Blood meal analysis of Anopheles vectors of simian malaria based on laboratory and field studies

Blood feeding and host-seeking behaviors of a mosquito play an imperative role in determining its vectorial capacity in transmitting pathogens. Unfortunately, limited information is available regarding blood feeding behavior of Anopheles species in Malaysia. Collection of resting Anopheles mosquitoes for blood meal analysis poses a great challenge especially for forest dwelling mosquitoes. Therefore, a laboratory-based study was conducted to evaluate the potential use of mosquitoes caught using human landing catch (HLC) for blood meal analysis, and subsequently to document blood feeding behavior of local Anopheles mosquitoes in Peninsular Malaysia. The laboratory-based experiment from this study revealed that mosquitoes caught using HLC had the potential to be used for blood meal analysis. Besides HLC, mosquitoes were also collected using manual aspirator and Mosquito Magnet. Overall, 47.4% of 321 field-caught Anopheles mosquitoes belonging to six species were positive for vertebrate host DNA in their blood meal. The most frequent blood meal source was human (45.9%) followed by wild boar (27.4%), dog (15.3%) and monkey (7.5%). Interestingly, only Anopheles cracens and Anopheles introlatus (Leucosphyrus Group) fed on monkey. This study further confirmed that members of the Leucosphyrus Group are the predominant vectors for knowlesi malaria transmission in Peninsular Malaysia mainly due to their simio-anthropophagic feeding behavior.

The Centres for Disease Control Light Trap (CDC LT) and The Human Decoy Trap (HDT) Compared to The Human Landing Catch (HLC) for Measuring Anopheles Biting in Rural Tanzania

BackgroundThe intensity of vector mosquito biting is an important measure for malaria epidemiology and control. The human landing catch (HLC) is an effective entomological surveillance tool, but is labour-intensive, expensive and raises safety issues. The Centres for Disease Control light trap (CDC LT) and the human decoy trap (HDT) are less costly and exposure-free alternatives. This study compared the CDC LT and HDT against the HLC for measuring Anopheles (An.) biting in rural Tanzania and assessed their suitability as HLC proxies.MethodsIndoor mosquito surveys using HLC and CDC LT and outdoor surveys using HLC and HDT were conducted in 2017 and in 2019 in Ulanga, Tanzania in 19 villages, with one trap per house per night. Species composition, sporozoite rates and the numbers of mosquitoes caught by different trap types were compared. Aggregating the data by village and month, the Bland-Altman approach was used to assess agreement. ResultsOverall, 66,807 Anopheles funestus and 14,606 An. arabiensis adult females were caught from 6,013 CDC LT, 339 indoor HLC, 136 HDT and 195 outdoor HLC collections. Overall, the CDC LT caught fewer malaria vectors than indoor HLC: An. arabiensis (Adjusted rate ratio (Adj.RR) =0.35 (95% confidence interval (CI):0.27-0.46)) and An. funestus (Adj.RR=0.63(95%CI:0.51-0.79)). HDT caught fewer malaria vectors than outdoor HLC: An. arabiensis (Adj.RR=0.04(95%CI:0.01-0.14)) and An. funestus (Adj.RR=0.10(95%CI:0.07-0.15)). The bias and variability of the ratios of geometric mean mosquitoes caught by CDC LT and HDT relative to HLC collections for the same village-month were dependent on mosquito densities. The relative efficacies of both CDC LT and HDT declined with mosquito abundance. The variability in the ratios was substantial for low HLC counts and decreased as mosquito abundance increased. CDCLT caught a higher proportion of infected An. arabiensis and An. funestus than HLC, and HDT caught no infected mosquitoes.ConclusionsIf caution is taken in appreciation of its limitations, the CDC LT is suitable for use in routine entomological surveys and may be preferable for measuring sporozoite rates for Afrotropical mosquitoes. Use of HLC is useful to estimate human exposure to mosquitoes for estimating Entomological Inoculation Rate (EIR). The present design of the HDT is not amenable for use to conduct large-scale entomological surveys.

