Indoor residual spraying of experimental huts in Cameroon highlights the potential of Fludora® Fusion to control wild pyrethroid-resistant malaria vectors.

Elevated resistance to pyrethroids in major malaria vectors has led to the introduction of novel insecticides including neonicotinoids. There is a fear that efficacy of these new insecticides could be impacted by cross-resistance mechanisms from metabolic resistance to pyrethroids. In this study, after evaluating the resistance to deltamethrin, clothianidin and mixture of clothianidin + deltamethrin in the lab using CDC bottle assays, the efficacy of the new IRS formulation Fludora® Fusion was tested in comparison to clothianidin and deltamethrin applied alone using experimental hut trials against wild free-flying pyrethroid-resistant Anopheles funestus from Elende and field An. gambiae collected from Nkolondom reared in the lab and released in the huts. Additionally, cone tests on the treated walls were performed each month for a period of twelve months to evaluate the residual efficacy of the sprayed products. Furthermore, the L1014F-kdr target-site mutation and the L119F-GSTe2 mediated metabolic resistance to pyrethroids were genotyped on a subset of mosquitoes from the EHT to assess the potential cross-resistance. All Anopheles species tested were fully susceptible to clothianidin and clothianidin + deltamethrin mixture in CDC bottle assay while resistance was noted to deltamethrin. Accordingly, Fludora® Fusion (62.83% vs 42.42%) and clothianidin (64.42% vs 42.42%) induced significantly higher mortality rates in EHT than deltamethrin (42.42%) against free flying An. funestus from Elende in month 1 (M1) and no significant difference in mortality was observed between the first (M1) and sixth (M6) months of the evaluation (P > 0.05). However, lower mortality rates were recorded against An. gambiae s.s from Nkolondom (mortality rates 50%, 45.56% and 26.68%). In-situ cone test on the wall showed a high residual efficacy of Fludora® Fusion and clothianidin on the susceptible strain KISUMU (> 12 months) and moderately on the highly pyrethroid-resistant An. gambiae strain from Nkolondom (6 months). Interestingly, no association was observed between the L119F-GSTe2 mutation and the ability of mosquitoes to survive exposure to Fludora® Fusion, whereas a trend was observed with the L1014F-kdr mutation. This study highlights that Fludora® Fusion, through its clothianidin component, has good potential of controlling pyrethroid-resistant mosquitoes with prolonged residual efficacy. This could be therefore an appropriate tool for vector control in several malaria endemic regions.

Two New Species of Annulotrema (Monogenea: Dactylogyridae) Gill Parasites of Brycinus macrolepidotus Valenciennes, 1849 from Nyong River, Cameroon.

PURPOSE: Research on gill monogeneans of Characiform has made it possible to find two new species belonging to the genus Annulotrema. The purpose of this paper is to carry out their morphological description. METHODS: Specimens of Brycinus macrolepidotus were captured in the Nyong River at Akonolinga. Each monogenean was mounted between slide and cover slip in a drop of glycerin ammonium picrate mixture. The sclerotized parts of their haptor and reproductive organs were then drawn and measured. RESULTS: Two new species of Annulotrema were described. Annulotrema ngombiensis n. sp. is morphologically close to Annulotrema tenuicirra Paperna, 1973 and Annulotrema pikei Price, Peebles and Bamford, 1969. However, the new species differs from these other two mainly by the characteristic well-marked terminal cap and the tubiform prostatic reservoir of the accessory part of its MCO, as well as by its dorsal bar with a characteristic triangular piece in the middle of the fork and its ventral bar without filaments. Annulotrema nkengfacki n. sp. is distinguished from all other species previously described as well as from its closest congeners, Annulotrema helicocirra Paperna, 1973 and Annulotrema bouixi Birgi, 1988 by the structure of its MCO whose accessory part is composed of a trapezoid structure surmounted by another which is forked-shaped and by its arched-shaped ventral bar with an inner lining. CONCLUSION: Brycinus macrolepidotus from Cameroon can harbour two species of Annulotrema, both described in the current study. This result contributes to the knowledge of the species diversity of this genus in the Nyong Basin.

Myxobolus makombensis n. sp. infection in African carp Labeobarbus batesii from the Makombè River, Cameroon: morphological and molecular characterization.

