Vector competence of Aedes aegypti in transmitting Chikungunya virus: effects and implications of extrinsic incubation temperature on dissemination and infection rates.

BACKGROUND: Aedes aegypti is a competent arthropod vector of chikungunya virus (CHIKV). The rate at which the virus disseminate in the vector is limited by temperature of their environment which can be an important determinant of geographical and seasonal limits to transmission by the arthropods in the tropics. This study investigated the vector competence of Ae. aegypti for CHIKV at ambient temperature of 32 and 26 °C (Coastal and Western Kenya respectively) reared at Extrinsic Incubation Temperature (EIT) of 32 and 26 °C that resembles those in the two regions. METHODS: Ae. aegypti eggs were collected from coastal and Western Kenya, hatched in the insectary and reared to F1 generation. Four-day old mosquitoes were exposed to CHIKV through a membrane feeding. They were then incubated in temperatures mimicking the mean annual temperatures for Trans-Nzoia (26 °C) and Lamu (32 °C). After every 7, 10 and 13 days post infection (DPI); one third of exposed mosquitoes were sampled and assayed for virus infection and dissemination. RESULTS: The midgut infection rates (MIR) of Ae. aegypti sampled from Coastal Region was significantly (p < 0.05) higher than those sampled from Western Kenya, with no statistical differences observed for the coastal Ae. aegypti at EIT 26 and at 32 °C. The MIR of Ae. aegypti from the Western Region was significantly (p < 0.05) affected by the EIT, with mosquito reared at EIT 32 °C exhibiting higher MIR than those reared at EIT 26 °C. There was a significant (p < 0.05) interactive effects of the region, EIT and DPI on MIR. The disseminated infection rates for the CHIKV in Ae. aegypti in the legs (DIR-L) was higher in mosquitoes sampled from Coast regardless of the EIT while those from Western Kenya, dissemination rates were significantly higher at higher EIT of 32 °C. CONCLUSIONS: Vector competence was higher in mosquito populations reared under high temperatures which weakens the midgut infection barrier. Hence, suggesting Lamu population is more susceptible to CHIKV therefore having a weaker mid gut infection barrier than the Trans Nzoia population. These underscores importance of examining the course of infection at various ambient temperatures and EIT between regions mosquito populations.

PCBs in fish and their cestode parasites in Lake Victoria.

Polychlorinated biphenyls (PCBs) are classified as persistent organic pollutants (POPs) regulated by the Stockholm Convention (2001). Although their production and use was stopped almost three decades ago, PCBs are environmental persistent, toxic, and bioaccumulate in biota. We assessed the levels of 7 PCB congeners (IUPAC nos. 28, 52, 101, 118, 138, 153, and 180) in sediment and fish (Oreochromis niloticus, Lates niloticus, and Rastrineobola argentea) and evaluated the potential of cestode fish endoparasite (Monobothrioides sp., Proteocephalaus sp., and Ligula intestinalis) as biomonitors of PCBs in Lake Victoria, Kenya. The median concentration of Σ7PCBs in sediments and fish were 2.2-96.3 µg/kg dw and 300-3,000 µg/kg lw, respectively. At all the sampling sites, CB138, CB153, and CB180 were the dominant PCB congeners in sediment and fish samples. Compared to the muscle of the piscine host, Proteocephalaus sp. (infecting L. niloticus) biomagnified PCBs ×6-14 while Monobothrioides sp. (infecting O. niloticus) biomagnified PCBs ×4-8. Meanwhile, L. intestinalis (infecting R. argentea) biomagnified PCBs ×8-16 compared to the muscle of unparasitized fish. We demonstrate the occurrence of moderate to high levels of PCB in sediments and fish in Lake Victoria. We also provide evidence that fish parasites bioaccumulate higher levels of PCBs than their piscine hosts and therefore provide a promising biomonitor of PCBs. We urge further a long-term study to validate the use of the above cestode fish parasites as biomonitoring tools for PCBs.

Effects of dietary administration of stinging nettle (Urtica dioica) on the growth performance, biochemical, hematological and immunological parameters in juvenile and adult Victoria Labeo (Labeo victorianus) challenged with Aeromonas hydrophila.

