologic treatment outcomes. METHODS: A total of 118 HIV env C2V3 sequence isolates generated previously from 59 Kenyan patients receiving highly active antiretroviral therapy (HAART) were examined for tropism and glycosylation patterns. For analysis of Potential N-linked glycosylation sites (PNGs), amino acid sequences generated by the NCBI's Translate tool were applied to the HIVAlign and the N-glycosite tool within the Los Alamos Database. Viral tropism was assessed using Geno2Pheno (G2P), WebPSSM and Phenoseq platforms as well as using Raymond's and Esbjörnsson's rules. Chi square test was used to determine independent variables association and ANOVA applied on scale variables. RESULTS: At respective False Positive Rate (FPR) cut-offs of 5% (p = 0.045), 10% (p = 0.016) and 20% (p = 0.005) for CXCR4 usage within the Geno2Pheno platform, HIV-1 subtype and viral tropism were significantly associated in a chi square test. Raymond's rule (p = 0.024) and WebPSSM (p = 0.05), but not Phenoseq or Esbjörnsson showed significant associations between subtype and tropism. Relative to other platforms used, Raymond's and Esbjörnsson's rules showed higher proportions of X4 variants, while WebPSSM resulted in lower proportions of X4 variants across subtypes. The mean glycosylation density differed significantly between subtypes at positions, N277 (p = 0.034), N296 (p = 0.036), N302 (p = 0.034) and N366 (p = 0.004), with HIV-1D most heavily glycosylated of the subtypes. R5 isolates had fewer PNGs than X4 isolates, but these differences were not significant except at position N262 (p = 0.040). Cell-associated isolates from virologic treatment success subjects were more glycosylated than cell-free isolates from virologic treatment failures both for the NXT (p = 0.016), and for all the patterns (p = 0.011). CONCLUSION: These data reveal significant associations of HIV-1 subtype diversity, viral co-receptor tropism, viral suppression and envelope glycosylation. These associations have important implications for designing therapy and vaccines against HIV. Heavy glycosylation and preference for CXCR4 usage of HIV-1D may explain rapid disease progression in patients infected with these strains.

Mitochondrial DNA variation of domestic sheep (Ovis aries) in Kenya.

The history of domestic sheep (Ovis aries) in Africa remains largely unknown. After being first introduced from the Near East, sheep gradually spread through the African continent with pastoral societies. The eastern part of Africa was important either for the first diffusion of sheep southward or for putative secondary introductions from the Arabian Peninsula or southern Asia. We analysed mitochondrial DNA control region sequences of 91 domestic sheep from Kenya and found a high diversity of matrilines from the widespread haplogroup B, whereas only a single individual from haplogroup A was detected. Our phylogeography analyses of more than 500 available mitochondrial DNA sequences also identified ancestral haplotypes that were probably first introduced in Africa and are now widely distributed. Moreover, we found no evidence of an admixture between East and West African sheep. The presence of shared haplotypes in eastern and ancient southern African sheep suggests the possible southward movement of sheep along the eastern part of Africa. Finally, we found no evidence of an extensive introduction of sheep from southern Asia into Africa via the Indian Ocean trade. The overall findings on the phylogeography of East African domestic sheep set the grounds for understanding the origin and subsequent movements of sheep in Africa. The richness of maternal lineages in Kenyan breeds is of prime importance for future conservation and breeding programmes.

The potential role of roaming dogs in establishing a geographically novel life cycle of taeniids (Echinococcus spp. and Taenia spp.) in a non-endemic area.

