Differential effects of Bacillus sphaericus strain 2362 on Culex quinquefasciatus and its competitor Culex cinereus in West Africa

The larval susceptibility to Bacillus sphaericus strain 2362 of the non-man-biting mosquito Culex cinereus and the urban filariasis vector Culex quinquefasciatus, two competitor mosquitoes in polluted habitats, was compared. In the laboratory, both species ingested a similar amount of B. sphaericus spores when fed c. 2 x 10(5) spores per ml for 30 min. However, in the same experiment, third-instar larvae of Cx quinquefasciatus were reduced by 98% at 24 h exposure while Cx cinereus larvae were only reduced by 6% at 72 h. In the field, preimaginal populations of Cx cinereus ingested, within a week, more than 99% of the applied spores, but showed no significant decrease through 14 days in cesspools treated at 10 g/m2 of a flowable concentrate of B. sphaericus 2362, containing 2 x 10(10) spores/g. It is proposed that specific biological control of Cx quinquefasciatus could result from appropriate treatment of breeding-sites with larvicidal B. sphaericus and competitive displacement by Cx cinereus or other mosquitoes with larvae that are more tolerant of B. sphaericus.     

Cross-resistance between strains of Bacillus sphaericus but not B. thuringiensis israelensis in colonies of the mosquito Culex quinquefasciatus

Two colonies of Culex quinquefasciatus Say (Diptera: Culicidae) were selected with Bacillus sphaericus strains C3-41 and IAB59 in the laboratory for 13 and 18 generations; they attained 145,000- and 48.3-fold resistance, respectively, in comparison with a susceptible laboratory colony (SLCq) and showed very high levels of cross-resistance (8500- to 145,000-fold) to B. sphaericus strains C3-41, 1593, 2297 and 2362. They were relatively susceptible to B. sphaericus strains LP1-G and 47-6B (only 0.8- to 2.8-fold tolerance), with 24.8- to 48.3-fold cross-resistance to strain IAB59. B. sphaericus-resistant mosquito colonies remained highly susceptible to B. thuringiensis israelensis, suggesting that B.t.i. would be of value in the management of B. sphaericus-resistant Cx. quinquefasciatus colonies. The demonstration of low or no cross-resistance of two selected resistant Cx. quinquefasciatus colonies to IAB59, LP1-G and 47-6B strains of B. sphaericus and the finding of a major 49 kDa protein in these strains suggest that there is likely to be another mosquitocidal factor in the three strains.     

Rapid habituation by mosquito larvae to predator kairomones

Larvae of some species of mosquitoes have been shown to respond to water‐borne kairomones from predators by reducing bottom‐feeding and replacing it with surface filter‐feeding, which uses less movement and is thus less likely to attract a predator. However, if no predator attack takes place, then it would be more efficient to use a risk allocation strategy of habituating their response depending on the predator and the overall risk. The larvae of Culiseta longiareolata Macquart live in temporary rain‐filled pools, where they are exposed to a high level of predation. Within one hour, they responded to kairomones from dragonfly or damselfly nymphs, or to the fish Aphanius, by significantly reducing bottom‐feeding activity. Continued exposure to the predator kairomones resulted in habituation of their response to damselflies, a slower habituation to fish, but no habituation to dragonflies even after 30 h. In contrast, the larvae of Culex quinquefasciatus Say normally live in highly polluted and thus anaerobic water, where the predation risk will be much lower. They also showed a significant reduction in bottom‐feeding after 1 h of exposure to predator kairomones but had completely habituated this response within 6 h of continuous exposure. Some species of mosquito larvae can thus show a very rapid habituation to predator kairomones, while others only habituate slowly depending on the predator and overall predation risk.     

