An In vitro and in silico investigation of the antitrypanosomal activities of the stem bark extracts of Anopyxis klaineana (Pierre) Engl.

African Trypanosomiasis caused by trypanosome parasites continues to be a major neglected health problem, particularly in developing countries. Current treatments are marked by serious side effects, low effectiveness, high toxicity, and drug resistance prompting the need to develop novel, safe, effective, and alternative antitrypanosomal compounds. Anopyxis klaineana is an ethnomedicinal plant used in West Africa to treat many ailments including protozoan diseases. In this study, we investigated the antitrypanosomal potential of stem bark extracts of A. klaineana through in vitro and in silico approaches. A. klaineana extracts were tested for their antitrypanosomal activities against Trypanosoma brucei parasite in vitro using Alamar blue assay. In addition, the antioxidant and cytotoxic activities were determined. LC-ESI-QTOF-MS was used to identify potential bioactive compounds present in the A. klaineana extracts. Bioactive compounds identified were subjected to molecular docking studies against Trypanosoma brucei's trypanothione reductase (TR) and Uridine Diphosphate Galactose 4'-Epimerase (UDP). The A. klaineana extracts (methanol, hexane, chloroform, and ethyl acetate) exhibited potential anti-trypanosomal activities with IC50 values of 21.25 ± 0.755,4.35 ± 0.166,2.57 ± 0.153 and 22.92 ± 2.321 µg/mL respectively. Moreover, the methanolic crude extracts showed moderate cytotoxicity against HepG2 and PNT2 cells, with IC50 values of 68.0 ± 2.05 and 78.7 ± 2.63 µg/mL respectively. LC-MS analysis revealed the presence of 24 bioactive compounds with 5 being druglike. Risperidone, Ranolazine, Dihydro-7-Desacetyldeoxygedunin, 6 beta-Hydroxytriamcinolone acetonide, and Dimethylmatairesinol were identified as novel potential inhibitors of TR and UDP with binding affinities of -10.4, -7.9, -8.7, -8.4 and -7.1 kcal/mol respectively against TR and -10.8, -8.4, -8.4, -7.6 and -8.1 respectively against UDP. This study indicates that A. klaineana has potential antitrypanosomal properties and therefore may have the potential to be developed as a therapeutic intervention for treating African trypanosomiasis.

A literature review of schistosomiasis in Ghana: a reference for bridging the research and control gap.

Schistosomiasis is endemic in most sub-Saharan African countries, including Ghana, where the need for effective control involving preventive chemotherapy was indicated by the WHO. Mass drug administration commenced in 2008 and has continued since then in Ghana, but the country remains highly endemic. Here, we review the literature on schistosomiasis to identify research and knowledge gaps potentially affecting disease control. A total of 100 Ghana-related schistosomiasis literature sources were reviewed, showing that most studies were conducted on epidemiology, control of transmission and diagnosis. By contrast, many aspects of this disease remain neglected, including livestock schistosomiasis and its zoonotic potential, recent distribution of disease vectors or widely overlooked genital schistosomiasis. Stratified by region, the highest number of studies focus on Greater Accra, while studies are limited or absent for several other regions. Although this review shows apparent progress in terms of schistosomiasis research and control, a considerable amount of work remains to achieve at least a reduction in the prevalence of the disease, which affects a significant proportion of the population. National epidemiological data based on a nationwide survey, integrated control and improved monitoring and evaluation must be ensured.

Microbial Safety of Beef Along Beef Value Chains in the Ashaiman Municipality of Ghana.

Food from animal sources continues to be a significant food safety hazard. This study determined the microbial quality and safety of beef along beef value chains with case studies in the Ashaiman Municipality of Ghana. Raw beef samples were collected from four slaughter slabs in the Ashaiman Municipality and analyzed using standard microbiological methods to determine the quality and prevalence of specific pathogens, including Salmonella species, Listeria monocytogenes (L. monocytogenes), and Brucella species, as well as Toxoplasma gondii (T. gondii), Cyclospora cayetanensis (C. cayetanensis), and Cryptosporidium parvum (C. parvum). Data regarding food safety knowledge and practices were collected and observed from stakeholders (cattle farmers, butchers, and beef retailers). Salmonella typhimurium was isolated from 7.5% (6/80) of the total raw beef samples. However, L. monocytogenes, Brucella spp., T. gondii, C. cayetanensis, and C. parvum were not isolated in this study. The mean level of microbial contamination of beef from the slaughter slabs/abattoir [5.2 Log10 colony-forming unit (CFU)/g] was not significantly different (p > 0.05) from the mean level observed at retail points (5.4 Log10 CFU/g). However, the mean coliform count of 4.3 Log10 CFU/g recorded at retail shops exceeded the permissible limits of 104 CFU/g (4 Log10 CFU/g) required by the Ghana Standards Authority for safety of meat and carcasses. Knowledge on food safety was at average level for butchers and retailers. Unhygienic practices and poor sanitary conditions at the abattoirs and retail shops observed could be the main contributing factors to microbial contamination of raw beef. Continuous education for meat handlers on issues of food safety and monitoring of slaughter activities will reduce the rate and level of contamination of beef.