In this study, we examined myxozoan infections of Labeobarbus batesii sampled from the Makombè River in Cameroon. Fish were infected with Myxobolus makombensis n. sp. in the gill filament and M. dibombensis in the fins. Mature myxospores of M. makombensis n. sp. are pyriform in frontal view and biconvex in lateral view, with a truncated and slightly narrow anterior end. Spore dimensions (mean ± SD, with range in parentheses) are 17.5 ± 0.22 (16.2-18.9) µm length, 13.4 ± 0.25 (12-14.9) µm width, and 7 ± 0.21 (6.7-7.5) µm thickness, and spores exhibit a conspicuous anterior intercapsular appendix of 4.4 ± 0.18 (3.9-5.5) µm length. Myxospores have 2 pyriform polar capsules of unequal size; the larger one is 9.8 ± 0. 22 (8.2-10.9) µm long × 4.7 ± 0.15 (3.5-5.2) µm wide, and the smaller one is 8.8 ± 0.22 (7-10) µm long × 4.3 ± 0.12 (3.5-5.2) µm wide. Polar filaments possess 10 to 11 coils in the large polar capsule and 8 to 10 coils in the small polar capsule. Phylogenetic analysis of SSU rDNA sequences showed clustering of M. makombensis n. sp. close to M. dibombensis recently reported from the fins of the same host within a clade composed exclusively of parasites infecting cyprinid fishes.

Morphological and Molecular Characterization of Myxobolus nkondjockei sp. nov. (Myxozoa: Myxobolidae), A Parasite of Labeo batesii Boulenger, 1911 (Teleostei: Cyprinidae) from Makombè River in Cameroon.

PURPOSE: Myxozoans are economically important group of metazoan parasites, which can cause diseases in a large variety of commercially important fishes. Increased knowledge on molecular features has shown that traditional descriptive characters may be misleading. Combination of both descriptive and molecular features is therefore necessary for an integrated taxonomic assessment. METHODS: Cyprinid Labeo batesii, sampled in the Makombè River at Nkondjock in Cameroon were examined for myxosporeans. Identification of parasite species was based on morphological and molecular sequence analyses of myxospores. Phylogenetic analysis was performed using maximum likelihood (ML) and Bayesian inference (BI) methods. RESULTS: The scales of L. batesii were infected by Myxobolus nkondjockei sp. nov Their mature myxospores are ovoid in frontal view and lenticular in lateral view, with two rounded ends. These myxospores measured 10.3 (10-10.9) µm length and 8.0 (7.3-8.5) µm width. Myxospores have two ovoid and equal sizes polar capsules. They measured 4.5 (4.0-5.0) µm in length and 2.4 (2-2.9) µm in width. Polar tubules were coiled in 4-5 turns perpendicular to the longitudinal axis of the polar capsules. Phylogenetic analysis of the 18S rDNA sequence show clustering of M. nkondjockei sp. n. close to an undetermined species Myxobolus sp. reported infecting gill lamellas of Labeo rohita from India. CONCLUSION: The morphological, molecular and phylogenetic data provided for M. nkondjockei sp. n. are solid basis for further identification of this myxozoan of which pathogenicity probably plays an economic role at culturing the hosts.

Local adaptation in thermal tolerance for a tropical butterfly across ecotone and rainforest habitats.

Thermal adaptation to habitat variability can determine species vulnerability to environmental change. For example, physiological tolerance to naturally low thermal variation in tropical forests species may alter their vulnerability to climate change impacts, compared with open habitat species. However, the extent to which habitat-specific differences in tolerance derive from within-generation versus across-generation ecological or evolutionary processes are not well characterized. Here we studied thermal tolerance limits of a Central African butterfly (Bicyclus dorothea) across two habitats in Cameroon: a thermally stable tropical forest and the more variable ecotone between rainforest and savanna. Second generation individuals originating from the ecotone, reared under conditions common to both populations, exhibited higher upper thermal limits (CTmax) than individuals originating from forest (∼3°C greater). Lower thermal limits (CTmin) were also slightly lower for the ecotone populations (∼1°C). Our results are suggestive of local adaptation driving habitat-specific differences in thermal tolerance (especially CTmax) that hold across generations. Such habitat-specific thermal limits may be widespread for tropical ectotherms and could affect species vulnerability to environmental change. However, microclimate and within-generation developmental processes (e.g. plasticity) will mediate these differences, and determining the fitness consequences of thermal variation for ecotone and rainforest species will require continued study of both within-generation and across-generation eco-evolutionary processes. This article has an associated First Person interview with the first author of the paper.

