The effect of climatic factors on the number of malaria cases in an inland and a coastal setting from 2011 to 2017 in the equatorial rain forest of Cameroon.

BACKGROUND: Weather fluctuation affects the incidence of malaria through a network of causuative pathays. Globally, human activities have ultered weather conditions over time, and consequently the number of malaria cases. This study aimed at determining the influence of humidity, temperature and rainfall on malaria incidence in an inland (Muyuka) and a coastal (Tiko) settings for a period of seven years (2011-2017) as well as predict the number of malaria cases two years after (2018 and 2019). METHODS: Malaria data for Muyuka Health District (MHD) and Tiko Health District (THD) were obtained from the Regional Delegation of Public Health and Tiko District Health service respectively. Climate data for MHD was obtained from the Regional Delegation of Transport while that of THD was gotten from Cameroon Development Coorporation. Spearman rank correlation was used to investigate the relationship between number of malaria cases and the weather variables and the simple seasonal model was used to forecast the number of malaria cases for 2018 and 2019. RESULTS: The mean monthly rainfall, temperature and relative humidity for MHD were 200.38 mm, 27.050C, 82.35% and THD were 207.36 mm, 27.57 °C and 84.32% respectively, with a total number of malaria cases of 56,745 and 40,160. In MHD, mean yearly humidity strongly correlated negatively with number of malaria cases (r = - 0.811, p = 0.027) but in THD, a moderate negative yearly correlation was observed (r = - 0.595, p = 0.159). In THD, the mean seasonal temperature moderately correlated (r = 0.599, p = 0.024) positively with the number of malaria cases, whereas MHD had a very weak negative correlation (r = - 0.174, p = 0.551). Likewise mean seasonal rainfall in THD moderately correlated (r = - 0.559, p = 0.038) negatively with malaria cases, contrary to MHD which showed a very weak positive correlation (r = 0.425, p = 0.130). The simple seasonal model predicted 6,842 malaria cases in Muyuka, for 2018 and same number for 2019, while 3167 cases were observed in 2018 and 2848 in 2019. Also 6,738 cases of malaria were predicted for MHD in 2018 likewise 2019, but 7327 cases were observed in 2018 and 21,735 cases in 2019. CONCLUSION: Humidity is the principal climatic variable that negatively influences malaria cases in MHD, while higher seasonal temperatures and lower seasonal rain fall significantly increase malaria cases in THD.

Genetic diversity and drug resistance surveillance of Plasmodium falciparum for malaria elimination: is there an ideal tool for resource-limited sub-Saharan Africa?

The intensification of malaria control interventions has resulted in its global decline, but it remains a significant public health burden especially in sub-Saharan Africa (sSA). Knowledge on the parasite diversity, its transmission dynamics, mechanisms of adaptation to environmental and interventional pressures could help refine or develop new control and elimination strategies. Critical to this is the accurate assessment of the parasite's genetic diversity and monitoring of genetic markers of anti-malarial resistance across all susceptible populations. Such wide molecular surveillance will require selected tools and approaches from a variety of ever evolving advancements in technology and the changing epidemiology of malaria. The choice of an effective approach for specific endemic settings remains challenging, particularly for countries in sSA with limited access to advanced technologies. This article examines the current strategies and tools for Plasmodium falciparum genetic diversity typing and resistance monitoring and proposes how the different tools could be employed in resource-poor settings. Advanced approaches enabling targeted deep sequencing is valued as a sensitive method for assessing drug resistance and parasite diversity but remains out of the reach of most laboratories in sSA due to the high cost of development and maintenance. It is, however, feasible to equip a limited number of laboratories as Centres of Excellence in Africa (CEA), which will receive and process samples from a network of peripheral laboratories in the continent. Cheaper, sensitive and portable real-time PCR methods can be used in peripheral laboratories to pre-screen and select samples for targeted deep sequence or genome wide analyses at these CEAs.

An evolutionary approach to identify potentially protective B cell epitopes involved in naturally acquired immunity to malaria and the role of EBA-175 in protection amongst denizens of Bolifamba, Cameroon.