Larvicidal potentials of leaf and bark extracts of Eucalyptus camaldulensis (Schlect) and Eucalyptus citriodora (Hook) on Culex quinquefasciatus (Say) larvae

The larvicidal activity of leaf and bark extracts of Eucalyptus camaldulensis and Eucalyptus citriodora on juveniles of Culex quinquefasciatus was determined. Larvae were successfully bred in the laboratory to obtain a “pure” colony. One hundred and sixty‐two batches of 25 larvae each were treated with 3.90, 15.63, 62.50, 250 and 1000 mg/L of plant extract in three replicates, for 24 h to record mortality and for up to 96 h to record feeding and fecundity. There was an increase in percentage mortality with increase in concentration of the petroleum ether plant extracts. There was a statistically significant difference (P < 0.05) between extracts used in the bioassay and mortality of larvae, but there was no significant difference (P > 0.05) between the two plant species. Statistical analysis of average mortality figures using Probit analysis for the leaf extracts of E. citriodora and E. camaldulensis, respectively, had LC₅₀ of 245.47 (26.89 ± 10.13 [SE]) and 316.23 (22.89 ± 8.95) for the crude extracts; 97.72 (44.95 ± 12.89) and 223.87 (29.17 ± 9.56) for the petroleum ether extracts; and 162.18 (30.7 ± 11.16) and 257.04 (26.47 ± 9.28) for the methanol extracts. The bark extracts of E. citriodora and E. camaldulensis, respectively, had LC₅₀ of 251.19 (27.78 ± 8.39) and 398.11 (20.7 ± 7.17) for methanol; and 302.0 (21.56 ± 8.11) and 630.96 (13.56 ± 6.4) for hexane. Both plant species showed anti‐feeding properties against larvae of Culex quinquefasciatus.     

Mosquitoes of Rarotonga,Cook Islands: A survey of breeding sites

Abstract

Two surveys of Rarotonga, Cook Islands (21°20^'S, 160°16^'W) were made to determine the mosquito fauna of the island, and to identify the habitats required for breeding by searching for larvae. The first survey was made during the “dry season” in May 2001, the second during the “wet season” in February 2002. The mosquito fauna comprised four species Culex (Culex) quinquefasciatus Say, Culex (Culex) annulirostris Skuse, Aedes (Stego‐myia) aegypti (Linnaeus) and Aedes (Stegomyia) polynesiensis Marks. Larvae of the Culex species were most often found in larger natural and artificial water bodies. The Aedes species bred in both natural and artificial containers of all sizes. Ae. polynesiensis was the most widespread species, using natural holes in all regions as well as artificial containers in the urban areas. Most larvae of Ae. aegypti were located in small artificial containers. The two Aedes species are the vectors of dengue fever on the island. Mosquito control during outbreaks should specifically target the artificial containers preferred by Aedes sp. for breeding habitats.     

Evaluation of factors for rapid development of Culex quinquefasciatus in belowground stormwater treatment devices

Water samples from 11 belowground stormwater treatment Best Management Practices (BMPs) were evaluated for their capacity to support rapid development of the West Nile virus (WNV) mosquito vector, Culex quinquefasciatus. The observed minimum development time from egg to pupa ranged from six to over 30 days. Concentrations of potential food resources (total suspended solids and the particulate organic matter in water samples) were significantly correlated to development times. In addition, the rate of immature mosquito development was both site‐dependent and variable in time, suggesting that factors favorable to rapid development were strongly influenced by watershed characteristics and seasonal changes in temperature. Measured temperatures in belowground BMPs suggest that these structures may remain amenable to WNV virus activity longer each year than sites aboveground.     

Efficacy of Esbiothrin mosquito coils at community level in northern Tanzania

Field trials with mosquito coils containing 0.15% Esbiothrin at Usa River settlement, northern Tanzania, during 1986 achieved a 52% reduction in biting rates of Anopheles arabiensis Patton and a 73% reduction of Culex quinquefasciatus Say mosquitoes. No mosquito biting activity occurred during the coil burning period which started at 20.00 hours daily and lasted for an average of 8.6 h. A sociological survey indicated that the coils were favourably accepted and produced no serious side-effects.     

Septic tanks as larval habitats for the mosquitoes Aedes aegypti and Culex quinquefasciatus in Playa‐Playita,Puerto Rico

