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Forest conservation by peripheral communities is one of the most advocated strategies for ensuring sustainable supply of forest goods and services. In Kenya, although conservation activities genera...

Summary There is limited information about the relative composition and health benefits of various honey consumed across Africa. This study aimed at estimating the bioactive constituents, in vitro radical scavenging and antibacterial activities of 16 kinds of honey obtained from different geographical locations in Kenya. Manuka 5 + honey was included for comparison. Some of the tested honey had biochemicals and bioactivities similar to or higher than Manuka 5 + honey. The honey exhibited DPPH radical scavenging ability, with several types of honey showing superior scavenging potential than Manuka 5 + honey, owing to their high phenol content. All types of honey inhibited the growth of E. coli and further showed a substantial amount of nonperoxide antimicrobial activity. The geographical origin of honey had an influence on its bioactive contents. Overall, these findings suggest that Kenyan honey has great therapeutic potential, and thus, its clinical application should not be overlooked., Kenya is rich in floral diversity that is unique from one geographical location to the other. This provides the honeybees an opportunity to process honey of varying composition. These honeys contain among other components, the phytochemicals like phenols, flavonoids, and other bioactives, just like manuka 5+ honey. Some of these bioactives contribute significantly to the antibacterial and antioxidant properties of honey. It can therefore be deduced that the consumption of Kenyan honey will be beneficial to human health.

The variegated coffee bug Antestiopsis thunbergii Gmelin is a key pest of Coffea arabica in East Africa. Although the bug feeds on various parts of the coffee plant, it has a strong preference for mature green berries which are essential for the bug to complete its life cycle, reproduce and enhance its longevity. To locate mature green coffee berries, we hypothesized that second instar nymphs, which are the most mobile and active feeding immature stage of the bug, must rely on key volatile compounds emitted by the host for recognition. We tested this hypothesis using behavioral and electrophysiological assays and chemical analysis. In olfactometer assays, the second instar nymphs were strongly attracted to volatiles emitted from mature green berries but avoided those from ripe coffee berries of C. arabica. Coupled gas chromatography–electroantennographic detection (GC/EAD) isolated five antenally active components from mature green berries volatiles, four of which were identified by coupled GC–mass spectrometry as toluene, anisole, methyl 3-ethyl-4-methylpentanoate and (5S,7S)-conophthorin. In concentration assays, in which second instar nymphs did not respond to toluene, they were strongly attracted to anisole, methyl 3-ethyl-4-methylpentanoate and (5S,7S)-conophthorin, and a blend from the three compounds at concentrations lower or equivalent to the natural volatile extract. Our results suggest that the blend from these three compounds allows host recognition in second instar nymphs of the variegated coffee bug and is a candidate kairomone for monitoring the pest in coffee plantations.

The present study was part of a larger surveillance effort to identify the determinants of African honey bee health, and, particularly, to detect honey bee pathogens across Kenya, where 160 colonies were examined from 32 apiaries (five colonies/apiary). From each colony, 20 individual foragers, nurse bees, worker pupae, and drone pupae were sampled separately. These were organized as 30 foragers, 32 nurse bees, 28 worker pupae, and 10 drone pupae pools. Nucleic acid was extracted from the pooled homogenates and tested using a panel of 18 different (RT-)PCR methods targeted at detectingPaenibacillus larvae,Melissococcus plutonius,Ascophaera apis,Aspergillusspp.,Nosema ceranae, N. apis,Deformed wing virus(DWV),Varroa destructor virus 1(VDV 1),Acute bee paralysis virus(ABPV),Sacbrood virus(SBV),Israeli acute paralysis virus(IAPV),Black queen cell virus(BQCV),Chronic bee paralysis virus(CBPV), andKashmir bee virus. All amplified bands were sequenced and compared to the GenBank database. VDV 1 was the most abundant virus at 50% prevalence in the 100 bee pools. It was closely followed by DWV at 44%. The others were BQCV (36%), SBV (14%), IAPV (9%), ABPV (8%), andN. ceranae(5%). The pathogens co-existed within apiaries. VDV 1 was present in 66% of the apiaries, DWV in 69%, BQCV in 69%, SBV in 28%, IAPV in 22%, ABPV in 19%, andN. ceranaein 13%. The study concludes that these pathogens should be incorporated in honey bee disease surveillance activities in the region.

