A Cross Sectional Study on the Bidirectional Interactions Between Leptospirosis and HIV Infection Among Patients from Maputo Central Hospital, Mozambique.

Introduction: This study aims to determine the baseline seroprevalence of leptospirosis, a zoonotic and neglected disease, in people living with HIV (PWH) in Maputo, Mozambique, and to evaluate the relationship between selected HIV-related factors that might influence risk of coinfection with leptospirosis, such as degree of immunosuppression, as assessed by CD4 cell count, World Health Organization (WHO) HIV/AIDS clinical stage and antiretroviral therapy (ART) intake. Methods: This was a descriptive cross-sectional analysis of 157 PWH, aged over 18 years old, admitted to the Maputo Central Hospital, in Maputo, Mozambique, between March 2020 and October 2021. The study participants were recruited as a convenience sample regardless of the reasons for their admission. We collected sociodemographic and clinical data, including ART and WHO HIV/AIDS clinical stage, and blood for CD4 cell count and detection of Leptospira IgG antibodies using a commercial Kit ab247199 Leptospira IgG ELISA (www.abcam.com/ab247199) with sensitivity and specificity of 100% and 97.3%, respectively. Laboratory testing was performed at the Faculty of Medicine, Eduardo Mondlane University and Laboratory of Clinical Analysis, in Maputo. Results: Participants were aged 18 to 72 years (median age 39 years; SD ± 10.5), the majority were female 100 (63.7%), from urban areas 138 (87.9%), with secondary-level education 80 (51%). The overall seroprevalence of Leptospira IgG antibodies was 40.1%. The median CD4 cell count was 385 cells/µl (02 to 2297; SD ± 378.47). Higher seroprevalence of Leptospira antibodies was found among participants with CD4 cell counts <250 cells/µl (54.8%), WHO HIV/AIDS stage IV (70.2%) and those on ART (92%), though there were no statistically significant differences between groups with and without Leptospira antibodies. Conclusion: Our study confirmed that Leptospira antibodies are highly prevalent in PWH in Maputo; however, Leptospira infection was not associated with the degree of immunosuppression, WHO HIV/AIDS clinical stage, or the use of ART. Our data support the need for routine screening for leptospirosis in PWH in Mozambique. Future studies are warranted to characterize the incidence and outcomes of symptomatic leptospirosis in this patient population and to identify circulating serovars and species in the country and region, as well as the implicated reservoirs.

The burden of T. solium cysticercosis and selected neuropsychiatric disorders in Mocuba district, Zambézia province, Mozambique.

BACKGROUND: Taenia solium (neuro-)cysticercosis, a neglected tropical disease, can be associated with epileptic seizures and other neuropsychiatric (= neurological and psychiatric) disorders. This study aimed to evaluate the association of T. solium cysticercosis with selected neuropsychiatric disorders and/or symptoms (chronic headache, epileptic seizures/epilepsy and psychosis) in Mocuba district, Mozambique. METHODOLOGY: Between March and May 2018, a cross-sectional study was conducted among 1,086 participants aged 2 years or above in Mocuba district, Zambézia province, central Mozambique, to assess the seroprevalence of human cysticercosis and risk factors for infection, as well as to explore its relation to selected neuropsychiatric disorders. Socio-demographic and clinical data were collected from each participant using a modified questionnaire designed by the Cysticercosis Working Group for Eastern and Southern Africa. Additionally, neuropsychiatric disorders, such as chronic headache, epileptic seizures/epilepsy and psychosis were assessed using four vignettes. T. solium antigen and cysticercosis IgG in serum were detected using both T. solium antigen B158/B60 enzyme linked immunosorbent assay (ELISA) and LDBIO Cysticercosis Western Blot, respectively. PRINCIPAL FINDINGS: Overall, 112/1,086 participants (10.3%) were sero-positive for T. solium antigen or antibodies. Prevalence of antibodies (6.6%; n = 72) was higher than of antigens (4.9%; n = 54). In the questionnaires, 530 (49.5%) of participants reported chronic headache, 293 (27%) had generalized epileptic seizures, 188 (18%) focal seizures and 183 (18.3%) psychosis. We found a statistically significant association between seropositivity for T. solium and chronic headache (p = 0.013). Additionally, increasing age (p = 0.03) was associated with Ag-ELISA seropositivity. CONCLUSIONS: Our study revealed that in Mocuba, T. solium cysticercosis is prevalent and associated with self-reported chronic headache. Additionally, in the study setting, the seroprevalence of cysticercosis increased with age. However, it is not associated with other neuropsychiatric disorders such epileptic seizures/epilepsy and psychosis. Future studies are needed to confirm the high burden of neuropsychiatric disorders and their possible etiology, including neurocysticercosis, using additional serological, molecular biological and radiological diagnostic tools, as well as in-depth clinical examinations.

