Impact of repeated mass ivermectin administration using a community directed approach on L. loa infection in Chrysops silacea of the rain forest and forest savanna of Cameroon.

Background: Loiasis is an endemic filarial infection in the rainforest zone of West and Central Africa. Repeated annual community-directed treatment with ivermectin (CDTI) delivered for several years to control onchocerciasis has been shown to reduce the prevalence and intensity of Loiasis in some Loa loa-Onchocerca volvulus co-endemic areas. However, the impact of these multiple rounds of CDTI on entomological indicators of loiasis transmission is not known, and was therefore assessed in this study in areas with contrasting histories of CDTI. Methods: The study was conducted in the East, North-west and South-west 1 CDTI project sites of Cameroon. Two communities per CDTI project were selected for fly collection and dissection. Ivermectin treatment coverage was documented in these areas, and this was correlated to Chrysops infection and infective rates. A total of 7029 female Chrysops were collected from 6 communities of the 3 CDTI projects (East, North-west, and South-west 1) and from 2 communities in a non-CDTI district (East). Results: Chrysops biting densities and parous rates were significantly reduced in the North-west and South-west sites post-CDTI, while in the East, biting densities were similar in non-CDTI and CDTI sites, with higher parous rates observed in the non-CDTI site. Infection and infective rates in the East non-CDTI site were 4.4% and 1.8% respectively, as compared to 3.3% and 1.3% in the CDTI site after 10 ivermectin rounds (there were no baseline data for the latter). In the North-west site, significant reductions in Chrysops infection and infective rates from 10.2% and 4.2% respectively, to 3.5% and 1.2 (after 9 rounds of ivermectin treatment), were recorded following CDTI. In the South-west, infection rate significantly increased from 1.74% to 2.8% and infective rate remained statistically unchanged after 14 rounds of CDTI (0.45% - 0.40%). Similar trends in Mean Head L3 were observed except in the East site where this indicator was similar in both CDTI and control sites. Only in the North-west site did monthly transmission potentials decrease significantly. Conclusion: This study demonstrated that the impact of repeated annual treatment with ivermectin for the control of onchocerciasis using community directed delivery approach on the entomological indicators of loiasis varies with bioecological zones. Community directed treatment with ivermectin induced a significant reduction in the entomological indicators of loiasis in the North-West project site which lies in forest savanna area. A non-significant decrease was observed in the East project site and in contrast, a significant increase was observed in the South-West 1 project site which both lies in the rainforest zones.

Geographical emergence of sulfadoxine-pyrimethamine drug resistance-associated P. falciparum and P. malariae alleles in co-existing Anopheles mosquito and asymptomatic human populations across Cameroon.

Malaria molecular surveillance remains critical in detecting and tracking emerging parasite resistance to anti-malarial drugs. The current study employed molecular techniques to determine Plasmodium species prevalence and characterize the genetic diversity of Plasmodium falciparum and Plasmodium malariae molecular markers of sulfadoxine-pyrimethamine resistance in humans and wild Anopheles mosquito populations in Cameroon. Anopheles mosquito collections and parasitological survey were conducted in villages to determine Plasmodium species infection, and genomic phenotyping of anti-folate resistance was accomplished by sequencing the dihydrofolate-reductase (dhfr) and dihydropteroate-synthase (dhps) genes of naturally circulating P. falciparum and P. malariae isolates. The malaria prevalence in Elende was 73.5% with the 5-15 years age group harboring significant P. falciparum (27%) and P. falciparum + P. malariae (19%) infections. The polymorphism breadth of the pyrimethamine-associated Pfdhfr marker revealed a near fixation (94%) of the triple-mutant -A16I51R59N108I164. The Pfdhps backbone mediating sulfadoxine resistance reveals a high frequency of the V431A436G437K540A581A613 alleles (20.8%). Similarly, the Pmdhfr N50K55L57R58S59S114F168I170 haplotype (78.4%) was predominantly detected in the asexual blood stage. In contrast, the Pmdhps- S436A437occured at 37.2% frequency. The combined quadruple N50K55L57R58S59S114F168I170_ S436G437K540A581A613 (31.9%) was the major circulating haplotype with similar frequency in humans and mosquitoes. This study highlights the increasing frequency of the P. malariae parasite mostly common in asymptomatic individuals with apparent P. falciparum infection. Interventions directed at reducing malaria transmission such as the scaling-up of SP are favoring the emergence and spread of multiple drug-resistant alleles between the human and mosquito host systems.