Increased ShTAL1 IgE responses post-Praziquantel treatment may be associated with a reduced risk to re-infection in a Ghanaian S. haematobium-endemic community.

BACKGROUND: Evidence from recent studies in Schistosoma mansoni-endemic areas show an age-associated immunity that is positively correlated with IgE titres to Schistosoma mansoni-specific tegumental allergen-like protein 1 (SmTAL1). The structural homology between SmTAL1 and the S. haematobium-specific TAL1 (ShTAL1) has been verified, yet it remains unclear whether similar age- and immune-associated trends characterize ShTAL1. This community-based intervention study was conducted to assess whether ShTAL1IgE responses post-treatment with praziquantel (PZQ) might be associated with a reduced risk to re-infection with S. haematobium. METHODOLOGY/PRINCIPAL FINDINGS: This study was conducted at Agona Abodom, Central Region, Ghana, and involved 114 participants aged 6 to 55 years. EDTA blood samples were collected at baseline and 7 weeks after PZQ treatment (Follow-up). Baseline and Follow-up titres of specific IgG1, IgG4, and IgE antibodies to the S. haematobium-specific adult worm antigen (ShAWA), the Sh-specific soluble egg antigen (ShSEA), and the Sh-specific tegumental-allergen-like 1 protein (ShTAL1) in plasma samples were measured using sandwich ELISA. Participants at both time points also provided stool and urine for helminth egg detection by microscopy. Prevalence of S. haematobium at baseline was 22.80%, and decreased to 3.50% at Follow-up. The egg reduction rate (ERR) was 99.87%. Overall plasma levels of ShTAL1-IgE increased 7 weeks post-PZQ treatment, and with increasing age; whiles S. haematobium infection prevalence and intensity decreased. For S. haematobium-infected participants who were egg-negative at Follow-up (N = 23), minimal median levels of ShTAL1-IgE were observed for all age groups prior to treatment, whilst median levels increased considerably among participants aged 12 years and older at Follow-up; and remained minimal among participants aged 11 years or less. In the univariate analysis, being aged 12 years or older implied an increased likelihood for ShTAL1-IgE positivity [12-14 years (cOR = 9.64, 95% CI = 2.09-44.51; p = 0.004); 15+ years (cOR = 14.26, 95% CI = 3.10-65.51; p = 0.001)], and this remained significant after adjusting for confounders [12-14 years (aOR = 22.34, 95% CI = 2.77-180.14; p = 0.004); ≥15 years (aOR = 51.82, 95% CI = 6.44-417.17; p < 0.001)]. Conversely, median ShTAL1-IgG4 titres were hardly detectible at Follow-up. CONCLUSIONS/SIGNIFICANCE: These findings demonstrate that increased IgE levels to ShTAL1 7 weeks after PZQ treatment could be associated with a reduced risk to re-infection, and adds to the large body of evidence suggesting a protective role of the treatment-induced ShTAL1 antigen in schistosomiasis infections. It was also quite clear from this work that apart from being persistently S. haematobium-positive, elevated ShTAL1-IgG4 levels at Follow-up could be indicative of susceptibility to re-infection. These outcomes have important implications in vaccine development, and in shifting the paradigm in mass chemotherapy programmes from a 'one-size-fits-all' approach to more sub-group-/participant-specific strategies in endemic areas.

The Incidence, Intensity, and Risk Factors for Soil Transmissible Helminthes Infections among Waste Handlers in a Large Coastal Periurban Settlement in Southern Ghana.