BACKGROUND: The search for a vaccine against malaria caused by Plasmodium falciparum has lasted for more than 100 years, with considerable progress in the identification of a number of vaccine candidates. The post-genomic era offers new opportunities for an expedited search using rational vaccine design and prioritization of key B-cell epitopes involved in natural acquired immunity. METHODS: Malaria vaccine candidate genes that have reached clinical trial were searched on an evolutionary relationship tree, to determine their level of lineage-specificity. Ten other genes with similar protein features and level of lineage specificity to the vaccine candidates were randomly selected, and computationally evaluated for the presence of B-cell epitopes. The protein fragment with maximum probability of putative epitopes were synthesized and used in an ELISA experiment to determine the presence of antibodies to these peptides, in the serum of malaria patients and healthy malaria uninfected inhabitants from a malaria endemic region (Bolifamba), alongside with a vaccine candidate EBA-175. RESULTS: Two peptide fragments of 25 and 30 amino acid length from PF3D7_1233400 and PF3D7_1437500 respectively, coded as PF4-123 and PF4-143 were shown to contain B-cell epitope(s). Total IgG antibodies to these peptides were not significantly different between sick and healthy participants, but cytophilic antibodies to these peptides were significantly higher in healthy participants (p < 0.03). Total IgG to the vaccine candidate EBA-175 was significantly higher in sick participants than in healthy participants, likewise cytophilic antibodies (p < 0.04). Antibodies to the peptides PF4-123 and PF4-143 correlated negatively (p = 0.025 and 0.008 and r = -0.291 and -0.345, respectively) to parasite load. Total IgG antibodies to EBA-175 showed a negative correlation to parasite load (r = -0.144), which was not significant (p = 0.276). Duration of stay in Bolifamba also negatively correlated with parasite load (p = 0.026, r = -0.419) and total IgG to PF4-143 was significantly associated with prolonged duration of stay in the locality of Bolifamba, Cameroon (p = 0.006, r = 0.361). CONCLUSIONS: The present study has identified two genes PF3D7_1233400 and PF3D7_1437500 containing peptide fragment (PF4-123 and PF4-143) with B-cell epitopes that are correlated with naturally acquired immunity to malaria. A pipeline has been developed for rapid identification of other B-cell epitopes involved in naturally acquired immunity.

The current status of malaria epidemiology in Bolifamba, atypical Cameroonian rainforest zone: an assessment of intervention strategies and seasonal variations.

BACKGROUND: Global malaria has been on the decline over the past decade due to expansion of interventions. The present study aimed at determining the current status of malaria epidemiology in the context of sustained interventions and seasonal variations in Bolifamba, which represents a typical semi-urban malaria endemic community in the Cameroonian rainforest. METHODS: A monthly cross-sectional survey was carried out in Bolifamba, a multi ethnic semi-urban locality on the eastern flanks of Mt Cameroon, for a year during which blood samples were collected from participants and examined for malaria parasites by microscopy. Correlation analysis of seasonal/monthly malaria prevalence was done with weather data from Ekona, a nearby village with a meteorological station. Intervention strategy such as use of Insecticide Treated Bed Net (ITBN) and risk factors such as duration of stay in the locality, age and housing type were also investigated. RESULTS: The results revealed a malaria prevalence of 38.3 % in the rainy season, which was significantly higher than 24.4 % observed in the dry season (P < 0.0001). A high prevalence of asymptomatic malaria which was more than double the prevalence of symptomatic malaria on a monthly basis was observed, 30.7 % vs 17.8 % in the rainy and dry season respectively (p < 0.0001) and asymptomatic malaria was significantly associated with anemia (p < 0.005). April was the peak month of malaria prevalence and coincided with peak periods of both asymptomatic and symptomatic malaria. The Plasmodium falciparum parasite rates in the 2- up to 10-years age group (PfPR(2-10)) was 40.8 %. The regular use of ITBN was significantly associated with low prevalence of 31.7 % as opposed to irregular or non-usage of ITBN 38.2 % (p < 0.05). Log of parasite load was found to initially increase to 2.49 with less than 5 years of stay, and decreased gradually with increasing duration of stay in the locality (p = 0.046). Climatic factors were significantly and positively associated with monthly malaria prevalence and the strongest predictors of malaria prevalence were rainfall and minimum temperature with r values of 0.563 and 0.6 respectively. CONCLUSIONS: The study highlights the role of seasonal change in modifying malaria prevalence during the year and the beneficial effect of ITBN. It also underscores a sublime problem of asymptomatic malaria associated with anemia, and indicates that partial immunity is acquired with prolonged stay in Bolifamba. This preliminary result is the basis of ongoing work to identify the antigens involved in acquired immunity.