INTRODUCTION: Cystic Echinococcosis (CE) is endemic in humans and livestock in many pastoral communities in Kenya. The distribution of the disease is enhanced by several factors, including livestock trade, which has allowed for the spread of CE to non-endemic areas such as western Kenya. Dogs' roaming behaviour, with consequent contamination of the environment with intestinal parasites, could then lead to parasite establishment. This study examined dogs' infection levels with taeniid eggs and their potential role in contaminating the environment with intestinal parasites. METHODOLOGY: We selected sixteen ruminant slaughterhouses in Busia and Bungoma Counties, and around each slaughterhouse we identified ten homesteads owning free-roaming dogs. We administered a questionnaire on dog management practices to the homestead owner and collected a faecal sample from the dog's rectum. In homesteads around 8 of the 16 slaughterhouses, we collared dogs with a GPS tracker to assess their movement patterns. The faecal samples were examined microscopically following zinc-chloride sieving-floatation technique for the presence of taeniid eggs and other canine intestinal parasites. Polymerase Chain Reaction - Restriction Fragment Length Polymorphism of NADH dehydrogenase subunit 1 gene and sequencing were used to confirm taeniid eggs identified during microscopy. Additionally, the Coproantigen-ELISA was used to detect the presence of taeniid antigen in a sub-set of the faecal samples. RESULTS: Helminths detected in the 155 dogs sampled included hookworms (n = 92; 59.4%), ascarids (n = 15; 9.7%), and taeniids (n = 1; 0.6%). Through Copro-PCR, 13 eggs extracted from the sample of the only taeniid infected dog were sequenced and identified as E. canadensis (G6/7) [n = 1], Taenia multiceps [n = 1], and Taenia serialis [n = 6]; the remaining were indeterminate. Of the 77 faecal samples tested for E. granulosus sensu lato (s. l.) with the Copro-ELISA, 64 (83.1%) were negative, 12 (15.6%) were positive, while 1 (1.3%) was suspicious. The dogs travelled a median of 13.5 km daily, and 28 dogs visited the slaughterhouses during the 5-day recording period. CONCLUSION: The results indicate a relatively high carriage of zoonotic parasites by free-roaming domestic dogs in western Kenya, which poses a risk to human and livestock populations. We report for the first time a domestic lifecycle of Echinococcus canadensis and Taenia multiceps in western Kenya, as well as a presumptive sylvatic cycle of coenurosis by T. serialis. We recommend an extensive and ongoing Copro-antigen survey of dog faeces, broader assessment of dog parasites with zoonotic potential, adherence to slaughterhouse management practices, and dog-ownership programmes to highlight the importance of deworming and restricted dog movements.

Factors affecting uptake of cervical cancer early detection measures among women in Thika, Kenya.

In this study, 50 in-depth interviews were carried out with women from the general population in Thika, Kenya. We explored awareness, attitudes, and behavior toward cervical cancer and screening measures among the women. The concept of the health belief model (HBM) was used to develop the topics for the in-depth interviews. Our findings highlight the lack of awareness of cervical cancer and the benefits of early detection measures as critical barriers that affect women's participation in screening programs. We provide a basis for designing programs that will be acceptable and accessible to a larger population, resulting in a reduced cervical cancer burden.

Nevirapine plasma concentration is associated with virologic failure and the emergence of drug-resistant mutations among HIV patients in Kenya: A cross sectional study.

This study aimed to determine the association between the plasma concentration of nevirapine (NVP) and clinical outcomes. In this cross-sectional study, sociodemographic and clinical data were collected from 233 HIV patients receiving NVP-based first-line antiretroviral therapy (ART) regimens in Nairobi, Kenya. The mean age was 41.2 (SD ±â€…11.9) years. Fifty-four (23.2%) patients had virological failure (>1000 copies/mL), whereas 23 (9.9%) were infected with drug-resistant HIV strains. Eleven patients had nucleoside reverse transcriptase inhibitor resistance mutations, including M184V and T215Y, whereas 22 had non-nucleoside reverse transcriptase inhibitor resistance mutations, including G190A, K103N, V106A, Y181C, A98G, and Y188L. The median NVP plasma concentration was 6180 ng/mL (IQR 4444-8843 ng/mL), with 38 (16.3%) patients having suboptimal NVP plasma levels of <3400 ng/mL. The majority 23 of the 38 (60.5%) patients with NVP Cmin < 3400 ng/mL were significantly infected with drug-resistant HIV virus (P = .001). In the multivariate analysis, the time taken to arrive at the ART clinic (ß -11.1, 95% CI -21.2 to -1.1; P = .031), higher HIV viral load (ß -2008, 95% CI -3370.7 to -645.3; P = .004), and the presence of HIV drug resistance mutation (ß 3559, 95% CI 2580.8-4537.2; P = .0001) were associated with NVP plasma concentration. A significant proportion of patients receiving the NVP-based regimen had supra- and sub-therapeutic plasma concentrations. Higher HIV viral load and the presence of HIV drug-resistant mutations are important factors associated with NVP plasma concentrations.