Adult Aedes aegypti (Linnaeus) (Diptera: Culicidae) were previously recovered from emergence traps on septic tanks in southeastern Puerto Rico. In this study we quantified immature mosquito abundance and its relationship with structural variables of the septic tanks and chemical properties of the water containing raw sewage. A miniaturized floating funnel trap was used to sample 89 septic tanks for larvae in the Puerto Rican community of Playa‐Playita. Aedes aegypti larvae were recovered from 18% of the sampled tanks (10.3 larvae per septic tank per day). Larval presence was positively associated with cracking of the septic tank walls and uncovered access ports. Larval abundance was positively associated with cracking of the septic tank walls and larger tank surface areas, and inversely associated with the total dissolved solids (TDS). Culex quinquefasciatus (Say) larvae were also recovered from 74% of the septic tanks (129.6 larvae per septic tank per day). Larval presence was negatively associated with TDS in the water and larval abundance was positively associated with cracking of the septic tank walls. A screened, plastic emergence trap was used to sample 93 septic tanks within the community for Ae. aegypti and Cx. quinquefasciatus adults. Aedes aegypti adults were recovered from 49% of the sampled tanks (8.7 adults per septic tank per day) and Cx. quinquefasciatus adults were recovered from 97% of the sampled tanks (155.5 adults per septic tank per day). Aedes aegypti adult presence was positively associated with cracking, uncapped openings and septic water pH. The Ae. aegypti adult counts were positively associated with cracking and inversely associated with TDS and conductivity. This study marks the first published record of the recovery of Ae. aegypti larvae from holding tanks containing raw sewage in the Caribbean region. Our study indicates that Ae. aegypti larvae are present in sewage water and that septic tanks have at least the potential to maintain dengue transmission during the dry season.     

Unusual productivity of Aedes aegypti in septic tanks and its implications for dengue control

Increased DEN-2 virus transmission in Puerto Rico during 2005 prompted the implementation of a rapid intervention programme to suppress Aedes aegypti (L.) (Diptera: Culicidae) emergence, which in turn lead to the discovery of previously unknown breeding sites underground. Initially, the following control measures were applied in Playa/Playita (PP), a town of 1,400 households, to all areas where the number of pupae per person exceeded the expected threshold for dengue transmission; all containers likely to be aquatic habitats were turned over and containers too large to turn were treated with 1 p.p.m. methoprene. The impact of these interventions was evaluated by comparing the number of resting adult mosquitoes (by backpack aspiration and sweepnetting in bedrooms) pre-intervention, with numbers at 3 and 5 weeks post-intervention, and by evaluating pupal density at 4 weeks post-intervention in PP and in a nearby town, Coqui (CO; 1500 households), which was not treated. The pre-intervention and post-intervention densities of resting Ae. aegypti adults were significantly larger in the intervention town, although the density of pupae in surface containers was low and similar in both towns at 4 weeks post-intervention. At 3 weeks post-intervention, the density of resting adults decreased by only 18% of pre-intervention levels, but returned to pre-intervention levels 5 weeks after treatment. By contrast, the density of resting adults in CO steadily decreased to 48% and 61%, at 3 and 5 weeks after the initial surveys, respectively. Geographical Information Systems identified significant clustering of adult mosquitoes, which led to the discovery of underground aquatic habitats (septic tanks) that were producing large numbers of Ae. aegypti and Culex quinquefasciatus (Say) in the treated town. We calculated that septic tanks could produce > 18 000 Ae. aegypti and approximately 170 000 Cx quinquefasciatus adults per day. Septic tanks are likely to be common and widespread in suburban and rural Puerto Rico, where, apparently, they can contribute significantly to the maintenance of island-wide dengue virus endemicity.     

Susceptibility of mosquitoes in central Taiwan to natural infections of Dirofilaria immitis

From October 1997 to September 1998, 3085 Culex quinquefasciatus (Say) (Diptera: Culicidae), 584 Cx. tritaeniorhynchus (Giles) (Diptera: Culicidae), 392 Cx. annulus (Theobald) (Diptera: Culicidae), 374 Aedes albopictus (Skuse) (Diptera: Culicidae) and 102 Armigeres subalbatus (Coquillet) (Diptera: Culicidae) were collected and examined for Dirofilaria immitis (Leidy) (Spirurida: Filariidae) infection. However, only Cx. quinquefasciatus and Ae. albopictus were infected, with a prevalence of 4.28% and 3.74%, respectively. The intensity of D. immitis found in Ae. albopictus (3.43 larvae/mosquito) was higher than that found in Cx. quinquefasciatus (2.89 larvae/mosquito). After being fed with canine blood containing 7500 microfilariae (mf) per mL, Cx. quinquefasciatus ingested approximately two times as many mf as Ae. albopictus (mean of 31.73 in comparison to 16.47). However, almost three times as many third-stage infective larvae developed in Ae. albopictus as in Cx. quinquefasciatus (mean of 3.25 as compared with 1.10), with a vector efficiency index (VEI) of 19.73 and 3.47, respectively. The results showed that Cx. quinquefasciatus and Ae. albopictus served as natural vectors of D. immitis in central Taiwan. Although Ae. albopictus was more efficient for heartworm transmission, Cx. quinquefasciatus may play a more prominent role on the transmission of dirofilariasis in central Taiwan.     