Background Small targets comprising panels of blue and insecticide-treated black netting material each 0.25 × 0.25 m have been shown to attract and kill Glossina fuscipes fuscipes Newstead, 1910 (Diptera: Glossinidae) thereby reducing its population density by over 90% in field trials. However, their attractive ability has not been fully exploited for sampling purposes. Therefore, in this study we assessed the effectiveness of using sticky small targets as sampling tools for G. f. fuscipes in western Kenya. We also determined the influence of colour on the landing response of female and male flies on sticky small targets. Methods Using a series of randomised block experiments, the numbers of tsetse flies caught with sticky small targets were compared with those caught with biconical traps. A negative binomial regression was used to model fly catches. Odds ratios as measures of association between the landing response on the blue or black panel of the sticky small target and the sex of flies were obtained from a multiple logistic regression. Results The results showed that sticky small targets caught 13.5 and 3.6 times more female and male tsetse flies than biconical traps (Z = 9.551, P < 0.0001 and Z = 5.978, P < 0.0001, respectively). Females had a 1.7 times likelihood of landing on the black panel than males (Z = 2.25, P = 0.025). Conclusion This study suggests that sticky small targets are an effective sampling tool for G. f. fuscipes. Therefore, we recommend the use of sticky small targets as an alternative to biconical traps for observational and experimental investigations of G. f. fuscipes.

Understanding the roles of volatile organic compounds (VOCs) in insect–plant interactions is a key component towards the development of safe pest management strategies and sustainable agriculture. Kairomones in unripe berries of Coffea arabica mediate host recognition of second instar nymphs of Antestia bugs, Antestiopsis thunbergii Gmelin, and are good candidates for the pest management. In the current study, we investigated the VOCs that contribute to deterring the pest from ripe berries. Behavioral assays showed that headspace volatiles collected from ripe berries were significantly avoided by second instar nymphs. A mixture of the ripe berry volatiles and unripe berries (known preferred diet) was also significantly avoided when tested against blank or unripe berries, thereby confirming that VOCs from ripe berries had altered the host recognition process and had an odor masking effect. Coupled gas chromatography/mass spectrometric (GC/MS) analysis of the headspace volatiles revealed a blend of chemicals of which ten elicited electrophysiological activity in antennae of second instar nymphs. Five of these compounds including; 3-hydroxy-2-butanone, 2-heptanone, 2-isopropyl-3-methoxypyrazine [IPMP], 2-isobutyl-3-methoxypyrazine [IBMP] and (E)-β-caryophyllene were identified as unique to the volatiles of ripe berries and they elicited avoidance behavior in second instar nymphs when tested singly and in a blend. In addition, their blend also inhibited responses of nymphs to a synthetic attractant blend (kairomone). Our results suggest that the blend of the five compounds can be exploited as repellents in the management of A. thunbergii by pushing the pest away from coffee plantations or interfering with the pest–host recognition process through masking of suitable host odors.

Pesticide residues in honey and pollen from Seychelles against a target of 108 pesticides using LC-MS/MS were analyzed. Fifteen pesticides were detected, at trace levels (