Description of Heteracris lecoqi Yetchom-Fondjo amp; Kekeunou sp. nov. and Heteracris hannai Wandji amp; Kekeunou sp. nov. and redescription of Heteracris guineensis (Krauss, 1890) (Orthoptera: Acrididae) with comments on its ecology in the Southern part of Cameroon.

The specimens studied were collected with sweep net and pitfall in the forests, agro-forests, herbaceous fallows, and crop fields of 14 localities in the southern part of Cameroon, from August 2015 to February 2018. The results show that Heteracris hannai sp. nov. and Heteracris lecoqi sp. nov. differ from previously known species of the same genus by the pattern of coloration and the details or shape of the phallic complex. H. lecoqi sp. nov. is characterized by light brownish body; male cercus with flattened, downcurved and obtuse apex; lophus strongly curved; interlophal space with V shape; apodeme of cingulum bow, convergent, with V-shape; valve of cingulum in lateral view longer than apical valve of penis; ramus in ventral view joined. H. hannai sp. nov. is distinct to other species by brown to grey body; male cercus with apex rounded, curved inside; lophus curved; interlophal space with U-shape; apodeme of cingulum thick, slightly parallel, with U shape; valve of cingulum in lateral view hardly longer than apical valve of penis; ramus in ventral view opened. The characteristics of H. guineensis are closer to H. hannai sp. nov. than to H. lecoqi sp. nov. H. lecoqi sp. nov. was collected only in the fallows while H. hannai sp. nov. was collected in the forests, agro-forests, fallows, and crop fields. Compared to H. guineensis, both new species are scarce in the natural vegetation and their distribution area is limited to two and four localities respectively for H. lecoqi sp. nov. and H. hannai sp. nov. All these three grasshopper's species were recorded as accidental species in all types of vegetation.

Geographic distribution and abundance of the Afrotropical subterranean scale insect Stictococcus vayssierei (Hemiptera: Stictococcidae), a pest of root and tuber crops in the Congo basin.

Stictococcus vayssierei is a major pest of root and tuber crops in central Africa. However, data on its ecology are lacking. Here we provide an updated estimate of its distribution with the aim of facilitating the sustainable control of its populations. Surveys conducted in nine countries encompassing 13 ecological regions around the Congo basin showed that African root and tuber scale was present in Cameroon, Central African Republic, Congo, Democratic Republic of Congo, Equatorial Guinea, Gabon and Uganda. It was not found on the sites surveyed in Chad and Nigeria. The pest occurred in the forest and the forest-savannah mosaic as well as in the savannah where it was never recorded before. However, prevalence was higher in the forest (43.1%) where cassava was the most infested crop, compared to the savannah (9.2%) where aroids (cocoyam and taro) were the most infested crops. In the forest habitat, the pest was prevalent in all but two ecological regions: the Congolian swamp forests and the Southern Congolian forest-savanna mosaic. In the savannah habitat, it was restricted to the moist savannah highlands and absent from dry savannahs. The scale was not observed below 277 m asl. Where present, the scale was frequently (87.1% of the sites) attended by the ant Anoplolepis tenella. High densities (>1000 scales per plant) were recorded along the Cameroon-Gabon border. Good regulatory measures within and between countries are required to control the exchange of plant materials and limit its spread. The study provides information for niche modeling and risk mapping.

Malaria Transmission around the Memve'ele Hydroelectric Dam in South Cameroon: A Combined Retrospective and Prospective Study, 2000⁻2016.