Larval habitat for the avian malaria vector Culex quinquefasciatus (Diptera: Culicidae) in altered mid‐elevation mesic‐dry forests in Hawai'i

Effective management of avian malaria (Plasmodium relictum) in Hawai'i's endemic honeycreepers (Drepanidinae) requires the identification and subsequent reduction or treatment of larval habitat for the mosquito vector, Culex quinquefasciatus (Diptera: Culicidae). We conducted ground surveys, treehole surveys, and helicopter aerial surveys from 2001–2003 to identify all potential larval mosquito habitat within two 100+ ha mesic‐dry forest study sites in Hawai'i Volcanoes National Park, Hawai'i; ‘Ainahou Ranch and Mauna Loa Strip Road. At ‘Ainahou Ranch, anthropogenic sites (43%) were more likely to contain mosquitoes than naturally occurring (8%) sites. Larvae of Cx. quinquefasciatus were predominately found in anthropogenic sites while Aedes albopictus larvae occurred less frequently in both anthropogenic sites and naturally‐occurring sites. Additionally, moderate‐size (≈20–22,000 liters) anthropogenic potential larval habitat had >50% probability of mosquito presence compared to larger‐ and smaller‐volume habitat (<50%). Less than 20% of trees surveyed at ‘Ainahou Ranch had treeholes and few mosquito larvae were detected. Aerial surveys at ‘Ainahou Ranch detected 56% (95% CI: 42–68%) of the potential larval habitat identified in ground surveys. At Mauna Loa Strip Road, Cx. quinquefasciatus larvae were only found in the rock holes of small intermittent stream drainages that made up 20% (5 of 25) of the total potential larval habitat. The volume of the potential larval habitat did not influence the probability of mosquito occurrence at Mauna Loa Strip Road. Our results suggest that Cx. quinquefasciatus abundance, and subsequently avian malaria, may be controlled by larval habitat reduction in the mesic‐dry landscapes of Hawai'i where anthropogenic sources predominate.     

Phenotypic variation of an alien species in a new environment: the body size and diet of American mink over time and at local and continental scales

Introduced species must adapt their ecology, behaviour, and morphological traits to new conditions. The successful introduction and invasive potential of a species are related to its levels of phenotypic plasticity and genetic polymorphism. We analysed changes in the body mass and length of American mink (Neovison vison) since its introduction into the Warta Mouth National Park, western Poland, in relation to diet composition and colonization progress from 1996 to 2004. Mink body mass decreased significantly during the period of population establishment within the study area, with an average decrease of 13% from 1.36 to 1.18 kg in males and of 16% from 0.83 to 0.70 kg in females. Diet composition varied seasonally and between consecutive years. The main prey items were mammals and fish in the cold season and birds and fish in the warm season. During the study period the proportion of mammals preyed upon increased in the cold season and decreased in the warm season. The proportion of birds preyed upon decreased over the study period, whereas the proportion of fish increased. Following introduction, the strictly aquatic portion of mink diet (fish and frogs) increased over time, whereas the proportion of large prey (large birds, muskrats, and water voles) decreased. The average yearly proportion of large prey and average‐sized prey in the mink diet was significantly correlated with the mean body masses of males and females. Biogeographical variation in the body mass and length of mink was best explained by the percentage of large prey in the mink diet in both sexes, and by latitude for females. Together these results demonstrate that American mink rapidly changed their body mass in relation to local conditions. This phenotypic variability may be underpinned by phenotypic plasticity and/or by adaptation of quantitative genetic variation. The potential to rapidly change phenotypic variation in this manner is an important factor determining the negative ecological impacts of invasive species. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 681–693.     

