Protein export into malaria parasite-infected erythrocytes: mechanisms and functional consequences.

Phylum Apicomplexa comprises a large group of obligate intracellular parasites of high medical and veterinary importance. These organisms succeed intracellularly by effecting remarkable changes in a broad range of diverse host cells. The transformation of the host erythrocyte is particularly striking in the case of the malaria parasite Plasmodium falciparum. P. falciparum exports hundreds of proteins that mediate a complex cellular renovation marked by changes in the permeability, rigidity, and cytoadherence properties of the host erythrocyte. The past decade has seen enormous progress in understanding the identity and function of these exported effectors, as well as the mechanisms by which they are trafficked into the host cell. Here we review these advances, place them in the context of host manipulation by related apicomplexans, and propose key directions for future research.

Enterocytozoon bieneusi and Encephalitozoon intestinalis (microsporidia) in HIV-positive patients in central Spain.

Microsporidia are fungi-related eukaryotic intracellular parasites that opportunistically infect immunocompromised individuals such as those infected by the human immunodeficiency virus (HIV). Among them, Enterocytozoon bieneusi and Encephalitozoon spp. are the most clinically relevant species. We investigated the occurrence and genetic diversity of microsporidial and protist infections in mostly immunocompetent HIV-positive patients in Madrid, Spain. A structured questionnaire was used to retrieve data on factors potentially associated with an increased risk of infection, including sexual attitudes and sex-risk behaviour. Faecal samples (n = 96) from 81 HIV-positive patients were collected and analysed by molecular (PCR and Sanger sequencing) methods. Two microsporidial pathogens were detected: Ent. bieneusi (2.5%, 95% CI: 0.3-8.6) and Enc.intestinalis (4.9%, 95% CI: 1.4-12.2). The two Ent. bieneusi isolates were identified as zoonotic genotype A. Among protists, Entamoeba dispar was the species most prevalently found (33.3%, 95% CI: 23.2-44.7), followed by Blastocystis spp. (19.8%, 95% CI: 11.7-30.1), Giardia duodenalis (13.6%, 95% CI: 7.0-23.0), and Cryptosporidium spp. and Entamoeba histolytica (2.5%, 95% CI: 0.3-8.6 each). Cyclospora cayetanensis and Cystoisospora belli were not detected. Subtypes ST1 (70.6%, 12/17) and ST3 (29.4%, 5/17) were identified within Blastocystis sp., sub-assemblages AII and BIII (50%, 1/2 each) within G. duodenalis, and Cry. parvum and canine-adapted Cry. canis (50%, 1/2 each) within Cryptosporidium spp. Microsporidial and protist parasites were frequent in well-controlled, mostly immunocompetent HIV-positive patients and should be included in diagnostic algorithms when diarrhoea is present.

Opportunistic microsporidial and protist intestinal infections were relatively common in well-controlled HIV-positive patients in Madrid, Spain. These agents should be suspected and appropriately diagnosed in HIV-positive patients presenting with diarrhoea regardless of their immunological status.

In vitro culture of Tritrichomonas foetus in bovine cervico-vaginal fluid.

Tritrichomonas foetus is the causative agent of bovine trichomonosis, a venereal disease that can lead to fetal loss. T. foetus proliferates in the vagina and cervix and invades the uterus and fetal cavities. It is not clear how T. foetus grows in the host or how its infection rarely demonstrable after 4 months, is controlled. Cervical vaginal mucus (CVM) is a protective barrier against potentially harmful microorganisms. Here, we demonstrate that bovine CVM is a medium in which this protozoan parasite can grow in vitro. T. foetus multiplied at different rates depending on the time of the estrous period from which the CVM was obtained. Growth rates were higher in CVM obtained 7-10 days after estrus.

Intestinal parasite infections associated with sociodemographic and seasonal factors in the Western Amazon.

