Plasmodium species: master renovators of their host cells

Plasmodium parasites, the causative agents of malaria, have developed elaborate strategies that they use to survive and thrive within different intracellular environments. During the blood stage of infection, the parasite is a master renovator of its erythrocyte host cell, and the changes in cell morphology and function that are induced by the parasite promote survival and contribute to the pathogenesis of severe malaria. In this Review, we discuss how Plasmodium parasites use the protein trafficking motif Plasmodium export element (PEXEL), protease-mediated polypeptide processing, a novel translocon termed the Plasmodium translocon of exported proteins (PTEX) and exomembranous structures to export hundreds of proteins to discrete subcellular locations in the host erythrocytes, which enables the parasite to gain access to vital nutrients and to evade the immune defence mechanisms of the host.

Ecology of urban malaria vectors in Niamey, Republic of Niger

BACKGROUND: Urbanization in African cities has major impact on malaria risk. Niamey, the capital of the Republic of Niger, is situated in the West African Sahel zone. The short rainy season and human activities linked with the Niger River influence mosquito abundance. This study aimed at deciphering the factors of distribution of urban malaria vectors in Niamey.

METHODS: The distribution of mosquito aquatic stages was investigated monthly from December 2002 to November 2003, at up to 84 breeding sites, throughout Niamey. An exploratory analysis of association between mosquito abundance and environmental factors was performed by a Principal Component Analysis and confirmed by Kruskall-Wallis non-parametric test. To assess the relative importance of significant factors, models were built for Anopheles and Culicinae. In a second capture session, adult mosquitoes were collected weekly with pyrethrum sprays and CDC light-traps from June 2008 to June 2009 in two differentiated urban areas chosen after the study's first step. Members of the Anopheles gambiae complex were genotyped and Anopheles females were tested for the presence of Plasmodium falciparum circumsporozoite antigens using ELISA.

RESULTS: In 2003, 29 % of 8420 mosquitoes collected as aquatic stages were Anopheles. They were significantly more likely to be found upstream, relatively close to the river and highly productive in ponds. These factors remained significant in regression and generalized linear models. The Culicinae were found significantly more likely close to the river, and in the main temporary affluent stream. In 2009, Anopheles specimens, including Anopheles gambiae s.l. (95 %), but also Anopheles funestus (0.6 %) accounted for 18 % of the adult mosquito fauna, with a large difference between the two sampled zones. Three members of the An. gambiae complex were found: Anopheles arabiensis, Anopheles coluzzii, and An. gambiae. Nineteen (1.3 %) out of 1467 females tested for P. falciparum antigen were found positive.

CONCLUSION: The study provides valuable update knowledge on malaria vector ecology and distribution in Niamey. The identification of spatial and environmental risk factors could pave the way to larval source management strategy and allow malaria vector control to focus on key zones for the benefit of the community.

Diseño, optimización y validación de sistemas de amplificación genómica para el diagnóstico de malaria y filariosis humanas