Comparison of carbon dioxide-baited trapping systems for sampling outdoor mosquito populations in Tanzania

For collecting mosquitoes (Diptera: Culicidae) the outdoor catching efficiency of four types of trapping devices baited with carbon dioxide (CO2, 300 ml/ min) was evaluated and compared in two areas of Tanzania. The types of traps employed were: the CDC miniature trap with the incandescent light bulb switched on or off; electric nets (ENT) and a Counterflow Geometry (CFG) trap. In Njage, southeast Tanzania, Anopheles gambiae Giles sensu stricto was the most abundant of the seven mosquito species obtained, comprising of 74.3% of the total number caught (n=2,171). In Muheza, north-east Tanzania, Culex quinquefasciatus Say was the predominant species (90.9%) among 1,080 caught. At both localities the CFG trap was superior to the CDC trap with light-on or light-off for sampling both An. gambiae and Cx. quinquefasciatus. Efficiency of the CFG trap and ENT were similar for sampling these species of mosquitoes (P > 0.05). However, ENT was superior to the CDC trap with light-off for collecting both species. Significantly more (P < 0.05) Cx. quinquefasciatus were obtained by the CDC trap with light-off than with light-on, especially outdoors. It is concluded that both ENT and the CFG are effective tools for sampling populations of An. gambiae and Cx. quinquefasciatus outdoors.     

Understanding the mechanism of Chikungunya virus vector competence in three species of mosquitoes

Chikungunya virus (CHIKV) is primarily transmitted by Aedes spp. mosquitoes. The present study investigated vector competence for CHIKV in Aedes aegypti and Aedes albopictus mosquitoes found in Madurai, South India. The role of receptor proteins on midguts contributing to permissiveness of CHIKV to Aedes spp. mosquitoes was also undertaken. Mosquitoes were orally infected with CHIKV DRDE‐06. Infection of midguts and dissemination to heads was confirmed by immunofluorescence assay at different time points. A plaque assay was performed from mosquito homogenates at different time points to study CHIKV replication. Presence of putative CHIKV receptor proteins on mosquito midgut epithelial cells was detected by virus overlay protein binding assay (VOPBA). The identity of these proteins was established using mass spectrometry. CHIKV infection of Ae. aegypti and Ae. albopictus midguts and dissemination to heads was observed to be similar. A plaque assay performed with infected mosquito homogenates revealed that CHIKV replication dynamics was similar in Aedes sp. mosquitoes until 28 days post infection. VOPBA performed with mosquito midgut membrane proteins revealed that prohibitin could serve as a putative CHIKV receptor on Aedes mosquito midguts, whereas an absence of CHIKV binding protein/s on Culex quinquefasciatus midguts can partially explain the non‐permissiveness of these mosquitoes to infection.     

Field evaluations of disposable sticky lures for surveillance of Aedes aegypti (Stegomyia aegypti) and Culex quinquefasciatus in Jakarta

From December 1997 to April 1998, disposable sticky lures (1608 lure days) were trialled in homes in north Jakarta, Indonesia as surveillance tools for Aedes aegypti (Stegomyia aegypti) (Diptera: Culicidae) and Culex quinquefasciatus (Diptera: Culicidae), referenced to indoor resting adult collections (92 × 10 min). The lures collected 89.4% of the total of 1339 Ae. aegypti and 92.1% of the total of 1272 Cx. quinquefasciatus collected by all methods. Because there were no significant differences with respect to numbers collected in bedrooms, living rooms and kitchens, bedrooms were selected for subsequent trials for reasons of convenience. The main trials involved a replicated complete block design with L‐lysine and sodium carbonate. Lures without attractant or with four different dilutions of L‐lysine collected 3.4–8.5 times more Ae. aegypti and 4.2–8.1 times more Cx. quinquefasciatus than were collected by mouth aspirator. Lures with or without dilutions of sodium carbonate collected 2.7–5.0 times more Ae. aegypti and 1.8–4.2 times more Cx. quinquefasciatus than aspirator collections. The precision associated with catches of sticky lures was better than that for aspirator collections. Although olfactants generally improved the numbers of mosquitoes collected, the differences in catch between lures with and without attractants were usually non‐significant. Any deficit in catch may be offset by increasing the surveillance period to ≥30 days to detect all four dengue serotypes from infected mosquitoes.     