Soil-transmissible helminthes (STH) infections are among the most common sanitation-related public health problems in poor periurban settlements of tropical regions of low- and middle-income countries. In Ghana, research studies documenting the incidence rate, intensity, and occupational risk factors of STH infections among adults are scanty. A prospective cohort study of 261 waste handlers was conducted to investigate this. Stool samples were collected after 90 and 180 days of treatment with albendazole (400 mg per dose). The geometric mean intensity of STH among waste handlers after 180 days of treatment was 2.8 eggs/gram (light intensity), with an incidence rate of 1.5%. The proportion of waste handlers with light intensity STH infections was 4.8%. The odds of STH infection among female waste handlers were 80% lower when compared with male waste handlers (aOR = 0.2; 95% CI: 0.0-0.8). Waste handlers who used rubber gloves when working were 80% (aOR = 0.2: 95% CI: 0.2-1.9) protected from STH infections compared with those who did not use gloves. Infections with STH among the 261 waste handlers significantly correlated with the type of waste handling activities (LR χ 2 = 15.3; p=0.033) with the highest proportion of infection found among transporters, 2 (40%). Waste handlers should receive periodic antihelminthic treatment, at least once every six months, practice adequate hand hygiene, and use suitable personal protective equipment during work.

Risk of faecal pollution among waste handlers in a resource-deprived coastal peri-urban settlement in Southern Ghana.

Resource-deprived coastal peri-urban settlements in Southern Ghana are characterized by indiscriminate solid waste disposal and open defecation practices. Persons engaged in waste handling in such communities perform their activities with little or no personal protective equipment. They are thus confronted with the risk of faecal pollution of the hands and other bodily parts. A mixed method approach was used to investigate 280 waste handlers performing different activities to estimate recent faecal pollution of their hands and to observe the utilization of personal protective equipment and sanitation/hygiene facilities during work. The log concentration of E. coli on hands of waste handlers after work (8.60 ± 4.20 CFU/hand, mean ± standard deviation) was significantly higher compared with the E. coli log concentration before work (2.95 ± 1.89 CFU/hand, mean ± standard deviation) (p<0.001). The odds of faecal pollution was significantly higher (aOR 4.2; 95% CI: 1.9-9.1) for workers aged 35 years and above compared with those less than 35 years; and for workers at public toilet facilities (aOR 3.0; 95% CI: 1.0-8.4) compared with those who worked for private waste handling companies. Female workers were, however, 60% less likely (aOR 0.4; 95% CI: 0.2-0.8) to experience faecal pollution of their hands compared with males. The workers had limited access to water and sanitation and hygiene facilities, and about one-fifth (n = 59; 21.1%) did not use personal protective equipment during work. Waste handlers should be provided and instructed in proper use of personal protective equipment, have access to sanitation facilities and adopt improved hygiene behaviour to avoid the risk of faecal pollution and associated disease risk.

Effective school-based preventive interventions for alcohol use in Africa: a systematic review.

BACKGROUND: Despite recognition of the risks of alcohol use and importance of prevention from an early age, the effectiveness of school-based interventions in Africa has not been clarified. OBJECTIVE: We aimed to identify effective school-based alcohol use prevention interventions in Africa. METHODS: We searched eight databases for peer-reviewed articles published until February 3, 2019 that reported on randomized controlled trials, cluster randomized controlled trials, controlled clinical trials, pre-post quasi-experimental studies, cohort studies, and case-control studies. The full-texts of relevant studies were searched. RESULTS: Four of 2797 papers met our eligibility criteria. All reported interventions targeted secondary school students in South Africa and were incorporated in the school curriculum. The interventions comprised multi-component activities with participatory and peer educational methods, and applied modified programs originally developed in the US. However, intervention effects were inconsistent among studies, although the interventions tended to have a positive effect on non-drinkers at baseline, with stronger effects in girls. CONCLUSION: interventions had positive effects on students that were non-drinkers at baseline, especially girls. Although we could not find robust evidence that school-based interventions changed attitudes, frequency/quantity of drinking, and intentions to use alcohol, one intervention showed an increase in students' alcohol refusal self-efficacy.

Oral activity of the antimalarial endoperoxide 6-(1,2,6,7-tetraoxaspiro[7.11]nonadec-4-yl)hexan-1-ol (N-251) against Leishmania donovani complex.

