Diversity, abundance of anopheline species, and malaria transmission dynamics in high-altitude areas of western Cameroon

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Several entomological studies have been conducted to describe malaria transmission in different eco-epidemiological settings in Cameroon; knowledge gaps persist, particularly in highland areas. This study aimed to characterize malaria vectors in three localities along an altitudinal gradient in the western region: Santchou (700 m), Dschang (1400 m), and Penka Michel (1500 m). Methods : Human landing catches were conducted from May to June 2023 from 6:00 pm to 9:00 am. Mosquitoes were sorted into genera, and all Anopheles species were identified using morphological taxonomic keys and species-specific Polymerase Chain reaction (PCR). Entomological indicators were assessed including species composition and abundance, biting behavior, infection rate, and entomological inoculation rate (EIR). Genomic DNA from the head and thoraces were tested for Plasmodium infection by real-time PCR. Results : 2,835 Anopheles mosquitoes were identified, including An. gambiae, An. coluzzii, An. funestus, An. leesoni, An. nili, and An. ziemanni , with An. gambiae being the most prevalent at all sites. The human-biting rate of An. gambiae s.l. was significantly higher (p-value < 0.001) in Penka Michel compared to Santchou and Dschang (45.25 b/h/n vs 3.1 b/h/n and 0.41 b/h/n), and appears to be the most infected vector, and infectious vector distribution is highly focal, with entomological inoculation rates 13-fold higher in Penka Michel compared to Santchou (1.11 vs 0.08ibites/human/night). P. falciparum was the dominant malaria parasite (67% at Santchou, 62% at Penka Michel), but P. malariae (30%) and P. ovale (1.21%) infections were also detected. Conclusion : The study highlights a difference in mosquito composition and host-seeking behavior with altitude and the need for continued surveillance to monitor vector populations and prevent potential malaria outbreaks in these highland areas. Anopheles diversity behavior transmission highland western Cameroon Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Background Malaria remains a major public health problem worldwide [1]. The latest WHO estimates point to 249 million cases and 608,000 deaths, with 94% of cases occurring in the African region. Children under 5 years and pregnant women represent the most vulnerable and affected groups. In addition to therapeutic care, considerable progress has been made in the implementation of vector control measures such as long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) [2,3]. The rapid spread of these strategies has so far been a major factor in the decline in malaria morbidity and mortality over the past two decades [3]. Malaria remains a major public health challenge in Cameroon, in 2023, Cameroon recorded 4.85 million cases of suspected malaria, of which over a million were severe, mainly in children under 5. Malaria contributed to 7.3% of hospital deaths [4]. The entire population of approximately 28 million people is at risk of malaria, with the East, Centre, and North regions being the most affected [5]. The intensification of free LLIN distribution has contributed substantially to reducing the malaria burden in the country, with an estimated population coverage of 80% [5–7]. However, the effectiveness of these control tools is threatened by several factors, including insecticide resistance, changes in the feeding and resting behavior of the main malaria vectors [8–12], and the spatiotemporal variation in malaria transmission across different ecological zones of the country. Cameroon is often known as “Africa in miniature” due to the diversity of its natural environment distributed across its 10 provinces: deserts, coasts, mountains, rainforests, and savannah, resulting in heterogenous conditions [13] that directly influence mosquito distribution, abundance, survival, and malaria parasite development. Variations in vector ecology across the landscape can lead to uneven exposure and transmission risks [14]. The lowlands are characterized by floods and swamps, promoting larvae survival and a greater abundance of vectors, resulting in high malaria prevalence [15–17]. In contrast, the highlands consist of hills and valleys, resulting in a sparse distribution of larval habitats and a lower malaria prevalence ( P. falciparum prevalence based on RDT positivity of 11-19%) [18]. The diverse malaria eco-epidemiological contexts and local vector ecologies call for tailored interventions for each transmission focus, as malaria epidemiology fluctuates over time and is correlated with the local success of control programs. The success of these control strategies therefore depends on up-to-date information on bionomic, and vector transmission patterns in different epidemiological contexts, particularly in regions at high risk of epidemics, including high-altitude areas. In recent years, mountainous regions of Africa have experienced an increase in the frequency and intensity of malaria epidemics. These areas, characterized by lower temperatures and humidity, present unique challenges for malaria transmission [19–22]. Unlike holoendemic lowland regions, highland areas are typically hypoendemic, making successful entomological inoculation more likely [23–25]. Despite the significance of the Bamileke plateau in western Cameroon (large population and role in the country's economic life), research on malaria transmission in these areas has been limited [26,27]. Moreover, recent reports of Plasmodium vivax infections in Dschang [28,29] further underscore the need for a comprehensive understanding of malaria dynamics in this region. To address this knowledge gap, we conducted a series of entomological and parasitological (unpublished data) surveys in three localities of the region. We aimed to assess the extent of malaria transmission and identify the primary vectors driving the disease, considering the region's diverse ecological settings and population movements. Materials and methods Description of the study area The study was carried out from May to June 2023 in three high-altitude localities in the Menoua division along an altitudinal transect in the Western region of Cameroon: Santchou (700m), Dschang (1400m) and Penka Michel (1500m) are located in a savannah landscape within the Guineo-Congolese bioclimatic domain, on the volcanic line of Cameroon [30,31] ( Fig. 1 ). Santchou (5°25'N; 9°58'E) is located at an altitude of 700m in the vast Mbô plain, bounded to the south by the Manengouba mountain and to the north by a sloping cliff, delimiting it from the Foréké village. The area is mesoendemic for malaria, with a prevalence of 36.19% [27]. Santchou covers an area of 335 km 2 and has a population ranging from 15,000 to around 25,000 and benefits from a dense hydrographic (the river Nkam and its tributaries) network and good rainfall. As a semi-urban city, Santchou has a low plain topography with a hygromorphic soil type. The climate is equatorial Guinean, characterized by four seasons of unequal duration: the long rainy season (mid-August to October); the short rainy season (March to June); the long dry season (mid-October to March); and the short dry season (June to mid-August). Average rainfall is 1,363.3 mm/year and annual temperature ranges from 20.83°C to 23.35°C [32]. The town of Dschang (5°25'N; 10°03'E) is an urban environment that lies at the summit of the Mbô cliffs on the southeastern slopes of the Bamboutos mountains. It is bounded on the south by the commune of Santchou and on the northeast by the Bamboutos mountains. It covers an area of 262 km 2 and has a population of 120,207 inhabitants. The topography consists of alternating hills and valleys crisscrossed by small watercourses. It is characterized by 2 seasons: a rainy season (eight months) from mid-March to mid-November and a dry season (four months) from mid-November to mid-March. The area is characterized by a mosaic of agricultural landscapes and is classified as hypoendemic for malaria, with a prevalence of 25.04% [27]. Average precipitation is 2,000 mm/year and the daily temperature range can exceed 23°C during the dry season which constitutes a particularity of this locality [33] Penka Michel (5°28'N; 10°15'E) landscape is marked by a shallow topography, with a population of 98,229 inhabitants and a population density of 383.7 inhabitants/km². The area has a relatively low malaria prevalence of 38.4% among children under five years old [34]. The relief is that of an undulating plateau with a flat bottom. As in Dschang (29 km from Penka Michel), the area is characterized by a sub-equatorial climate, with distinct wet and dry seasons. The rains peak in August-September when up to 345.1 mm of rainfall can be recorded monthly. Temperatures are relatively low and constant due to the high altitude, with an annual temperature range of only 26°C [35]. These diverse ecological settings, ranging from lowland to highland, provide an ideal opportunity to study the influence of environmental factors on malaria transmission dynamics. This study was conducted following the fourth distribution campaign of Long-Lasting Insecticidal Net (LLIN) in the Western region of Cameroon in 2022. Figure 1 : Map showing the study sites in the Western Region of Cameroon. Mosquito collections and processing Households were randomly selected in each site to assess malaria transmission using the human-landing catch (HLC) approach [36] from 6:00 pm to 9:00 am. In Dschang, HLCs were carried out in 10 villages in four health zones (Bafou, Fialah-foreke, Foto, and Siteu). Four and three villages were selected respectively for mosquito collection in Santchou and Penka Michel. During each survey, adult mosquitoes were collected indoors (living room or bedroom) and outdoors (outdoor shelter) in randomly selected homes, with the distance between two houses being at least 100m. HLCs were carried out during 11 nights in Dschang with a total number of 166 men/nights, 5 nights in the Santchou district with a total number of 78 men/nights, and 4 nights in the Penka Michel district with a total number of 50 men/nights leveraging trained volunteers. To minimize collectors’ bias due to the variability of attractiveness and collection skills, volunteers changed positions every two hours between indoor and outdoor stations and rotated through the houses. Each collector was provided with a flashlight and a bag containing hemolysis tubes with cotton. Mosquitoes landing on the legs of the collectors were captured using haemolysis tubes and stored in a labeled bag. At each sampling station, bags containing mosquitoes were collected hourly for morphological identification. Anopheles larval collection and identification of adult mosquitoes Anopheles larval surveys were carried out in all the water collections identified in the study areas. The pre-imaginal stages of anopheles present in these sites were collected using the Dipping method [37] and brought back to the insectarium for rearing. On arrival at the insectarium, the larvae were transferred to tanks containing water from the lodge, according to their stage of development, and then fed with TetraMinBaby®. The resulting pupae were transferred to small cups and placed in adult-rearing cages until they emerged. Mosquitoes collected were sorted by genus ( Culex, Anopheles, Aedes, Mansonia, and others) and species level using specific identification keys [38]. Anopheles mosquitoes were then dissected into head-thorax, and abdomen, preserved individually in Eppendorf tubes containing silica gel for further molecular analysis. Laboratory processing of mosquitoes DNA extraction, molecular identification of Anopheles species Genomic DNA was extracted from the head-thorax and legs using a 2% CTAB (cetyltrimethylammonium bromide) solution according to Collins et al [39]. Genomic DNA was amplified by multiplex Polymerase Chain Reaction (PCR) and targeted the intergenic spacer of the ribosomal DNA (rDNA IGS) and internal transcribed spacer region 2 (ITS2) of An. gambiae s.l. and An. funestus s.l , respectively [40,41]. Subsequently, restriction fragment length polymorphism (RFLP) was performed for An. gambiae s.l speciation [40]. Detection of Plasmodium-infected mosquitoes Genomic DNA extracted from Anopheles head-thorax was tested for the presence of Plasmodium spp. by real-time PCR according to Mangold et al [42]. The primer sequences target a polymorphic fragment of the small subunit of 18S ribosomal RNA (18S rRNA), enabling the speciation of P. falciparum, P. vivax, P. ovale, and P. malariae. Statistical analysis Data were recorded in a Microsoft Excel spreadsheet (Office 2016) and imported to R studio software (San Diego, CA, USA) for analysis. Entomological indices (including vector diversity, relative abundance, aggressiveness, biting cycle and behavior, endophagy, and infection rate (IR) were