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.

Mycobacterium tuberculosis is the causative agent of tuberculosis in the southern ecological zones of Cameroon, as shown by genetic analysis.

BACKGROUND: Tuberculosis (TB) is a major cause of mortality and suffering worldwide, with over 95% of TB deaths occurring in low- and middle-income countries. In recent years, molecular typing methods have been widely used in epidemiological studies to aid the control of TB, but this usage has not been the case with many African countries, including Cameroon. The aims of the present investigation were to identify and evaluate the diversity of the Mycobacterium tuberculosis complex (MTBC) isolates circulating in two ecological zones of Cameroon, seven years after the last studies in the West Region, and after the re-organization of the National TB Control Program (NTBCP). These were expected to shed light also on the transmission of TB in the country. The study was conducted from February to July 2009. During this period, 169 patients with symptomatic disease and with sputum cultures that were positive for MTBC were randomly selected for the study from amongst 964 suspected patients in the savannah mosaic zone (West and North West regions) and the tropical rainforest zone (Central region). After culture and diagnosis, DNA was extracted from each of the MTBC isolates and transported to the BecA-ILRI Hub in Nairobi, Kenya for molecular analysis. METHODS: Genetic characterization was done by mycobacterial interspersed repetitive unit-variable number tandem repeat typing (MIRU-VNTR) and Spoligotyping. RESULTS: Molecular analysis showed that all TB cases reported in this study were caused by infections with Mycobacterium tuberculosis (98.8%) and Mycobacterium africanum (M. africanum) (1.2%) respectively. We did not detect any M. bovis. Comparative analyses using spoligotyping revealed that the majority of isolates belong to major clades of M. tuberculosis: Haarlem (7.6%), Latin American-Mediterranean (34.4%) and T clade (26.7%); the remaining isolates (31.3%) where distributed among the minor clades. The predominant group of isolates (34.4%) corresponded to spoligotype 61, previously described as the "Cameroon family. Further analysis based on MIRU-VNTR profiles had greater resolving power than spoligotyping and defined additional genotypes in the same spoligotype cluster. CONCLUSION: The molecular characterization of MTBC strains from humans in two ecological regions of Cameroon has shown that M. tuberculosis sensu stricto is the predominant agent of TB cases in the zones. Three decades ago, TB was reported to be caused by M. africanum in 56.0% of cases. The present findings are consistent with a major shift in the prevalence of M. tuberculosis in Cameroon.

Antimalarial drug interactions of compounds isolated from Kigelia africana (Bignoniaceae) and their synergism with artemether, against the multidrug-resistant W2mef Plasmodium falciparum strain.

For decades, drug resistance has been the major obstacle in the fight against malaria, and the search for new drugs together with the combination therapy constitutes the major approach in responding to this situation. The present study aims at assessing the in vitro antimalarial activity of four compounds isolated from Kigelia africana stem bark (atranorin - KAE1, specicoside - KAE7, 2ß,3ß,19α-trihydroxy-urs-12-20-en-28-oic acid - KAE3, and p-hydroxy-cinnamic acid - KAE10) and their drug interactions among themselves and their combination effects with quinine and artemether. The antiplasmodial activity and drug interactions were evaluated against the multidrug-resistant W2mef strain of Plasmodium falciparum using the parasite lactate dehydrogenase assay. Three of the four compounds tested were significantly active against W2mef: specicoside (IC(50) = 1.02 ± 0.17 µM), 2ß,3ß,19α-trihydroxy-urs-12-en-28-oic acid (IC(50) = 1.86 ± 0.15 µM) and atranorin (IC(50) = 1.78 ± 0.18 µM), whereas p-hydroxy-cinnamic acid showed a weak activity (IC(50) = 12.89 ± 0.87 µM). A slight synergistic effect was observed between atranorin and 2ß,3ß,19α-trihydroxy-urs-12-en-28-oic acid (Combination index, CI = 0.82) whereas the interaction between specicoside and p-hydroxy-cinnamic acid were instead antagonistic (CI = 2.67). All the three compounds showed synergistic effects with artemether, unlike the slight antagonistic interactions of atranorin and 2ß,3ß,19α-trihydroxy-urs-12-en-28-oic acid in combination with quinine. K. africana compounds are therefore likely to serve as leads in the development of new partner drugs in artemether-based combination therapy.