Displacement rates of tsetse affect performance of targets during vector control. Fly size, one of the indicators of population structure usually obtained from wing measurement, is among the determinants of displacement rates. Although recovery of tsetse in previous intervention areas has been widely reported, the population structure of tsetse that recover is rarely evaluated despite being associated with displacements rates. Previously, intervention trials had reduced tsetse densities by over 90% from3 flies/trap/day to1fly/trap/day on Big Chamaunga and Manga islands of Lake Victoria in western Kenya. In this study, we assessed the recovery in densities of Glossina fuscipes fuscipes on the two islands and evaluated the effects vector control might have on the population structure. A before and after intervention study was undertaken on four islands of Lake Victoria in western Kenya; Small and Big Chamaunga, Manga and Rusinga Islands, two of which tsetse control intervention had previously been undertaken. Three years after intervention average G. f. fuscipes catches in biconical traps were estimated on each island. Wing centroid size (CS) (a measurement of fly size) and shape, indicators of the population structure of flies from the four islands were compared using geometric morphometric analyses. CS and shape of available female but not male tsetse wings obtained before the intervention trial on Big and Small Chamaunga islands were compared with those from the same islands after the intervention trial. G. f. fuscipes apparent density on the previous intervention islands were9 flies/trap/day. Irrespective of sex, wing shape did not isolate tsetse based on their islands of origin. The fly size from Big and Small Chamaunga did not differ significantly before intervention trials (P = 0.728). However, three years after the intervention flies from Big Chamaunga were significantly smaller than those from Small Chamaunga (P 0.003). Further, there was an increase in the divergence of wing morphology between flies collected from Big Chamaunga and those from Small Chamaunga after tsetse control. In conclusion, even though populations are not isolated, vector control could influence the population structure of tsetse by exerting size and wing morphology differential selection pressures. Therefore, we recommend further studies to understand the mechanism behind this as it may guide future vector control strategies.

Bee keeping is indispensable to global food production. It is an alternate income source, especially in rural underdeveloped African settlements, and an important forest conservation incentive. However, dwindling honeybee colonies around the world are attributed to pests and diseases whose spatial distribution and influences are not well established. In this study, we used remotely sensed data to improve the reliability of pest ecological niche (EN) models to attain reliable pest distribution maps. Occurrence data on four pests (Aethina tumida, Galleria mellonella, Oplostomus haroldi and Varroa destructor) were collected from apiaries within four main agro-ecological regions responsible for over 80% of Kenya’s bee keeping. Africlim bioclimatic and derived normalized difference vegetation index (NDVI) variables were used to model their ecological niches using Maximum Entropy (MaxEnt). Combined precipitation variables had a high positive logit influence on all remotely sensed and biotic models’ performance. Remotely sensed vegetation variables had a substantial effect on the model, contributing up to 40.8% for G. mellonella and regions with high rainfall seasonality were predicted to be high-risk areas. Projections (to 2055) indicated that, with the current climate change trend, these regions will experience increased honeybee pest risk. We conclude that honeybee pests could be modelled using bioclimatic data and remotely sensed variables in MaxEnt. Although the bioclimatic data were most relevant in all model results, incorporating vegetation seasonality variables to improve mapping the ‘actual’ habitat of key honeybee pests and to identify risk and containment zones needs to be further investigated.

Background: The small hive beetle (SHB), Aethina tumida is an invasive pest of the honey bee. Although no previous methods have led to its successful management, yeast inoculated pollen baited-traps have showed promise as quick monitoring tools. In this study, we evaluated the role of olfaction in SHB response to Apicure®, an essential oil-based biopesticide that has shown potential for the management of honey bee pests and diseases. Methods: Volatiles from Apicure® were collected using super Q adsorbent traps. Subsequent analysis was done using Gas chromatography- mass spectrophotometry (GC-MS) to ascertain the components of Apicure®. The selectivity and sensitivity of antennal receptors of A. tumida adults to the volatile compounds were determined using behavioral assays and Gas Chromatography-Electroantennodetection (GC-EAD). Results: GC-MS analysis showed that Apicure® consists of 40 compounds. GC-EAD analysis isolated 11 compounds that elicited antennal response with the SHB. Of these, linalool, camphor, geraniol and α-terpineol were confirmed to be strongly repellant, while limonene was attractive to SHB in dual-choice olfactometer assays. Conclusion: Our results demonstrate that the major components in Apicure® are mainly repellants thus prospective in disrupting the host recognition by the SHB. The product therefore can be up-scaled for the management of SHB.