Epidemiology of malaria, schistosomiasis, and geohelminthiasis amongst children 3-15 years of age during the dry season in Northern Cameroon.

BACKGROUND: The double burden of malaria and helminthiasis in children poses an obvious public health challenge, particularly in terms of anemia morbidity. While both diseases frequently geographically overlap, most studies focus on mono-infection and general prevalence surveys without molecular analysis. The current study investigated the epidemiological determinants of malaria, schistosomiasis, and geohelminthiasis transmission among children in the North Region of Cameroon. METHODOLOGY: School and pre-school children aged 3-15 year-of-age were enrolled from three communities in March 2021 using a community cross-sectional design. Capillary-blood samples were obtained, and each was examined for malaria parasites using rapid-diagnostic-test (RDT), microscopy, and PCR while hemoglobin level was measured using a hemoglobinometer. Stool samples were analyzed for Schistosoma mansoni, S. guineensis, and soil-transmitted-helminthiasis (STH) infections using the Kato Katz method, and urine samples were assessed for the presence of S. haematobium eggs (including hybrids) using the standard urine filtration technique. RESULT: A malaria prevalence of 56% (277/495) was recorded by PCR as opposed to 31.5% (156/495) by microscopy and 37.8% (186/495) by RDT. Similarly, schistosomiasis was observed at prevalence levels of up to 13.3% (66/495) overall [S. haematobium (8.7%); S. mansoni (3.8%); mixed Sh/Sm (0.6%); mixed Sh/Sm/Sg (0.2%). Both infections were higher in males and the 3-9 year-of-age groups. A high frequency of PCR reported P. falciparum mono-infection of 81.9% (227/277) and mixed P. falciparum/P. malariae infection of 17.3% (48/277) was observed. Malaria-helminths co-infections were observed at 13.1% (65/495) with marked variation between P. falciparum/S. haematobium (50.8%, 33/65); P. falciparum/S. mansoni (16.9%, 11/65) and P. falciparum/Ascaris (9.2%, 6/65) (χ2 = 17.5, p = 0.00003). Anemia prevalence was 32.9% (163/495), categorically associated with P. falciparum (45.8%, 104/227), Pf/Sh (11.5%, 26/227), and Pf/Sm (3.9%, 9/227) polyparasitism. CONCLUSION: Polyparasitism with malaria and helminth infections is common in school-aged children despite periodic long-lasting insecticide-treated nets (LLINs) distribution and regular school-based praziquantel (for schistosomiasis) and albendazole (for STH) campaigns. Co-existence of Plasmodium parasites and helminths infections notably Schistosoma species among children may concurrently lead to an increase in Plasmodium infection with an enhanced risk of anemia, highlighting the necessity of an integrated approach for disease control interventions.

Efficacy of Trapping Methods in the Collection of Eretmapodites (Diptera: Culicidae) Mosquitoes in an Afrotropical Rainforest Region, South western Cameroon.

Very little data exist on the biology of an afrotropical rainforest mosquito Eretmapodites (Er.) in a world undergoing dramatic changes due to deforestation. The aim was to assess the efficacy of different trapping methods in the collection of Er. mosquito in forested area. This was a longitudinal study involving collection of mosquitoes for over two years. Multiple collection methods (grouped into two categories), were used; i) net baited and un-baited traps to collect adults, ii) techniques that target immature stages subsequently reared to adults. All males were identified by genitalia dissection. Five thousand seven hundred and four mosquitoes representing 11 genera among which 2,334 Er. were identified. Mosquito abundance was highest in the net traps (n = 1276 (56.4%)) and sweep nets (n = 393(17.4%)) respectively. The abundance was highest in green colored net traps (435(34.09%)) with significant value of χ2= 40.000, P < 0.001 and in pigeons baited traps (473 (37.06%)) with significant value of χ2= 42.000, P = 0.003. The diversity ranges from H' = 2.65; DS = 0.84; SR = 24; ACE = 24.77 in sweep net to H' = 0; DS = 0; SR = 1; ACE = 1 in rock pool among males mosquitoes. While for females, H = 1.14; DS = 0.71; SR = 5; ACE = 5.16, in sweep net to H = 0; DS = 0; SR = 1; ACE = 1 in rock pool, tarpaulin, resting cage. Net traps, bamboo pot, and sweep netting are efficient in collecting high abundance of forest mosquitoes in the Talanagaye rainforest.

Prevalence, antimicrobial susceptibility pattern and associated risk factors for urinary tract infections in pregnant women attending ANC in some integrated health centers in the Buea Health District.