Factors Driving COVID-19 Vaccine Hesitancy in Cameroon and Their Implications for Africa: A Comparison of Two Cross-Sectional Studies Conducted 19 Months Apart in 2020 and 2022.

Many efficacious COVID-19 vaccines have been approved for general use but their ability to control the disease is being undermined by slow uptake. Resources are needed to persuade people to obtain a COVID-19 vaccine. Here we compare this present study and a previous one to assess the impact of the Cameroon government's policy and efforts to reduce COVID-19 vaccine hesitancy after one year of implementation. After obtaining ethical clearance and informed consent, 6732 participants completed a questionnaire about COVID-19 vaccine hesitancy and acceptance. It was observed that the government's policies and efforts reduced COVID-19 vaccine hesitancy significantly, but this was not enough to ensure the herd immunity necessary to control the disease. The risk factors associated with vaccine hesitancy were the consumption of traditional herbal remedies; living in an urban setting; being female, jobless or a student; working in the education sector; being a politician/policy maker/administrator, engineer or technician; medium income; no education/primary school/secondary/high school/professional training; and working in the informal sector. In contrast, people who were male, healthcare personnel, high-income earners, participants who do not consume traditional herbal remedies, infected or knowing someone who has been infected by COVID-19, and having a chronic illness or comorbidity, were associated with COVID-19 vaccine acceptance. Participants also gave several reasons they were either hesitant or willing to take the vaccine. A more rigorous surveillance system is needed to systematically monitor drivers of vaccine hesitancy, establish tailored interventions promoting vaccine acceptance, and evaluate the impact of these interventions.

Intermittent preventive treatment with Sulphadoxine-Pyrimethamine (IPTp-SP) is associated with protection against sub-microscopic P. falciparum infection in pregnant women during the low transmission dry season in southwestern Cameroon: A Semi - longitudinal study.

The current guidelines for malaria prevention and control during pregnancy in Africa is predicated on the prevention of infection and/or disease through intermittent preventive treatment in pregnancy (IPTp), insecticide-treated nets (ITNs) and effective malaria case diagnosis and management. Concerns that increasing SP resistance in some areas of SSA may have compromised IPTp-SP efficacy prompted this contemporaneous study, designed to assess the prevalence and risk factors of sub-microscopic infection in parturient women during the low transmission season in Mutengene, a rapidly growing semi-urban area in Southwest Region, Cameroon. Pregnant women originally reporting for the establishment of antenatal clinic care during the dry season were followed-up to term and their pregnancy outcomes recorded. About 2 ml of venous blood was collected for malaria diagnosis using PfHRP2/pLDH malaria rapid diagnostic kit and light microscopy. DNA was extracted from dried blood spots by the Chelex-100 method and the Plasmodium falciparum status detected by nested PCR amplification of the 18SrRNA gene using specific predesigned primers. Of the 300 women enrolled, the proportion of malaria parasite infected as determined by microscopy, RDT and PCR was 12.9%, 16.4% and 29.4% respectively, with 39.9% overall infected with P. falciparum by microscopy and/or RDT and/or PCR and a very low-density infection, averaging 271 parasites per microliter of blood. About 25.0% (68/272) of women who were negative by microscopy were positive by PCR (submicroscopic P. falciparum infection), with primigravidae and IPTp-SP non usage identified as independent risk factors for submicroscopic P. falciparum parasitaemia while fever history (aOR = 4.83, 95% CI = 1.28-18.22, p = 0.020) was associated with risk of malaria parasite infection overall. IPTp-SP use (p = 0.007) and dosage (p = 0.005) significantly influenced whether or not the participant will be malaria parasite negative or carry submicroscopic or microscopic infection. Although Infant birthweight and APGAR score were independent of the mother's P. falciparum infection and submicroscopic status, infant's birthweight varied with the gravidity status (p = 0.001) of the mother, with significantly lower birthweight neonates born to primigravidae compared to secundigravidae (p = 0.001) and multigravidae (p = 0.003). Even in holo-endemic dry season, there exists a large proportion of pregnant women with very low density parasitaemia. IPTp-SP seems to be relevant in controlling submicroscopic P. falciparum infections, which remains common in pregnant women, and are hard to diagnose, with potentially deleterious consequences for maternal and fetal health. Future studies should be carried out in hyperendemic malaria foci where the parasitemia levels are substantially higher in order to confirm the efficacy of IPTp-SP.