We investigated effects of dietary administration of stinging nettle (Urtica dioica) on growth performance, biochemical, hematological and immunological parameters in juvenile and adult Victoria Labeo (Labeo victorianus) against Aeromonas hydrophila. Fish were divided into 4 groups and fed for 4 and 16 weeks with 0%, 1%, 2% and 5% of U. dioica incorporated into the diet. Use of U. dioica in the diet resulted in improved biochemical, hematological and immunological parameters. Among the biochemical parameters; plasma cortisol, glucose, triglyceride and cholesterol decreased while total protein and albumin in fish increased with increasing dietary inclusion of U. dioica. Among the haematology parameters: red blood cell (RBC), white blood cell (WBC) counts, haematocrit (Htc), mean cell haemoglobin (MCH), mean cell haemoglobin concentration (MCHC) and netrophiles increased with increasing dietary inclusion levels of U. dioica, some depending on the fish age. Serum immunoglobulins, lysozyme activity and respiratory burst were the main immunological parameters in the adult and juvenile L. victorianus measured and they all increased with increasing herbal inclusion of U. dioica in the diet. Dietary incorporation of U. dioica at 5% showed significantly higher relative percentage survival (up to 95%) against A. hydrophila. The current results demonstrate that using U. dioica can stimulate fish immunity and make L. victorianus more resistant to bacterial infection (A. hydrophila).

Symptomatic malaria diagnosis overestimate malaria prevalence, but underestimate anaemia burdens in children: results of a follow up study in Kenya.

BACKGROUND: The commonly accepted gold standard diagnostic method for detecting malaria is a microscopic reading of Giemsa-stained blood films. However, symptomatic diagnosis remains the basis of therapeutic care for the majority of febrile patients in malaria endemic areas. This study aims to compare the discrepancy in malaria and anaemia burdens between symptomatic diagnosed patients with those diagnosed through the laboratory. METHODS: Data were collected from Western Kenya during a follow-up study of 887 children with suspected cases of malaria visiting the health facilities. In the laboratory, blood samples were analysed for malaria parasite and haemoglobin levels. Differences in malaria prevalence between symptomatic diagnosis and laboratory diagnosis were analysed by Chi-square test. Bayesian probabilities were used for the approximation of the malaria and anaemia burdens. Regression analysis was applied to: (1) determine the relationships between haemoglobin levels, and malaria parasite density and (2) relate the prevalence of anaemia and the prevalence of malaria. RESULTS: The prevalence of malaria and anaemia ranged from 10% to 34%, being highest during the rainy seasons. The predominant malaria parasite was P. falciparum (92.3%), which occurred in higher density in children aged 2‒5 years. Fever, high temperature, sweating, shivering, vomiting and severe headache symptoms were associated with malaria during presumptive diagnosis. After conducting laboratory diagnosis, lower malaria prevalence was reported among the presumptively diagnosed patients. Surprisingly, there were no attempts to detect anaemia in the same cohort. There was a significant negative correlation between Hb levels and parasite density. We also found a positive correlation between the prevalence of anaemia and the prevalence of malaria after laboratory diagnosis indicating possible co-occurrence of malaria and anaemia. CONCLUSION: Symptomatic diagnosis of malaria overestimates malaria prevalence, but underestimates the anaemia burden in children. Good clinical practice dictates that a laboratory should confirm the presence of parasites for all suspected cases of malaria.

Monitoring exposure to heavy metals among children in Lake Victoria, Kenya: environmental and fish matrix.

This study used hair and nails to biomonitor heavy metals (Pb, Cd, Cr and Cu) from geological source and exposure through regular fish consumption among children in Lake Victoria, Kenya. Concentration of Pb and Cu in water reflected anthropogenic pathways, while Cd and Cr reflected accumulation from the catchment basin. Higher concentration of heavy metals in the nails samples than the hair samples suggested longer term exposure. The estimated intake of Cd and Cr from fish in one site associated with high concentration of the metals from geological source was appreciably above the respective recommended daily allowance, signifying possible health risks to humans. Significant correlations between Pb, Cd and Cu in hair, nails and heavy metals from fish consumed suggested fish consumption as possible pathway of heavy metals in humans. Possible health risks from heavy metals were likely due to consumption of higher quantities of fish and from geological basins.

Effectiveness of option B highly active antiretroviral therapy (HAART) prevention of mother-to-child transmission (PMTCT) in pregnant HIV women.

BACKGROUND: Ensuring that no baby is born with HIV is an essential step towards achieving an AIDS-free generation. To achieve this, strategies that decouple links between childbirth and HIV transmission are necessary. Traditional forms of prevention of mother-to-child transmission of HIV (PMTCT), has been recommended. Recognizing the importance and challenges of combination of methods to achieve rapid PMTCT, the World Health Organization (WHO) recommended option B Highly Active Antiretroviral Therapy (HAART) for all HIV-positive pregnant women. This study aimed to evaluate the effectiveness of the HAART in PMTCT. A cohort of HIV-infected pregnant women in Kenya were obtained from the DREAM Center, Nairobi. The study participants underwent adherence counselling and Option B of HAART [Nevirapine(NVP) + Lamivudine + Zidovudine] at the fourth week of gestation followed by an intravenous NVP administration intrapartum and postpartum NVP syrup to the respective infants for six weeks. Absolute pre-HAART and post-HAART CD4 counts and viral loads counts were determined. Comparison of the CD4 counts and viral loads before and after administration of HAART were done using Wilcoxon's Matched Pairs Signed-Ranks Test. FINDINGS: The mean absolute CD4 cell counts in mothers after administration of HAART was significantly higher (Z = 15.664, p < 0.001) than before the administration of HAART). Also the viral load of the mothers significantly (Z = 11.324, p < 0.001) reduced following HAART treatment. Following the HAART administration in mothers, up to 90% of children were confirmed to be HIV negative. CONCLUSION: Administration of HAART to mothers and children demonstrated an effective mechanism of PMTCT. However, other aspects of HAART such as adherence, costs, mothers behaviour during HAART, and the child feeding programme during the therapy should further be evaluated and ascertained how they can affect the overall efficacy of option B HAART in PMTCT.