Intestinal parasites are a constant public health problem in the Amazon region, with a high prevalence of cases related to poor sanitary conditions. We investigated the sociodemographic and seasonal factors associated with human intestinal parasite infections in an area of the Western Amazon, Brazil, from September 2017 to August 2019. Data were collected using a database available at the Diagnostic Support Centre (Centro de Apoio ao Diagnóstico, CAD) of the Municipality of Rio Branco, on positive diagnoses for intestinal parasites. Among the 53,200 samples analysed, 18.3% (n = 9712) were positive. Of these, 96.4% (n = 9363) and 3.6% (n = 349) were protozoan and helminthic infections, respectively. Males showed higher odds ratio (OR) for Enterobius vermicularis infection (OR: 2.3) and giardiasis (OR: 1.9) and lower OR for Endolimax nana (OR: 0.9) and Entamoeba coli (OR: 0.9) infections. Individuals aged ≥ 15 presented higher OR for Strongyloides stercoralis (OR: 3.4), hookworms (OR: 2.3), and almost all protozoan infections than younger individuals. In the dry season, the OR for hookworms (OR: 1.5), Iodamoeba butschlii (OR: 1.4), and Endolimax nana (OR: 1.3) infections was higher than that in the rainy season, including a high chance of polyparasitism (OR: 1.6). We concluded that there was a significant difference between the different types of intestinal parasites, particularly protozoa, with high OR in the dry season and for certain groups.

Parasitism as a lifestyle: Ultimate intimacy between Apicomplexan protozoans and metazoan hosts.

Editorial: The Apicomplexa parasite Toxoplasma gondii glides on substrate with a helical path and releases material that forms a trail behind. The helical microtubules (green) periodically compress and relax, acting as spring force by coupling with the myosin motor (red).

Prevalence and Risk Factors for Intestinal Protozoan Infections in Children with Cancer in Taif, Western Saudi Arabia.

BACKGROUND: Cancer is a category of diseases that cause an individual's immune system to become suppressed. In a case-control study, the current study aims to detect the frequency of intestinal parasites and related risk factors in children with cancer. METHODS: Stool samples were collected from 178 children with cancers (cases) and 150 cancer-free children (controls) who sought treatment for diarrheal episodes at nearby hospitals. Samples were processed by direct smear examination, concentration technique, permanent staining by Lugol's iodine, modified Ziehl-Neelsen, modified trichrome, and chromotrope 2R stains. RESULTS: The overall prevalence of intestinal parasites was 7.3% (24/328), with non-statistically significant differences between cases (7.8%; 14/178) and controls (6.6%; 10/150). Children with leukemia had a higher infection rate (9%; 9/100) than children with lymphoma (6.9%; 3/43) or solid tumors (5.7%; 2/35). Blastocystis sp. (3.3%) was the most common intestinal parasite found in cases, followed by Cryptosporidium sp. (2.2%), Giardia lamblia (1.6%), and Microsporidia sp. (0.5%). For all parasites, no statistical difference was found between the two groups. (p > 0.05). Male gender, young age, non-bottled water use, travel to parasite-endemic areas, living in an urban area, and infrequent hand washing were all associated with intestinal parasitosis, with non-statistical significance observed between the two groups. In children with cancer, intestinal parasites were found to be significantly associated with chronic (p = 0.04) and severe (p = 0.03) diarrhea. CONCLUSIONS: Children with cancer, particularly those with hematological cancers, should be screened for intestinal parasites on a regular basis and treated for their overall health.

Antiprotozoal agents: How have they changed over a decade?

Neglected tropical diseases are a diverse group of communicable diseases that are endemic in low- or low-to-middle-income countries located in tropical and subtropical zones. The number and availability of drugs for treating these diseases are low, the administration route is inconvenient in some cases, and most of them have safety, efficacy, or adverse/toxic reaction issues. The need for developing new drugs to deal with these issues is clear, but one of the most drastic consequences of this negligence is the lack of interest in the research and development of new therapeutic options among major pharmaceutical companies. Positive changes have been achieved over the last few years, although the overall situation remains alarming. After more than one decade since the original work reviewing antiprotozoal agents came to light, now it is time to question ourselves: How has the scenario for the treatment of protozoal diseases such as malaria, leishmaniasis, human African trypanosomiasis, and American trypanosomiasis changed? This review covers the last decade in terms of the drugs currently available for the treatment of these diseases as well as the clinical candidates being currently investigated.

Mass mortality in endangered fan mussels Pinna nobilis (Linnaeus 1758) caused by co-infection of Haplosporidium pinnae and multiple Vibrio infection in Çanakkale Strait, Turkey.