calculated and presented as descriptive statistics. The chi-square test was used to compare mosquito density between sites. T-test and two-way ANOVA followed by Bonferroni’s multiple comparison tests were used to compare indoor and outdoor biting rates within the sites. The level of statistical significance was set at p-value ≤ 0.05. Biodiversity was assessed using two common diversity indices: the Shannon-Weaver index (H') [43] and the Simpson index (λ) [44]. These indices consider both the number of species (species richness) and the relative abundance of each species (evenness) in a sample. The similarity between the species communities at the three sites (Santchou, Dschang, and Penka Michel) was assessed using two ecological metrics: the General Jaccard Index [45], calculated as GJI% = d / (a + b + c - 2d) , where: a = number of species unique to community Santchou; b = number of species unique to community Dschang; c = number of species common to all the three communities; d = number of species unique of Penka Michel and the Renyi diversity profile (H α ) [46]. The entomological inoculation rate (EIR) was calculated using the following formula: EIR=IR×HBR , where IR= rate of female anopheles testing positive for Plasmodium spp infection, HBR= the human-biting rate expressed as the ratio of the number of anopheles mosquitoes captured by HLC to the total number of men by night. Results Diversity and specific richness of Culicidae fauna along the transect A total of 7,935 mosquitoes belonging to six genera were collected over 294 person-nights. The distribution of collected mosquito genera by sampling site is shown in T able 1 . Mosquitoes of the genus Culex spp were predominant (40.9%; n=3,245/7,935), while the genus Anopheles accounted for 39.82% (n= 3,160/7,935) . In addition to these genera , Aedes, Mansonia, Coquillettidia, and Eretmapodie were also collected, representing respectively 5.11% (n= 406/7,935), 14.13% (n= 1,122/7,935), 0.05% (n= 4/7,935) and 0.01% (n= 1/7,935). Overall, more mosquito was collected in Penka Michel (41.11%, n=3,262/7,935) compared to Dschang (33.23%, n=2,637/7,935) and Santchou (25.62%, n=2,033/7,935) and the difference in the distribution of mosquitoes between sites was statistically significant (p-value=0.0001, X 2 =9671, ddl=10). The genus Anopheles was more prevalent in Penka Michel (86.91%, n = 2,835) while in Dschang Culex spp represented 94.25% (n= 2,493) of the collection, and Santchou, Mansonia spp was predominant at 55.2%, (n= 1,119). Analysis of the Shannon-Weaver diversity indices (H') reveals a clear pattern: Santchou boasts the highest diversity (H' = 1.028), followed by Penka Michel (H' = 0.41) and Dschang (H' = 0.263). By using the Simpson diversity index (D), where lower values indicate higher diversity. Santchou (D = 0.219) has the lowest D value, followed by Penka Michel (D = 0.904) and lastly Dschang (D = 0.992). These findings suggest a relatively even distribution of species abundance in Santchou, while Dschang is dominated by Culex mosquitoes and Penka Michel by Anopheles . Further analysis using Jaccard's similarity index (60%) indicates a moderate level of similarity among the three communities. This observation is corroborated by the Renyi alpha biodiversity profiles depicted in Fig 2 . These profiles suggest that while each community likely harbors a dominant species, other species are present in sufficient numbers to contribute to overall diversity. Table 1: frequency and diversity of Culicidae fauna in Dschang, Santchou, and Penka Michel Species Santchou Dschang Penka Michel N % N % N % An. gambiae s.l 242 11.90 71 2.70 2,308 70.75 An. funestus s.l - - - - 503 15.42 An. ziemanni 4 0.20 7 0.27 24 0.74 An. nili s.l 3 0.15 - - - - Ae.africanus 0 0 51 1.93 320 9.81 Ae. simpsoni 0 0 3 0.11 0 0 Ae. aegypti 2 0.09 2 0.07 1 0.03 Ae. palpalis 4 0.19 5 0.18 0 0 Ae. albopictus 0 0 3 0.11 1 0.03 Aedes spp. 2 0.09 1 0.03 5 0.15 Culex spp. 652 32.07 2,493 94.54 100 3.07 Mansonia spp. 1,119 55.04 3 0.11 0 0 Coquilletidia spp. 4 0.2 0 0 0 0 Eretmapodie spp. 1 0.05 0 0 0 0 Total 2,033 100 2,637 100 3,262 100 Species richness 6 4 3 Shannon_H 1.028 0.263 0.41 Simpson_D 0.219 0.992 0.904 Jaccard 60% N= number; %= percentage; p-value=0.0001, chi-square=9671, ddl=10 Figure 2: Diversity profiles based on Rényi's H-alpha series between the three sites Anopheles species composition and abundance A total of 3,448 Anopheles mosquitoes were collected during the entomological survey, comprising 3,160 adults and 288 immature stages (larvae and pupae). Immature stages were primarily found in stagnant water pools. The limited availability of suitable anopheles breeding sites likely contributed to the lower number of larvae collected (75 in Dschang, 98 in Santchou, and 115 in Penka Michel). All immature stages were identified morphologically. An. gambiae s.l. was the only species found in breeding sites. Among the 3,160 adult mosquitoes, four species were morphologically identified: An. gambiae sl. (82.88%) , An. funestus s.l. (15.92%), An. nili (0.09%) and An. ziemanni (1.11%). Fig.3.A shows the proportion of Anopheles species collected at each site throughout the study. An. gambiae s.l. was the predominant vector in all sites representing 82.9% (2,619/3,160), while An. ziemanni, which is considered a secondary malaria vector represented only 1.61%, 9.21%, and 0.85% in Santchou, Dschang, and Penka Michel, respectively. Molecular speciation revealed that An. gambiae and An. coluzzii were the only members of the An. gambiae complex identified in the three study sites. An. gambiae was the main malaria vector, with 87.6% (n=298/340), 90.1% (n=131/144), and 95.8% (n=365/381), respectively, in Santchou, Dschang, and Penka Michel. For a subset of 176 An. funestus s.l . analyzed, 89.2% (n=157/176) were An. funestus s.s. and 10.8% (n=19/176) were An. leesoni ( Fig. 3. B ) No significant difference was recorded in the diversity of different anopheline mosquitoes across the study area. However, based on Shannon Weaver diversity indices for Anopheles species (Santchou H’=0.99, Dschang H’=0.25, and Penka Michel H’=0.33), Santchou stands out as the site with the greatest species diversity, harboring a rich array of species with a relatively balanced distribution of abundances. Figure 3: Anopheles fauna and abundance according to the study site; A: Global species composition B: Species composition of Anopheles sibling species; a=species composition within the An. gambiae complex in Santchou; b=species composition within the An. gambiae complex in Dschang; c=species composition within the An. gambiae complex in Penka Michel; d=species composition within the An. funestus group in Penka Michel. Anopheles biting behavior and cycle. Anopheles mosquitoes were collected outdoors and indoors and the relative aggressiveness (bites/human/night) of the dominant vectors An. gambiae s.s. and An. funestus s.s. is shown in Fig.4 . Anopheles mosquitoes bit both outdoors and indoors, with a significant difference observed in Dschang (64.47% vs 35.53%; p-value=0.039), Santchou (59.84% vs 40.16%; p-value=0.196), and Penka Michel, (54.50% vs 45.50%; p-value=0.577), with the tendency towards exophagy in Dschang. According to the biting cycle ( Fig. 5 ), the major malaria vectors were captured both indoors and outdoors during all collection periods (from 6:00 p.m. to 9:00 a.m.). However, there were notable differences across sites. In Santchou, An. gambiae s.l. biting activity varied throughout the night, starting to increase gradually at 08:00 p.m., peaking between 10:00 to 11:00 p.m. both indoors and outdoors, and a gradual decrease until 09:00 a.m. In Dschang, mosquito bites occurred mostly outdoors with a peak between 2:00 to 3:00 a.m. outdoors and between 11 p.m. to 12:00 p.m. indoors. In Penka Michel, An. gambiae s.l. biting activity was intense in the first half of the night, peaking between 10:00-11:00 p.m. outdoors and between 12:00 p.m.-1:00 a.m. indoors. For An. funestus s.l , the biting activity increased progressively, with high biting activity in the early hours of the morning, between 2:00 a.m.-3:00 a.m. indoors. Figure 4: An. gambiae s.l. and An. funestus s.l. biting behavior in Santchou, Dschang, and Penka Michel Figure 5: Biting cycles of An. gambiae s.l. and An. funestus s.l. collected indoors and outdoors in Santchou, Dschang, and Penka Michel. Plasmodium infection rate Of the 720 Anopheles mosquitoes tested by real-time PCR for the presence of Plasmodium infections. Only 19 (2.7%) of the mosquitoes were found to be infected by Plasmodium . These mosquitoes were from three species ( An. gambiae, An. funestus, and An. ziemanni ). In Santchou, the overall infection rate was 2.4% (6/249), with An. gambiae carrying mono-infections of P. falciparum at 66.67% (4/6) and P. malariae at 33.33% (2/6), respectively. In Dschang, no infected specimen was detected among the 76 mosquitoes analyzed. In Penka Michel, the overall infection rate was 3.2% (13/392). An. gambiae, An. funestus, and An. ziemanni tested positive for P. falciparum at 61.54% (8/13;) , P. malariae at 30.76% (4/13;), and P. ovale at 7.7% (1/13;). Human biting rate and entomological inoculation rate The entomological inoculation rate (EIR) in each study site is presented in Table 2 . An. gambiae s.l. was the most aggressive species with a human biting rate (HBR) of 3.1, 0.41, and 45.25 bites per human per night (b/h/n) respectively in Santchou, Dschang, and Penka Michel (p-value=0.0001). The global EIR for Penka Michel was 1.84 infective bites per human per night (ib/h/n), 23 times higher than in Santchou (0.08ib/h/n). The level of malaria transmission varies between Anopheles species and sites; An. gambiae appears to be the most competent malaria vector with an estimated EIR of 0.08ib/p/n in Santchou, and 1.1ib/p/n in Penka Michel, followed by An. funestus with 0.47 ib/p/n. The contribution of An. ziemanni in the transmission was recorded in Penka Michel, with 0.04 ib/p/n. Table 2: The entomological inoculation rate (EIR) for each site. Sites Mosquito species Tested Positive IR (%) HBR (b/h/n) EIR (ib/h/n) Santchou An. gambiae s.s. 242 6 2.5 3.1 0.08 An. funestus s.l. 0 0 0 0 0 An. ziemanni 4 0 0 0.05 0 An. nili s.l. 3 0 0 0.03 0 Global 249 6 2.41 3.19 0.077 Dschang An. gambiae s.s. 69 0 0 0.41 0 An. funestus s.l. 0 0 0 0 0 An. ziemanni 7 0 0 0.04 0 An. nili s.l. 0 0 0 0 0 Global 76 0 0 0.45 0 Penka Michel An. gambiae s.s. 285 7 2.45 45.25 1.11 An. funestus s.l. 83 4 4.82 9.86 0.47 An. ziemanni 24 2 8.33 0.47 0.04 An. nili s.l. 0 0 0 0 0 Global 392 13 3.32 55.58 1.84 IR (Infection Rate); EIR (Entomological Inoculation Rate); HBR (Human Biting Rate); p-value˃0.05 Discussion Entomological monitoring of malaria vectors plays a critical role in understanding their biology, ecology, and spatial distribution [47]. This knowledge is essential for designing targeted control strategies and deploying the most effective tools. This study was focused specifically on the highlands of Cameroon, where malaria control remains a challenge. Following the mass distribution of Long-lasting insecticidal nets (LLINs) as a vector control method, this study provides valuable data on key entomological parameters of malaria transmission in this region. Six Anopheles vectors were identified: An. gambiae s.s , An. coluzzii , An. funestus s.s , An. leesoni , An. nili , and An. ziemanni . This observed diversity is consistent with the varied aquatic habitats present in the study area, including permanent and temporary water bodies such as streams, lakes, rivers, and puddles, which provide suitable breeding grounds for these malaria vectors. However, a notable decline in overall species diversity compared to a previous study by Tchuinkam et al . [31] suggests potential shifts in the local mosquito population; these findings align with previous studies in the same region [48,49]. Climate change and associated environmental alterations are likely contributing factors to these changes, as reported for other Cameroonian epidemiological contexts [7]. The presence of these vectors, particularly the highly anthropophilic and competent An. gambiae s.s. and An. funestus s.s suggest their critical role in the persistence of malaria transmission in the study area. Among the Anopheles species identified, An. gambiae was the most prevalent across all three study sites. This dominance aligns with previous findings in Cameroon's West Region [31,50,51]. Its adaptability to highland environments is also well documented [5,52]. An. funestus , another major vector, was only found in Penka Michel at 1500m, representing 17.7% of the anopheline population collected. Only three specimens of An. nili s.l . were collected in Santchou and An. ziemanni was present at very low densities. This uneven distribution of Anopheles species likely stems from several factors, with temperature differences playing a potential role. During the rainy season, the temperatures are lower at Penka Michel and Dschang