Contribution of soil, water and food consumption to metal exposure of children from geological enriched environments in the coastal zone of Lake Victoria, Kenya.

Geologically enriched environments may contain high concentrations of some metals. In areas where industrial exposures remain superficial, children may be exposed to these geological metals through soil, drinking water and consumption of food locally grown. The aim of this study was to assess the contribution of soil, water and food consumption to cadmium (Cd), chromium (Cr), copper (Cu), zinc (Zn), cobalt (Co) and titanium (Ti) body burden of children in the geologically metal rich coastal zone of Lake Victoria, Kenya. We evaluated the relationship between the metal body burden of children by analyzing the hair samples and the estimated metal intake through exposure to soil, drinking water, and food consumption. The body burdens of all metals in children at the exposed sites were high, suggesting environmental exposure. Most of the foods consumed resulted in metal intake below the maximum recommended daily intake. However, the net contribution of all food items jointly exceeded the recommended maximum daily intake at some of the exposure sites. Our results further demonstrated that the highest intake of metals occurred via ingestion of high quantities of the cyprinid fish Rastrineobola argentea. Positive linear relationships between the metal body burden in children and estimated daily intake were observed for Cd, Cr, Ti in children consuming high quantity of R. argentea; for Zn and Co in children highly exposed to soils and consuming high quantity of Brassica oleraceae, while Co burden in the body was associated with high consumption of maize meal. This study demonstrated that exposure to soil and consumption of higher quantities of some foods contributed to the increased metal body burdens of children in a geologically exposed region.

Combination therapy using Pentostam and Praziquantel improves lesion healing and parasite resolution in BALB/c mice co-infected with Leishmania major and Schistosoma mansoni.

BACKGROUND: Most natural host populations are exposed to a diversity of parasite communities and co-infection of hosts by multiple parasites is commonplace across a diverse range of systems. Co-infection with Leishmania major and Schistosoma mansoni may have important consequences for disease development, severity and transmission dynamics. Pentavalent antimonials and Praziquantel (PZQ) have been relied upon as a first line of treatment for Leishmania and Schistosoma infections respectively. However, it is not clear how combined therapy with the standard drugs will affect the host and parasite burden in concomitance. The aim of the current study was to determine the efficacy of combined chemotherapy using Pentostam and PZQ in BALB/c mice co-infected with L. major and S. mansoni. METHODS: The study used BALB/c mice infected with L. major and S. mansoni. A 3 × 4 factorial design with three parasite infection groups (Lm, Sm, Lm + Sm designated as groups infected with L. major, S. mansoni and L. major + S. mansoni, respectively) and four treatment regimens [P, PZQ, P + PZQ and PBS designating Pentostam®(GlaxoSmithKline UK), Praziquantel (Biltricide®, Bayer Ag. Leverkusen, Germany), Pentostam + Praziquantel and Phosphate buffered saline] as factors was applied. In each treatment group, there were 10 mice. Lesion development was monitored for 10 weeks. The parasite load, body weight, weight of the spleen and liver were determined between week 8 and week 10. RESULTS: Chemotherapy using the first line of treatment for L. major and S. mansoni reduced the lesion size and parasite loads but did not affect the growth response, spleen and liver. In the co-infected BALB/c mice, the use of Pentostam or PZQ did not result in any appreciable disease management. However, treatment with P + PZQ resulted in significantly (p < 0.05) larger reduction of lesions, net increase in the body weight, no changes in the spleen and liver weight and reduced Leishman-Donovan Units (LDU) and worm counts than BALB/c mice treated with Pentostam or PZQ alone. CONCLUSIONS: The present study demonstrated that the combined first line of treatment is a more effective strategy in managing co-infection of L. major and S. mansoni in BALB/c mice.

Element profiles in hair and nails of children reflect the uptake from food and the environment.

