Life-history attributes of juvenile Anopheles gambiae s.s. in central Uganda; implications for malaria control interventions.

Malaria is among the leading causes of death in Uganda, and Anopheles gambiae sensu stricto (s.s.) is the predominant vector. Although current vector control interventions have greatly reduced the malaria burden, the disease persists. New interventions are needed in order to eradicate them. Evaluation of new tools will require the availability of well-characterized test vector populations. Juvenile An. gambiae s.s. from Kibbuye and Kayonjo-derived populations were characterized under semi-field and laboratory conditions, given that various vector traits, including abundance and fitness are dependent on development profiles at this life stage. Ten replicates comprising 30 first instar larvae each were profiled for various life-history attributes (egg hatching, larval development time, larval survivorship, pupal weight and pupation rate). All parameters were similar for the two sites under laboratory conditions. However, the similarities or differences between field and laboratory development were parameter-specific. Whereas, larval survivorship and pupal weight were similar across seasons and laboratory in colonies from both sites, in the semi-field settings, pupation rate and larval survivorship differed between seasons in both sites. In addition, the average larval development time during the wet season was longer than that of the laboratory for both sites. Availability of mirror field sites is important for future tool evaluations.

House design and risk of malaria, acute respiratory infection and gastrointestinal illness in Uganda: A cohort study.

House construction is rapidly modernizing across Africa but the potential benefits for human health are poorly understood. We hypothesised that improvements to housing would be associated with reductions in malaria, acute respiratory infection (ARI) and gastrointestinal illness in an area of low malaria endemicity in Uganda. Data were analysed from a cohort study of male and female child and adult residents (n = 531) of 80 randomly-selected households in Nagongera sub-county, followed for 24 months (October 4, 2017 to October 31, 2019). Houses were classified as modern (brick walls, metal roof and closed eaves) or traditional (all other homes). Light trap collections of mosquitoes were done every two weeks in all sleeping rooms. Every four weeks, we measured malaria infection (using microscopy and qPCR to detect malaria parasites), incidence of malaria, ARI and gastrointestinal illness. We collected 15,780 adult female Anopheles over 7,631 nights. We collected 13,277 blood samples of which 10.2% (1,347) were positive for malaria parasites. Over 958 person years we diagnosed 38 episodes of uncomplicated malaria (incidence 0.04 episodes per person-year at risk), 2,553 episodes of ARI (incidence 2.7 episodes per person-year) and 387 episodes of gastrointestinal illness (incidence 0.4 episodes per person-year). Modern houses were associated with a 53% lower human biting rate compared to traditional houses (adjusted incidence rate ratio [aIRR] 0.47, 95% confidence interval [CI] 0.32-0.67, p<0.001) and a 24% lower incidence of gastrointestinal illness (aIRR 0.76, 95% CI 0.59-0.98, p = 0.04) but no changes in malaria prevalence, malaria incidence nor ARI incidence. House improvements may reduce mosquito-biting rates and gastrointestinal illness among children and adults. For the health sector to leverage Africa's housing modernization, research is urgently needed to identify the healthiest house designs and to assess their effectiveness across a range of epidemiological settings in sub-Saharan Africa.

Identification and characterization of immature Anopheles and culicines (Diptera: Culicidae) at three sites of varying malaria transmission intensities in Uganda.