The presence of pollutants in honey can influence honey bee colony performance and devalue its use for human consumption. Using liquid chromatography tandem mass spectrometry (LC-MS/MS), various clean-up methods were evaluated for efficient determination of multiclass pesticide contaminants in honey. The selected clean-up method was optimized and validated and then applied to perform a preliminary study of commercial honey samples from Africa. The most efficient method was primary-secondary amine (PSA) sorbent which was significantly different from the others (P

Defining the structures and locations of the glycans attached on secreted proteins and virus envelope proteins is important in understanding how glycosylation affects their biological properties. Glycopeptide mass spectrometry (MS)-based analysis is a very powerful, emerging approach to characterize glycoproteins, in which glycosylation sites and the corresponding glycan structures are elucidated in a single MS experiment. However, to date there is not a consensus regarding which mass spectrometric platform provides the best glycosylation coverage information. Herein, we employ two of the most widely used MS approaches, online high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC/ESI-MS) and offline HPLC followed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), to determine which of the two approaches provides the best glycosylation coverage information of a complex glycoprotein, the group M consensus HIV-1 envelope, CON-S gp140DeltaCFI, which has 31 potential glycosylation sites. Our results highlight differences in the informational content obtained between the two methods such as the overall number of glycosylation sites detected, the numbers of N-linked glycans present at each site, and the type of confirmatory information obtained about the glycopeptide using MS/MS experiments. The two approaches are quite complementary, both in their coverage of glycopeptides and in the information they provide in MS/MS experiments. The information in this study contributes to the field of mass spectrometry by demonstrating the strengths and limitations of two widely used MS platforms in glycoprotein analysis.

Mass spectral analysis is an increasingly common method used to characterize glycoproteins. When more than one glycosylation site is present on a protein, obtaining MS data of glycopeptides is a highly effective way of obtaining glycosylation information because this approach can be used to identify not only what the carbohydrates are but also at which glycosylation site they are attached. Unfortunately, this is not yet a routine analytical approach, in part because data analysis can be quite challenging. We are developing strategies to simplify this analysis. Presented herein is a novel mass spectrometry technique that identifies the peptide moiety of either sulfated, sialylated, or both sialylated and sulfated glycopeptides. This technique correlates product ions in collision-induced dissociation (CID) experiments of suspected glycopeptides to a peptide composition using a newly developed web-based tool, GlycoPep ID. After identifying the peptide portion of glycopeptides with GlycoPep ID, the process of assigning the rest of the glycopeptide composition to the MS data is greatly facilitated because the "unknown" portion of the mass assignment that remains can be directly attributed to the carbohydrate component. Several examples of the utility and reliability of this method are presented herein.

Recently, there has been multi-agency promotion of entomophagy as an environmentally-friendly source of food for the ever increasing human population especially in the developing countries. However, food quality and safety concerns must first be addressed in this context. We addressed these concerns in the present study using the edible stink bug Encosternum delegorguei, which is widely consumed in southern Africa. We analysed for mycotoxins, and health beneficials including antioxidants, amino acids and essential fatty acids using liquid chromatography coupled to quadrupole time of flight mass spectrometry (LC-Qtof-MS) and coupled gas chromatography (GC)-MS. We also performed proximate analysis to determine nutritional components. We identified the human carcinogen mycotoxin (aflatoxin B1) at low levels in edible stink bugs that were stored in traditonally woven wooden dung smeared baskets and gunny bags previously used to store cereals. However, it was absent in insects stored in clean zip lock bags. On the other hand, we identified 10 fatty acids, of which 7 are considered essential fatty acids for human nutrition and health; 4 flavonoids and 12 amino acids of which two are considered the most limiting amino acids in cereal based diets. The edible stink bug also contained high crude protein and fats but was a poor source of minerals, except for phosphorus which was found in relatively high levels. Our results show that the edible stink bug is a nutrient- and antioxidant-rich source of food and health benefits for human consumption. As such, use of better handling and storage methods can help eliminate contamination of the edible stink bug with the carcinogen aflatoxin and ensure its safety as human food.