BACKGROUND: Urinary tract infections (UTIs) are the second most frequent infections after respiratory tract infections that affect humans, with over 150 million cases per year. The anatomy of the female urinary tract predisposes them to UTIs than men. More so, physiological and hormonal changes during pregnancy put pregnant woman at risk of UTIs. Untreated UTI(s) in pregnancy can be detrimental to both the mother and child causing preterm labour, low birth weight and pyelonephritis. The situation is worrisome because the infection can be asymptomatic. This study investigated the prevalence and risk factors for UTIs, diagnostic potential of dipstick analyses and antimicrobial susceptibility of uropathogens from pregnant women attending ANC in some Integrated Health Centers (IHCs) in Buea Health District (BHD). METHODS: A structured questionnaire was administered to consenting pregnant women at participating IHCs to collect data on demographic characteristics, risk factors and symptoms of UTI. Urine samples were collected for dipstick analysis and culture. Antibiograms were performed on the isolates by the disc diffusion method. A bivariate analysis was performed to investigate the association of the risk factors to UTI. Chi square (χ2) test, odds ratios with corresponding 95% confidence intervals were used to compare statistics and test for associations at a significant level of p ≤ 0.05. RESULTS: Of the 287 participants recruited, 89(31%) were positive for UTI. There were 150 women with no symptoms of which 43(28.7%) were positive for UTI. E. coli was the most frequent (43.2%) of the organisms implicated in bacteriuria. There was no significant association between the risk factors studied and UTI. Isolates were most sensitive to ciprofloxacin (73.5%) and gentamycin (73.5%) and resistant to ceftriaxone (70.6%) and tetracycline (64.7%). Nitrite test was highly specific (100%) for the diagnosis of UTI while leucocyte esterase was more sensitive (48.3%) than specific (44.9%). CONCLUSIONS: The prevalence of UTI in BHD was high. In conformity with previous findings in same area, there were no risk factors associated with UTI. We recommended a longitudinal study with a larger sample size to follow up the women to term in order to determine the gravity of this infection on pregnancy outcomes.

Onchocerca ochengi male worms implanted in SCID mice and Gerbil: Relationship between microfilaridermia status of cows, nodular worm viability and fertility and worm survival in the rodents.

BACKGROUND: Current treatment options for onchocerciasis are sub-optimal, prompting research and development of a safe cure (macrofilaricide). Onchocerca ochengi, a parasite of cattle, is used as a close surrogate for the human parasite O. volvulus in a murine model for pre-clinical screening of macrofilaricides. Skin from naturally infected cattle have been used in previous studies as a reliable source of parasite material. However, there is limited knowledge on how source-related factors such as the microfilaridermia status of the cattle, the nodule load and nodular worm viability may affect survival of male O. ochengi worms implanted in the rodent hosts. Such relationships were investigated in this study. METHODS: Dermal tissue and nodules were obtained from Gudali cattle, dissected and cultured to obtain migrating microfilariae (mf) and male worms. Emerged male worms were implanted into SCID mice and Gerbils (Meriones unguiculatus) and recovery rates were determined upon 42 days post implantation. Finally, nodules were processed for histology and embryogram analyses to assess the nodular worm viability and fertility, respectively. RESULTS: Of the 69 cattle sampled, 24 (34.8%) were mf+ and 45 (65.2%) were mf-. The mean nodule loads were 180.5 ± 117.7 (mf+) and 110.6 ± 102.7 (mf-) (p = 0.0186). The mean male worm harvest from nodules were 76.8 ± 120.3 and 47.2 ± 33.4 (p = 0.2488) for mf+ and mf- cattle, respectively. The number of male worms per 100 nodules were 57/100 and 46/100 nodules for mf+ and mf- cows, respectively. Female worms from nodules of mf- cows had higher counts of both normal and abnormal embryos with higher proportions of dead nodular worms evinced by histology compared to those from mf+ cows. A total of 651 worms were implanted into mice and gerbils, out of which 129 (19.81%) were recovered. Logistic regression analysis indicated that the microfilaridermia status of the cattle (presence of mf) (OR = 4.3319; P = 0.001) is the single most important predictor of the success of male worm recovery after implantation into rodents. CONCLUSION: Microfilaridermic cattle provide a promising source of adult O. ochengi. Male worms from this group of cattle have a better success rate of survival in a murine implant model. Nevertheless, in the programmatic point of view, amicrofilaridermic Gudali cattle would still constitute an important source of O. ochengi male worms with relatively good viability after implantation into rodents.