Las enfermedades parasitarias o parasitosis son un conjunto de enfermedades infecciosas producidas por protozoos, helmintos, e incluso artrópodos. La enfermedad parasitaria más importante es la malaria que está incluida en la lista de enfermedades de la pobreza. Otras enfermedades parasitarias han sido incluidas en las denominadas enfermedades olvidadas o desatendidas (NTD: Neglected Tropical Diseases) entre las que se encuentran las filariosis linfática y onchocercosis. La malaria está causada por el género Plasmodium (protozoos apicomplexo) Las especies que pueden causar la infección en humanos son: P. falciparum. P. vivax, P. malariae, P. ovale y P. knowlesi. Las filarias son nematodos finos y largos, parásitos de la sangre, la linfa y los tejidos subcutáneos y conectivos que producen en el humano la filariosis. Su transmisión se produce por insectos hematófagos (mosquitos y moscas) que actúan como vectores. Las especies de filarias de interés clínico para los humanos son Wuchereria. bancrofti, Brugia. malayi y B. timori (filariosis linfática), Onchocerca volvulus, Loa. loa y Mansonella streptocerca (filarias dérmicas), y Mansonella perstans y M. ozzardi (mansonelosis). Todas ellas presentan estadios larvales, conocidos como microfilarias (L1), que circulan en sangre o en tejido subcutáneo que son las formas infectivas para los vectores. En el Laboratorio de Malaria & otras Parasitosis Emergentes se ha desarrollado una PCR en tiempo real para malaria (Malaria RT-PCR), una Nested-PCR para filarias (Nested-Filaria PCR) y una PCR en tiempo real para filarias (RT-Filaria-PCR) como Sistemas de Análisis Múltiple para la detección de varias especies de plasmodios y varias especies de filarias en muestras de cualquier índole como indicador que los Sistemas de Análisis Múltiple son comparativamente superiores a los métodos de detección individual y a la microscopía sin perder sensibilidad y especificidad. Todos los métodos desarrollados han dado muy buenos resultados en cuanto a sensibilidad y especificidad frente a los métodos tradicionales, de tal manera que hoy en día se usan en el Laboratorio de Malaria & otras Parasitosis Emergentes como métodos de referencia, planteando la posibilidad de usar el método de las filarias para un estudio actualizado de la distribución y prevalencia de las filarias en las zonas endémicas.

Evaluation of five commercial methods for the extraction and purification of DNA from human faecal samples for downstream molecular detection of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp

High quality, pure DNA is required for ensuring reliable and reproducible results in molecular diagnosis applications. A number of in-house and commercial methods are available for the extraction and purification of genomic DNA from faecal material, each one offering a specific combination of performance, cost-effectiveness, and easiness of use that should be conveniently evaluated in function of the pathogen of interest. In this comparative study the marketed kits QIAamp DNA stool mini (Qiagen), SpeedTools DNA extraction (Biotools), DNAExtract-VK (Vacunek), PowerFecal DNA isolation (MoBio), and Wizard magnetic DNA purification system (Promega Corporation) were assessed for their efficacy in obtaining DNA of the most relevant enteric protozoan parasites associated to gastrointestinal disease globally. A panel of 113 stool specimens of clinically confirmed patients with cryptosporidiosis (n = 29), giardiasis (n = 47) and amoebiasis by Entamoeba histolytica (n = 3) or E. dispar (n = 10) and apparently healthy subjects (n = 24) were used for this purpose. Stool samples were aliquoted in five sub-samples and individually processed by each extraction method evaluated. Purified DNA samples were subsequently tested in PCR-based assays routinely used in our laboratory. The five compared methods yielded amplifiable amounts of DNA of the pathogens tested, although performance differences were observed among them depending on the parasite and the infection burden. Methods combining chemical, enzymatic and/or mechanical lysis procedures at temperatures of at least 56 °C were proven more efficient for the release of DNA from Cryptosporidium oocysts.

Characterisation of PTEX88, a component of the Plasmodium translocon

This thesis explored the role of PTEX88, a putative component of the malaria translocon involved in trafficking parasite proteins into host red blood cells. Reverse genetic approaches were employed to demonstrate PTEX88 plays an auxiliary role in translocon function but has an impact on parasite virulence and immune evasion.

Estudio de las modificaciones conformacionales y su influencia en la actividad inmunológica de péptidos derivados de las familias de las proteínas spect-1 y -2, siap-1 y -2 de plasmodium falciparum