Dam constructions are considered a great concern for public health. The current study aimed to investigate malaria transmission in the Nyabessan village around the Memve'ele dam in South Cameroon. Adult mosquitoes were captured by human landing catches in Nyabessan before and during dam construction in 2000-2006 and 2014-2016 respectively, as well as in the Olama village, which was selected as a control. Malaria vectors were morphologically identified and analyzed for Plasmodium falciparum circumsporozoite protein detection and molecular identification of Anopheles (A.) gambiae species. Overall, ten malaria vector species were identified among 12,189 Anopheles specimens from Nyabessan (N = 6127) and Olama (N = 6062), including A. gambiae Giles (1902), A. coluzzii Coetzee (2013), A. moucheti Evans (1925), A. ovengensis Awono (2004), A. nili Theobald (1903), A. paludis Theobald (1900), A. zieanni, A. marshallii Theobald (1903), A. coustani Laveran (1900), and A. obscurus Grünberg (1905). In Nyabessan, A. moucheti and A. ovengensis were the main vector species before dam construction (16-50 bites/person/night-b/p/n, 0.26-0.71 infective bites/person/night-ib/p/n) that experienced a reduction of their role in disease transmission in 2016 (3-35 b/p/n, 0-0.5 ib/p/n) (p < 0.005). By contrast, the role of A. gambiae s.l. and A. paludis increased (11-38 b/p/n, 0.75-1.2 ib/p/n) (p < 0.01). In Olama, A. moucheti remained the main malaria vector species throughout the study period (p = 0.5). These findings highlight the need for a strong vector-borne disease surveillance and control system around the Memve'ele dam.

Morphological and molecular characterization of Myxobolus dibombensis sp. n. (Myxozoa: Myxobolidae), a parasite of the African carp Labeobarbus batesii (Teleostei: Cyprinidae) from Dibombe River, Cameroon.

Myxobolus dibombensis sp. n. (Cnidaria: Myxosporea: Bivalvulida) is described from the fins of the African carp, Labeobarbus batesii, based on morphological and molecular data. Prevalence of infection was 51.9% (67/129). Ovoid to spherical cyst-like plasmodia were found in the intrasegmental region and among the fin rays. No pathological changes were found in the fish host tissue surrounding the cyst-like plasmodia. Mature myxospores were ovoid in frontal view and lenticular in lateral view, with slightly truncated anterior and rounded posterior ends. Myxospores measured 16.8 (15.8-18.0) µm long and 11.4 (10.0-13.0) µm wide. There was a triangular intercapsular appendix measuring 3.8 (2.6-4.5) µm long. Polar capsules were ovoid and slightly unequal in size, occupying approximately one-third of the myxospore length. The larger polar capsule measured 7 (6-8) µm long and 3.6 (3-4) µm wide, while the smaller one measured 5.8 (4.8-7.0) µm long and 3 (2-4) µm wide. The larger polar capsule contained nine to 11 filament coils, whereas the smaller one contained seven to nine coils. SSU rDNA gene sequence of M. dibombensis sp. n. did not match any sequences available in the GenBank. The similarity with available Myxobolus spp. sequences ranged from 65 to 81%. The novel species clustered with M. algonquinensis, which infects the cyprinid Luxilus cornutus from Canada.

Seasonal Polyphenism in Bicyclus dorothea (Lepidoptera: Nymphalidae) Across Different Habitats in Cameroon.

Many organisms exhibit changes in phenotypic traits as a response to seasonal environmental variation. We investigated the role of habitat in generating seasonal polyphenism in different populations of the light bush brown butterfly Bicyclus dorothea (Cramer, 1779) (Lepidoptera: Nymphalidae) in Cameroon. Butterflies were caught during the wet and dry seasons across four localities representing two distinct habitats, namely forest and ecotone (forest-savanna transition zone) over a 2-yr period (2015-2016). We found distinct variation in the wing pattern characteristics of butterflies in response to seasonality and habitat. Specifically we observed that: 1) all wing characters are not seasonally plastic in B. dorothea; 2) populations from ecotone tend to be more variable, with individuals exhibiting wings with large spots during the wet season and very reduced spots in the dry season while in forest populations, individuals exhibit wings with large spots during the wet season, but in the dry season, spots are not as greatly reduced as their ecotone counterparts; 3) this polyphenism in B. dorothea alternated consistently during the wet and dry seasons over the 2 yr of sampling. Bicyclus species have become a textbook example of seasonal polyphenism while this study extends this model system to the unique forest-ecotone gradient of Central Africa and demonstrates the complexity of seasonal forms in different habitats.