Hypericum lanceolatum (Hypericaceae) as a potential source of new anti-malarial agents: a bioassay-guided fractionation of the stem bark.

BACKGROUND: Malaria is a major public health threat in Africa, and traditional medicine continues to play a key role in its control especially in rural areas. A bioassay-guided fractionation was carried out in order to evaluate the anti-malarial potential and the safety of the methanol extract of the Hypericum lanceolatum stem bark. METHODS: The anti-plasmodial activity was assayed by the lactate dehydrogenase method (pLDH) against the multidrug-resistant W2mef laboratory strain, and a field isolate (SHF4) of Plasmodium falciparum. Cytotoxicity tests were carried out using the LLC-MK2 monkey kidney epithelial cells. RESULTS: Five compounds were isolated from the most active and least cytotoxic ethylacetate sub-extract: betulinic acid (HLT1), 2,2',5,6'-tetrahydroxybenzophenone (HLT2), 5-hydroxy-3-methoxyxanthone (HLT3), 3-hydroxy-5-methoxyxanthone (HLT4) and HLT0 (yet to be identified). Three of the tested compounds presented significant anti-plasmodial activities (with 50% inhibitory concentration, IC50 < 5 µM), with 5-hydroxy-3-methoxyxanthone exerting the highest activity, followed by HLT0 and betulinic acid. All the compounds with significant anti-plasmodial activity were non-cytotoxic, except betulinic acid which showed a 50% cytotoxic concentration, CC50 of 25 µg/mL. CONCLUSIONS: These findings justify the use of H. lanceolatum stem bark as anti-malarial by traditional healers of Western Cameroon, and could constitute a good basis for further studies towards development of new drug candidates or phytomedicines for malaria.

In vitro antiplasmodial activity and cytotoxicity of crude extracts and compounds from the stem bark of Kigelia africana (Lam.) Benth (Bignoniaceae).

In order to assess the potential of the stem bark of Kigelia africana (Lam.) Benth as source of new anti-malarial leads, n-hexane and ethyl acetate (EtOAc) extracts and four compounds isolated from the stem bark were screened in vitro against the chloroquine-resistant W-2 and two field isolates of Plasmodium falciparum using lactate dehydrogenase assay. The products were also tested for their cytotoxicity on LLC/MK2 monkey kidney cells. The EtOAc extract exhibited a significant antiplasmodial activity (IC(50) = 11.15 µg/mL on W-2; 3.91 and 4.74 µg/mL on field CAM10 and SHF4 isolates, respectively), whereas the n-hexane fraction showed a weak activity (IC(50) = 73.78 µg/mL on W-2 and 21.85 µg/mL on SHF4). Three out of the four compounds showed good activity against all the three different parasite strains (IC(50) <5 µM). Specicoside exhibited the highest activity on W-2 (IC(50) = 1.54 µM) followed by 2ß, 3ß, 19α-trihydroxy-urs-12-en-28-oic acid (IC(50) = 1.60 µM) and atranorin (IC(50) = 4.41 µM), while p-hydroxycinnamic acid was the least active (IC(50) =53.84 µM). The EtOAc extract and its isolated compounds (specicoside and p-hydroxycinnamic acid) were non-cytotoxic (CC(50) > 30 µg/mL), whereas the n-hexane extract and two of its products, atranorin and 2ß, 3ß, 19α-trihydroxy-urs-12-en-28-oic acid showed cytotoxicity at high concentrations, with the last one being the most toxic (CC(50) = 9.37 µg/mL). These findings justify the use of K. africana stem bark as antimalaria by traditional healers of Western Cameroon, and could constitute a good basis for further studies towards development of new leads or natural drugs for malaria.

Assessment of Vaccine Hesitancy to a COVID-19 Vaccine in Cameroonian Adults and Its Global Implication.