SPECT-1 y -2 y SIAP-1 y -2 son proteínas pertenecientes al esporozoíto de Plasmodium falciparum causante de la malaria más agresiva en los humanos. Estas proteínas están involucradas en el paso del parásito a través de las células del hospedero humano y en la invasión del hepatocito, haciéndolas blancos atractivos para ser estudiadas. Péptidos conservados de alta capacidad de unión (cHABPs) a células HeLa y HepG2 derivados de estas moléculas son no inmunogénicos porque son incapaces de generar una respuesta inmunitaria, pero son claves para el parásito porque cumplen una función importante durante la infección del hospedero humano. En este trabajo se encontró que algunos cHABPs pertenecientes a las proteínas SPECT-1 y -2, están posiblemente involucrados con la unión y formación de poros sobre la membrana de las células hospederas, ayudando al esporozoíto a abrirse paso través de las células del hospedero. Por otro lado, con el fin de cambiar el comportamiento inmunológico de cHABPs derivados de SPECT-1 y -2 y SIAP-1 y -2, se obtuvieron nuevos péptidos mediante el reemplazo de aminoácidos críticos por otros residuos cuya masa molecular sea similar, pero diferente en su polaridad. En este trabajo se reporta que dichas modificaciones promovieron cambios en la estructura secundaria (determinada por DC o 1H-RMN) de los péptidos modificados (mHABPs) cuando se comparó con la estructura de los cHABPs nativos; adicionalmente, estos mHABPs invirtieron su comportamiento inmunológico convirtiéndose en péptidos inmunogénicos inductores de anticuerpos. Lo que permite establecer la existencia de una relación entre la estructura que adoptan estos mHABPs con su actividad inmunológica. Además, algunos de los mHABPs estudiados aquí, pueden ser candidatos a ser incluidos en la vacuna contra la malaria químicamente sintetizada multi-epitope y multi-estadio que se está desarrollando en la Fundación Instituto de Inmunología de Colombia (FIDIC).

Geographical patterns of the species richness of helminth parasites of moles (Talpa spp.) in Spain: Separating the effects of sampling effort from those of other conditioning factors

We analysed the viscera of 534 moles (Ta l p a spp.) from 30 of the 47 provinces of peninsular Spain, including 255 individuals of T. europaea from eight provinces, 154 individuals of T. occidentalis from 20 provinces, and 125 unidentified Ta l p a individuals from two provinces. We identified their helminth parasites and determined parasite species richness. We related parasite species richness with sampling effort using both a linear and a logarithmic function. We then performed stepwise linear regressions to predict mole parasite species richness from a small set of selected predictor variables that included sampling effort. We applied the resulting models to forecast T. euro p a e a, T. occidentalis, and Ta l p a spp. parasite species richness in all provinces with recorded host presence, assuming different levels of sampling eff o r t . F i n a l l y, we used partial regression analysis to partition the variation explained by each of the selected variables in the models. We found that mole parasite species richness is strongly conditioned by sampling effort, but that other factors such as cropland area and environmental disturbance have significant independent effects.

A study on monogenean parasites from the gills of some cichlids(Pisces: Cichlidae) from the Brazilian Amazon.

The aim of this study was to investigate monogenean fauna in gills of Geophagus camopiensis, Pterophyllum scalare, Satanoperca jurupari,and Satanoperca acuticeps in a tributary from the Amazon River system in Brazil. A total of 2,148 monogenean specimens were collected from140 fish examined from March 2012 to March 2013, and 84.3% of these fish were parasitized by 1 or more species. Such monogeneans were:Sciadicleithrum geophagi, Sciadicleithrum juruparii, Gussevia spiralocirra and Gyrodactylus sp. However, only G. camopiensis was parasitizedby more than 1 species of monogenean, while S. jurupari and S. acuticeps were parasitized by the same species. Prevalence, mean intensity andmean abundance varied among host species and the highest levels of infection were by G. spiralocirra followed by S. geophagi, both parasiteswith aggregated dispersion. Abundance of monogeneans was not influenced by the size of the host. In G. camopiensis, the infection levels byS. geophagi did not vary during the rainy or drainage seasons. This is the first study on monogenean infections for G. camopiensis and S. acuticeps.