compared to the lowland site (Santchou) might explain some variation in vector densities. Previous studies have shown that cooler temperatures can delay larval development and increase mortality [53–55]. However, Penka Michel, despite having a similar climate to Dschang and located at roughly the same altitude, exhibited densities of An. gambiae thirty-three times higher. These observations suggest that climatic influence may not fully explain the discrepancies in vector densities. The hilly topography of Penka Michel likely plays a significant role. This geography promotes water accumulation and retention, creating more abundant and stable breeding sites for Anopheles species (swamps, rivers, lakes). Balls et al. (2004) demonstrated a link between flat-bottomed highland valleys, prone to water accumulation, and increased malaria risk in Tanzania. Similarly, other studies [56,55,52] highlight altitude, topography, and land use as significant environmental factors impacting malaria vector abundance in highland regions [57]. The observed differences in vector densities between Penka Michel and Dschang, despite similar altitudes, can be also attributed to factors such as urbanization. Dschang, with its higher level of urbanization, exhibited a significantly higher abundance of Culex spp (alone accounted for 76.8% of the total Culex spp. collected). This finding aligns with previous observations in the city [49,48], which reported a high abundance of larval habitats and populations of Culex spp. The Culex genus is an indicator of urbanization [48], whose predominance in urban areas could be attributed to factors such as intense human activity (economic and industrial), and inadequate environmental management practices, which can create suitable breeding sites [58,59]. While Culex spp. dominated the urban landscape, and in rural areas, the distribution changes. Aedes spp. and Mansonia spp. were also present, reaching 10.3% and 55.2% in Penka Michel and Santchou, respectively. These genera represent a substantial public health concern due to their established roles as vectors for various diseases in Africa [60]. Aedes spp., for instance, are notorious for transmitting arboviruses such as dengue and yellow fever, while Culex spp., Anopheles spp., and Mansonia spp. are implicated in the transmission of lymphatic filariasis [36–38]. Specific to our study, we identified four Aedes species: Ae. simpsoni , Ae. albopictus , Ae. aegypti , and Ae. africanus . These species have been associated with the transmission of dengue and other arboviral diseases in various regions [64–66], underscoring the potential public health risks posed by their presence in the study area. Consistent with previous studies in the west region [31,52,51], this study identified the presence of both An. gambiae s.s. and An. coluzzii , sibling species within the An. gambiae s.l . complex, co-existing along the altitudinal transect (sympatry). An. gambiae s.s . is known to be more adapted to semi-urban environments, preferring sunlit larval habitats. Conversely, An. coluzzii typically thrives in urbanized areas with artificial, polluted, and wetter breeding sites [51,67,68]. Furthermore, the An. funestus group taxa found exclusively at Penka Michel were primarily composed of An. funestus s.s . and An. leesoni . Recently the epidemiological role of An. leesoni in malaria transmission was demonstrated in certain regions of Cameroon [69]. An interesting observation was the difference in An. gambiae s.l . biting behavior across the altitudinal transect. Compared to previous studies [31] reporting endophagic behavior (biting indoors) in the same area, this study found exophagic behavior (feeding outdoors) in Dschang and exophilic behavior (resting outdoors) in Santchou. This shift could be attributed to behavioral changes in response to intensified vector control efforts, particularly the widespread distribution of LLINs: on this date, the household coverage rate in Milda is 100 (unpublished data). Similar trends have been observed in recent Cameroonian studies [10,52] and across Africa [70–72]. However, continuous entomological monitoring is crucial to understand better these mosquito species' spatial and behavioral variations in the study sites. In Penka Michel, both An. gambiae and An. funestus exhibited primarily endophagic behavior. This might be linked to the high mosquito nuisance levels, prompting residents to spend more nights indoors, which influences mosquito-biting behavior. Nevertheless, further investigations are needed to assess the sensitivity of these mosquito populations to the effects of insecticide-treated nets. Taken as a whole, An. gambiae was aggressive throughout the night, both indoors and outdoors, with peak biting activity occurring between 10:00 pm and 3:00 am. This extended biting period could be linked to the relatively low long-lasting insecticidal net (LLIN) usage in the western region (58.7%), falling below the 80% coverage recommended by the WHO [73]. These findings align with previous studies, which reported peak biting activity for both species occurring late at night (between 00:00 pm and 6:00 am) [74,50,51]. It is important to acknowledge the limitations of the study. While we have assessed the impact of LLINs on malaria transmission, we were unable to fully evaluate the distribution and actual use of LLINs in our study sites. Further research is needed to address these gaps. Overall, malaria transmission intensity across the study sites appeared low, with an entomological inoculation rate (EIR) ranging from 0.08 to 1.84 infected bites per human per night. Interestingly, An. gambiae , which was the most prevalent mosquito species biting exhibited the highest infection rate, followed by An. funestus and An. ziemanni . These findings contrast with the high EIR reported by Tchuinkam in 2010 [31] in the same area. This difference could potentially be attributed to climatic changes observed in recent years, where such variations have been shown in laboratory studies to affect survival and reproductive capacity [10]. Among the Plasmodium species identified, P. falciparum was the dominant parasite, accounting for up to 63% of infections in both Santchou and Penka Michel. This aligns with findings from other studies [25,75–77] and confirms the primary epidemiological role of P. falciparum in the region. Infections with P. malariae and P. ovale were also detected in infected vectors ( An. gambiae , An. funestus, and An. ziemanni ). The presence of non-falciparum malaria parasites in these locations highlights the importance of including these parasites in control programs to achieve successful malaria elimination efforts. While no P. vivax infections were detected in the analyzed mosquito samples, this doesn't necessarily exclude its presence in this region. Seminal studies have reported P. vivax cases in Duffy-negative individuals residing in the western Cameroon region (Dschang, Santchou), with prevalence ranging from 0.5% to 35% [28,29]. Furthermore, another study identified P. vivax infections within the An. coluzzii vector population in Tibati (Adamaoua region) [78]. This finding suggests the potential circulation of the parasite among the human population and its possible transmission by Anopheles mosquitoes. Additionally, Ossè and collaborators reported cases of P. vivax infection in vectors An. gambiae and An. coluzzii from Benin [79]. Conclusion We report on the variation in the abundance and dynamics of malaria vector populations in the West Cameroon highlands. Six Anopheles species were present in the study area, all known to be malaria vectors in Cameroon. An. gambiae was the most widespread species in the sites, followed by An. funestus although the distribution of the latter was highly focal. The uneven distribution of anopheline species within the study area further confirms that the presence of these species varies according to the micro and macro-environmental differences present in the bio-ecological zones, even at the same altitude. An. gambiae has shown a preference for biting outdoors rather than indoors in the lowland plain and on the mountain plateau, which could compromise malaria control strategies using mainly LLINs. Therefore, the implementation of additional tools (i.e., larviciding, integrated management, and environmental management) to combat these outdoor-biting mosquitoes must be considered. Abbreviations CTAB: Cetyltrimethyl ammonium bromide ; EIR: Entomological inoculation rate ; IR: Infection rate ; IRS: Insecticide residual spraying ; LLINs: Long-lasting insecticidal nets ; HBR: Human-biting rate ; PCR-RFLP: PCR-restriction fragment length polymorphism ; rRNA: Ribosomal ribonucleic acid ; PNLP: National malaria control program Declarations Ethics approval The study was approved by Cameroon's National Ethics Committee for Human Health Research (CNERSH, n° 2022/12/1507/CE/CNERSH/SP). The study was conducted in collaboration with the various local health authorities. Free and informed consent was obtained from volunteers (for mosquito collection) and family heads through individual interviews and group meetings. Presumptive malaria treatment was administered free of charge to volunteers before and after HLC collections. Consent for publication Consent to publish has been obtained from all included persons in the study. Conflicts of interests The authors declare no competing interests. The findings and conclusions expressed herein are those of the authors and do not necessarily represent the official position of the NIAID or NIH. Authors contributions JJJ, IAB, RRD, and SEN conceived the research; BCK, AGBT, II, RRD, and SEN designed the study protocol. BCK, AGBT, II, COJ, MM, PKT, ZRPH, SJW, LMA, and BFT carried out the field and laboratory assays. RRD, and SEN supervised field collection. JJJ, IAB, RRD, and SEN contributed reagents/materials. BCK, AGBT, II, LMA, JAMM, RRD, and SEN analyzed and interpreted data. BCK and SEN drafted the manuscript. BCK, AGBT, JJAM, II, COJ, MM, PKT, ZRPH, SJW, CTN, LMA, BFT, JMS, JBP, JTL, JJJ, IAB, RRD, and SEN reviewed and helped write the manuscript. All authors made intellectual input to this study. All authors read and approved the final manuscript. Funding This study was supported by the National Institute for Allergy and Infectious Diseases (NIAID) as part of the National Institutes of Health (NIH) in the United States (R01AI165537 to JJJ, SEN, and RRD). Availability of data and materials Data are archived and available on request from the corresponding author Acknowledgments We would like to thank the residents and mosquito collectors at the various sites who agreed to take part in this study, as well as their respective district authorities for their collaboration and support. References World malaria report 2023 [Internet]. [cited 2024 Mar 18]. Available from: https://www.who.int/teams/global-malaria-programme/reports/world-malaria-report-2023 World malaria report 2022 [Internet]. [cited 2024 Mar 22]. Available from: https://www.who.int/teams/global-malaria-programme/reports/world-malaria-report-2022 Bhatt S, Weiss DJ, Cameron E, Bisanzio D, Mappin B, Dalrymple U, et al. 