Sex differentiation studies relating to releaser aggregation pheromones of the desert locust,Schistocerca gregaria

Behavioural responses of nymphs and adults in the gregarious phase of the desert locust,Schistocerca gregaria (Forskal) (Orthoptera: Acrididae) were investigated in a single-chamber bioassay system to a choice of two columns of air, one permeated with airborne volatiles emanating from either sex of nymphs or adults and the other untreated. There was no sexual differentiation in the production of or response to nymphal volatiles. Young adults of either sex did not produce a stimulus with significant activity. Of the older adults, only the males produced the aggregation stimulus to which both sexes were equally responsive. Charcoal-trapped volatiles from the two sexes of nymphs and adults evoked similar aggregation responses. Antennae of the older adults showed significantly higher EAG responses than those of fifth instar nymphs to all four volatile collections, of which volatiles from older adult males were the most stimulatory and evoked the highest EAG amplitudes.     

Afrotropical sand fly-host plant relationships in a leishmaniasis endemic area,Kenya

The bioecology of phlebotomine sand flies is intimately linked to the utilization of environmental resources including plant feeding. However, plant feeding behavior of sand flies remains largely understudied for Afrotropical species. Here, using a combination of biochemical, molecular, and chemical approaches, we decipher specific plant-feeding associations in field-collected sand flies from a dry ecology endemic for leishmaniasis in Kenya. Cold-anthrone test indicative of recent plant feeding showed that fructose positivity rates were similar in both sand fly sexes and between those sampled indoors and outdoors. Analysis of derived sequences of the ribulose-1,5-bisphosphate carboxylase large subunit gene (rbcL) from fructose-positive specimens implicated mainly Acacia plants in the family Fabaceae (73%) as those readily foraged on by both sexes of Phlebotomus and Sergentomyia. Chemical analysis by high performance liquid chromatography detected fructose as the most common sugar in sand flies and leaves of selected plant species in the Fabaceae family. Analysis of similarities (ANOSIM) of the headspace volatile profiles of selected Fabaceae plants identified benzyl alcohol, (Z)-linalool oxide, (E)-β-ocimene, p-cymene, p-cresol, and m-cresol, as discriminating compounds between the plant volatiles. These results indicate selective sand fly plant feeding and suggest that the discriminating volatile organic compounds could be exploited in attractive toxic sugar- and odor- bait technologies control strategies.     

Monitoring Aethina tumida (Coleoptera: Nitidulidae) with baited bottom board traps: occurrence and seasonal abundance in honey bee colonies in Kenya

The population dynamics of the honey bee pest Aethina tumida Murray (small hive beetle) have been studied in the United States with flight and Langstroth hive bottom board traps baited with pollen dough inoculated with a yeast Kodamaea ohmeri associated with the beetle. However, little is known about the population dynamics of the beetle in its native host range. Similarly baited Langstroth hive bottom board traps were used to monitor the occurrence and seasonal abundance of the beetle in honey bee colonies at two beekeeping locations in Kenya. Trap captures indicated that the beetle was present in honey bee colonies in low numbers all year round, but it was most abundant during the rainy season, with over 80% trapped during this period. The survival of larvae was tested in field releases under dry and wet soil conditions, and predators of larvae were identified. The actvity and survival of the beetle were strongly influenced by a combination of abiotic and biotic factors. Larval survival was higher during wet (28%) than dry (1.1%) conditions, with pupation occurring mostly at 0-15 cm and 11-20 cm, respectively, beneath the surface soil during these periods. The ant Pheidole megacephala was identified as a key predator of larvae at this site, and more active during the dry than wet seasons. These observations imply that intensive trapping during the rainy season could reduce the population of beetles infesting hives in subsequent seasons especially in places where the beetle is a serious pest.     

Plant growth-promoting rhizobacteria do not pose any deleterious effect on cowpea and detectable amounts of ethylene are produced

Summary The use of trap crops such as cowpea could reduce the effects of the root parasitic weed, Striga hermonthica and its subsequent constraints on the growth of cereals. Certain bacteria could augment the trap crop stimulatory effect. We studied the effect of three bacteria introduced to the rhizosphere of three cowpea varieties at planting. Number of days to cowpea flowering was noted and at harvest, data were collected on pod characteristics and biomass. Means of data subjected to ANOVA were compared using Tukey’s Studentized Range Test. We analysed bacterial headspace volatiles for ethylene by gas chromatography and gas chromatography–mass spectrometry. Bacterial type significantly influenced the cowpea varieties with better performance over the non-inoculated control. Average pod weight (g) with bacterial treatment was 37.97 for Enterobacter sakazakii 8MR5, 34.38 for Pseudomonas 44MS8 and 27.46 for Pseudomonas 10M3. Non-inoculated control had an average weight of 20.98 g. Bacteria promoted a significant increase in pod weight (≥30.89%), fresh biomass (≥24.22%), and improved pod number (≥20.54%) and pod wall thickness (≥7.33%) with no deleterious effect on plant health. Ethylene released by the bacteria ranged from trace concentrations in Pseudomonas sp. to 210 nmoles/10⁸ c. f. u./ml in Ent. sakazakii 8MR5.     