Anopheline Diversity in Indonesia: An Evaluation of Animal-Baited Sampling Techniques

Indonesia has rich Anopheline (Diptera: Culicidae) mosquito species living in various types of ecosystems. The study was conducted to profile and compare Anopheles diversity, equitability, and dominance in various ecosystems using different animal-based sampling techniques. The present study analyzed a subset of data collected from a nation-wide vector and animal reservoirs survey in 2016. Analyses were restricted to three ecosystem types (forest, nonforest, and coastal areas) in Java and Sumatera Islands. A total of 5,477 Anopheles were collected by using animal-baited (n = 1,909) and animal-baited trap nets (n = 1,978), consisting of 23 Anopheline species. Overall, Anopheles vagus was the most abundant species, followed by An. subpictus and An. barbirostris. Among the three ecosystems, the forest had a higher diversity index (H′ = 1.98), but each ecosystem has its specific predominant species. Compared with the animal-baited method, the Anopheles abundance collected by animal-baited trap nets was two-fold higher. Ecosystem, elevation, and sampling methods were associated with the abundance of female Anopheles (P-value < 0.001). Our findings revealed that Anopheles were found in a different ecosystem, indicating the potential of malaria transmission. This suggests that improved malaria vector surveillance is essential in all types of ecosystem. Furthermore, the study suggested that animal-baited trap nets could be used as the standard method of outdoor resting sampling in Indonesia in addition to the traditional human landing collection approach.

Larvae ecology and adult activity of Aedes mariae (Diptera: Culicidae) in a touristic rock-pool area of the Balearic Islands (Western Mediterranean)

Mosquitoes are vectors of several diseases of medical concern such as malaria or dengue and can also negatively affect tourism and the life-quality of the neighbourhood. The species Aedes mariae (Sergent and Sergent, 1903) is a poorly studied mosquito that breeds in rock-pools of the Mediterranean coast. General Linear Mixed Models (GLMM) were used to determine drivers affecting the presence and abundance of this species. Abiotic and biotic factors were recorded in rock-pools with the presence of Ae. mariae sub-adults across a supralittoral area of Majorca Island (Balearic Islands, Spain) from July 2018 to June 2019. We tested how abiotic factors affected the presence of larvae, while the biotic factors were used to check their effect on larvae abundance. human landing collection was also conducted to assess the adult activity of this species. Valuable data were recorded to improve our knowledge about the bioecology of Ae. mariae in a touristic area of the island of Majorca. Salinity and pH were the most explanatory variables for the presence of Ae. mariae larvae. The presence of Posidonia oceanica (L.) Delile 1813 leaves negatively affected the abundance of Ae. mariae larvae while the presence of other fauna enhanced it. Adult females of Ae. mariae were active for 26 min after sunset in June and its host-seeking activity decreased during autumn months. Control methods against this species should be focussed on rock-pools and planning treatments according to tides, waves and precipitation.

Bio-ecological study of Culex quinquefasciatus as a potential vector of Japanese encephalitis in some provinces in Indonesia

Culex quinquefasciatus is a mosquito known as Japanese encephalitis (JE) vector in several regions in Indonesia. The bioecological study is essential to optimize the vector control of JE. The purpose of the study was to obtain the ecological information of Cx. quinquefasciatus in 15 provinces in Indonesia: Aceh, West Sumatra, Lampung, Bangka Belitung, Banten, West and East Java, West and South Kalimantan, North and Southeast Sulawesi, East and West Nusa Tenggara, Maluku, and North Maluku. Mosquitoes were collected using the human landing catches (HLCs), light trap, and morning resting collection. The larva survey was conducted in potential habitats of Cx. quinquefasciatus. The mosquito was detected for the JE virus using the polymerase chain reaction (PCR) method. The distribution of Cx. quinquefasciatus was recorded using a GPS tool and visualized using Global Mapper. The results showed that Cx. quinquefasciatus in 15 provinces had similar behavior. The Cx. quinquefasciatus sucked blood indoors and was found throughout the night. Peak density of Cx. quinquefasciatus varies by province. Cx. quinquefasciatus breeding places are found in most ecosystems in various altitudes. Based on PCR examination, the JE virus has not been detected, thus lowering the potential for JE transmission in some provinces in Indonesia.

Anopheles drivers of persisting malaria transmission in Guna Yala, Panamá: an operational investigation