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TCHUENKAM","email":"","orcid":"","institution":"University of Dschang","correspondingAuthor":false,"prefix":"","firstName":"Pacôme","middleName":"K.","lastName":"TCHUENKAM","suffix":""},{"id":401621828,"identity":"917b544b-2338-4ea0-9d44-3910a4e4c188","order_by":5,"name":"Zachary R. POPKIN-HALL","email":"","orcid":"","institution":"University of North Carolina at Chapel Hill","correspondingAuthor":false,"prefix":"","firstName":"Zachary","middleName":"R.","lastName":"POPKIN-HALL","suffix":""},{"id":401621829,"identity":"7565a8f1-3011-4a31-85c8-1a9848351161","order_by":6,"name":"Mariama MBOUH","email":"","orcid":"","institution":"University of Dschang","correspondingAuthor":false,"prefix":"","firstName":"Mariama","middleName":"","lastName":"MBOUH","suffix":""},{"id":401621830,"identity":"0651bc0c-6522-4f1b-9cea-cc133aa5357b","order_by":7,"name":"Jean Arthur MBIDA MBIDA","email":"","orcid":"","institution":"University of Douala","correspondingAuthor":false,"prefix":"","firstName":"Jean","middleName":"Arthur MBIDA","lastName":"MBIDA","suffix":""},{"id":401621831,"identity":"babe1047-d068-415a-a38c-f152ec08dd09","order_by":8,"name":"Charlène Tina NANSSONG","email":"","orcid":"","institution":"University of Douala","correspondingAuthor":false,"prefix":"","firstName":"Charlène","middleName":"Tina","lastName":"NANSSONG","suffix":""},{"id":401621832,"identity":"9bf8f9db-2875-4925-8b8f-dea02ec053ce","order_by":9,"name":"Luc Marcel ABATE","email":"","orcid":"","institution":"MIVEGEC, Université Montpellier, IRD, CNRS","correspondingAuthor":false,"prefix":"","firstName":"Luc","middleName":"Marcel","lastName":"ABATE","suffix":""},{"id":401621833,"identity":"7fa7d758-a635-43e2-9114-a85ee06f9629","order_by":10,"name":"Clément Janvier ONGUENE","email":"","orcid":"","institution":"Centre Pasteur du Cameroun","correspondingAuthor":false,"prefix":"","firstName":"Clément","middleName":"Janvier","lastName":"ONGUENE","suffix":""},{"id":401621834,"identity":"7494b204-be8a-4913-97e9-7c550c645943","order_by":11,"name":"Brigitte FOTSO TUMAMO","email":"","orcid":"","institution":"Centre Pasteur du Cameroun","correspondingAuthor":false,"prefix":"","firstName":"Brigitte","middleName":"FOTSO","lastName":"TUMAMO","suffix":""},{"id":401621835,"identity":"0b9fbd32-05d9-47c9-a7c6-5214426dffc9","order_by":12,"name":"Jacob M. SADLER","email":"","orcid":"","institution":"University of North Carolina at Chapel Hill","correspondingAuthor":false,"prefix":"","firstName":"Jacob","middleName":"M.","lastName":"SADLER","suffix":""},{"id":401621836,"identity":"50d5152b-9e00-497e-90f8-d376c2e18119","order_by":13,"name":"Jonathan B. PARR","email":"","orcid":"","institution":"University of North Carolina at Chapel Hill","correspondingAuthor":false,"prefix":"","firstName":"Jonathan","middleName":"B.","lastName":"PARR","suffix":""},{"id":401621837,"identity":"d78bf17b-a8ad-4b61-a1b2-18e0d6dc5028","order_by":14,"name":"Jessica T. LIN","email":"","orcid":"","institution":"University of North Carolina at Chapel Hill","correspondingAuthor":false,"prefix":"","firstName":"Jessica","middleName":"T.","lastName":"LIN","suffix":""},{"id":401621838,"identity":"1d1ce6cc-3ba4-4b99-a80c-fa1608f8fc30","order_by":15,"name":"Jonathan J. JULIANO","email":"","orcid":"","institution":"University of North Carolina at Chapel Hill","correspondingAuthor":false,"prefix":"","firstName":"Jonathan","middleName":"J.","lastName":"JULIANO","suffix":""},{"id":401621839,"identity":"72f1c5f9-74d1-4bfd-9dd8-0721e059f09c","order_by":16,"name":"Innocent Ali MBULLI","email":"","orcid":"","institution":"University of Dschang","correspondingAuthor":false,"prefix":"","firstName":"Innocent","middleName":"Ali","lastName":"MBULLI","suffix":""},{"id":401621840,"identity":"de4da4e3-dc1a-4bb3-bd7f-0bdfc40243a2","order_by":17,"name":"Rhoel R. DINGLASAN","email":"","orcid":"","institution":"University of Florida","correspondingAuthor":false,"prefix":"","firstName":"Rhoel","middleName":"R.","lastName":"DINGLASAN","suffix":""},{"id":401621841,"identity":"c8372d52-d8f2-4f2f-a043-575a016aa2b7","order_by":18,"name":"Sandrine Eveline NSANGO","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA40lEQVRIie3OsQrCMBCA4ZPKubR2TajGVwgEFLEP0y6dMoujIGQTV5+kOBYKulRcddPFTVAQ0UG0Rec2o2D+4ZIhHxcAk+l38210JwnUxvndSrRIxJpkGXwIBlokFQwk1yM9Nz0eriMrVJDdyGPhM0BnX0r686gnWhmGyprGdJpFArDBSwnfQtejys63rGPiqDQcd1T5x/hmdcsJyYk80mdBECtIIrv0orhAkOg5WmQrhx5kAUOyFIO2igRWks0qpvfRy+7MJofdSfnMrSJFlv0566SYGgCgdv/Ss85rk8lk+r/eRbpB9PUZ6WQAAAAASUVORK5CYII=","orcid":"","institution":"University of Bertoua","correspondingAuthor":true,"prefix":"","firstName":"Sandrine","middleName":"Eveline","lastName":"NSANGO","suffix":""}],"badges":[],"createdAt":"2024-12-01 13:38:07","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5558659/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5558659/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s12936-025-05480-w","type":"published","date":"2025-08-06T15:57:59+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":73723768,"identity":"50d5944e-97a2-4e44-8b2d-5ff251fbf97e","added_by":"auto","created_at":"2025-01-14 03:35:11","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":466914,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMap showing the study sites in the Western Region of Cameroon.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5558659/v1/2a07ad908eede03ce6dfd1dc.png"},{"id":73723770,"identity":"1440a4f1-bad8-44cd-aab3-8df513b84865","added_by":"auto","created_at":"2025-01-14 03:35:11","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":55443,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDiversity profiles based on Rényi's H-alpha series between the three sites\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5558659/v1/a6687fd0513f54691556d9e2.png"},{"id":73723769,"identity":"c8a3bf26-99a0-4b51-b65b-c4d6392de2b1","added_by":"auto","created_at":"2025-01-14 03:35:11","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":116887,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAnopheles fauna and abundance according to the study site; A: Global species composition B: Species composition of\u003c/strong\u003e\u003cem\u003e\u003cstrong\u003e Anopheles \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003esibling species; a=species composition within the\u003c/strong\u003e\u003cem\u003e\u003cstrong\u003e An. gambiae \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003ecomplex in Santchou; b=species composition within the\u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAn. gambiae \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003ecomplex in Dschang; c=species composition within the\u003c/strong\u003e\u003cem\u003e\u003cstrong\u003e An. gambiae \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003ecomplex in Penka Michel; d=species composition within the \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAn. funestus \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003egroup in Penka Michel.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5558659/v1/e260a20c7874eca06c8fcd07.png"},{"id":73723772,"identity":"d8ba9969-a68d-4e36-acb2-d7ffa857cf7a","added_by":"auto","created_at":"2025-01-14 03:35:11","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":59614,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eAn. gambiae s.l.\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e and \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAn. funestus s.l.\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e biting behavior in Santchou, Dschang, and Penka Michel\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5558659/v1/936df6adb105d252eb30d778.png"},{"id":73723779,"identity":"2e17b57c-1c05-40e4-a264-84bf72e0fb46","added_by":"auto","created_at":"2025-01-14 03:35:12","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":147078,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eBiting cycles of An. gambiae s.l. and An. funestus s.l. collected indoors and outdoors in Santchou, Dschang, and Penka Michel.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-5558659/v1/9175ba332aa875869fa60c57.png"},{"id":88814206,"identity":"d5a47dab-06a9-42a2-8234-61878c691d2a","added_by":"auto","created_at":"2025-08-11 16:08:30","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2636003,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5558659/v1/f9301fe3-f7ee-4088-9a4f-96a01166e9b1.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Diversity, abundance of anopheline species, and malaria transmission dynamics in high-altitude areas of western Cameroon","fulltext":[{"header":"Background","content":"\u003cp\u003eMalaria remains a major public health problem worldwide [1]. The latest WHO estimates point to 249 million cases and 608,000 deaths, with 94% of cases occurring in the African region. Children under 5 years and pregnant women represent the most vulnerable and affected groups. In addition to therapeutic care, considerable progress has been made in the implementation of vector control measures such as long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) [2,3]. The rapid spread of these strategies has so far been a major factor in the decline in malaria morbidity and mortality over the past two decades [3].\u003c/p\u003e\n\u003cp\u003eMalaria remains a major public health challenge in Cameroon, in 2023, Cameroon recorded 4.85 million cases of suspected malaria, of which over a million were severe, mainly in children under 5. Malaria contributed to 7.3% of hospital deaths [4]. The entire population of approximately 28 million people is at risk of malaria, with the East, Centre, and North regions being the most affected [5]. The intensification of free LLIN distribution has contributed substantially to reducing the malaria burden in the country, with an estimated population coverage of 80% [5\u0026ndash;7]. However, the effectiveness of these control tools is threatened by several factors, including insecticide resistance, changes in the feeding and resting behavior of the main malaria vectors [8\u0026ndash;12], and the spatiotemporal variation in malaria transmission across different ecological zones of the country. Cameroon is often known as \u0026ldquo;Africa in miniature\u0026rdquo; due to the diversity of its natural environment distributed across its 10 provinces: deserts, coasts, mountains, rainforests, and savannah, resulting in heterogenous conditions [13] that directly influence mosquito distribution, abundance, survival, and malaria parasite development.\u003c/p\u003e\n\u003cp\u003eVariations in vector ecology across the landscape can lead to uneven exposure and transmission risks [14]. The lowlands are characterized by floods and swamps, promoting larvae survival and a greater abundance of vectors, resulting in high malaria prevalence [15\u0026ndash;17]. In contrast, the highlands consist of hills and valleys, resulting in a sparse distribution of larval habitats and a lower malaria prevalence (\u003cem\u003eP. falciparum\u003c/em\u003e prevalence based on RDT positivity of 11-19%) [18]. The diverse malaria eco-epidemiological contexts and local vector ecologies call for tailored interventions for each transmission focus, as malaria epidemiology fluctuates over time and is correlated with the local success of control programs.\u003c/p\u003e\n\u003cp\u003eThe success of these control strategies therefore depends on up-to-date information on bionomic, and vector transmission patterns in different epidemiological contexts, particularly in regions at high risk of epidemics, including high-altitude areas. In recent years, mountainous regions of Africa have experienced an increase in the frequency and intensity of malaria epidemics. These areas, characterized by lower temperatures and humidity, present unique challenges for malaria transmission [19\u0026ndash;22]. Unlike holoendemic lowland regions, highland areas are typically hypoendemic, making successful entomological inoculation more likely [23\u0026ndash;25]. Despite the significance of the Bamileke plateau in western Cameroon (large population and role in the country\u0026apos;s economic life), research on malaria transmission in these areas has been limited [26,27]. Moreover, recent reports of \u003cem\u003ePlasmodium vivax\u003c/em\u003e infections in Dschang [28,29] further underscore the need for a comprehensive understanding of malaria dynamics in this region. To address this knowledge gap, we conducted a series of entomological and parasitological (unpublished data) surveys in three localities of the region. We aimed to assess the extent of malaria transmission and identify the primary vectors driving the disease, considering the region\u0026apos;s diverse ecological settings and population movements.\u003c/p\u003e"},{"header":"Materials and methods ","content":"\u003cp\u003e\u003cstrong\u003eDescription of the study area\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was carried out from May to June 2023 in three high-altitude localities in the Menoua division along an altitudinal transect in the Western region of Cameroon: Santchou (700m), Dschang (1400m) and Penka Michel (1500m) are located in a savannah landscape within the Guineo-Congolese bioclimatic domain, on the volcanic line of Cameroon [30,31]\u0026nbsp;(\u003cstrong\u003eFig. 1\u003c/strong\u003e).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSantchou (5\u0026deg;25\u0026apos;N; 9\u0026deg;58\u0026apos;E) is located at an altitude of 700m in the vast Mb\u0026ocirc; plain, bounded to the south by the Manengouba mountain and to the north by a sloping cliff, delimiting it from the For\u0026eacute;k\u0026eacute; village. The area is mesoendemic for malaria, with a prevalence of 36.19%\u0026nbsp;[27]. Santchou covers an area of 335 km\u003csup\u003e2\u003c/sup\u003e and has a population ranging from 15,000 to around 25,000 and benefits from a dense hydrographic (the river Nkam and its tributaries) network and good rainfall. As a semi-urban city, Santchou has a low plain topography with a hygromorphic soil type. The climate is equatorial Guinean, characterized by four seasons of unequal duration: the long rainy season (mid-August to October); the short rainy season (March to June); the long dry season (mid-October to March); and the short dry season (June to mid-August). Average rainfall is 1,363.3 mm/year and annual temperature ranges from 20.83\u0026deg;C to 23.35\u0026deg;C [32].