Assessment of Parasitic Contamination of Lettuce and Cabbages Sold in Selected Markets in Maputo City, Mozambique.

Background: In Mozambique, vegetables are the main source of nutrients and they are consumed raw or cooked. Their consumption, especially in their raw form, might be a source of contamination, which may take place during production and/or transportation as well as in the markets. In this study we aimed to assess the profile and frequency of parasites in lettuces and cabbages sold in some selected markets in Maputo city, the capital of Mozambique. Methods: A cross sectional study was conducted between February and March 2018, in 10 selected markets from Maputo City. A total of 100 combined samples of lettuces and cabbages were analyzed using a spontaneous sedimentation method. Data such as gender and age of the vendors and about handling of vegetables at the point of sale were also collected. Results: Out of the samples, 84 (84%) were contaminated with parasites (86% of lettuces and 82% of cabbages). The analyses performed revealed the presence of E. coli (66%), S. stercoralis (40%), E. histolytica (20%), A. duodenalis (6%), S. haematobium (4%), A. lumbricoides (2%), S. mansoni (2%) and T. trichiura (1%). Two markets, Janete and Fajardo showed contamination in all samples. A Chi square analysis revealed a significant association between the contaminated cabbages and the market where they were obtained. The majority of vendors (57%) do not wash their hands before handling vegetables. Most of the vendors had tables to place vegetables (72%) and those tables are cleaned before display (86%). None of the differences observed in each risk factor studied were statistically significant. Conclusion: The present study highlights that contamination of raw vegetables with pathogenic parasites in markets might represent a vector for transmission of intestinal and water- borne parasites to consumers. Future studies should investigate the extent of vegetable contamination at each stage of the supply chain from the farm to consumers, as well as its relationship to human parasitic infection and its consequences. Good hygienic practices of farmers, vendors and consumers should be enforced to break the contamination chain. Whenever possible, hydroponic cultivation should be encouraged to avoid the influence of highly contaminated soils.

Neurocysticercosis in Epileptic Children: An Overlooked Condition in Mozambique, Challenges in Diagnosis, Management and Research Priorities.

Previous studies suggest that neurocysticercosis (NCC), the most common cause of acute symptomatic seizures (ASS), epilepsy and other neuropsychiatric disorders, typically presents with a solitary lesion and focal seizures in children from places where cysticercosis is endemic. We report a series of 3 patients, aged 7 to 11 years, with a history of epilepsy and or recurrent headache referred from Mocuba to the Quaternary Central Hospital in Quelimane, Zambeze Province, Mozambique, an area endemic for cysticercosis. Clinical history and examination, blood chemistry and hemogram screening, serological testing for Cysticercus antigens and antibodies detection, and a computerized tomography (CT) scan, were performed. NCC was confirmed in all 3 patients, based on criteria defined by Del Bruto. Two confirmed cases tested positive for antigen (Ag) by enzyme-linked immunosorbent assay (ELISA) with CT lesions in different stages of parasite evolution. Headache/encephalopathy was present in all patients. This case series of children with epilepsy confirms for the first time the presence of NCC in children from Zambezia province, an east-central region of Mozambique. Further, NCC should be included in the differential diagnosis of children with ASS, epilepsy and other neuropsychiatric disorders. Future studies should be targeted to the identification of biomarkers to support the diagnosis of NCC, given the limited availability of imaging tools and limited value of serological assays for the diagnosis and management of NCC.