BACKGROUND: Over the last two decades, there has been remarkable progress in malaria control in sub-Saharan Africa, due mainly to the massive deployment of long-lasting insecticidal nets and indoor residual spraying. Despite these gains, it is clear that in many situations, additional interventions are needed to further reduce malaria transmission. The World Health Organization (WHO) has promoted the Integrated Vector Management (IVM) approach through its Global Vector Control Response 2017-2030. However, prior roll-out of larval source management (LSM) as part of IVM, knowledge on ecology of larval aquatic habitats is required. METHODS: Aquatic habitats colonized by immature Anopheles and culicines vectors were characterized at three sites of low, medium and high malaria transmission in Uganda from October 2011 to June 2015. Larval surveys were conducted along transects in each site and aquatic habitats described according to type and size. Immature Anopheles, culicines and pupae from the described habitats were sampled using standard dipping methods to determine larval and pupae densities. Larvae were identified as anopheline or culicine, and counted. Pupae were not identified further. Binary logistic regression analysis was used to identify factors associated with the presence of immature Anopheles and culicines in each site. RESULTS: A total of 1205 larval aquatic habitats were surveyed and yielded a total of 17,028 anopheline larvae, 26,958 culicine larvae and 1189 pupae. Peaks in larval abundance occurred in all sites in March-May and August-October coinciding with the rainy seasons. Anopheles larvae were found in 52.4% (n = 251) of aquatic habitats in Tororo, a site of high transmission, 41.9% (n = 536) of habitats in Kanungu, a site with moderate malaria transmission, and 15.8% (n = 418) in Jinja, a site with low malaria transmission. The odds of finding larvae was highest in rice fields compared to pools in both Tororo (odds ratio, OR = 4.21, 95% CI 1.22-14.56, p = 0.02) and Kanungu (OR = 2.14, 95% CI 1.12-4.07, p = 0.02), while in Jinja the odd were highest in containers (OR = 4.55, 95% CI = 1.09-19.14, p = 0.03). In Kanungu, larvae were less likely to be found in containers compared to pools (OR = 0.26, 95% CI 0.09-0.66, p = 0.008) and river fringe (OR = 0.19, 95% CI 0.07-0.52, p = 0.001). Medium sized habitats were associated with high odds of finding larvae compared to small habitats (OR = 3.59, 95% CI 1.18-14.19, p = 0.039). CONCLUSIONS: These findings show that immature Anopheles and culicines were common in areas of high and moderate transmission but were rare in areas of low transmission. Although immature Anopheles and culicines were found in all types of water bodies, they were most common in rice fields and less common in open drains and in river fringes. Methods are needed to reduce the aquatic stages of anopheline mosquitoes in human-made habitats, particularly rice fields.

Impact of vector control interventions on malaria transmission intensity, outdoor vector biting rates and Anopheles mosquito species composition in Tororo, Uganda.

BACKGROUND: Long-lasting insecticidal nets (LLINs) and indoor residual spraying of insecticide (IRS) are widely recommended for the prevention of malaria in endemic regions. Data from human landing catches provide information on the impact of vector control on vector populations. Here, malaria transmission indoors and outdoors, before and after mass deployment of LLINs and IRS in Uganda was compared. METHODS: The study took place in Tororo district, a historically high transmission area where universal LLIN distribution was conducted in November 2013 and May 2017 and 6 rounds of IRS implemented from December 2014 to July 2018. Human landing catches were performed in 8 houses monthly from October 2011 to September 2012 (pre-intervention period) and every 4 weeks from November 2017 to October 2018 (post-intervention period). Mosquitoes were collected outdoors from 18:00 to 22:00 h and indoors from 18:00 to 06:00 h. Female Anopheles were tested for the presence of Plasmodium falciparum sporozoites and species identification performed using gross dissection and polymerase chain reaction (PCR). RESULTS: The interventions were associated with a decline in human biting rate from 19.6 to 2.3 female Anopheles mosquitoes per house per night (p < 0.001) and annual entomological inoculation rate from 129 to 0 infective bites per person per year (p < 0.001). The proportion of mosquitoes collected outdoors increased from 11.6 to 49.4% (p < 0.001). Prior to the interventions the predominant species was Anopheles gambiae sensu stricto (s.s.), which comprised an estimated 76.7% of mosquitoes. Following the interventions, the predominant species was Anopheles arabiensis, which comprised 99.5% of mosquitoes, with almost complete elimination of An. gambiae s.s. (0.5%). CONCLUSIONS: Mass distribution of LLINs and 6 rounds of IRS dramatically decreased vector density and sporozoite rate resulting in a marked reduction in malaria transmission intensity in a historically high transmission site in Uganda. These changes were accompanied by a shift in vector species from An. gambiae s.s. to An. arabiensis and a relative increase in outdoor biting.

First detection of Paenibacillus larvae the causative agent of American Foulbrood in a Ugandan honeybee colony.

Paenibacillus larvae is a highly contagious and often lethal widely distributed pathogen of honeybees, Apis mellifera but has not been reported in eastern Africa to date. We investigated the presence of P. larvae in the eastern and western highland agro-ecological zones of Uganda by collecting brood and honey samples from 67 honeybee colonies in two sampling occasions and cultivated them for P. larvae. Also, 8 honeys imported and locally retailed in Uganda were sampled and cultivated for P. larvae. Our aim was to establish the presence and distribution of P. larvae in honeybee populations in the two highland agro-ecological zones of Uganda and to determine if honeys that were locally retailed contained this lethal pathogen. One honeybee colony without clinical symptoms for P. larvae in an apiary located in a protected area of the western highlands of Uganda was found positive for P. larvae. The strain of this P. larvae was genotyped and found to be ERIC I. In order to compare its virulence with P. larvae reference strains, in vitro infection experiments were conducted with carniolan honeybee larvae from the research laboratory at Ghent University, Belgium. The results show that the virulence of the P. larvae strain found in Uganda was at least equally high. The epidemiological implication of the presence of P. larvae in a protected area is discussed.