\u003c/p\u003e\n\u003cp\u003eThe town of Dschang (5\u0026deg;25\u0026apos;N; 10\u0026deg;03\u0026apos;E) is an urban environment that lies at the summit of the Mb\u0026ocirc; cliffs on the southeastern slopes of the Bamboutos mountains. It is bounded on the south by the commune of Santchou and on the northeast by the Bamboutos mountains. It covers an area of 262 km\u003csup\u003e2\u003c/sup\u003e and has a population of 120,207 inhabitants. The topography consists of alternating hills and valleys crisscrossed by small watercourses. It is characterized by 2 seasons: a rainy season (eight months) from mid-March to mid-November and a dry season (four months) from mid-November to mid-March. The area is characterized by a mosaic of agricultural landscapes and is classified as hypoendemic for malaria, with a prevalence of 25.04% [27]. Average precipitation is 2,000 mm/year and the daily temperature range can exceed 23\u0026deg;C during the dry season which constitutes a particularity of this locality [33]\u0026nbsp;\u003c/p\u003e\n\u003cp\u003ePenka Michel (5\u0026deg;28\u0026apos;N; 10\u0026deg;15\u0026apos;E) landscape is marked by a shallow topography, with a population of 98,229 inhabitants and a population density of 383.7 inhabitants/km\u0026sup2;. The area has a relatively low malaria prevalence of 38.4% among children under five years old\u0026nbsp;[34]. The relief is that of an undulating plateau with a flat bottom. As in Dschang (29 km from Penka Michel), the area is characterized by a sub-equatorial climate, with distinct wet and dry seasons. The rains peak in August-September when up to 345.1 mm of rainfall can be recorded monthly. Temperatures are relatively low and constant due to the high altitude, with an annual temperature range of only 26\u0026deg;C\u0026nbsp;[35].\u003c/p\u003e\n\u003cp\u003eThese diverse ecological settings, ranging from lowland to highland, provide an ideal opportunity to study the influence of environmental factors on malaria transmission dynamics. This study was conducted following the fourth distribution campaign of Long-Lasting Insecticidal Net (LLIN) in the Western region of Cameroon in 2022.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFigure\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003cstrong\u003e: Map showing the study sites in the Western Region of Cameroon.\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMosquito collections and processing\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHouseholds were randomly selected in each site to assess malaria transmission using the human-landing catch (HLC) approach\u0026nbsp;[36]\u0026nbsp;from 6:00 pm to 9:00 am.\u003c/p\u003e\n\u003cp\u003eIn Dschang, HLCs were carried out in 10 villages in four health zones (Bafou, Fialah-foreke, Foto, and Siteu). Four and three villages were selected respectively for mosquito collection in Santchou and Penka Michel.\u003c/p\u003e\n\u003cp\u003eDuring each survey, adult mosquitoes were collected indoors (living room or bedroom) and outdoors (outdoor shelter) in randomly selected homes, with the distance between two houses being at least 100m. HLCs were carried out during 11 nights in Dschang with a total number of 166 men/nights, 5 nights in the Santchou district with a total number of 78 men/nights, and 4 nights in the Penka Michel district with a total number of 50 men/nights\u0026nbsp;leveraging trained volunteers. To minimize collectors\u0026rsquo; bias due to the variability of attractiveness and collection skills, volunteers changed positions every two hours between indoor and outdoor stations and rotated through the houses. Each collector was provided with a flashlight and a bag containing hemolysis tubes with cotton. Mosquitoes landing on the legs of the collectors were captured using haemolysis tubes and stored in a labeled bag. At each sampling station, bags containing mosquitoes were collected hourly for morphological identification.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAnopheles larval collection and identification of adult mosquitoes\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAnopheles larval surveys were carried out in all the water collections identified in the study areas. The pre-imaginal stages of anopheles present in these sites were collected using the Dipping method\u0026nbsp;[37]\u0026nbsp;and brought back to the insectarium for rearing. On arrival at the insectarium, the larvae were transferred to tanks containing water from the lodge, according to their stage of development, and then fed with TetraMinBaby\u0026reg;. The resulting pupae were transferred to small cups and placed in adult-rearing cages until they emerged.\u003c/p\u003e\n\u003cp\u003eMosquitoes collected were sorted by genus (\u003cem\u003eCulex, Anopheles, Aedes, Mansonia,\u0026nbsp;\u003c/em\u003eand others) and species level using specific identification keys [38]. \u003cem\u003eAnopheles\u003c/em\u003e mosquitoes were then dissected into head-thorax, and abdomen, preserved individually in Eppendorf tubes containing silica gel for further molecular analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eLaboratory processing of mosquitoes\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDNA extraction, molecular identification of \u003cem\u003eAnopheles\u003c/em\u003e species\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGenomic DNA was extracted from the head-thorax and legs using a 2% CTAB (cetyltrimethylammonium bromide) solution according to Collins \u003cem\u003eet al\u0026nbsp;\u003c/em\u003e[39]. Genomic DNA was amplified by multiplex Polymerase Chain Reaction (PCR) and targeted the intergenic spacer of the ribosomal DNA (rDNA IGS) and internal transcribed spacer region 2 (ITS2) of \u003cem\u003eAn. gambiae s.l.\u0026nbsp;\u003c/em\u003eand \u003cem\u003eAn. funestus s.l\u003c/em\u003e, respectively [40,41]. Subsequently, restriction fragment length polymorphism (RFLP) was performed for \u003cem\u003eAn. gambiae s.l\u003c/em\u003e speciation [40].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDetection of \u003cem\u003ePlasmodium-infected\u003c/em\u003e mosquitoes\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGenomic DNA extracted from \u003cem\u003eAnopheles\u003c/em\u003e head-thorax was tested for the presence of \u003cem\u003ePlasmodium\u003c/em\u003e \u003cem\u003espp.\u003c/em\u003e by real-time PCR according to Mangold \u003cem\u003eet al\u0026nbsp;\u003c/em\u003e[42]. The primer sequences target a polymorphic fragment of the small subunit of 18S ribosomal RNA (18S rRNA), enabling the speciation of \u003cem\u003eP.\u003c/em\u003e\u003cem\u003e\u0026nbsp;falciparum, P. vivax, P. ovale,\u0026nbsp;\u003c/em\u003eand \u003cem\u003eP. malariae.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were recorded in a Microsoft Excel spreadsheet (Office 2016) and imported to R studio software (San Diego, CA, USA) for analysis. Entomological indices (including vector diversity, relative abundance, aggressiveness, biting cycle and behavior, endophagy, and infection rate (IR) were calculated and presented as descriptive statistics. The chi-square test was used to compare mosquito density between sites. T-test and two-way ANOVA followed by Bonferroni\u0026rsquo;s multiple comparison tests were used to compare indoor and outdoor biting rates within the sites. The level of statistical significance was set at p-value \u0026le; 0.05.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBiodiversity was assessed using two common diversity indices: the Shannon-Weaver index (H\u0026apos;) [43] and the Simpson index (\u0026lambda;) [44]. These indices consider both the number of species (species richness) and the relative abundance of each species (evenness) in a sample.\u003c/p\u003e\n\u003cp\u003e\u003cimg 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\" width=\"679\" height=\"55\"\u003e\u003c/p\u003e\n\u003cp\u003eThe similarity between the species communities at the three sites (Santchou, Dschang, and Penka Michel) was assessed using two ecological metrics: the General Jaccard Index [45], calculated as \u003cem\u003eGJI% = d / (a + b + c - 2d)\u003c/em\u003e, where: a = number of species unique to community Santchou; b = number of species unique to community Dschang; c = number of species common to all the three communities; d = number of species unique of Penka Michel and the Renyi diversity profile (H\u003csub\u003e\u0026alpha;\u003c/sub\u003e) [46].\u003c/p\u003e\n\u003cp\u003e\u003cimg src=\"data:image/png;base64,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\" width=\"567\" height=\"85\"\u003e\u003c/p\u003e\n\u003cp\u003eThe entomological inoculation rate (EIR) was calculated using the following formula:\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eEIR=IR\u0026times;HBR\u003c/em\u003e,\u003cem\u003e\u0026nbsp;\u003c/em\u003ewhere IR= rate of female anopheles testing positive for \u003cem\u003ePlasmodium spp\u0026nbsp;\u003c/em\u003einfection, HBR= the human-biting rate expressed as the ratio of the number of anopheles mosquitoes captured by HLC to the total number of men by night.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eDiversity and specific richness of Culicidae\u003cem\u003e\u0026nbsp;\u003c/em\u003efauna along the transect\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA\u0026nbsp;total of 7,935\u0026nbsp;mosquitoes\u0026nbsp;belonging to six genera were collected over 294 person-nights.\u0026nbsp;The distribution of collected mosquito genera by sampling site is shown in T\u003cstrong\u003eable 1\u003c/strong\u003e. Mosquitoes of the genus \u003cem\u003eCulex spp\u0026nbsp;\u003c/em\u003ewere predominant (40.9%; n=3,245/7,935), while the genus \u003cem\u003eAnopheles\u0026nbsp;\u003c/em\u003eaccounted for 39.82% (n= 3,160/7,935)\u003cem\u003e.\u0026nbsp;\u003c/em\u003eIn\u003cem\u003e\u0026nbsp;\u003c/em\u003eaddition to these genera\u003cem\u003e,\u0026nbsp;\u003c/em\u003e\u003cem\u003eAedes, Mansonia,\u0026nbsp;\u003c/em\u003e\u003cem\u003eCoquillettidia,\u0026nbsp;\u003c/em\u003eand \u003cem\u003eEretmapodie\u0026nbsp;\u003c/em\u003ewere also collected, representing respectively 5.11% (n= 406/7,935), 14.13% (n= 1,122/7,935), 0.05% (n= 4/7,935) and 0.01% (n= 1/7,935). Overall,\u0026nbsp;more mosquito was collected in Penka Michel (41.11%, n=3,262/7,935) compared to Dschang (33.23%, n=2,637/7,935) and Santchou (25.62%, n=2,033/7,935) and the difference in the distribution of mosquitoes between sites was statistically significant (p-value=0.0001, X\u003csup\u003e2\u003c/sup\u003e=9671, ddl=10). The genus \u003cem\u003eAnopheles\u0026nbsp;\u003c/em\u003ewas more prevalent in\u003cem\u003e\u0026nbsp;\u003c/em\u003ePenka Michel (86.91%, n\u003cem\u003e=\u0026nbsp;\u003c/em\u003e2,835) while in Dschang \u003cem\u003eCulex spp\u0026nbsp;\u003c/em\u003erepresented 94.25% (n= 2,493) of the collection, and Santchou, \u003cem\u003eMansonia spp\u0026nbsp;\u003c/em\u003ewas predominant at 55.2%, (n= 1,119).\u003c/p\u003e\n\u003cp\u003eAnalysis of the Shannon-Weaver diversity indices (H\u0026apos;) reveals a clear pattern: Santchou boasts the highest diversity (H\u0026apos; = 1.028), followed by Penka Michel (H\u0026apos; = 0.41) and Dschang (H\u0026apos; = 0.263). By using the Simpson diversity index (D), where lower values indicate higher diversity. \u0026nbsp; Santchou (D = 0.219) has the lowest D value, followed by Penka Michel (D = 0.904) and lastly Dschang (D = 0.992). These findings suggest a relatively even distribution of species abundance in Santchou, while Dschang is dominated by \u003cem\u003eCulex\u003c/em\u003e mosquitoes and Penka Michel by \u003cem\u003eAnopheles\u003c/em\u003e. Further analysis using Jaccard\u0026apos;s similarity index (60%) indicates a moderate level of similarity among the three communities. This observation is corroborated by the Renyi alpha biodiversity profiles depicted in \u003cstrong\u003eFig 2\u003c/strong\u003e. These profiles suggest that while each community likely harbors a dominant species, other species are present in sufficient numbers to contribute to overall diversity.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1: frequency and diversity of\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eCulicidae\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;fauna in Dschang, Santchou, and Penka Michel\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"619\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpecies\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 18.4707%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSantchou\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 18.9176%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDschang\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 17.1301%;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePenka Michel\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eN\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eN\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eN\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. gambiae s.l\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e242\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e11.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e2.