Visceral leishmaniasis (VL) is a major problem worldwide and causes significant morbidity and mortality. Existing drugs against VL have limitations, including their invasive means of administration long duration of treatment regimens. There are also concerns regarding increasing treatment relapses as well as the identification of resistant clinical strains with the use of miltefosine, the sole oral drug for VL. There is, therefore, an urgent need for new alternative oral drugs for VL. In the present study, we show the leishmanicidal effect of a novel, oral antimalarial endoperoxide N-251. In our In vitro studies, N-251 selectively and specifically killed Leishmania donovani D10 amastigotes with no accompanying toxicity toward the host cells. In addition, N-251 exhibited comparable activities against promastigotes of L. donovani D10, as well as other L. donovani complex parasites, suggesting a wide spectrum of activity. Furthermore, even after a progressive infection was established in mice, N-251 significantly eliminated amastigotes when administered orally. Finally, N-251 suppressed granuloma formation in mice liver through parasite death. These findings indicate the therapeutic effect of N-251 as an oral drug, hence suggest N-251 to be a promising lead compound for the development of a new oral chemotherapy against VL.

Agro-ecological zone and farm diversity are factors associated with haemoglobin and anaemia among rural school-aged children and adolescents in Ghana.

Understanding contextual risk factors for haemoglobin (Hb) status and anaemia of rural school-aged children (SAC) and adolescents is critical in developing appropriate interventions to prevent anaemia. We analysed secondary data from the baseline of an impact evaluation of the Ghana School Feeding Programme to determine the severity of anaemia and contextual factors associated with anaemia and Hb status among rural SAC (6-9 years; n = 323) and adolescents (10-17 years; n = 319) in Ghana. We used regression models with variable selection based on backward elimination in our analyses. The mean Hb was 113.8 ± 13.1 g/L, and the overall prevalence of anaemia was 52.3%, being 55.1% and 49.5% among SAC and adolescents, respectively. We identified child's age (ß = 2.21, P < 0.001); farm diversity score (ß = 0.59, P = 0.036); and agro-ecological zone (P trend <0.001) as the main predictors of Hb of SAC. Household asset index (P trend = 0.042) and agro-ecological zone (P trend <0.001) were predictors of Hb in adolescents. Agro-ecological zone and age were predictors of anaemia, but the effect of age was only significant for girls and not boys (prevalence odds ratio [POR] = 1.35, 95% CI [1.04, 1.76] vs. POR = 1.14, 95% CI [0.88, 1.46]). SAC in households with maize stock were less likely to be anaemic (POR = 0.55, 95% CI [0.32, 0.97]). Household dietary diversity score (ß = 0.59, P = 0.033) was associated with Hb status for the full sample only. Anaemia is a severe public health problem among SAC and adolescents in rural Ghana irrespective of sex. Farm diversity score, availability of maize stock in the household, household asset index, and agro-ecological zone were the main predictors of Hb and anaemia among the rural SAC and adolescents.

Low Microfilaremia Levels in Three Districts in Coastal Ghana with at Least 16 Years of Mass Drug Administration and Persistent Transmission of Lymphatic Filariasis.

Ghana has been implementing mass drug administration (MDA) of ivermectin and albendazole for the elimination of lymphatic filariasis (LF) since the year 2000, as part of the Global Programme to Eliminate Lymphatic Filariasis (GPELF). It was estimated that 5⁻6 years of treatment would be sufficient to eliminate the disease. Tremendous progress has been made over the years, and treatment has stopped in many disease endemic districts. However, despite the successful implementation of MDA, there are districts with persistent transmission. In this study we assessed the epidemiology of LF in three adjoining districts that have received at least 16 years of MDA. The assessments were undertaken one year after the last MDA. 1234 adults and 182 children below the age of 10 years were assessed. The overall prevalence of circulating filarial antigen in the study participants was 8.3% (95% CI: 6.9⁻9.9), with an estimated microfilaria prevalence of 1.2%. The microfilarial intensity in positive individuals ranged from 1 to 57 microfilariae/mL of blood. Higher antigen prevalence was detected in males (13.0%; 95% CI: 10.3⁻16.2) compared to females (5.5%; 95% CI: 4.1⁻7.2). The presence of infection was also highest in individuals involved in outdoor commercial activities, with the risks of infection being four- to five-fold higher among farmers, fishermen, drivers and artisans, compared to all other occupations. Using bednets or participating in MDA did not significantly influence the risk of infection. No children below the age of 10 years were found with infection. Detection of Wb123 antibodies for current infections indicated a prevalence of 14.4% (95% CI: 8.1⁻23.0) in antigen-positive individuals above 10 years of age. No antibodies were detected in children 10 years or below. Assessment of infection within the An. gambiae vectors of LF indicated an infection rate of 0.9% (95% CI: 0.3⁻2.1) and infectivity rate of 0.5% (95% CI: 0.1⁻1.6). These results indicate low-level transmission within the districts, and suggest that it will require targeted interventions in order to eliminate the infection.