Factors influencing the prevalence and infestation levels of Varroa destructor in honeybee colonies in two highland agro-ecological zones of Uganda.

Varroa mites are ecto-parasites of honeybees and are a threat to the beekeeping industry. We identified the haplotype of Varroa mites and evaluated potential factors that influence their prevalence and infestation levels in the eastern and western highland agro-ecological zones of Uganda. This was done by collecting samples of adult worker bees between December 2014 and September 2015 in two sampling moments. Samples of bees were screened for Varroa using the ethanol wash method and the mites were identified by molecular techniques. All DNA sequences obtained from sampled mite populations in the two zones were 100 % identical to the Korean Haplotype (AF106899). Mean mite prevalence in the apiaries was 40 and 53 % for the western and eastern zones, respectively, during the first sampling. Over the second sampling, mean mite prevalence increased considerably in the western (59 %) but not in the eastern (51 %) zone. Factors that were associated with Varroa mite infestation levels include altitude, nature of apiary slope and apiary management practices during the first sampling. Our results further showed that Varroa mites were spreading from lower to higher elevations. Feral colonies were also infested with Varroa mites at infestation levels not significantly different from those in managed colonies. Colony productivity and strength were not correlated to mite infestation levels. We recommend a long-term Varroa mite monitoring strategy in areas of varying landscape and land use factors for a clear understanding of possible changes in mite infestation levels among African honeybees for informed decision making.

Temporal patterns in Saturnidae (silk moth) and Sphingidae (hawk moth) assemblages in protected forests of central Uganda.

Forest-dependent biodiversity is threatened throughout the tropics by habitat loss and land-use intensification of the matrix habitats. We resampled historic data on two moth families, known to play central roles in many ecosystem processes, to evaluate temporal changes in species richness and community structure in three protected forests in central Uganda in a rapidly changing matrix. Our results show some significant declines in the moth species richness and the relative abundance and richness of forest-dependent species over the last 20-40 years. The observed changes in species richness and composition among different forests, ecological types, and moth groups highlight the need to repeatedly monitor biodiversity even within protected and relatively intact forests.

Biting patterns and seasonality of Anopheles gambiae sensu lato and Anopheles funestus mosquitoes in Kamuli District, Uganda.

BACKGROUND: We investigated the biting patterns and seasonal abundances of Anopheles gambiae s.l. and An. funestus mosquitoes in Kamuli District, Uganda. METHODS: Hourly indoor and outdoor catches of human biting mosquitoes were sampled from 19.00 to 07.00 hours for four consecutive nights each month using bed net traps in forty-eight houses randomly selected from Bugabula county where insecticide-treated bed nets (ITNs) had been used for at least five years and Budiope county where ITNs had not been used. The indoor and outdoor human-biting fractions, time of biting of the anophelines and climatic data were recorded from January to December 2010. Data were analysed using Multi-way analysis of variance, Kruskal-wallis rank sum test and Pearson correlation. The number of mosquitoes caught biting humans and resting indoors, the indoor and outdoor human biting densities and biting rates during different hours of the night, and mosquito abundances for a twelve-month sampling period in both zones are reported. RESULTS: Approximately four times more Anopheles mosquitoes were caught biting humans in Budiope County than in the Bugabula zone, with An. gambiae s. l. catches exceeding those of An. funestus. In both zones, peak night biting occurred between 23.00 and 05.00 hours. The majority of bites occurred between 03.00 and 06.00 hours for both Anopheles gambiae s. l. and funestus group. Outdoor biting densities of Anopheles gambiae s. l. exceeded the indoor biting densities throughout the night in both zones, while the indoor and outdoor human biting densities of An. funestus group were apparently equal. The outdoor and indoor human biting rates were similar in both zones. In Bugabula county, the abundance of An. gambiae s.l. was rainfall-dependent, while the An. funestus group could thrive with or without rain fall. In Budiope county, both An. gambiae s.l. and An. funestus mosquitoes thrived all year round regardless of the amount of rainfall. CONCLUSION: Considering the biting patterns, and seasonal abundances exhibited by Anopheles gambiae s.l. and An. funestus mosquitoes in Kamuli district, intensive use of ITNs combined with indoor residual spraying, environmental management and improved house designs in the context of integrated vector management may be the appropriate vector control strategy.