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e2,308\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e70.75\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. funestus s.l\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e503\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e15.42\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. ziemanni\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e0.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e0.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. nili s.l\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e0.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eAe.africanus\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e1.93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e320\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e9.81\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eAe. simpsoni\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eAe. aegypti\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e0.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eAe. palpalis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e0.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e0.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eAe. albopictus\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eAedes spp.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e0.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0.15\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eCulex spp.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e652\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e32.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e2,493\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e94.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e3.07\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eMansonia spp.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e1,119\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e55.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eCoquilletidia spp.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cem\u003eEretmapodie spp.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2,033\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.6395%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e100\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.2354%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2,637\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9.6822%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e100\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3,262\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 8.4906%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e100\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003eSpecies richness\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 18.4707%;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 18.9176%;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 17.1301%;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003eShannon_H\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 18.4707%;\"\u003e\n \u003cp\u003e1.028\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 18.9176%;\"\u003e\n \u003cp\u003e0.263\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 17.1301%;\"\u003e\n \u003cp\u003e0.41\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003eSimpson_D\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 18.4707%;\"\u003e\n \u003cp\u003e0.219\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 18.9176%;\"\u003e\n \u003cp\u003e0.992\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" style=\"width: 17.1301%;\"\u003e\n \u003cp\u003e0.904\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 22.3436%;\"\u003e\n \u003cp\u003eJaccard\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" style=\"width: 54.5184%;\"\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp;60%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eN= number; %= percentage; p-value=0.0001, chi-square=9671, ddl=10\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFigure 2: Diversity profiles based on R\u0026eacute;nyi\u0026apos;s H-alpha series between the three sites\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eAnopheles\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;species composition\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eand abundance\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA total of 3,448 Anopheles mosquitoes were collected during the entomological survey, comprising 3,160 adults and 288 immature stages (larvae and pupae). Immature stages were primarily found in stagnant water pools. The limited availability of suitable anopheles breeding sites likely contributed to the lower number of larvae collected (75 in Dschang, 98 in Santchou, and 115 in Penka Michel). All immature stages were identified morphologically. \u003cem\u003eAn. gambiae s.l.\u003c/em\u003e was the only species found in breeding sites.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAmong the 3,160 adult mosquitoes, four species were morphologically identified: \u003cem\u003eAn. gambiae sl. (82.88%)\u003c/em\u003e, \u003cem\u003eAn. funestus s.l. (15.92%), An. nili (0.09%)\u0026nbsp;\u003c/em\u003eand \u003cem\u003eAn. ziemanni\u003c/em\u003e (1.11%). \u003cstrong\u003eFig.3.A\u003c/strong\u003e shows the proportion of \u003cem\u003eAnopheles\u003c/em\u003e species collected at each site throughout the study. \u003cem\u003eAn. gambiae s.l.\u003c/em\u003e was the predominant vector in all sites representing 82.9% (2,619/3,160), while \u003cem\u003eAn. ziemanni,\u003c/em\u003e which is considered a secondary malaria vector represented only 1.61%, 9.21%, and 0.85% in Santchou, Dschang, and Penka Michel, respectively.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMolecular speciation revealed that \u003cem\u003eAn. gambiae\u0026nbsp;\u003c/em\u003eand \u003cem\u003eAn. coluzzii\u0026nbsp;\u003c/em\u003ewere the only members of the \u003cem\u003eAn. gambiae\u0026nbsp;\u003c/em\u003ecomplex identified in the three study sites. \u003cem\u003eAn. gambiae\u0026nbsp;\u003c/em\u003ewas the\u003cem\u003e\u0026nbsp;\u003c/em\u003emain malaria vector, with 87.6% (n=298/340), 90.1% (n=131/144), and 95.8% (n=365/381), respectively, in Santchou, Dschang, and Penka Michel. For a subset of 176 \u003cem\u003eAn. funestus s.l\u003c/em\u003e. analyzed, 89.2% (n=157/176) were \u003cem\u003eAn. funestus s.s.\u0026nbsp;\u003c/em\u003eand 10.8% (n=19/176) were \u003cem\u003eAn. leesoni\u0026nbsp;\u003c/em\u003e(\u003cstrong\u003eFig. 3. B\u003c/strong\u003e)\u003c/p\u003e\n\u003cp\u003eNo significant difference was recorded in the diversity of different anopheline mosquitoes across the study area. However, based on Shannon Weaver diversity indices for \u003cem\u003eAnopheles\u003c/em\u003e species (Santchou H\u0026rsquo;=0.99, Dschang H\u0026rsquo;=0.25, and Penka Michel H\u0026rsquo;=0.33), Santchou stands out as the site with the greatest species diversity, harboring a rich array of species with a relatively balanced distribution of abundances.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFigure 3:\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;Anopheles fauna and abundance according to the study site; A: Global species composition B: Species composition of\u003cem\u003e\u0026nbsp;Anopheles\u0026nbsp;\u003c/em\u003esibling species; a=species composition within the\u003cem\u003e\u0026nbsp;An. gambiae\u0026nbsp;\u003c/em\u003ecomplex in Santchou; b=species composition within the\u003cem\u003e\u0026nbsp;An. gambiae\u0026nbsp;\u003c/em\u003ecomplex in Dschang; c=species composition within the\u003cem\u003e\u0026nbsp;An. gambiae\u0026nbsp;\u003c/em\u003ecomplex in Penka Michel; d=species composition within the \u003cem\u003eAn. funestus\u0026nbsp;\u003c/em\u003egroup in Penka Michel.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eAnopheles\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003ebiting behavior and cycle.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eAnopheles\u003c/em\u003e mosquitoes were collected outdoors and indoors and the relative aggressiveness (bites/human/night) of the dominant vectors \u003cem\u003eAn. gambiae s.s.\u003c/em\u003e and \u003cem\u003eAn. funestus s.s.\u003c/em\u003e is shown in \u003cstrong\u003eFig.4\u003c/strong\u003e. \u003cem\u003eAnopheles\u003c/em\u003e mosquitoes bit both outdoors and indoors, with a significant difference observed in Dschang (64.47% \u003cem\u003evs\u003c/em\u003e 35.53%; p-value=0.039), Santchou (59.84% \u003cem\u003evs\u003c/em\u003e 40.16%; p-value=0.196), and Penka Michel, (54.50% \u003cem\u003evs\u003c/em\u003e 45.50%; p-value=0.577), with the tendency towards exophagy in Dschang.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAccording to the biting cycle (\u003cstrong\u003eFig. 5\u003c/strong\u003e), the major malaria vectors were captured both indoors and outdoors during all collection periods (from 6:00 p.m. to 9:00 a.m.). However, there were notable differences across sites. In Santchou, \u003cem\u003eAn. gambiae s.l.\u003c/em\u003e biting activity varied throughout the night, starting to increase gradually at 08:00 p.m., peaking between 10:00 to 11:00 p.m. both indoors and outdoors, and a gradual decrease until 09:00 a.m. In Dschang, mosquito bites occurred mostly outdoors with a peak between 2:00 to 3:00 a.m. outdoors and between 11 p.m. to 12:00 p.m. indoors. In Penka Michel, \u003cem\u003eAn. gambiae s.l.\u0026nbsp;\u003c/em\u003ebiting activity was intense in the first half of the night, peaking between 10:00-11:00 p.m. outdoors and between 12:00 p.m.-1:00 a.m. indoors. For \u003cem\u003eAn. funestus s.l\u003c/em\u003e, the biting activity increased progressively, with high biting activity in the early hours of the morning, between 2:00 a.m.-3:00 a.m. indoors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFigure 4: \u003cem\u003eAn. gambiae s.l.\u003c/em\u003e and \u003cem\u003eAn. funestus s.l.\u003c/em\u003e biting behavior in Santchou, Dschang, and Penka Michel\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFigure 5:\u003c/strong\u003e \u003cstrong\u003eBiting cycles of\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eAn. gambiae s.l.\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eand\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eAn. funestus s.l.\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003ecollected indoors and outdoors in Santchou, Dschang, and Penka Michel.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003ePlasmodium\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;infection rate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOf the 720 \u003cem\u003eAnopheles\u003c/em\u003e mosquitoes tested by real-time PCR for the presence of \u003cem\u003ePlasmodium\u003c/em\u003e infections. Only 19 (2.7%) of the mosquitoes were found to be infected by \u003cem\u003ePlasmodium\u003c/em\u003e. These mosquitoes were from three species (\u003cem\u003eAn. gambiae, An. funestus,\u003c/em\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003eand \u003cem\u003eAn. ziemanni\u003c/em\u003e). \u0026nbsp;In Santchou, the overall infection rate was 2.4% (6/249), with \u003cem\u003eAn. gambiae\u003c/em\u003e carrying mono-infections of \u003cem\u003eP. falciparum\u003c/em\u003e at 66.67% (4/6) and \u003cem\u003eP. malariae\u0026nbsp;\u003c/em\u003eat 33.33% (2/6), respectively. \u0026nbsp;In Dschang, no infected specimen was detected among the 76 mosquitoes analyzed. \u0026nbsp;In Penka Michel, the overall infection rate was 3.2% (13/392). \u003cem\u003eAn. gambiae, An. funestus,\u0026nbsp;\u003c/em\u003eand \u003cem\u003eAn. ziemanni\u003c/em\u003e tested positive for \u003cem\u003eP. falciparum at\u0026nbsp;\u003c/em\u003e61.54% (8/13;)\u003cem\u003e, P. malariae\u0026nbsp;\u003c/em\u003eat\u003cem\u003e\u0026nbsp;\u003c/em\u003e30.76%\u003cem\u003e\u0026nbsp;\u003c/em\u003e(4/13;), and\u003cem\u003e\u0026nbsp;P. ovale\u003c/em\u003e at 7.7% (1/13;).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHuman biting rate and entomological inoculation rate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe entomological inoculation rate (EIR) in each study site is presented in \u003cstrong\u003eTable 2\u003c/strong\u003e. \u003cem\u003eAn. gambiae s.l.\u003c/em\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003ewas the most aggressive species with a human biting rate (HBR) of 3.1, 0.41, and 45.25 bites per human per night (b/h/n) respectively in Santchou, Dschang, and Penka Michel (p-value=0.0001). The global EIR for Penka Michel was 1.84 infective bites per human per night (ib/h/n), 23 times higher than in Santchou (0.08ib/h/n). The level of malaria transmission varies between \u003cem\u003eAnopheles\u003c/em\u003e species and sites; \u003cem\u003eAn. gambiae\u003c/em\u003e appears to be the most competent malaria vector with an estimated EIR of 0.08ib/p/n in Santchou, and 1.1ib/p/n in Penka Michel, followed by\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cem\u003eAn. funestus\u0026nbsp;\u003c/em\u003ewith 0.47 ib/p/n. The contribution of \u003cem\u003eAn. ziemanni\u0026nbsp;\u003c/em\u003ein the transmission was recorded in Penka Michel, with 0.04 ib/p/n.