Perigestational suppression of weight gain with central leptin gene therapy results in lower weight F1 generation

The efficacy of central leptin therapy on weight homeostasis through various phases of reproduction, pregnancy outcome and postnatal, prepubertal and pubertal growth of offspring was assessed. Enhanced leptin transgene expression after a single intracerebroventricular injection of recombinant adeno-associated virus vector encoding the leptin gene (rAAV-lep) decreased calorie intake and weight in adult nulliparous female rats. rAAV-lep treated rats conceived normally, displayed unremarkable pregnancy rate, parturition and delivered normal sized litters. Significantly lower weight was maintained through gestation, lactation, and post-lactation periods. The maintenance of a modest weight reduction was accompanied by voluntarily reduced calorie intake, increased thermogenic energy expenditure, decreased adiposity as reflected by drastically reduced leptin levels, and suppressed insulin and insulin-like growth factor 1 levels through lactation and post-lactation in rAAV-lep treated dams. The offspring at birth weighed significantly less than those of controls and this lower weight range was sustained during postnatal, prepubertal, pubertal and adult (3 months old) periods, contemporaneous with metabolic circulating hormones in the normal range. For the first time we show the persistent efficacy of central leptin gene therapy to suppress weight gain through all phases of reproduction, lactation and post-lactation in dams and reveal the potential imprinting link to producing lower weight in the F1 generation.     

Population Genetics of Two Key Mosquito Vectors of Rift Valley Fever Virus Reveals New Insights into the Changing Disease Outbreak Patterns in Kenya

Rift Valley fever (RVF) outbreaks in Kenya have increased in frequency and range to include northeastern Kenya where viruses are increasingly being isolated from known (Aedes mcintoshi) and newly-associated (Ae. ochraceus) vectors. The factors contributing to these changing outbreak patterns are unclear and the population genetic structure of key vectors and/or specific virus-vector associations, in particular, are under-studied. By conducting mitochondrial and nuclear DNA analyses on >220 Kenyan specimens of Ae. mcintoshi and Ae. ochraceus, we uncovered high levels of vector complexity which may partly explain the disease outbreak pattern. Results indicate that Ae. mcintoshi consists of a species complex with one of the member species being unique to the newly-established RVF outbreak-prone northeastern region of Kenya, whereas Ae. ochraceus is a homogeneous population that appears to be undergoing expansion. Characterization of specimens from a RVF-prone site in Senegal, where Ae. ochraceus is a primary vector, revealed direct genetic links between the two Ae. ochraceus populations from both countries. Our data strongly suggest that unlike Ae. mcintoshi, Ae. ochraceus appears to be a relatively recent, single ''introduction'' into Kenya. These results, together with increasing isolations from this vector, indicate that Ae. ochraceus will likely be of greater epidemiological importance in future RVF outbreaks in Kenya. Furthermore, the overall vector complexity calls into question the feasibility of mosquito population control approaches reliant on genetic modification.     

Central leptin gene therapy blocks ovariectomy-induced adiposity

Objective: In this study, we tested the hypothesis that insufficiency of leptin restraint in the hypothalamus is responsible for promoting weight gain and adiposity after ovariectomy (ovx). Whether increasing leptin transgene expression can overcome the diminution in leptin restraint was evaluated in ovx rats. Research Methods and Procedures: Enhanced leptin or green fluorescent protein (GFP; control) transgene expression was induced by a single intracerebroventricular injection of recombinant adeno‐associated viral vector encoding either leptin gene (rAAV‐lep) or GFP gene (rAAV‐GFP; control) in acutely and chronically ovx rats. Body weight and food intake responses were monitored weekly. White adipose tissue (WAT) mass and serum levels of WAT‐derived hormones, leptin, and adiponectin were analyzed at termination of the experiments. Results and Discussion: An increase in leptin transgene expression in the hypothalamus initiated soon after ovx blocked hyperphagia and body weight gain and markedly suppressed WAT mass and adipokines, leptin, and adiponectin. Similar suppression of weight gain and adiposity and serum leptin and adiponectin levels after intracerebroventricular rAAV‐lep injection in chronically ovx rats were observed concomitant with unchanged daily food intake. These findings are consistent with the hypothesis that in the absence of ovarian steroids, the existent insufficiency of leptin restraint at the hypothalamic level can be overcome with ectopic leptin expression, thereby reinstating central control on weight and adiposity.     