Since the outbreak of COVID-19 in December 2019, no global consensus treatment has been developed and generally accepted for the disease. However, eradicating the disease will require a safe and efficacious vaccine. In order to prepare for the eventual development of a safe and efficacious COVID-19 vaccine and to enhance its uptake, it is imperative to assess vaccine hesitancy in Cameroonians. After obtaining ethical clearance from the Institutional Review Board of the University of Buea, a questionnaire was administered (May-August 2020) to consenting adults either online or in person. A qualitative thematic analysis was done to analyze the participants' answers to the open questions. A deductive approach was used, that is, the codes and patterns according to the World Health Organization (WHO) Strategic Advisory Group of Experts (SAGE) Working Group Matrix of Determinants of vaccine hesitancy. The number of consenting adult Cameroonians who completed the questionnaire were 2512 (Two thousand five hundred and twelve). Vaccine hesitancy to a COVID-19 vaccine was 84.6% in Cameroonians. Using the WHO recommended Matrix of Determinant of Vaccine hesitancy, the most prominent determinants observed in this study were: Communication and Media Environment, Perception of pharmaceutical industry, Reliability and/or source of vaccine and cost. Most Cameroonians agree that even though there are benefits of a clinical trial, they will prefer it should be done out of the continent and involving African scientists for eventual acceptance and uptake. The concerns of safety, efficacy and confidence has to be addressed using a Public Engagement approach if a COVID-19 vaccine has to be administered successfully in Africa or Cameroon specifically. Since this study was carried out following WHO standards, its result can be compared to those of other studies carried out in different cultural settings using similar standards.

Investigating the risk factors for seroprevalence and the correlation between CD4+ T-cell count and humoral antibody responses to Toxoplasma gondii infection amongst HIV patients in the Bamenda Health District, Cameroon.

BACKGROUND: Toxoplasmosis is caused by an obligate intracellular tissue protozoan parasite, Toxoplasma gondii that infect humans and other warm-blooded animals. Transmission to humans is by eating raw or inadequately cooked infected meat or through ingestion of oocysts that cats have passed in faeces. Studies have shown life-threatening and substantial neurologic damage in immunocompromised patients; however, 80% of humans remain asymptomatic. The aim of this study was to determine the seroprevalence of Toxoplasma gondii infection in HIV positive patients and the risk factors associated with the infection, and to investigate the correlation between CD4+ T-cell count and toxoplasma specific antibodies as possible predictors of each other amongst HIV patients in the Bamenda Health District of the North West Region of Cameroon. METHODS: A cross-sectional study was conducted, in which 325 HIV patients were recruited for administration of questionnaire, serological diagnosis of T. gondii and measurement of CD4+ T-cell count. Bivariate and multivariate logistic regression was used to identify risk factors associated with T. gondii infection while the linear regression was used to investigate the relationship between CD4+ T-cell count and antibody levels against T. gondii. RESULTS: The findings showed that, majority (45.8%) of HIV patients suffered from chronic (IgG antibody) infection, and 6.5% from acute (IgM and IgM/IgG antibody) toxoplasma infection. The overall sero-prevalence of T. gondii infection amongst HIV patients was 50.5%. On the whole, 43 men (45.7%) and 127 women (55%) presented with anti- T. gondii antibodies; however, there was no significant difference amongst males and females who were positive to T. gondii infection (p = 0.131). Marital status (p = 0.0003), contact with garden soil (p = 0.0062), and garden ownership (p = 0.009), were factors that showed significant association with T. gondii infection. There was no significant difference (p = 0.909) between the mean CD4+ T-cell count of HIV patients negative for toxoplasma infection (502.7 cells/mL), chronically infected with T. gondii (517.7 cells/mL) and acutely infected with T. gondii (513.1 cells/mL). CD4+ T-cell count was neither a predictor of IgM antibody titer (r = 0.193, p = 0.401), nor IgG antibody titer (r = 0.149, p = 0.519) amongst HIV patients acutely infected with T. gondii. CONCLUSION: The findings from this study underscore the need to implement preventive and control measures to fight against T. gondii infection amongst HIV patients in the Bamenda Health District.

Analysis of the Role of TpUB05 Antigen from Theileria parva in Immune Responses to Malaria in Humans Compared to Its Homologue in Plasmodium falciparum the UB05 Antigen.