Socio-Behavioral Risk Factors Associated with Cryptosporidiosis in HIV/AIDS Patients Visiting the HIV Referral Clinic at Cape Coast Teaching Hospital, Ghana.

OBJECTIVE: To identify the socio-behavioral risk factors associated with cryptosporidiosis among HIV/AIDS patients with chronic diarrhea symptoms visiting the HIV referral clinic at Cape Coast Teaching Hospital, Ghana. METHODS: A cross-sectional study was conducted among 50 HIV/AIDS patients with recurrent diarrhea. Questionnaires were administered to collect social and behavioral risk factors associated with Cryptosporidium and other opportunistic protozoan parasitic infections in HIV patients. Stool samples were collected for the diagnosis of enteric protozoan pathogens using modified Ziehl-Neelsen and acid-fast staining methods. CD4+ cells counts of study subjects were obtained from patients clinical records. The data obtained were analyzed using Pearson chi-square and multivariate-adjusted statistics tool on SPSS 16 for Windows. RESULTS: Twenty-seven (54%) of the subjects were infected with enteric protozoan pathogens. The prevalences of Cryptosporidium, Cyclospora and Microsporidium infections were 46%, 32% and 16%, respectively. Cryptosporidium infection was significantly associated with drinking water (×2=13.528, p<0.001), Cyclospora was associated with the type of drinking water (×2=14.931, p<0.001) and toilet facilities used by the study subjects (×2=12.463, p<0.01), whiles Microsporidium infection was associated with hand washing behavior (×2=12.463, p<0.01). Enteric protozoans were frequently encountered among subjects with CD4+ T-cell count <200 cells/mm3. However, coinfection of Cyclospora spp & Cryptosporidium spp was not observed in CD4+ cell count <200 and >500 cells/mm3. Multivariate analysis showed that the risk factor for Cryptosporidium infection among HIV/AIDS patients was the source of drinking water (pipe borne water 76.2% prevalence: sachet water 25%; OR=0.10, 95%CI: 0.03-0.39, p<0.001). CONCLUSION: We report the risk factor for exposure of Cryptosporidium infection among HIV/AIDS patients for the first time in Ghana. The contamination of drinking water by protozoan parasites should be a public health concern. These results provide the stepping block to understand the transmission dynamics of Cryptosporidium and other opportunistic pathogens in HIV/AIDS infected patients in Ghana.

Gastrointestinal helminths in farmers and their ruminant livestock from the Coastal Savannah zone of Ghana.

To identify the gastrointestinal helminths of veterinary, zoonotic and public health importance in farmers and their ruminant livestock in Ghana, faecal samples were collected from 95 farmers and their livestock (cattle = 328, sheep = 285 and goats = 217) and examined by microscopy and/or molecular techniques. Overall, 21 farmers tested positive for at least one gastrointestinal helminth, 80.9% of which were single infections and 19.0% co-infections. The parasites identified in the farmers consisted of hookworms (n = 13) (9 were Necator americanus and the other 4 could not be amplified by PCR), Trichostrongylus spp. (n = 9), Schistosoma mansoni (n = 1), Schistosoma haematobium (n = 1) and Diphyllobothrium latum (n = 1). In livestock, strongylid nematodes were dominant (56.6%), followed by Paramphistomum spp. (16.9%), Dicrocoelium spp. (7.1%), Thysaniezia spp. (5.8%), Trichuris spp. (3.3%), Moniezia spp. (3.1%), Fasciola spp. (2.8%), Toxocara spp. (1.1%) and Schistosoma spp. (0.2%). Genotyping of Trichostrongylus spp. in the farmer's stools identified six T. colubriformis similar to T. colubriformis detected in cattle, sheep and goats in the study, two Trichostrongylus spp. with 98.3% and 99.2% genetic similarity to T. probolurus respectively and one Trichostrongylus spp. which showed 96.6% similarity to both T. probolurus and T. rugatus. Trichostrongylus axei was also identified in cattle, sheep and goats. This is the first molecular characterisation of Trichostrongylus spp. in Ghana and the species identified in the present study suggests zoonotic transmission from cattle, sheep and goats. Further studies involving larger numbers of farmers and their household members are essential to understand the transmission dynamics and impact of these parasites on farming communities in Ghana.