Fine scale spatial urban land cover factors associated with adult mosquito abundance and risk in Tucson,Arizona

It is currently unclear what role microhabitat land cover plays in determining the seasonal spatial distribution of Aedes aegypti and Culex quinquefasciatus, disease vectors of dengue and West Nile Virus, respectively, in Tucson, AZ. We compared mosquito abundance to sixteen land cover variables derived from 2010 NAIP multispectral data and 2008 LiDAR height data. Mosquitoes were trapped with 30–9 traps from May to October of 2010 and 2011. Variables were extracted for five buffer zones (10–50 m radii at 10 m intervals) around trapping sites. Stepwise regression was performed to determine the best scale for observation and the influential land cover variables. The 30 m radius buffer was determined to be the best for observing the land cover‐mosquito abundance relationship. Ae. aegypti presence was positively associated with structure and medium height trees and negatively associated with bare earth; Cx. quinquefasciatus presence was positively associated with pavement and medium height trees and negatively associated with shrubs. These findings emphasize vegetation, impervious surfaces, and soil influences on mosquito presence in an urban setting. Lastly, the land cover‐mosquito abundance relationships were used to produce risk maps of seasonal presence that highlight high risk areas in Tucson, which may be useful for focusing mosquito control program actions.     

Haemocyte population and ultrastructural changes during the immune response of the mosquito Culex quinquefasciatus to microfilariae of Wuchereria bancrofti

Abstract Haemocytes circulating in the haemolymph protect insects against pathogens that enter the haemocoel. Changes in haemocyte morphology and differences in haemocyte counts during the immune response of Culex quinquefasciatus Say (Diptera: Culicidae) to microfilariae of Wuchereria bancrofti (Cobbold) (Spirurida: Onchocercidae) were investigated in the present study. The mean number of total haemocytes was significantly elevated in infected mosquitoes (P < 0.001), reaching a peak on the third day post‐infection. Differential counts show that mean numbers of prohaemocytes, plasmatocytes, granular cells and oenocytoids increased significantly after infection with microfilariae granulocytes compared to the control and näive groups of Cx. quinquefasciatus (P < 0.05). Changes in proportional counts of haemocytes were also analysed in haemolymph perfusates of Cx. quinquefasciatus infected with W. bancrofti. On the first day post‐infection, infected mosquitoes showed an increase in the proportion of prohaemocytes (18.8% compared to 9.6% for the control) and of oenocytoids (7.1% compared to 4.7% control); however, they exhibited lower levels of plasmatocytes (36.6% compared to 42.1% control) and granular cells (36.1% compared to 41.4% control). On day 14 post‐infection, similar changes were observed for these haemocyte types, except that the proportion of granular cells was significantly greater than the control (41.2% compared to 31.3% control). Although an enhancement of prohaemocyte numbers was observed, this cellular type did not show any ultrastructural alteration. On the other hand, granular cells, plasmatocytes and oenocytoids presented morphological alterations indicative of innate immunological activation in mosquitoes infected with W. bancrofti.     

Mosquito control in Dar es Salaam.

Abstract. In preparation for a trial of polystyrene beads and pyriproxyfen for the control of Culex quinquefasciatus mosquitoes, surveys of their breeding were carried out in two contrasting areas of Dar es Salaam, Mikocheni and Ilala, during the dry season.
Sanitation structures (latrines, soakage pits, septic tanks and cess pits) were the most prolific breeding places, totalling 780 in Mikocheni and 1544 in Ilala. Those in Mikocheni were estimated to contain about 1.4 times more mosquito pupae, per site, than in such structures in Ilala. This was both because a higher proportion of sites contained visible water and because sites with water were more likely to contain pupae in Mikocheni. The relative importance of the different types of structure, in terms of productivity, was the same in both areas. Although septic tanks and cess pits made up only 10.5% of the on-site sanitation structures in Ilala, they contained 53% of the total number of pupae in enclosed sites; they were therefore particularly important targets for treatment with polystyrene beads. A survey during the rainy season of sites in Ilala revealed little change in the proportion that were wet, or in the frequency of breeding in those with visible water.
The number, type and area of open breeding sites varied greatly between the two study areas. In Mikocheni the area of open breeding sites was 100 times greater than in Ilala, with 97% of the 13 ,000 m² being flooded grassland. In Ilala all but four of the sixty-six open breeding sites were puddles of sullage water derived from bathrooms. Different strategies for control in open breeding sites are required in the two areas of Dar es Salaam.