Findings of a community screening programme for human cystic echinococcosis in a non-endemic area.

Cystic Echinococcosis (CE) is a zoonosis caused by infection with the larval stages of the taeniid cestodes of the species complex Echinococcus granulosus sensu lato. It is prevalent among transhumant communities in East Africa, including those residing in northern Kenya. The movement of livestock from these regions of high incidence to areas of low incidence creates an indirect risk of disease spill-over to humans. To assess possible establishment of the CE life cycle outside known endemic regions, we used a portable ultrasound scanner to screen for the presence of human CE in Bungoma County of western Kenya, an area which imports substantial numbers of cattle for slaughter from neighbouring pastoralist regions. Eight sentinel sites were purposively selected based on their proximity to slaughterhouses handling animals introduced from pastoralist regions, and necessary permissions to conduct the study were sought. Regression analyses were conducted to identify risk factors associated with the presence of abdominal and cystic lesions (CL). In total, 1002 participants were screened; of these, 654 (65.3%) were female and the median age was 43. Farming (n = 403; 43.4%) was the most frequent occupation, followed by professional (i.e. on regular salary) (n = 215; 23.1%), and business (n = 207; 22.3%) categories. Sixty-seven participants (6.7%) had abnormal ultrasound findings, of these, 7 (1.1%) had simple liver cysts/CL, as per WHO classification. As such, their outcome was inconclusive and they were not put on treatment but advised to attend follow-up investigations in a referral health facility. Other abnormal findings included splenomegaly (n = 14), ovarian cysts (n = 14), uterine fibroids (n = 10), polycystic kidneys (n = 6), and benign prostatic hyperplasia (n = 6). Age was unconditionally associated with the presence of presumptive CL. These results contribute to CE baseline data while providing insights on the implementation of ultrasound diagnosis in the field, as recommended by the WHO for targeted control of echinococcosis by 2030.

Antimicrobial resistance profiles and genetic basis of resistance among non-fastidious Gram-negative bacteria recovered from ready-to-eat foods in Kibera informal housing in Nairobi, Kenya.