Anti-erythrocyte antibodies may contribute to anaemia in Plasmodium vivax malaria by decreasing red blood cell deformability and increasing erythrophagocytosis.

2016

Clypeotheca, a new skeletal structure in scleractinian corals : a potential stress indicator

Physiological responses to environmental stress are increasingly well studied in scleractinian corals. This work reports a new stress-related skeletal structure we term clypeotheca. Clypeotheca was observed in several livecollected common reef-building coral genera and a two to three kya subfossil specimen from Heron Reef, Great Barrier Reef and consists of an epitheca-like skeletal wall that seals over the surface of parts of the corallum in areas of stress or damage. It appears to form from a coordinated process wherein neighboring polyps and adjoining coenosarc seal themselves off from the surrounding environment as they contract and die. Clypeotheca forms from inward skeletal centripetal growth at the edges of corallites and by the merging of flange-like outgrowths that surround individual spines over the surface of the coenosteum. Microstructurally, the merged flanges are similar to upsidedown dissepiments and true epitheca. Clypeotheca is interpreted primarily as a response to stress that may help protect the colony from invasion of unhealthy tissues by parasites or disease by retracting tissues in areas that have become unhealthy for the polyps. Identification of skeletal responses of corals to environmental stress may enable the frequency of certain types of environmental stress to be documented in past environments. Such data may be important for understanding the nature of reef dynamics through intervals of climate change and for monitoring the effects of possible anthropogenic stress in modern coral reef habitats.

Sewage disposal and wildlife health in Antarctica

Sewage and its microbiology, treatment and disposal are important to the topic of Antarctic wildlife health because disposal of untreated sewage effluent into the Antarctic marine environment is both allowed and commonplace. Human sewage contains enteric bacteria as normal flora, and has the potential to contain parasites, bacteria and viruses which may prove pathogenic to Antarctic wildlife. Treatment can reduce levels of micro-organisms in sewage effluent, but is not a requirement of the Environmental Protocol to the Antarctic Treaty (the Madrid Protocol). In contrast, the deliberate release of non-native organisms for any other reason is prohibited. Hence, disposal of sewage effluent to the marine environment is the only activity routinely undertaken in Antarctica knowing that it will likely result in the release of large numbers of potentially non-native species. When the Madrid Protocol was negotiated, the decision to allow release of untreated sewage effluent was considered the only pragmatic option, as a prohibition would have been costly, and may not have been achievable by many Antarctic operators. In addition, at that time the potential for transmission of pathogens to wildlife from sewage was not emphasised as a significant potential risk. Since then, the transmission of disease-causing agents between species is more widely recognised and it is now timely to consider the risks of continued discharge of sewage effluent in Antarctica and whether there are practical alternatives.

The future for public health

What is the future for public health in the twenty first century? Can we glean an idea about the future of public health from its past? As Winston Churchill once said ‘the further backward you look, the further forward you can see’. What then can we see in the history of public health that gives us an idea of where public health might be headed in the future? In the twentieth century there was substantial progress in public health in Australia. These improvements were brought about through a number of factors. In part, improvements were due to improved knowledge about the natural history of disease and its treatment. Added to this knowledge was a shifting focus from legislative measures to protect health, to the emergence of improved promotion and prevention strategies and a general improvement in social and economic conditions for people living in countries like Australia. The same could not, however, be said for poorer countries, many of whom have the most fundamental of sanitary and health protection issues still to deal with. For example, in sub-Saharan Africa and Russia, the decline in life expectancy may be an aberration or it may be related to a range of interconnected factors. In Russia, factors such as alcoholism, violence, suicide, accidents and cardiovascular disease could be contributing to the falling life expectancy (McMichael & Butler 2007). In sub-Saharan Africa, a range of issues such as HIV/AIDS, poverty, malaria, tuberculosis, undernutrition, totally inadequate infrastructure, gender inequality, conflict and violence, political taboos and a complete lack of political will, have all contributed to a dramatic drop in life expectancy (McMichael & Butler 2007).