The present study evaluated if the element profiles in hair and nails of children from the Lake Victoria region in Kenya reflect uptake from food and the environment. To this purpose, the element concentrations in hair and nails were related to element concentrations measured in food items, water, and soil. The highest intake of most elements occurred via ingestion of the fish Rastrineobola argentea, which is consumed in higher quantities than other fish and food items. Element concentrations in hair and nails were correlated to food and soil in element-specific patterns. Multivariate analysis combining principal component analysis and multidimensional scaling enabled us to cluster individuals from specific sites and to relate differences between sites to food-consumption patterns and environmental exposures. Site-specific differences in macroelement concentrations among the children were attributed to patterns of food consumption, while those in microelement concentrations reflected differences in geochemical background. It is concluded that the simultaneous analysis of elements in human hair and nails allows separation of populations based on food consumption and geochemical background.

Parasites modify sub-cellular partitioning of metals in the gut of fish.

Infestation of fish by parasites may influence metal accumulation patterns in the host. However, the subcellular mechanisms of these processes have rarely been studied. Therefore, this study determined how a cyprinid fish (Rastrineobola argentea) partitioned four metals (Cd, Cr, Zn and Cu) in the subcellular fractions of the gut in presence of an endoparasite (Ligula intestinalis). The fish were sampled along four sites in Lake Victoria, Kenya differing in metal contamination. Accumulation of Cd, Cr and Zn was higher in the whole body and in the gut of parasitized fish compared to non-parasitized fish, while Cu was depleted in parasitized fish. Generally, for both non-parasitized and parasitized fish, Cd, Cr and Zn partitioned in the cytosolic fractions and Cu in the particulate fraction. Metal concentrations in organelles within the particulate fractions of the non-parasitized fish were statistically similar except for Cd in the lysosome, while in the parasitized fish, Cd, Cr and Zn were accumulated more by the lysosome and microsomes. In the cytosolic fractions, the non-parasitized fish accumulated Cd, Cr and Zn in the heat stable proteins (HSP), while in the parasitized fish the metals were accumulated in the heat denatured proteins (HDP). On the contrary, Cu accumulated in the HSP in parasitized fish. The present study revealed specific binding of metals to potentially sensitive sub-cellular fractions in fish in the presence of parasites, suggesting interference with metal detoxification, and potentially affecting the health status of fish hosts in Lake Victoria.

Dynamics of metal uptake and depuration in a parasitized cyprinid fish (Rastrineobola argentea).

Infestation of fish by endoparasites may potentially influence metal uptake and elimination by the host. We quantified the metal uptake rate constant (k(u)) and efflux rate constants (k(e)) of radiolabeled Cd and Co in the cyprinid fish Rastrineobola argentea experimentally infected with the parasite Ligula intestinalis. During 24h, the accumulation of Cd and Co increased linearly with no evident steady state in uninfected fish, infected fish and in the parasite. Following aqueous exposures, the k(u) for Cd in parasites was about 3× higher than that of infected fish and 6× higher than for the uninfected fish. The k(u) for Co was up to 15× higher in the parasites than that of infected fish and 7.5× higher than for the uninfected fish. The k(e) for excretion of Cd were consistently higher for the uninfected fish than for the infected fish and also higher for uninfected fish than the parasite. The k(e) for Co for the uninfected fish was 1.4-2.0× lower than in the infected fish, but higher for parasites compared to uninfected fish (1.3-2.3×). Pulse-chase feeding experiments with radiolabeled copepods showed that Cd assimilation efficiency from food was higher in infected fish, while Co was assimilated more effectively by uninfected fish. The observed differences in metal dynamics between infected and uninfected R. argentea in the laboratory concord with differences in metal concentrations measured in natural populations in Lake Victoria. Our findings provide evidence that L. intestinalis infection enhances Cd accumulation, but depletes the essential Co in the cyprinid fish R. argentea. We conclude that the combined stress of parasites and pollution changes metal risks to fish hosts in a metal specific manner.

Metal specific partitioning in a parasite-host assemblage of the cestode Ligula intestinalis and the cyprinid fish Rastrineobola argentea.

When evaluating metal accumulation patterns in parasite-host assemblages species specific metal requirements should be taken into account. The aim of the present study was therefore to determine the metal specific partitioning in a parasite-host assemblage of the cestode Ligula intestinalis and the cyprinid fish Rastrineobola argentea and to determine the effect of the parasites on the metal balance of the fish. To this purpose the host-parasite assemblage was analysed for several metals at sites in the coastal zone of Lake Victoria differing in metal contamination. Our results showed that some elements (Ca, Sr, and Mg) reflected the physiological differences of bone formation and ionic balance and pointed to physiological disturbances of infested R. argentea. Other essential metals including Cu and Co were subject of element competition between fish and parasite, while only a micro-element (Cr) and a non-essential metal (Cd) displayed a partitioning with high concentration in the parasite. The present study clearly demonstrated the impact of the large cestodes on their small fish hosts and it is concluded that the partitioning of metals in the assemblage of R. argentea and L. intestinalis is subject to metal specific mechanisms for essential and non-essential elements.