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2: The entomological inoculation rate (EIR) for each site.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"711\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSites\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMosquito species\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTested\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePositive\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eIR (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHBR (b/h/n)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eEIR (ib/h/n)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"5\" valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSantchou\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. gambiae s.s.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e242\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e3.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. funestus s.l.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. ziemanni\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. nili s.l.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003eGlobal\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e249\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e2.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e3.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0.077\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"5\" valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDschang\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. gambiae s.s.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e0.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. funestus s.l.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. ziemanni\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e0.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. nili s.l.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003eGlobal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e0.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"5\" valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePenka Michel\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. gambiae s.s.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e285\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e2.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e45.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e1.11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. funestus s.l.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e4.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e9.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0.47\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. ziemanni\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e8.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e0.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0.04\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003e\u003cem\u003eAn. nili s.l.\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003eGlobal\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e392\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 81px;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 82px;\"\u003e\n \u003cp\u003e3.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 101px;\"\u003e\n \u003cp\u003e55.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 99px;\"\u003e\n \u003cp\u003e1.84\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eIR (Infection Rate); EIR (Entomological Inoculation Rate); HBR (Human Biting Rate); p-value˃0.05\u003c/strong\u003e\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eEntomological monitoring of malaria vectors plays a critical role in understanding their biology, ecology, and spatial distribution [47]. This knowledge is essential for designing targeted control strategies and deploying the most effective tools. This study was focused specifically on the highlands of Cameroon, where malaria control remains a challenge. Following the mass distribution of Long-lasting insecticidal nets (LLINs) as a vector control method, this study provides valuable data on key entomological parameters of malaria transmission in this region.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSix \u003cem\u003eAnopheles\u003c/em\u003e vectors were identified: \u003cem\u003eAn. gambiae s.s\u003c/em\u003e, \u003cem\u003eAn. coluzzii\u003c/em\u003e, \u003cem\u003eAn. funestus s.s\u003c/em\u003e, \u003cem\u003eAn. leesoni\u003c/em\u003e, \u003cem\u003eAn. nili\u003c/em\u003e, and \u003cem\u003eAn. ziemanni\u003c/em\u003e. This observed diversity is consistent with the varied aquatic habitats present in the study area, including permanent and temporary water bodies such as streams, lakes, rivers, and puddles, which provide suitable breeding grounds for these malaria vectors. However, a notable decline in overall species diversity compared to a previous study by Tchuinkam et \u003cem\u003eal\u003c/em\u003e. [31] suggests potential shifts in the local mosquito population; these findings align with previous studies in the same region [48,49]. Climate change and associated environmental alterations are likely contributing factors to these changes, as reported for other Cameroonian epidemiological contexts [7]. The presence of these vectors, particularly the highly anthropophilic and competent \u003cem\u003eAn. gambiae s.s.\u003c/em\u003e and \u003cem\u003eAn. funestus s.s\u003c/em\u003e suggest their critical role in the persistence of malaria transmission in the study area.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAmong the \u003cem\u003eAnopheles\u003c/em\u003e species identified, \u003cem\u003eAn. gambiae\u003c/em\u003e was the most prevalent across all three study sites. This dominance aligns with previous findings in Cameroon\u0026apos;s West Region [31,50,51]. Its adaptability to highland environments is also well documented [5,52]. \u003cem\u003eAn. funestus\u003c/em\u003e, another major vector, was only found in Penka Michel at 1500m, representing 17.7% of the anopheline population collected. Only three specimens of \u003cem\u003eAn. nili s.l\u003c/em\u003e. were collected in Santchou and \u003cem\u003eAn. ziemanni\u003c/em\u003e was present at very low densities. This uneven distribution of \u003cem\u003eAnopheles\u003c/em\u003e species likely stems from several factors, with temperature differences playing a potential role. During the rainy season, the temperatures are lower at Penka Michel and Dschang compared to the lowland site (Santchou) might explain some variation in vector densities. Previous studies have shown that cooler temperatures can delay larval development and increase mortality [53\u0026ndash;55]. However, Penka Michel, despite having a similar climate to Dschang and located at roughly the same altitude, exhibited densities of \u003cem\u003eAn. gambiae\u0026nbsp;\u003c/em\u003ethirty-three times higher. These observations suggest that climatic influence may not fully explain the discrepancies in vector densities. The hilly topography of Penka Michel likely plays a significant role. This geography promotes water accumulation and retention, creating more abundant and stable breeding sites for \u003cem\u003eAnopheles\u003c/em\u003e species (swamps, rivers, lakes). Balls \u003cem\u003eet al.\u003c/em\u003e (2004) demonstrated a link between flat-bottomed highland valleys, prone to water accumulation, and increased malaria risk in Tanzania. Similarly, other studies [56,55,52] highlight altitude, topography, and land use as significant environmental factors impacting malaria vector abundance in highland regions [57].\u003c/p\u003e\n\u003cp\u003eThe observed differences in vector densities between Penka Michel and Dschang, despite similar altitudes, can be also attributed to factors such as urbanization. Dschang, with its higher level of urbanization, exhibited a significantly higher abundance of \u003cem\u003eCulex\u003c/em\u003e spp (alone accounted for 76.8% of the total \u003cem\u003eCulex\u003c/em\u003e spp. collected). This finding aligns with previous observations in the city [49,48], which reported a high abundance of larval habitats and populations of \u003cem\u003eCulex\u0026nbsp;\u003c/em\u003espp. The \u003cem\u003eCulex\u003c/em\u003e genus is an indicator of urbanization [48], whose predominance in urban areas could be attributed to factors such as intense human activity (economic and industrial), and inadequate environmental management practices, which can create suitable breeding sites [58,59]. While \u003cem\u003eCulex\u003c/em\u003e spp. dominated the urban landscape, and in rural areas, the distribution changes. \u003cem\u003eAedes\u003c/em\u003e spp. and \u003cem\u003eMansonia\u003c/em\u003e spp. were also present, reaching 10.3% and 55.2% in Penka Michel and Santchou, respectively. These genera represent a substantial public health concern due to their established roles as vectors for various diseases in Africa [60]. \u003cem\u003eAedes\u003c/em\u003e spp., for instance, are notorious for transmitting arboviruses such as dengue and yellow fever, while \u003cem\u003eCulex\u003c/em\u003e spp., \u003cem\u003eAnopheles\u003c/em\u003e spp., and \u003cem\u003eMansonia\u003c/em\u003e spp. are implicated in the transmission of lymphatic filariasis [36\u0026ndash;38]. Specific to our study, we identified four \u003cem\u003eAedes\u003c/em\u003e species: \u003cem\u003eAe. simpsoni\u003c/em\u003e, \u003cem\u003eAe. albopictus\u003c/em\u003e, \u003cem\u003eAe. aegypti\u003c/em\u003e, and \u003cem\u003eAe. africanus\u003c/em\u003e. These species have been associated with the transmission of dengue and other arboviral diseases in various regions [64\u0026ndash;66], underscoring the potential public health risks posed by their presence in the study area.\u003c/p\u003e\n\u003cp\u003eConsistent with previous studies in the west region [31,52,51], this study identified the presence of both \u003cem\u003eAn. gambiae s.s.\u003c/em\u003e and \u003cem\u003eAn. coluzzii\u003c/em\u003e, sibling species within the \u003cem\u003eAn. gambiae s.l\u003c/em\u003e. complex, co-existing along the altitudinal transect (sympatry). \u003cem\u003eAn. gambiae s.s\u003c/em\u003e. is known to be more adapted to semi-urban environments, preferring sunlit larval habitats. Conversely, \u003cem\u003eAn. coluzzii\u003c/em\u003e typically thrives in urbanized areas with artificial, polluted, and wetter breeding sites [51,67,68]. Furthermore, the \u003cem\u003eAn. funestus\u0026nbsp;\u003c/em\u003egroup taxa found exclusively at Penka Michel were primarily composed of \u003cem\u003eAn. funestus s.s\u003c/em\u003e. and \u003cem\u003eAn. leesoni\u003c/em\u003e. Recently the epidemiological role of \u003cem\u003eAn. leesoni\u0026nbsp;\u003c/em\u003ein malaria transmission was demonstrated in certain regions of Cameroon [69].\u003c/p\u003e\n\u003cp\u003eAn interesting observation was the difference in \u003cem\u003eAn. gambiae s.l\u003c/em\u003e. biting behavior across the altitudinal transect. Compared to previous studies [31] reporting endophagic behavior (biting indoors) in the same area, this study found exophagic behavior (feeding outdoors) in Dschang and exophilic behavior (resting outdoors) in Santchou. This shift could be attributed to behavioral changes in response to intensified vector control efforts, particularly the widespread distribution of LLINs: on this date, the household coverage rate in Milda is 100 (unpublished data). Similar trends have been observed in recent Cameroonian studies [10,52] and across Africa [70\u0026ndash;72]. However, continuous entomological monitoring is crucial to understand better these mosquito species\u0026apos; spatial and behavioral variations in the study sites. In Penka Michel, both \u003cem\u003eAn. gambiae\u003c/em\u003e and \u003cem\u003eAn. funestus\u003c/em\u003e exhibited primarily endophagic behavior. This might be linked to the high mosquito nuisance levels, prompting residents to spend more nights indoors, which influences mosquito-biting behavior. Nevertheless, further investigations are needed to assess the sensitivity of these mosquito populations to the effects of insecticide-treated nets.\u003c/p\u003e\n\u003cp\u003eTaken as a whole, \u003cem\u003eAn. gambiae\u0026nbsp;\u003c/em\u003ewas aggressive throughout the night, both indoors and outdoors, with peak biting activity occurring between 10:00 pm and 3:00 am. This extended biting period could be linked to the relatively low long-lasting insecticidal net (LLIN) usage in the western region (58.7%), falling below the 80% coverage recommended by the WHO [73]. These findings align with previous studies, which reported peak biting activity for both species occurring late at night (between 00:00 pm and 6:00 am) [74,50,51]. It is important to acknowledge the limitations of the study. While we have assessed the impact of LLINs on malaria transmission, we were unable to fully evaluate the distribution and actual use of LLINs in our study sites. Further research is needed to address these gaps.