Differential susceptibility of Onchocerca volvulus microfilaria to ivermectin in two areas of contrasting history of mass drug administration in Cameroon: relevance of microscopy and molecular techniques for the monitoring of skin microfilarial repopulation within six months of direct observed treatment.

BACKGROUND: Ivermectin is an excellent microfilaricide against Onchocerca volvulus. However, in some regions, long term use of ivermectin has resulted in sub-optimal responses to the treatment. More data to properly document the phenomenon in various contexts of ivermectin mass drug administration (IVM-MDA) is needed. Also, there is a need to accurately monitor a possible repopulation of skin by microfilariae following treatment. Skin snip microscopy is known to have a low sensitivity in individuals with light infections, which can be the case following treatment. This study was designed with two complementary objectives: (i) to assess the susceptibility of O. volvulus microfilariae to ivermectin in two areas undergoing IVM-MDA for different lengths of time, and (ii) to document the repopulation of skin by the O. volvulus microfilariae following treatment, using 3 independent diagnostic techniques. METHOD: Identified microfilaridermic individuals were treated with ivermectin and re-examined after 1, 3, and 6 months using microscopy, actin real-time PCR (actin-qPCR) and O-150 LAMP assays. Susceptibility to ivermectin and trends in detecting reappearance of skin microfilariae were determined using three techniques. Microscopy was used as an imperfect gold standard to determine the performance of actin-qPCR and LAMP. RESULTS: In Bafia with over 20 years of IVM-MDA, 11/51 (21.6%) direct observe treated microfilaridemic participants were still positive for skin microfilariae after 1 month. In Melong, with 10 years of IVM-MDA, 2/29 (6.9%) treated participants were still positive. The microfilarial density reduction per skin biopsy within one month following treatment was significantly lower in participants from Bafia. In both study sites, the molecular techniques detected higher proportions of infected individuals than microscopy at all monitoring time points. LAMP demonstrated the highest levels of sensitivity and real-time PCR was found to have the highest specificity. CONCLUSION: Patterns in skin mirofilariae clearance and repopulation were established. O. volvulus worms from Bafia with higher number of annual MDA displayed a lower clearance and higher repopulation rate after treatment with ivermectin. Molecular assays displayed higher sensitivity in monitoring O. volvulus microfilaridemia within six months following treatment.

Effect of deforestation on prevalence of avian haemosporidian parasites and mosquito abundance in a tropical rainforest of Cameroon.

Habitat change caused by deforestation can modify the interactions of many biotic and abiotic factors, and in turn influence patterns of diseases in wild birds. Whether deforestation directly or indirectly affects the prevalence of avian haemosporidian parasites through their hosts and/or vectors is still not well understood. We sampled understory bird communities (insectivorous, frugivorous, granivorous and nectarivorous birds) and mosquitoes in three habitats showing a gradient of deforestation (pristine forest, fragmented forest, and young palm oil plantation), to assess the effects of habitat changes on avian haemosporidian (Plasmodium and Haemoproteus) prevalence and its relationship to bird feeding guilds and mosquito abundance. Blood samples of 845 individual birds belonging to 85 species and 27 families were collected in the three habitat types and screened using microscopy and PCR. Plasmodium infections were detected in 136 individuals (16.09%) and varied significantly among habitat types while Haemoproteus infections were detected in 98 individuals (11.60%) and did not vary significantly among habitat types. However, the prevalence of Plasmodium and Haemoproteus in bird feeding groups varied significantly among habitats. Nectarivorous and granivorous birds had the highest Plasmodium and Haemoproteus prevalence, respectively. The abundance of mosquitoes varied significantly among habitat types and the prevalence of Plasmodium significantly and positively correlated with mosquito abundance in fragmented forest. This study highlights the importance of host and mosquito determinants in the transmission dynamics of avian Plasmodium and Haemoproteus infections following habitat changes. Selective logging favored an increase in the prevalence of Plasmodium in insectivores, the prevalence of Haemoproteus in nectarivores and the abundance of female mosquitoes while, the establishment of the palm oil plantation favored an increase in the prevalence of Plasmodium in granivores and Haemoproteus in nectarivores. Species feeding behavior is also an important determinant to consider for a better understanding of patterns of parasite infections in a changing environment.

Mapping the geographical distribution of podoconiosis in Cameroon using parasitological, serological, and clinical evidence to exclude other causes of lymphedema.