OBJECTIVE: This cross-sectional study conducted in Kibera, Kenya, sought to gain insights on relative microbial contamination levels of popular unprocessed food types, determine antimicrobial resistance (AMR) burden and the carriage of integrons that are essential elements for spreading antimicrobial resistance genes (ARG). Foods analysed consisted of cooked vegetables (kale, cabbage, and nightshades), boiled cereal foods (beans, rice, and Githeri, which is a mixture of beans and maize), meat, Omena fish (fried silver cyprinids), and Ugali (a product of simmered maize flour in boiled water). RESULTS: The analysis detected contamination levels exceeding 2×104 c.f.u. ml-1 in 106 (38 %) of the 281 ready-to-eat foods analysed. The majority of food types had microbial contaminations of between 4.0×104 and 2.3×106 c.f.u. ml-1. Kale was the most contaminated with a mean of 2.3×106 c.f.u. ml-1, while Omena was the least contaminated with 4.0×104 c.f.u. ml-1. Foods sold close to open sewage and refuse sites were more contaminated than those sold in relatively 'cleaner' settings (P <0.0001, O.R 0.1162, C.I 0.1162-0.120). A total of 405 bacterial isolates were recovered and included; Klebsiella spp 116 (29 %), Escherichia coli 104 (26 %), Enterobacter agglomerans 88 (22 %), Proteus mirabilis 30 (7 %), Salmonella spp 28 (7 %), Citrobacter freundii 27 (7 %) and Serratia marcescens 12 (3 %). Imipenem (IPM, 100 %) was the most effective antimicrobial agent, followed by cefepime (98 %). Ampicillin (AMP, 33 %), trimethoprim (TMP, 27 %), and sulfamethoxazole (SMX, 23 %) on the other hand, were the least effective antimicrobials. The analysis also found ten isolates (2 %) that had co-resistance to third-generation cephalosporins, fluoroquinolone (CIP), quinolones (NAL) and aminoglycosides (GEN); hereby we refer to this phenotype as the ßFQA. The prevalence of multidrug-resistant (MDR) strains was 23 % (93), while that of extended-spectrum ß-lactamases (ESBL) producing strains was 4 % (17). The bla TEM was the most prevalent (55 %) ß-lactamase (bla) gene among the screened 93 MDR-strains. Carriage of class one integrons (intI1) was more common (23 %) than intl2 (3 %) among these MDR-strains. Bacterial diversity analysis using the GTG5-PCR found no significant clusters for analysed E. coli and K. pneumoniae, suggesting recovered isolates were genetically diverse and not due to non-clonal expansion. The findings of this study are an indication that contaminated foods can be a reservoir for enteric pathogens and a source of AMR strains.

Comparative Analysis of SLA-1, SLA-2, and DQB1 Genetic Diversity in Locally-Adapted Kenyan Pigs and Their Wild Relatives, Warthogs.

Swine leukocyte antigen (SLA) plays a central role in controlling the immune response by discriminating self and foreign antigens and initiating an immune response. Studies on SLA polymorphism have demonstrated associations between SLA allelic variants, immune response, and disease resistance. The SLA polymorphism is due to host-pathogen co-evolution resulting in improved adaptation to diverse environments making SLA a crucial genomic region for comparative diversity studies. Although locally-adapted African pigs have small body sizes, they possess increased resilience under harsh environmental conditions and robust immune systems with reported tolerance to some diseases, including African swine fever. However, data on the SLA diversity in these pigs are not available. We characterized the SLA of unrelated locally-adapted domestic pigs from Homa Bay, Kenya, alongside exotic pigs and warthogs. We undertook SLA comparative diversity of the functionally expressed SLA class I (SLA-1, SLA-2) and II (DQB1) repertoires in these three suids using the reverse transcription polymerase chain reaction (RT-PCR) sequence-based typing (SBT) method. Our data revealed higher genetic diversity in the locally-adapted pigs and warthogs compared to the exotic pigs. The nucleotide substitution rates were higher in the peptide-binding regions of the SLA-1, SLA-2, and DQB1 loci, indicative of adaptive evolution. We obtained high allele frequencies in the three SLA loci, including some breed-specific private alleles, which could guide breeders to increase their frequency through selection if confirmed to be associated with enhanced resilience. Our study contributes to the growing body of knowledge on genetic diversity in free-ranging animal populations in their natural environment, availing the first DQB1 gene data from locally-adapted Kenyan pigs.

Archaeal diversity in the haloalkaline Lake Elmenteita in Kenya.

A non-culture approach was used to study the archaeal diversity in Lake Elmenteita, Kenya. Five different sampling points were selected randomly within the lake. Wet sediments and water samples were collected from each sampling point. In addition, dry mud cake was collected from three points where the lake had dried. DNA was extracted from these samples and the 16S rRNA genes were amplified using primers described to be Domain-specific for Archaea. Eleven clone libraries were constructed using PCR-amplified 16S rRNA genes. A total of 1,399 clones were picked and analysed via ARDRA. 170 ARDRA patterns were unique and the respective clones were selected for sequencing. 149 clones gave analysable sequences. BLAST analysis showed that 49 belong to the Domain Archaea while the others were either chimera or affiliated to eukaryotic taxa. Comparative sequence analysis of archaeal clones affiliated them to a wide range of genera. The order Halobacteriales was represented by members of the genera Natronococcus, Halovivax, Halobiforma, Halorubrum, and Halalkalicoccus. The highest percentage (46%) of the clones, however, belonged to uncultured members of the Domain Archaea in the order Halobacteriales. The results show that the archaeal diversity in the lake could be higher than previously reported.