Background Though most of Panamá is free from malaria, localized foci of transmission persist, including in the Guna Yala region. Government-led entomological surveillance using an entomological surveillance planning tool (ESPT) sought to answer programmatically-relevant questions that would enhance the understanding of both local entomological drivers of transmission and gaps in protection that result in persisting malaria transmission to guide local vector control decision-making. Methods The ESPT was used to design a sampling plan centered around the collection of minimum essential indicators to investigate the relevance of LLINs and IRS in the communities of Permé and Puerto Obaldía, Guna Yala, as well as to pinpoint any remaining spaces and times where humans are exposed to Anopheles bites (gaps in protection). Adult Anopheles were collected at three time points via human landing catches (HLCs), CDC Light Traps (LT), and pyrethrum spray catches (PSCs) during the rainy and dry seasons. Mosquitoes were identified to species via molecular methods. Insecticide susceptibility testing of the main vector species to fenitrothion was conducted. Results In total, 7537 adult Anopheles were collected from both sites. Of the 493 specimens molecularly confirmed to species, two thirds (n = 340) were identified as Nyssorhynchus albimanus, followed by Anopheles aquasalis. Overall Anopheles human biting rates (HBRs) were higher outdoors than indoors, and were higher in Permé than in Puerto Obaldía: nightly outdoor HBR ranged from 2.71 bites per person per night (bpn) (Puerto Obaldía), to 221.00 bpn (Permé), whereas indoor nightly HBR ranged from 0.70 bpn (Puerto Obaldía) to 81.90 bpn (Permé). Generally, peak biting occurred during the early evening. The CDC LT trap yields were significantly lower than that of HLCs and this collection method was dropped after the first collection. Pyrethrum spray catches resulted in only three indoor resting Anopheles collected. Insecticide resistance (IR) of Ny. albimanus to fenitrothion was confirmed, with only 65.5% mortality at the diagnostic time. Conclusion The early evening exophagic behaviour of Anopheles vectors, the absence of indoor resting behaviours, and the presence of resistance to the primary intervention insecticide demonstrate limitations of the current malaria strategy, including indoor residual spraying (IRS) and long-lasting insecticidal nets (LLINs), and point to both gaps in protection and to the drivers of persisting malaria transmission in Guna Yala. These findings highlight the need for continued and directed entomological surveillance, based on programmatic questions, that generates entomological evidence to inform an adaptive malaria elimination strategy.

Coverage, Usage, Physical Integrity, and Bio-efficacy of Olyset Nets in the Plateau Region, South Benin Following the 2011 Nationwide Distribution

The present study investigated in 8 villages of the Plateau region the coverage, usage, physical integrity, and bio-efficacy of the Olyset nets distributed nationwide by the Benin's National Malaria Control Programme in July 2011. The questionnaire administered as well as the observations made in the households allowed estimating the coverage and usage rates of the 2011 Olyset nets. While their physical integrity was assessed through standard WHO methodology, their bio-efficacy was evaluated through gas chromatography, and WHO cone testing performed with the Kisumu susceptible strain. Mosquito collections through human landing catches (HLCs) were also performed in torn nets to assess if a loss of protection of sleepers occurred as the nets fabric integrity got more damaged. Nine months postdistribution, the coverage and usage rates of the 2011 Olyset nets were 67.4% (95% CI: 65.8–68.9) and 73.3% (95% CI: 70.7–75.8) respectively. About 28% of the 2011 Olyset nets were torn. A drastic drop of the insecticide quantity on the fibers of the nets [from 7.08 µg (95% CI: 5.74–8.42) to 0.2 µg (95% CI: 0.01–0.38)] as well as mortality rates <80% were observed with most nets evaluated. Moreover, the biting rates of An. gambiae s.l. (Diptera: Culicidae) inside torn nets increased in line with their fabric integrity loss. These data support the conclusion that future deployment of nets in the field must be strengthened by community sensitization on their correct use in order to postpone as much as possible appearance of holes and loss of insecticidal activity and encourage repairing of torn nets.

“Slash and Clear”, a Community-Based Vector Control Method to Reduce Onchocerciasis Transmission by Simulium sirbanum in Maridi, South Sudan: A Prospective Study

Background: High ongoing Onchocerca volvulus transmission was recently documented in Maridi County, South Sudan. To complement community-directed treatment with ivermectin (CDTI) as the main onchocerciasis control strategy, we initiated a community-based vector control method “slash and clear” at the Maridi dam, a Simulium damnosum s.l. breeding site, to reduce O. volvulus transmission. Methods: Simulium damnosum s.l. biting rates were collected before and during the twenty months following the “slash and clear” intervention using the human landing catches. Black flies were dissected to measure parity rates before and twelve months after the intervention. Larvae and pupae of S. damnosum s.l. were collected from the dam for morphological and chromosomal characterization to identify the cytospecies involved. Results: Biting rates of S. damnosum s.l. close to the Maridi dam spillway decreased by >90% post-“slash and clear” for more than six months. Twelve months after the “slash and clear” intervention, the reduction in biting rates was still at <50% (p = 0.0007). Parity rates reduced from 13% pre-“slash and clear” (November 2019) to 5.6% post-“slash and clear” (November 2020). Larvae collected from the dam were identified as Simulium sirbanum. Conclusion: The “slash and clear” method was found to be an effective and cheap community-based method to reduce black fly biting rates caused by S. sirbanum. When repeated at least annually together with a high CDTI coverage, this intervention has the potential to considerably accelerate onchocerciasis elimination.