BACKGROUND: Podoconiosis is a non-filarial elephantiasis, which causes massive swelling of the lower legs. It was identified as a neglected tropical disease by WHO in 2011. Understanding of the geographical distribution of the disease is incomplete. As part of a global mapping of podoconiosis, this study was conducted in Cameroon to map the distribution of the disease. This mapping work will help to generate data on the geographical distribution of podoconiosis in Cameroon and contribute to the global atlas of podoconiosis. METHODS: We used a multi-stage sampling design with stratification of the country by environmental risk of podoconiosis. We sampled 76 villages from 40 health districts from the ten Regions of Cameroon. All individuals of 15-years old or older in the village were surveyed house-to-house and screened for lymphedema. A clinical algorithm was used to reliably diagnose podoconiosis, excluding filarial-associated lymphedema. Individuals with lymphoedema were tested for circulating Wuchereria bancrofti antigen and specific IgG4 using the Alere Filariasis Test Strips (FTS) test and the Standard Diagnostics (SD) BIOLINE lymphatic filariasis IgG4 test (Wb123) respectively, in addition to thick blood films. Presence of DNA specific to W. bancrofti was checked on night blood using a qPCR technique. PRINCIPAL FINDINGS: Overall, 10,178 individuals from 4,603 households participated in the study. In total, 83 individuals with lymphedema were identified. Of the 83 individuals with lymphedema, two were found to be FTS positive and all were negative using the Wb123 test. No microfilaria of W. bancrofti were found in the night blood of any individual with clinical lymphedema. None were found to be positive for W. bancrofti using qPCR. Of the two FTS positive cases, one was positive for Mansonella perstans DNA, while the other harbored Loa loa microfilaria. Overall, 52 people with podoconiosis were identified after applying the clinical algorithm. The overall prevalence of podoconiosis was found to be 0.5% (95% [confidence interval] CI; 0.4-0.7). At least one case of podoconiosis was found in every region of Cameroon except the two surveyed villages in Adamawa. Of the 40 health districts surveyed, 17 districts had no cases of podoconiosis; in 15 districts, mean prevalence was between 0.2% and 1.0%; and in the remaining eight, mean prevalence was between 1.2% and 2.7%. CONCLUSIONS: Our investigation has demonstrated low prevalence but almost nationwide distribution of podoconiosis in Cameroon. Designing a podoconiosis control program is a vital next step. A health system response to the burden of podoconiosis is important, through case surveillance and morbidity management services.

Molecular typing reveals substantial Plasmodium vivax infection in asymptomatic adults in a rural area of Cameroon.

BACKGROUND: Malaria in Cameroon is due to infections by Plasmodium falciparum and, to a lesser extent, Plasmodium malariae and Plasmodium ovale, but rarely Plasmodium vivax. A recent report suggested "Plasmodium vivax-like" infections around the study area that remained unconfirmed. Therefore, molecular and antigenic typing was used to investigate the prevalence of P. vivax and Duffy in asymptomatic adults resident in Bolifamba. METHODS: A cross-sectional study was conducted from July 2008 to October 2009. The status of all parasite species was determined by nested PCR in 269 blood samples collected. The P. falciparum and P. vivax anti-MSP/CSP antibody status of each subject was also determined qualitatively by a rapid card assay. Parasite DNA was extracted from a sample infected with three parasite species, purified and sequenced. The Duffy antigen status of 12 subjects infected with P. vivax was also determined by sequencing. In silico web-based tools were used to analyse sequence data for similarities and matches to reference sequences in public DNA databases. RESULTS: The overall malaria parasite prevalence in 269 individuals was 32.3% (87) as determined by PCR. Remarkably, 14.9% (13/87) of infections were caused either exclusively or concomitantly by P. vivax, established both by PCR and microscopic examination of blood smears, in individuals both positive (50%, 6/12) and negative (50%, 6/12) for the Duffy receptor. A triple infection by P. falciparum, P. vivax and P. malariae, was detected in one infected individual. Anti-MSP/CSP antibodies were detected in 72.1% (194/269) of samples, indicating high and continuous exposure to infection through mosquito bites. DISCUSSION: These data provide the first molecular evidence of P. vivax in Duffy positive and negative Cameroonians and suggest that there may be a significant prevalence of P. vivax infection than expected in the study area. Whether the P. vivax cases were imported or due to expansion of a founder effect was not investigated. Notwithstanding, the presence of P. vivax may complicate control efforts if these parasites become hypnozoitic or latent as the liver stage. CONCLUSIONS: These data strongly suggest that P. vivax is endemic to the south-west region of Cameroon and should be taken into account when designing malaria control strategies.