Further evaluation of recombinant Tsol-p27 by enzyme-linked immunoelectrotransfer blot for the serodiagnosis of cysticercosis in pigs from Mozambique.

BACKGROUND: Porcine cysticercosis has a negative impact on human health and the meat industry, as it makes infected meat unaproprieted for consuption and it is the main etiology of epileptic seizures in developing countries. There are multiple serological assays that use crude antigens with high sensitivity and specificity for the diagnosis of both porcine and human cysticercosis. Nonetheless, antigen preparation is time-consuming, needs a well-equipped laboratory and trained personnel and places those manipulating the meat at great risk for infection. New serodiagnostic approaches to the diagnosis of porcine and human cysticercosis have been directed towards the development of recombinant deoxyribonucleic acid technology for the generation of synthetic proteins that can serve as simplified, low-cost and harmless substitutes for native antigens. The aim of the present study was to further evaluate the recombinant Tsol-p27 protein as a target molecule in immunoassays for the serodiagnosis of porcine cysticercosis. From these data, we hoped to develop recommendations regarding its use in the serodiagnosis of porcine cysticercosis. RESULTS: We studied a panel of 83 naturally infected pig sera from Angónia District, Mozambique, an endemic area for porcine and human cysticercosis. These sera were previously tested by antigen enzyme-linked immunosorbent assay (Ag-ELISA) to detect antigens of T. solium. The serum panel was processed by enzyme-linked immunoelectrotransfer blot (EITB) assay against the recombinant Tsol-p27 protein and the Ag-ELISA assay results were used to compare and evaluate the performance of Tsol-p27 for the diagnosis of cysticercosis. Out of 83 sera, 24 (29.0%) were positive for Tsol-p27 and 59 (71%) were negative in the same assay. From the 37 sera that tested positive to Ag-ELISA, 11 (13.3%) were positive to Tsol-p27, while from 46 sera that tested negative to Ag-ELISA, 33 (39.7%) also tested negative to Tsol-p27. The sensitivity and specificity of Tsol-p27 was 29.7% and 71.7%, respectively, while the positive predictive value and negative predictive value were 45.8% and 55.9%, respectively, as calculated using Medcalc® version 15.0 software (MedCalc Software, Ostend, Belgium). CONCLUSION: While Tsol-p27 recombinant protein might be suitable for testing human sera, its performance in pigs is not acceptable, so other recombinant proteins should be evaluated alone or multiplexed.

Further characterization of Tsol-p27 as a diagnostic antigen in sub-Saharan Africa.

Commercial antigens used to diagnose human neurocysticercosis are obtained from either a soluble parasite extract or a parasite-derived glycoprotein fraction. The aim of the present study was to identify antigenic proteins as potential diagnostic candidates in Mozambique. Soluble proteins from Taenia solium cysticerci were separated by two-dimensional electrophoresis and blotted onto nitrocellulose membranes. Subtracted hybridization was performed with serum samples obtained from patients with neurocysticercosis (NCC) and from a NCC-negative control group. Six antigenic proteins were identified and sequenced by liquid chromatography-mass spectrometry. Among these we found Tsol-p27, which was previously identified as a diagnostic candidate in a study conducted in Nicaragua, Central America. Here, we evaluated Tsol-p27 and the antigen cC1 as potential recombinant diagnostic reagents, and also investigated the localization and partial function of Tsol-p27. Immunoblotting demonstrated that Tsol-p27 was recognized by all 10 serum samples from NCC-positive individuals, whereas cC1 was identified by only five of the 10 positive sera. None of the antigens were recognized by negative control sera. Despite the limited number of serum samples evaluated in this study, the results suggest that Tsol-p27 can be a suitable candidate for diagnosis of human NCC, not only in Central America but also in sub-Saharan Africa.