Plasmid profiling and incompatibility grouping of multidrug resistant Salmonella enterica serovar Typhi isolates in Nairobi, Kenya.

OBJECTIVES: Plasmids harbour antibiotic resistance genes which contribute to the emergence of multidrug resistant pathogens. We detected the presence of plasmids in multidrug resistant Salmonella enterica serovar Typhi (S. Typhi) isolates from our previous study and consequently determined their incompatibility groups and possibility of conjugation transmission. Plasmids were extracted from 98 multidrug resistant S. Typhi isolates based on alkaline lysis technique. Plasmid incompatibility grouping was established by PCR replicon typing using 18 pairs of primers to amplify FIA, FIB, FIC, HI1, HI2, I1-Iγ, L/M, N, P, W, T, A/C, K, B/O, X, Y, F and FIIA replicons. Antibiotic resistance phenotypes were conjugally transferred from S. Typhi isolates with plasmids to Escherichia coli K12F strain devoid of plasmids. RESULTS: Approximately 79.6% of the MDR S. Typhi isolates were related to the existence of plasmids. We detected 93.6% of plasmids belonging to incompatibility (Inc) group HI1. The other incompatibility groups identified included IncFIC (16.7%), IncP (1.3%), and IncI1 (1.3%) which appeared together with Inc HI1. MDR S. Typhi isolated carried a homologous plasmid of incompatibility group HI1 most of which transferred the resistance phenotypes of ampicillin, tetracycline and chloramphenicol to the transconjugants.

Plasma nevirapine concentrations predict virological and adherence failure in Kenyan HIV-1 infected patients with extensive antiretroviral treatment exposure.

Treatment failure is a key challenge in the management of HIV-1 infection. We conducted a mixed-model survey of plasma nevirapine (NVP) concentrations (cNVP) and viral load in order to examine associations with treatment and adherence outcomes among Kenyan patients on prolonged antiretroviral therapy (ART). Blood plasma was collected at 1, 4 and 24 hours post-ART dosing from 58 subjects receiving NVP-containing ART and used to determine cNVP and viral load (VL). Median duration of treatment was 42 (range, 12-156) months, and 25 (43.1%) of the patients had virologic failure (VF). cNVP was significantly lower for VF than non- VF at 1hr (mean, 2,111ng/ml vs. 3,432ng/ml, p = 0.003) and at 4hr (mean 1,625ng/ml vs. 3,999ng/ml, p = 0.001) but not at 24hr post-ART dosing. Up to 53.4%, 24.1% and 22.4% of the subjects had good, fair and poor adherence respectively. cNVP levels peaked and were > = 3µg.ml at 4 hours in a majority of patients with good adherence and those without VF. Using a threshold of 3µg/ml for optimal therapeutic nevirapine level, 74% (43/58), 65.5% (38/58) and 86% (50/58) of all patients had sub-therapeutic cNVP at 1, 4 and 24 hours respectively. cNVP at 4 hours was associated with adherence (p = 0.05) and virologic VF (p = 0.002) in a chi-square test. These mean cNVP levels differed significantly in non-parametric tests between adherence categories at 1hr (p = 0.005) and 4hrs (p = 0.01) and between ART regimen categories at 1hr (p = 0.004) and 4hrs (p<0.0001). Moreover, cNVP levels correlated inversely with VL (p< = 0.006) and positively with adherence behavior. In multivariate tests, increased early peak NVP (cNVP4) was independently predictive of lower VL (p = 0.002), while delayed high NVP peak (cNVP24) was consistent with increased VL (p = 0.033). These data strongly assert the need to integrate plasma concentrations of NVP and that of other ART drugs into routine ART management of HIV-1 patients.