In vitro antiprotozoan activity and mechanisms of action of selected Ghanaian medicinal plants against Trypanosoma, Leishmania, and Plasmodium parasites.

Trypanosomiasis, leishmaniasis, and malaria are protozoan infections of public health importance with thousands of new cases recorded annually. Control of these infection(s) with existing chemotherapy is limited by drug toxicity, lengthy parenteral treatment, affordability, and/or the emergence of resistant strains. Medicinal plants on the other hand are used in the treatment of various infectious diseases although their chemical properties are not fully evaluated. In this study, we screened 112 crude extracts from 72 selected Ghanaian medicinal plants for anti-Trypanosoma, anti-Leishmania, and anti-Plasmodium activities in vitro and investigated their mechanisms of action. Twenty-three extracts from 20 plants showed significant antiprotozoan activity against at least 1 of 3 protozoan parasites screened with IC50 values less than 20 µg/ml. Eleven extracts showed high anti-Trypanosoma activity with Bidens pilosa whole plant and Morinda lucida leaf extracts recording the highest activities. Their IC50 (selectivity index [SI]) values were 5.51 µg/ml (35.00) and 5.96 µg/ml (13.09), respectively. Nine extracts had high anti-Leishmania activity with Annona senegalensis and Cassia alata leaf extracts as the most active. Their IC50 (SI) values were 10.8 µg/ml (1.50) and 10.1 µg/ml (0.37), respectively. Six extracts had high anti-Plasmodium activity with the leaf and stem-bark extracts of Terminalia ivorensis recording the highest activity. Their IC50 (SI) values were 7.26 µg/ml (129.36) and 17.45 µg/ml (17.17), respectively. Only M. lucida at 25 µg/ml induced significant apoptosis-like cell death in Trypanosoma parasites. Anti-Leishmania active extracts induced varying morphological changes in Leishmania parasites such as multiple nuclei and/or kinetoplast, incomplete flagella division, or nuclear fragmentation. Active extracts may be potential sources for developing new chemotherapy against these infections.

An Evaluation of Treatment Outcomes in a Cohort of Clients on the DOTS Strategy, 2012-2016.

We present, for the first time, an evaluation of treatment outcomes in a cohort at a TB referral centre in the Central Region of Ghana. Of the 213 clients placed on DOTS, 59.2% (126/213) were sputum smear-positive. An overall cure rate of 90.2% (51.6% cured + 37.6% completed) and a death rate of 8.5% (18/213) were estimated. Of the number of clients who died, 5.7% (12/213) were males (χ2 = 2.891, p = 0.699; LR = 3.004, p = 0.699). Deaths were only recorded among clients who were > 19 years old (χ2 = 40.319, p = 0.099; LR = 41.244, p = 0.083). Also, 0.9% (2/213) was lost to follow-up, while 1.4% (3/213) had treatment failure. In total, 13.6% (7.0%, 15/213 males, and 6.6%, 14/213 females) of clients who were placed on DOTS were HIV seropositive. Ages of 40-49 years had the highest number, 13/213 (6.1%), infected with HIV, though the difference among the remaining age groups was not statistically significant (χ2 = 9.621, p = 0.142). Furthermore, 7.0% (15/213) had TB/HIV coinfection. Out of them, 9 were cured and 5 died at home, while 1 had treatment failure. Tuberculosis/HIV infection prevention advocacy and interventions that address sociodemographic determinants of unfavourable treatment outcomes are urgently required to augment national efforts towards control.

Insights into the mode of action of 1,2,6,7-tetraoxaspiro [7.11] nonadecane (N-89) against adult Schistosoma mansoni worms.