Sri Lanka in global medical research: a scientific analysis of the Sri Lankan research output during 2000-2009

Background Scientific research is an essential component in guiding improvements in health systems. There are no studies examining the Sri Lankan medical research output at international level. The present study evaluated the Sri Lankan research performance in medicine as reflected by the research publications output between years 2000-2009. Methods This study was based on Sri Lankan medical research publication data, retrieved from the SciVerse Scopus® from January 2000 to December 2009. The process of article selection was as follows: Affiliation - 'Sri Lanka' or 'Ceylon', Publication year - 'January 2000 to December 2009' and Subject area - 'Life and Health Sciences'. The articles identified were classified according to disease, medical speciality, institutions, major international collaborators, authors and journals. Results Sri Lanka's cumulative medical publications output between years 2000-2009 was 1,740 articles published in 160 different journals. The average annual publication growth rate was 9.1%. Majority of the articles were published in 'International' (n = 950, 54.6%) journals. Most articles were descriptive studies (n = 611, 35.1%), letters (n-345, 19.8%) and case reports (n = 311, 17.9%). The articles were authored by 148 different Sri Lankan authors from 146 different institutions. The three most prolific local institutions were Universities of; Colombo (n = 547), Kelaniya (n = 246) and Peradeniya (n = 222). Eighty four countries were found to have published collaborative papers with Sri Lankan authors during the last decade. UK was the largest collaborating partner (n = 263, 15.1%). Malaria (n = 75), Diabetes Mellitus (n = 55), Dengue (n = 53), Accidental injuries (n = 42) and Lymphatic filariasis (n = 40) were the major diseases studied. The 1,740 publications were cited 9,708 times, with an average citation of 5.6 per paper. The most cited paper had 203 citations, while there were 597 publications with no citations. The Sri Lankan authors' contribution to the global medical research output during the last decade was only 0.086%. Conclusion The Sri Lankan medical research output during the last decade is only a small fraction of the global research output. There it is a necessity to setup an enabling environment for research, with a proper vision, support, funds and training. In addition, collaborations across the region need to be strengthened to face common regional health challenges. Keywords: Sri Lanka, Medical research, Publication, Analysis

Incidence and geographic distribution of endemic Burkitt lymphoma in northern Uganda revisited

Endemic Burkitt lymphoma (BL) is etiologically associated with Epstein-Barr virus (EBV) and ecologically linked to Plasmodium falciparum malaria. However, these infections imperfectly correlate with BL epidemiology. To obtain recent epidemiological data, we studied district- and county-specific BL incidence and standardized incidence ratios using data collected from 1997 through 2006 at Lacor Hospital in northern Uganda, where studies were last done more than 30 years ago. Among 500 patients, median age was 6 years (inter-quartile range 5-8) and male-to-female ratio was 1.8:1. Among those known, most presented with abdominal (56%, M: F 1.4:1) vs. only facial tumors (35%, M: F 3.0:1). Abdominal tumors occurred in older (mean age: 7.0 vs. 6.0 years; p<0.001) and more frequently in female children (68% vs. 50%; OR 2.2, 95% CI 1.5-3.5). The age-standardized incidence was 2.4 per 100,000, being 0.6 in 1-4 year olds, 4.1 in 5-9 year olds and 2.8 in 10-14 year olds and varied 3-4-fold across districts. The incidence was lower in districts that were far from Lacor and higher in districts that were close to Lacor. While districts close to Lacor were also more urbanized, the incidence was higher in the nearby perirural areas. We highlight high BL incidence and geographic variation in neighboring districts in northern Uganda. While distance from Lacor clearly influenced the patterns, the incidence was lower in municipal than in surrounding rural areas. Jaw tumors were characterized by young age and male gender, but presentation has shifted away from facial to mostly abdominal. Keywords: Africa, cancer, malaria, Epstein-Barr virus, clustering, epidemiology