Despite the amount of resources deployed and the technological advancements in molecular biology, vaccinology, immunology, genetics, and biotechnology, there are still no effective vaccines against malaria. Immunity to malaria is usually seen to be species- and/or strain-specific. However, there is a growing body of evidence suggesting the possibility of the existence of cross-strain, cross-species, and cross-genus immune responses in apicomplexans. The principle of gene conservation indicates that homologues play a similar role in closely related organisms. The homologue of UB05 in Theileria parva is TpUB05 (XP_763711.1), which has been tested and shown to be associated with protective immunity in East Coast fever. In a bid to identify potent markers of protective immunity to aid malaria vaccine development, TpUB05 was tested in malaria caused by Plasmodium falciparum. It was observed that TpUB05 was better at detecting antigen-specific antibodies in plasma compared to UB05 when tested by ELISA. The total IgG raised against TpUB05 was able to block parasitic growth in vitro more effectively than that raised against UB05. However, there was no significant difference between the two study antigens in recalling peripheral blood mononuclear cell (PBMC) memory through IFN-γ production. This study suggests, for the first time, that TpUB05 from T. parva cross-reacts with UB05 from P. falciparum and is a marker of protective immunity in malaria. Hence, TpUB05 should be considered for possible development as a potential subunit vaccine candidate against malaria.

Detection, purification and characterisation of a secretory alkaline phosphatase from Onchocerca species.

An Onchocerca secretory alkaline phosphatase (E.C. 3.1.3.1) of molecular weight 90 kDa when in crude extract, but which dimerises to about 180 kDa upon purification, was detected, purified and characterised. The enzyme was found to be secreted by both O. ochengi and O. volvulus worms. It was shown to be of Onchocerca origin by Western blotting with bovine onchocerciasis sera and by its time-dependent release in cultures. The O. ochengi enzyme was purified to near homogeneity by a combination of polyethylene glycol precipitation, DEAE-cellulose chromatography and preparative electrophoresis. About 0.96 mg of the active enzyme was purified from 48.4 mg of the crude parasite-released products, giving a purification fold of 71.45 and a yield of 8.7%. The purified enzyme exhibited a typical Michaelis-Menten kinetics with optimum activity on p-nitrophenylphosphate (p-NPP) at pH 10.2. Its apparent K(m) for p-NPP was 0.56+/-0.03 mM and it required Mg(2+) and dithiothreitol (DTT) for stability throughout its purification. Sodium dodecyl sulphate at 2% (w/v) did not inhibit the enzyme activity, but apparently stabilised it during freezing. Inorganic phosphate inhibited the enzyme competitively with an apparent inhibition constant (K(i)) of 3.33+/-0.04 mM, whereas l-phenylalanine inhibited it in a mixed way with a K(i) of 3.18+/-0.03 mM. While contributing to the understanding of metabolism in Onchocerca, the present apparently unique enzyme which is likely to serve in the nutrition of the parasite could be further characterised as a macrofilaricide target or diagnostic marker in onchocerciasis.

Malaria vectors and transmission dynamics in coastal south-western Cameroon.

BACKGROUND: Malaria is a major public health problem in Cameroon. Unlike in the southern forested areas where the epidemiology of malaria has been better studied prior to the implementation of control activities, little is known about the distribution and role of anophelines in malaria transmission in the coastal areas. METHODS: A 12-month longitudinal entomological survey was conducted in Tiko, Limbe and Idenau from August 2001 to July 2002. Mosquitoes captured indoors on human volunteers were identified morphologically. Species of the Anopheles gambiae complex were identified using the polymerase chain reaction (PCR). Mosquito infectivity was detected by the enzyme-linked immunosorbent assay and PCR. Malariometric indices (plasmodic index, gametocytic index, parasite species prevalence) were determined in three age groups (<5 yrs, 5-15 yrs, >15 yrs) and followed-up once every three months. RESULTS: In all, 2,773 malaria vectors comprising Anopheles gambiae (78.2%), Anopheles funestus (17.4%) and Anopheles nili (7.4%) were captured. Anopheles melas was not anthropophagic. Anopheles gambiae had the highest infection rates. There were 287, 160 and 149 infective bites/person/year in Tiko, Limbe and Idenau, respectively. Anopheles gambiae accounted for 72.7%, An. funestus for 23% and An. nili for 4.3% of the transmission. The prevalence of malaria parasitaemia was 41.5% in children <5 years of age, 31.5% in those 5-15 years and 10.5% in those >15 years, and Plasmodium falciparum was the predominant parasite species. CONCLUSION: Malaria transmission is perennial, rainfall dependent and An. melas does not contribute to transmission. These findings are important in the planning and implementation of malaria control activities in coastal Cameroon and West Africa.

Malaria in pregnant Cameroonian women: the effect of age and gravidity on submicroscopic and mixed-species infections and multiple parasite genotypes.