Responses of the ambrosia beetle <Emphasis Type="Italic">Xylosandrus compactus</Emphasis> (Coleoptera: Curculionidea: Scolytinae) to volatile constituents of its symbiotic fungus <Emphasis Type="Italic">Fusarium solani</Emphasis> (Hypocreales: Nectriaceae)

Xylosandrus compactus is a polyphagus pest that cultivates a symbiotic fungus Fusarium solani in tunnels of host plants for food and is a major threat to coffee production in East Africa. We hypothesized that the female X. compactus, which is the only sex capable of flying to attack its hosts, is attracted to volatiles from F. solani. We investigated responses of females to volatiles released by fungal cultures and bioactive components identified in the fungal volatiles. In Y-tube olfactometer assays, ~68% of females were attracted to volatiles emitted from F. solani over clean air. Bioactive compounds were identified in the fungal volatiles by coupled gas chromatography (GC)/electroantennographic detection (EAD) and GC/mass spectrometric analyses as methyl isovalerate and 2,3-butanediol. We also identified ethanol, a known attractant of X. compactus, using solid phase microextraction captured fungal volatiles analyzed by GC/MS. In field trapping trials, we compared captures of females in plastic bottle traps baited with a range of doses of methyl isovalerate, 2,3-butanediol, and blends of the two compounds, with similar traps baited with solvent only and ethanol. Females were caught by all the baited traps at all the concentrations tested except traps baited with solvent only. Trap captures were however 14–37-fold lower in traps baited with single components and the blends than those baited with ethanol. The possible use of these components as a tool for kairomonal monitoring of populations of X. compactus is discussed.     

Survival rate,blood feeding habits and sibling species composition of Aedes simpsoni complex: Implications for arbovirus transmission risk in East Africa

Aedes simpsoni complex has a wide distribution in Africa and comprises at least three described sub-species including the yellow fever virus (YFV) vector Ae. bromeliae. To date, the distribution and relative contributions of the sub-species and/or subpopulations including bionomic characteristics in relation to YF transmission dynamics remain poorly studied. In this study conducted in two areas with divergent ecosystems: peri-urban (coastal Rabai) and rural (Rift Valley Kerio Valley) in Kenya, survival rate was estimated by parity in Ae. simpsoni s.l. mosquitoes sampled using CO₂-baited BG Sentinel traps. We then applied PCR targeting the nuclear internal transcribed spacer 2 (ITS2), region followed by sequencing and phylogenetic analytics to identify the sibling species in the Ae. simpsoni complex among parous and blood fed cohorts. Our results show that Ae. bromeliae was the most dominant sub-species in both areas, exhibiting high survival rates, human blood-feeding, and potentially, high vectorial capacity for pathogen transmission. We document for the first time the presence of Ae. lilii in Kenya and potentially yet-to-be described species in the complex displaying human feeding tendencies. We also infer a wide host feeding range on rodents, reptile, and domestic livestock besides humans especially for Ae. bromeliae. This feeding trend could likely expose humans to various zoonotic pathogens. Taken together, we highlight the utility of genotype-based analyses to generate precision surveillance data of vector populations for enhanced disease risk prediction and to guide cost-effective interventions (e.g. YF vaccinations).     

Potential of the Desert Locust Schistocerca gregaria (Orthoptera: Acrididae) as an Unconventional Source of Dietary and Therapeutic Sterols

Insects are increasingly being recognized not only as a source of food to feed the ever growing world population but also as potential sources of new products and therapeutic agents, among which are sterols. In this study, we sought to profile sterols and their derivatives present in the desert locust, Schistocerca gregaria, focusing on those with potential importance as dietary and therapeutic components for humans. Using coupled gas chromatography-mass spectrometry (GC-MS), we analyzed and compared the quantities of sterols in the different sections of the gut and tissues of the locust. In the gut, we identified 34 sterols which showed a patchy distribution, but with the highest composition in the foregut (55%) followed by midgut (31%) and hindgut (14%). Fed ad libitum on wheat seedlings, five sterols unique to the insect were detected. These sterols were identified as 7-dehydrocholesterol, desmosterol, fucosterol, (3β, 5α) cholesta-8, 14, 24-trien-3-ol, 4, 4-dimethyl, and (3β, 20R) cholesta-5, 24-dien-3, 20-diol with the first three having known health benefits in humans. Incubation of the fore-, mid- and hindgut with cholesterol-[4-¹³C] yielded eight derivatives, three of these were detected in the gut of the desert locust after it had consumed the vegetative diet but were not detected in the diet. Our study shows that the desert locust ingests phytosterols from a vegetative diet and, amplifies and metabolizes them into derivatives with potential salutary benefits and we discuss our findings in this context.