Control of morbidity associated with schistosomiasis via chemotherapy largely relies on the drug praziquantel. Repeated therapy with praziquantel has created concerns about the possible selection of resistant worms and necessitated the search for novel drugs to treat schistosomiasis. Here, a murine model was infected with Schistosoma mansoni and treated with oral 1,2,6,7-tetraoxaspiro [7.11] nonadecane (N-89), which caused a significant reduction in fecundity and egg burden and reduced morbidity when administered at 5-weeks post-infection. The analysis showed that the mode of action occurred through the ingestion of activated N-89 by the worms, and that there was no direct external effect on the S. mansoni worms. Ultrastructural analysis of the treated worms showed disruptions in the gut lumen and the presence of large volumes of material, suggestive of undigested blood meals or red blood cells. In addition, there were reduced vitelline cells in female worms and damage to sub-tegmental musculature in male worms. Eggs recovered from the treated mice showed both damage to the eggs and the production of immature eggs. Expression of mRNA responsible for gut and digestive function and egg production was also significantly affected by N-89 treatment, whereas control genes for musculature showed no significant changes. Thus, N-89 drastically affected the total digestive function and egg production of S. mansoni worms. Physiological processes requiring heme uptake such as egg production and eggshell formation were subsequently affected, suggesting that the compound could be a possible therapeutic drug candidate for schistosomiasis control.

Schistosoma Egg Antigen Induces Oncogenic Alterations in Human Prostate Cells.

Schistosomiasis is a neglected tropical disease that affects 200 million people and accounts for 100,000 deaths annually. In endemic geographical areas, schistosomiasis has been implicated as an etiological agent in the pathogenesis of bladder, colorectal, and renal carcinoma largely due to Schistosoma eggs in tissues that comes with chronic infection. Several studies have also reported cases of association between Schistosoma infection and prostate cancer. The possible causal association is however poorly understood. We hypothesized in this study that infection of the prostate cells with Schistosoma spp promotes cancer. Urine samples from individuals living in Galilea, a schistosomiasis endemic community in the Ga South District of Ghana, were collected and screened for Schistosoma infection via microscopy and multiplex PCR. Soluble egg antigens (SEA) were prepared from Schistosoma egg-positive urine samples and assessed for the ability to induce cancer-like phenotypes including excessive proliferation, oxidative stress (reduced glutathione (GSH) depletion), and diminished apoptosis in cultured human prostate (PNT2) cells. Molecular analysis revealed infecting schistosome species to be S. haematobium and S. mansoni. Prostate cell proliferation was significantly induced by 12.5 µg/ml SEA (p = 0.029). Also, SEA dose-dependently depleted cellular GSH. Flow cytometric analysis and fluorescence staining revealed that SEA dose-dependently diminished apoptosis, significantly, in prostate cells. Findings of this study suggest that schistosome infection may play a role in the pathogenesis of prostate cancer. In vivo studies are however needed to confirm this association.

Molecular characterization of Cryptosporidium and Giardia in farmers and their ruminant livestock from the Coastal Savannah zone of Ghana.

Cryptosporidium and Giardia are major causes of diarrhoea in developing countries including Ghana, however, nothing is known about the species and subtypes of Cryptosporidium and Giardia in farmers and their ruminant livestock in this country. A total of 925 faecal samples from humans (n=95), cattle (n=328), sheep (n=217) and goats (n=285), were screened for Cryptosporidium and Giardia by quantitative PCR (qPCR) at the 18S rRNA and glutamate dehydrogenase (gdh) loci respectively. Cryptosporidium positives were typed by sequence analysis of 18S and 60kDa glycoprotein (gp60) loci amplicons. Giardia positives were typed at the triose phosphate isomerase (tpi), beta-giardin (bg) and gdh loci. The prevalence of Cryptosporidium and Giardia by qPCR was 8.4% and 10.5% in humans, 26.5% and 8.5% in cattle, 34.1% and 12.9% in sheep, and 33.3% and 12.3% in goat faecal samples, respectively. G. duodenalis assemblages A and B were detected in humans and assemblage E was detected in livestock. Cryptosporidium parvum was the only species identified in humans; C. andersoni, C. bovis, C. ryanae and C. ubiquitum were identified in cattle; C. xiaoi, C. ubiquitum and C. bovis in sheep; and C. xiaoi, C. baileyi and C. parvum in goats. This is the first molecular study of Cryptosporidium and Giardia in livestock in Ghana. The identification of zoonotic species and the identification of C. parvum subtype IIcA5G3q in livestock, which has previously been identified in children in Ghana, suggests potential zoonotic transmission. Further studies on larger numbers of human and animal samples, and on younger livestock are required to better understand the epidemiology and transmission of Cryptosporidium and Giardia in Ghana.