Rapid and high throughput molecular identification of diverse mosquito species by high resolution melting analysis.

Mosquitoes are a diverse group of invertebrates, with members that are among the most important vectors of diseases. The correct identification of mosquitoes is paramount to the control of the diseases that they transmit. However, morphological techniques depend on the quality of the specimen and often unavailable taxonomic expertise, which may still not be able to distinguish mosquitoes among species complexes (sibling and cryptic species). High resolution melting (HRM) analyses, a closed-tube, post-polymerase chain reaction (PCR) method used to identify variations in nucleic acid sequences, has been used to differentiate species within the Anopheles gambiae and Culex pipiens complexes. We validated the use of PCR-HRM analyses to differentiate species within Anopheles and within each of six genera of culicine mosquitoes, comparing primers targeting cytochrome b ( cyt b), NADH dehydrogenase subunit 1 (ND1), intergenic spacer region (IGS) and cytochrome c oxidase subunit 1 ( COI) gene regions. HRM analyses of amplicons from all the six primer pairs successfully differentiated two or more mosquito species within one or more genera ( Aedes ( Ae. vittatus from Ae. metallicus), Culex ( Cx. tenagius from Cx. antennatus, Cx. neavei from Cx. duttoni, cryptic Cx. pipiens species), Anopheles ( An. gambiae s.s. from An. arabiensis) and Mansonia ( Ma. africana from Ma. uniformis)) based on their HRM profiles. However, PCR-HRM could not distinguish between species within Aedeomyia ( Ad. africana and Ad. furfurea), Mimomyia ( Mi. hispida and Mi. splendens) and Coquillettidia ( Cq. aurites, Cq. chrysosoma, Cq. fuscopennata, Cq. metallica, Cq. microannulatus, Cq. pseudoconopas and Cq. versicolor) genera using any of the primers. The IGS and COI barcode region primers gave the best and most definitive separation of mosquito species among anopheline and culicine mosquito genera, respectively, while the other markers may serve to confirm identifications of closely related sub-species. This approach can be employed for rapid identification of mosquitoes.

Variation in the response of eastern and southern Africa provenances of Faidherbia albida (Delile A. Chev) seedlings to water supply: A greenhouse experiment.

Rural communities value Faidherbia albida in farming systems and pastoralism. Faidherbia albida provides products such as medicine, fodder, fuel, wood, food and services such as shade, soil fertility and nutrient cycling. Excessive browsing by animals, branch lopping and pod harvesting, have critically reduced the natural regeneration in some areas which exposes it to challenges due to dependence upon natural regeneration. The objective of this research was to evaluate response of Faidherbia albida provenances from eastern (Taveta Wangingombe) and southern Africa (Lupaso, Kuiseb Manapools) to different watering regimes to aid in selection of provenances for domestication. The observed difference in growth was analyzed to determine whether they are genetic or environmentally induced. Genotype  [Formula: see text]  interaction were significant at ([Formula: see text]0.001, p[Formula: see text]0.05) in seedling height, diameter and leaf numbers. Seedling height (r=0.94 p=0.001) recorded the highest correlation coefficient among all the growth variables analyzed. The growth variation was greater for seedling height than that of diameter and leaf numbers (h[Formula: see text]=0.97). Hierarchical cluster analysis grouped the provenances into three clusters with cluster iii consisting of Taveta, Kuiseb and Lupaso while cluster ii and i composed of Wangingombe and Manapools respectively. Manapools recorded the highest genetic distance from Taveta, Kuiseb and Lupaso at 84.55 units. Wangingombe and Manapools are closely related genetically at a distance of 7.32. The maximum inter-cluster distance between cluster i and iii indicated wider genetic diversity between the provenances in these clusters and selection should be from this clusters for hybridization program to achieve novel breeds.