Polymerase chain reaction (PCR)-based methods were used to investigate malaria in pregnant women residing in Yaounde, Cameroon. Microscopy and species-specific PCR-based diagnosis show that at delivery 82.4% of the women were infected with Plasmodium falciparum (27.5% blood-smear positive and 54.9% submicroscopic infections). The prevalence of P. malariae and P. ovale was 7.6% and 2.5%, respectively, with 9.4% infected with more than one species. Based on genotyping of the merozoite surface protein 1 (msp-1) and msp-2 alleles, the mean number of genetically different P. falciparum parasites in peripheral blood was 3.4 (range = 1-9) and 3.5 (range 1-8) in the placenta. Plasmodium falciparum detected by microscopy and PCR as well as mixed-species infections were significantly higher in women < or = 20 years old and paucigravidae, but maternal anemia was associated only with microscopic detection of parasites. Neither submicroscopic infections nor number of parasite genotypes decreased significantly with age or gravidity. Thus, pregnancy-associated immunity helps reduce malaria to submicroscopic levels, but does not reduce the number of circulating parasite genotypes.

Sensitive and specific serodiagnosis of onchocerciasis with recombinant hybrid proteins.

Onchocerciasis remains a major health hazard in many tropical countries. However, the existing tools for diagnosis of the disease have limitations, particularly regarding the detection of low level or early infections. To design an optimized reagent, we exploited the high antibody reactivity of patient sera against the Onchocerca volvulus proteins Ov20 and Ov33, which have been described as highly sensitive and specific immunodiagnostic reagents for producing hybrid proteins. The construct OvH2 was composed of Ov20 fused to Ov33, while OvH3 consisted of the C-terminus of Ov20 linked to Ov33. When these constructs were tested with sera from patients with onchocerciasis and control sera, OvH2 showed a sensitivity of 98.5% and a specificity of 97.7% and OvH3 showed a sensitivity of 98.5% and a specificity of 95.35%. All non-responders were from Ecuador. These results surpass those of existing single recombinant antigens, suggesting that our hybrid proteins combined the sensitivity of the two parent proteins. Tests based on OvH2 should prove suitable for monitoring onchocerciasis control programs and individual diagnosis.

Detection of Rickettsia africae in patients and ticks along the coastal region of Cameroon.

Rickettsia africae was identified in seven (6%) of 118 patients with acute fevers of unknown etiology proven not to be malaria or typhoid fever from clinics along the coastal region of Cameroon by polymerase chain reaction (PCR) amplification and sequencing of the citrate synthase (gltA) and outer membrane protein A (ompA) genes of Rickettsia. The majority (71%) of the patients were female. Clinical manifestations included fever (100%), headache (71%), myalgia (71%), arthralgia (43%), pulmonary involvement (29%), and diffuse rash (14%). Moreover, R. africae was detected by PCR amplification and sequence analysis of the gltA and ompA genes in 62 (75%) of 83 adult Amblyomma variegatum ticks collected from cattle in the same region. These results confirm the presence of a previously unrecognized infectious disease in the indigenous Cameroonian population, as well as extend the established range of R. africae.

Sensitive and specific serodiagnosis of river blindness using Onchocerca ochengi antigens.

Despite the large-scale application of ivermectin for the treatment of human onchocerciasis, this filarial disease has remained a public health hazard, requiring rapid and sensitive diagnostic methods for more efficient management. Several serological methods based on homologous native or recombinant diagnostic antigens have been developed. However, these antigens have remained scarce in the endemic areas. We report herein the preparation and evaluation of an Onchocerca ochengi total worm extract together with its low molecular weight (< 31 kDa) and 14.4 kDa antigens for the diagnosis of human onchocerciasis. The 14.4 kDa antigen and the low molecular weight antigen fraction were identified by Western blot experiments. The antigens were employed for the detection of IgG4 in sera of 43 onchocerciasis patients, 35 normal Africans, 15 schistosomiasis patients, 11 ascariasis patients, 9 loaisis patients and 3 healthy Europeans. The diagnostic specificity values were 94.5, 87.5 and 100%, while the sensitivities were 90.7, 80 and 76.6% using the worm extract, low molecular weight antigen fraction and the 14.4 kDa antigen, respectively. We conclude that total worm extracts of O. ochengi can conveniently replace recombinant antigen cocktails in poor settings that do not have access to the latter.