Associations between Polymorphisms within Interferon Gamma and Tumor Necrosis Factor Genes and Toxoplasma Retinochoroiditis in Ghanaian Patients.

PURPOSE: To evaluate associations between IFN-γ +874T/A and TNF-α-308G/A polymorphism with the development of Toxoplasma retinochoroiditis. METHODS: By ARMS-PCR, a cross-sectional genetic study involving 30 patients with Toxoplasma retinochoroiditis and 87 controls was carried out. RESULTS: IFN-γ +874: by comparing with the AA genotype, individuals with the TT genotype had a 3.4 odds ratio (OR); AT had a 1.6 OR; and the T allele had a higher OR (1.6), indicating a likely susceptibility of IFN-γ +874T to the infection, though the overall distribution was not significant (p = 0.259). For TNF-α-308G/A, individuals with both GA and AA genotypes had lower ORs when compared with the GG genotype, implying the A allele could confer protection against Toxoplasma ocular lesions. CONCLUSIONS: IFN-γ +874T allele may increase the risk of ocular lesions in Toxoplasma infection. The principle of natural selection seems to also play a role. The less common TNF-308A allelic form could be protective against the development of Toxoplasma ocular infection.

In vitro anti-Leishmania activity of tetracyclic iridoids from Morinda lucida, benth.

Leishmaniasis is an infectious disease transmitted by the sand fly. It is caused by over 20 different species of Leishmania and has affected over 14 million people worldwide. One of the main forms of control of leishmaniasis is chemotherapy, but this is limited by the high cost and/or toxicity of available drugs. We previously found three novel compounds with an iridoid tetracyclic skeleton to have activity against trypanosome parasites. In this study, we determined the activity of the three anti-trypanosome compounds against Leishmania using field strain, 010, and the lab strain Leishmania hertigi. The minimum inhibitory concentration (MIC) of the compounds against 010 was determined by microscopy while the IC50 of compounds against L. hertigi was determined by fluorescence-activated cell sorting with Guava viacount analysis. We found two of the three compounds, molucidin and ML-F52, to have anti-Leishmania activity against both strains. The fluor-microscope observation with DAPI stain revealed that both Molucidin and ML-F52 induced abnormal parasites with two sets of nucleus and kinetoplast in a cell, suggesting that compounds might inhibit cytokinesis in Leishmania parasites. Molucidin and ML-F52 might be good lead compounds for the development of new anti-Leishmania chemotherapy.

Indication of Risk of Mother-to-Child Toxoplasma gondii Transmission in the Greater Accra Region of Ghana.

Objectives Congenital infection with Toxoplasma gondii is known to result in neurological and brain disorders including ophthalmic disorders later in life. Research in Ghana revealed high sero-prevalence among pregnant women and eye patients. This study determines the risk of congenital transmission of T. gondii infection in Accra, Ghana. Methods One hundred consented pregnant women aged 18-45 years (mean 29.85 ± 5.76) participated. Venous blood and tissue samples were taken from the maternal side of each placenta after delivery. Cord blood samples were also taken after they were separated from the infants. Finger-prick blood was taken from infants of participating women at 2 or 6 weeks post-natal. ELISA was used to detect T. gondii antibodies in all blood samples while Nested-PCR was used to detect T. gondii DNA from placental tissues. Data was analysed using SPSS v. 16. Results Overall, 37.6 % of maternal blood, 39.5 % of umbilical cord blood, and 57.5 % of post-natal infant blood were positive for anti-T. gondii IgG. No anti-T. gondii IgM was detected in any of those samples. Toxoplasma gondii DNA was detected in 39.8 % of placental tissue samples. Strong association was observed in the occurrence of placental T. gondii DNA and anti-T. gondii IgG positive women (ø = 0.810, p < 0.00001) as well as high Relative risk shown in the likelihood of foetal exposure to infection in latently-infected women (RR 10.39; CI 4.47-24.17; p < 0.00001). Conclusions for Practice The presence of anti-T. gondii IgG antibodies only, and T. gondii DNA in placental tissues indicate the women might have been infected early during the pregnancy, placing about 39.8 % of the babies at risk. These results can strongly influence policy to screen and treat pregnant women for T. gondii infection.