PURPOSE: Pinna nobilis (fan mussel) is one of the most important endemic bivalve molluscs in the Mediterranean and mass mortality events were observed in these mussels in recent years. In this study, we report mass mortalities caused by Haplosporidium pinnae, which has been spreading in the Mediterranean for 3 years, and reached the Çanakkale Strait, which is the entrance of the Marmara and the Black Sea. MATERIAL AND METHODS: Field observations during sampling and subsequent histopathological, biochemical, genetic, and microbiological analyses were carried out. RESULTS: These analyses showed that H. pinnae infection spread among the natural beds of P. nobilis, causing severe tissue damage and oxidative stress. Our phylogenetic analyses suggested that the parasite spread through the Mediterranean much faster than thought. The results showed that vibriosis originating from Vibrio coralliilyticus, Vibrio tubiashii, Vibrio mediterranei, and Vibrio hispanicus, acted together with H. pinnae in infected individuals and caused death. CONCLUSION: It is highly probable that the spread of H. pinnae to the Sea of Marmara and the Black Sea may occur earlier than expected, and it was concluded that mass deaths were caused by co-infection with H. pinnae and a geographically specific marine pathogen that can infect P. nobilis populations.

Avian haemosporidia in native and invasive sparrows at an Afrotropical region.

Bio-invasions are a major threat to biodiversity and ecosystems globally and may contribute to the proliferation of emerging infectious diseases. We examined the prevalence and phylogenetic diversity of avian haemosporidian parasites infecting the non-native house sparrows (Passer domesticus) and the native southern grey-headed sparrows (Passer diffusus). Blood samples from 104 sparrows (74 house sparrows and 30 southern grey-headed sparrows) mist-netted inside and around the Kruger National Park were used. Genomic DNA was extracted from each blood sample and subjected to nested PCR analyses, Sanger sequencing and phylogenetic analyses. Overall, 35.57% (37/104) of the birds sampled were infected with at least one haemosporidian parasites. Southern grey-headed sparrows had a higher parasite prevalence (60%) than house sparrows (24.3%). A total of 16 parasite lineages were identified, of which eight were novel lineages. Whereas Haemoproteus spp. showed the highest lineage diversity, Leucocytozoon spp. were the most prevalent parasites, albeit with significant differences between sparrow species. A single Plasmodium sp. infection was recorded in a southern grey-headed sparrow. In support of the enemy release hypothesis, we found that prevalence on non-native house sparrows was lower than prevalence recorded in their region of origin and also that they were infected only by indigenous parasites lineages.

Evolution of vector transmitted parasites by host switching revealed through sequencing of Haemoproteus parasite mitochondrial genomes.

Parasite species evolve by switching to new hosts, cospeciating with their current hosts, or speciating on their current hosts. Vector transmitted parasites are expected to speciate by host switching, but confirming this hypothesis has proved challenging. Parasite DNA can be difficult to sequence, thus well resolved parasite phylogenies that are needed to distinguish modes of parasite speciation are often lacking. Here, we studied speciation in vector transmitted avian haemosporidian parasites in the genus Haemoproteus and their warbler hosts (family Acrocephalidae). We overcome the difficulty of generating parasite genetic data by combining nested long-range PCR with next generation sequencing to sequence whole mitochondrial genomes from 19 parasite haplotypes confined to Acrocephalidae warblers, resulting in a well-supported parasite phylogeny. We also generated a well-supported host phylogeny using five genes from published sources. Our phylogenetic analyses confirm that these parasites have speciated by host switching. We also found that closely related host species shared parasites which themselves were not closely related. Sharing of parasites by closely related host species is not due to host geographic range overlap, but may be the result of phylogenetically conserved host immune systems.

Activity-Based Protein Profiling for the Study of Parasite Biology.