\u003c/p\u003e\n\u003cp\u003eOverall, malaria transmission intensity across the study sites appeared low, with an entomological inoculation rate (EIR) ranging from 0.08 to 1.84 infected bites per human per night. Interestingly, \u003cem\u003eAn. gambiae\u003c/em\u003e, which was the most prevalent mosquito species biting exhibited the highest infection rate, followed by \u003cem\u003eAn. funestus\u0026nbsp;\u003c/em\u003eand \u003cem\u003eAn. ziemanni\u003c/em\u003e. These findings contrast with the high EIR reported by Tchuinkam in 2010\u0026nbsp;[31] in the same area. This difference could potentially be attributed to climatic changes observed in recent years, where such variations have been shown in laboratory studies to affect survival and reproductive capacity [10].\u003c/p\u003e\n\u003cp\u003eAmong the \u003cem\u003ePlasmodium\u003c/em\u003e species identified, \u003cem\u003eP. falciparum\u003c/em\u003e was the dominant parasite, accounting for up to 63% of infections in both Santchou and Penka Michel. This aligns with findings from other studies [25,75\u0026ndash;77] and confirms the primary epidemiological role of \u003cem\u003eP. falciparum\u003c/em\u003e in the region. Infections with \u003cem\u003eP. malariae\u003c/em\u003e and \u003cem\u003eP. ovale\u003c/em\u003e were also detected in infected vectors (\u003cem\u003eAn. gambiae\u003c/em\u003e, \u003cem\u003eAn. funestus,\u0026nbsp;\u003c/em\u003eand\u003cem\u003e\u0026nbsp;An. ziemanni\u003c/em\u003e). The presence of non-falciparum malaria parasites in these locations highlights the importance of including these parasites in control programs to achieve successful malaria elimination efforts.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWhile no \u003cem\u003eP. vivax\u003c/em\u003e infections were detected in the analyzed mosquito samples, this doesn\u0026apos;t necessarily exclude its presence in this region. Seminal studies have reported \u003cem\u003eP. vivax\u003c/em\u003e cases in Duffy-negative individuals residing in the western Cameroon region (Dschang, Santchou), with prevalence ranging from 0.5% to 35% [28,29]. Furthermore, another study identified \u003cem\u003eP. vivax\u003c/em\u003e infections within the \u003cem\u003eAn. coluzzii\u003c/em\u003e vector population in Tibati (Adamaoua region) [78]. This finding suggests the potential circulation of the parasite among the human population and its possible transmission by \u003cem\u003eAnopheles\u003c/em\u003e mosquitoes. Additionally, Oss\u0026egrave; and collaborators reported cases of \u003cem\u003eP. vivax\u003c/em\u003e infection in vectors \u003cem\u003eAn. gambiae\u003c/em\u003e and \u003cem\u003eAn. coluzzii\u003c/em\u003e from Benin [79].\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eWe report on the variation in the abundance and dynamics of malaria vector populations in the West Cameroon highlands. Six \u003cem\u003eAnopheles\u003c/em\u003e species were present in the study area, all known to be malaria vectors in Cameroon. \u003cem\u003eAn. gambiae\u0026nbsp;\u003c/em\u003ewas the most widespread species in the sites, followed by \u003cem\u003eAn. funestus\u0026nbsp;\u003c/em\u003ealthough the distribution of the latter was highly focal. The uneven distribution of anopheline species within the study area further confirms that the presence of these species varies according to the micro and macro-environmental differences present in the bio-ecological zones, even at the same altitude. \u003cem\u003eAn. gambiae has\u0026nbsp;\u003c/em\u003eshown a preference for biting outdoors rather than indoors in the lowland plain and on the mountain plateau, which could compromise malaria control strategies using mainly LLINs. Therefore, the implementation of additional tools (i.e., larviciding, integrated management, and environmental management) to combat these outdoor-biting mosquitoes must be considered.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003e\u003cstrong\u003e\u003cem\u003eCTAB:\u0026nbsp;\u003c/em\u003e\u003c/strong\u003eCetyltrimethyl ammonium bromide\u003cstrong\u003e; \u003cem\u003eEIR:\u0026nbsp;\u003c/em\u003e\u003c/strong\u003eEntomological inoculation rate\u003cstrong\u003e; \u003cem\u003eIR:\u0026nbsp;\u003c/em\u003e\u003c/strong\u003eInfection rate\u003cstrong\u003e; \u003cem\u003eIRS:\u0026nbsp;\u003c/em\u003e\u003c/strong\u003eInsecticide residual spraying\u003cstrong\u003e; \u003cem\u003eLLINs:\u003c/em\u003e\u003c/strong\u003e Long-lasting insecticidal nets\u003cstrong\u003e; \u003cem\u003eHBR:\u0026nbsp;\u003c/em\u003e\u003c/strong\u003eHuman-biting rate\u003cstrong\u003e\u003cem\u003e; PCR-RFLP:\u0026nbsp;\u003c/em\u003e\u003c/strong\u003ePCR-restriction fragment length polymorphism\u003cstrong\u003e\u003cem\u003e; rRNA:\u0026nbsp;\u003c/em\u003e\u003c/strong\u003eRibosomal ribonucleic acid\u003cstrong\u003e\u003cem\u003e; PNLP:\u0026nbsp;\u003c/em\u003e\u003c/strong\u003eNational malaria control program\u0026nbsp;\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was approved by\u0026nbsp;Cameroon\u0026apos;s National Ethics Committee for Human Health Research (CNERSH, n\u0026deg; 2022/12/1507/CE/CNERSH/SP). The study was conducted in collaboration with the various local health authorities. Free and informed consent was obtained from volunteers (for mosquito collection) and family heads through individual interviews and group meetings. Presumptive malaria treatment was administered free of charge to volunteers before and after HLC collections.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eConsent to publish has been obtained from all included persons in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests. The findings and conclusions expressed herein are those of the authors and do not necessarily represent the official position of the NIAID or NIH.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors contributions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eJJJ, IAB, RRD, and SEN conceived the research; BCK, AGBT, II, RRD, and SEN designed the study protocol. BCK, AGBT, II, COJ, MM, PKT, ZRPH, SJW, LMA, and BFT carried out the field and laboratory assays. RRD, and SEN supervised field collection. JJJ, IAB, RRD, and SEN contributed reagents/materials. BCK, AGBT, II, LMA, JAMM, RRD, and SEN analyzed and interpreted data. BCK and SEN drafted the manuscript. BCK, AGBT, JJAM, II, COJ, MM, PKT, ZRPH, SJW, CTN, LMA, BFT, JMS, JBP, JTL, JJJ, IAB, RRD, and SEN reviewed and helped write the manuscript. All authors made intellectual input to this study. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was supported by the National Institute for Allergy and Infectious Diseases (NIAID) as part of the National Institutes of Health (NIH) in the United States (R01AI165537 to JJJ, SEN, and RRD).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData are archived and available on request from the corresponding author\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe would like to thank the residents and mosquito collectors at the various sites who agreed to take part in this study, as well as their respective district authorities for their collaboration and support.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eWorld malaria report 2023 [Internet]. 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Insects. 2023 Feb 25;14(3):231. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"malaria-journal","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"malj","sideBox":"Learn more about [Malaria Journal](http://malariajournal.biomedcentral.com/)","snPcode":"12936","submissionUrl":"https://submission.nature.com/new-submission/12936/3","title":"Malaria Journal","twitterHandle":"@malariajournal","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Anopheles, diversity, behavior, transmission, highland, western Cameroon","lastPublishedDoi":"10.21203/rs.3.rs-5558659/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5558659/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e: Assessing vector bionomics is crucial to improving vector control strategies. Several entomological studies have been conducted to describe malaria transmission in different eco-epidemiological settings in Cameroon; knowledge gaps persist, particularly in highland areas. This study aimed to characterize malaria vectors in three localities along an altitudinal gradient in the western region: Santchou (700 m), Dschang (1400 m), and Penka Michel (1500 m).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e: Human landing catches were conducted from May to June 2023 from 6:00 pm to 9:00 am. Mosquitoes were sorted into genera, and all \u003cem\u003eAnopheles\u003c/em\u003e species were identified using morphological taxonomic keys and species-specific Polymerase Chain reaction (PCR). Entomological indicators were assessed including species composition and abundance, biting behavior, infection rate, and entomological inoculation rate (EIR). Genomic DNA from the head and thoraces were tested for \u003cem\u003ePlasmodium\u003c/em\u003einfection by real-time PCR.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e: 2,835 Anopheles mosquitoes were identified, including \u003cem\u003eAn. gambiae, An. coluzzii, An. funestus, An. leesoni, An. nili, \u003c/em\u003eand\u003cem\u003e An. ziemanni\u003c/em\u003e, with \u003cem\u003eAn. gambiae\u003c/em\u003e being the most prevalent at all sites. The human-biting rate of \u003cem\u003eAn. gambiae s.l.\u003c/em\u003e was significantly higher (p-value \u0026lt; 0.001) in Penka Michel compared to Santchou and Dschang (45.25 b/h/n vs 3.1 b/h/n and 0.41 b/h/n), and appears to be the most infected vector, and infectious vector distribution is highly focal, with entomological inoculation rates 13-fold higher in Penka Michel compared to Santchou (1.11 vs 0.08ibites/human/night). \u003cem\u003eP. falciparum\u003c/em\u003e was the dominant malaria parasite (67% at Santchou, 62% at Penka Michel), but \u003cem\u003eP. malariae\u003c/em\u003e (30%) and \u003cem\u003eP. ovale \u003c/em\u003e(1.21%) infections were also detected.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e: The study highlights a difference in mosquito composition and host-seeking behavior with altitude and the need for continued surveillance to monitor vector populations and prevent potential malaria outbreaks in these highland areas.\u003c/p\u003e","manuscriptTitle":"Diversity, abundance of anopheline species, and malaria transmission dynamics in high-altitude areas of western Cameroon","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-01-14 03:35:07","doi":"10.21203/rs.3.rs-5558659/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-03-01T14:50:19+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-02-01T00:31:26+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-01-22T11:54:48+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"300750850374268102375709237449779722624","date":"2025-01-18T11:16:53+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"225053577468243570472308274411938763965","date":"2025-01-13T16:35:23+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"328900450941705049902085013452680814243","date":"2025-01-13T16:20:35+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-01-13T11:11:14+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-12-03T14:06:03+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-12-03T14:06:01+00:00","index":"","fulltext":""},{"type":"submitted","content":"Malaria Journal","date":"2024-12-01T13:29:21+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"malaria-journal","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"malj","sideBox":"Learn more about [Malaria Journal](http://malariajournal.biomedcentral.com/)","snPcode":"12936","submissionUrl":"https://submission.nature.com/new-submission/12936/3","title":"Malaria Journal","twitterHandle":"@malariajournal","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b7ad1fa4-4dc7-4f8c-bfd6-b15772782591","owner":[],"postedDate":"January 14th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-08-11T16:03:45+00:00","versionOfRecord":{"articleIdentity":"rs-5558659","link":"https://doi.org/10.1186/s12936-025-05480-w","journal":{"identity":"malaria-journal","isVorOnly":false,"title":"Malaria Journal"},"publishedOn":"2025-08-06 15:57:59","publishedOnDateReadable":"August 6th, 2025"},"versionCreatedAt":"2025-01-14 03:35:07","video":"","vorDoi":"10.1186/s12936-025-05480-w","vorDoiUrl":"https://doi.org/10.1186/s12936-025-05480-w","workflowStages":[]},"version":"v1","identity":"rs-5558659","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5558659","identity":"rs-5558659","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}