The HIV-1 envelope (Env) is a key determinant in mediating viral entry and fusion to host cells and is a major target for HIV vaccine development. While Env is typically about 50% glycan by mass, glycosylation sites are known to evolve, with some glycosylation profiles presumably being more effective at facilitating neutralization escape than others. Thus, characterizing glycosylation patterns of Env and native virions and correlating glycosylation profiles with infectivity and Env immunogenicity are necessary first steps in designing effective immunogens. Herein, we describe a mass spectrometry-based strategy to determine HIV-1 Env glycosylation patterns and have compared two mammalian cell expressed recombinant Env immunogens, one a limited immunogen and one that induces cross-clade neutralizing antibodies. We have used a glycopeptide-based mass mapping approach to identify and characterize Env's glycosylation patterns by elucidating which sites are utilized and what type of glycan motif is present at each glycosylation site. Our results show that the immunogens displayed different degrees of glycosylation as well as a different characteristic set of glycan motifs. Thus, these techniques can be used to (1) define glycosylation profiles of recombinant Env proteins and Env on mature virions, (2) define specific carbohydrate moieties at each glycosylation site, and (3) determine the role of certain carbohydrates in HIV-1 infectivity and in modulation of Env immunogenicity.

Follicle stimulating hormone (FSH) is one of the important hormones that regulate gonadal functions. This hormone is glycosylated, and the glycans greatly influence the biological properties. In the present study the negatively charged glycopeptides of equine and human pituitary follicle stimulating hormone (eFSH and hFSH) have been characterized in a glycosylation site-specific manner using FT-ICR-MS and Edman sequencing. The characteristic pattern of glycan distribution at each glycosylation site has been deduced and compared between horse and human FSH preparations. The data suggest that site-specific differences exist between glycoforms of human and equine FSH. For instance, except for one site in the beta subunit (Asn7) of hFSH all other sites in both species have sulfated glycoforms. Also, glycoforms at Asn52 of hFSH are all complex type, whereas in eFSH, both complex and hybrid structures exist at this site. There is also a higher percentage of sulfated glycans in the latter site compared to the former. This is the first study that characterizes the glycans from this hormone in a glycosylation site-specific manner, and these data can be used to begin correlative studies between glycosylation structure and hormone function.

Structural analysis of sulfated glycans is essential in understanding their biological significance. Here, we present a new approach to characterize sulfated glycans present on glycoproteins. The analysis is performed on glycopeptides, so information about the sulfated species is obtained in a glycosylation site-specific manner. This method employs an ion-pairing reagent to stabilize the SO3 group of the glycopeptide, allowing useful information to be obtained during MS/MS experiments. The amount of structural information obtained from (+)ESI-MS/MS of the ion-pair complexes for sulfated glycopeptides of equine thyroid stimulating hormone is compared with information obtained by (−)ESI-MS/MS of the underivatized, sulfated glycopeptides. The results indicate that this new method provides detailed insights into the sequence, branching, and type of N-glycans present, compared to analysis via (−)ESI-MS/MS.

We demonstrate a method that enhances the mass spectral signal of mono- and disulfated glycopeptides, present in glycoproteins that contain many other nonsulfated glycoforms. This method utilizes the tripeptide Lys-Lys-Lys as an ion-pairing reagent to complex selectively to sulfated species, and enhance their ion signal. The method is applied to the analysis of glycopeptides released from the enzymatic digestion of ovine luteinizing hormone. In this analysis, a disulfated glycopeptide is identified that was previously not detectable by MS assays, and a monosulfated glycoform, present at less than 1% abundance, is identified without any separation or enrichment of these species prior to analysis. In addition to enhancing the ion signal of sulfated glycopeptides, the ion-pairing technique is useful in obtaining structural information about the sulfated species.