Parasites exist within most ecological niches, often transitioning through biologically and chemically complex host environments over the course of their parasitic life cycles. While the development of technologies for genetic engineering has revolutionised the field of functional genomics, parasites have historically been less amenable to such modification. In light of this, parasitologists have often been at the forefront of adopting new small-molecule technologies, repurposing drugs into biological tools and probes. Over the last decade, activity-based protein profiling (ABPP) has evolved into a powerful and versatile chemical proteomic platform for characterising the function of enzymes. Central to ABPP is the use of activity-based probes (ABPs), which covalently modify the active sites of enzyme classes ranging from serine hydrolases to glycosidases. The application of ABPP to cellular systems has contributed vastly to our knowledge on the fundamental biology of a diverse range of organisms and has facilitated the identification of potential drug targets in many pathogens. In this chapter, we provide a comprehensive review on the different forms of ABPP that have been successfully applied to parasite systems, and highlight key biological insights that have been enabled through their application.

Intravital microscopy: Imaging host-parasite interactions in the brain.

Intravital fluorescence microscopy (IVM) is a powerful technique for imaging multiple organs, including the brain of living mice and rats. It enables the direct visualisation of cells in situ providing a real-life view of biological processes that in vitro systems cannot. In addition, to the technological advances in microscopy over the last decade, there have been supporting innovations in data storage and analytical packages that enable the visualisation and analysis of large data sets. Here, we review the advantages and limitations of techniques predominantly used for brain IVM, including thinned skull windows, open skull cortical windows, and a miniaturised optical system based on microendoscopic probes that can be inserted into deep tissues. Further, we explore the relevance of these techniques for the field of parasitology. Several protozoan infections are associated with neurological symptoms including Plasmodium spp., Toxoplasma spp., and Trypanosoma spp. IVM has led to crucial findings on these parasite species, which are discussed in detail in this review.

Co-opting oxylipin signals in microbial disease.

Oxylipins, or oxygenated lipids, are universal signalling molecules across all kingdoms of life. These molecules, either produced by microbial pathogens or their mammalian host, regulate inflammation during microbial infection. In this review, we summarise current literature on the biosynthesis pathways of microbial oxylipins and their biological activity towards mammalian cells. Collectively, these studies have illustrated how microbial pathogens can modulate immune rsponse and disease outcome via oxylipin-mediated mechanisms.

Molecular prevalence of intestinal parasites infections in children with diarrhea in Franceville, Southeast of Gabon.

BACKGROUND: Pediatric diarrhea caused by a range of pathogens, including intestinal parasites, is one of main causes of death among children under 5 years of age. The distribution of these parasitic infections overlaps in many environmental, socioeconomic and epidemiological settings. Their distribution and prevalence varies from region to region. In the current study, we assess the prevalence of intestinal parasites among pediatric patients with syndromic diarrheal disease living in Franceville, Gabon. METHODS: A cross-sectional study conducted in the Amissa Bongo Regional Hospital and Chinese-Gabonese Friendship Hospital in Franceville, between November 2016 and August 2017, enrolled a total of 100 diarrheic children between 0 and 180 months of age. Parasite detection in stool samples was performed using molecular diagnostic by PCR. Difference in means were tested by Student's t test and ANOVA while principal component analysis was used to determine the correlation between parasite distributions and age groups. RESULTS: The overall prevalence of intestinal parasite infection was 61% (61/100). Hymenolepis sp and Cryptosporidium hominis/parvum were the most common parasites (31 and 19%, respectively), followed by Encephalitozoon intestinalis (15%), Trichuris trichiura (4%), Dientamoeba fragilis (4%), and Enterocytozoon bieneusi (2%). The polyparasitism rate was 19.7%, with 83.3% double and 16.7% triple infections. Protozoan infections (66.7%) were more prevalent than helminths infections (33.3%). Seasonal association of the circulation of intestinal parasite was statistically significant (p = 0.03). Correlations between different parasites was also observed. CONCLUSION: The prevalence of intestinal parasitic infections is highest in diarrheic pediatric children. The prevalence of parasitic infections indicates that protozoa and helminths are the most common parasites in the Franceville environment. This study reinforces the importance of routine examination of diarrheic stool samples for the diagnostic of intestinal parasites. Further analyses are required to better understand the local epidemiology and risk factors associated with the transmission of intestinal parasites in Franceville, Gabon. KEYSWORDS: diarrhea, children, intestinal parasitic infections, molecular diagnostic, Franceville, Gabon.

Prevalence and associated risk factors of human intestinal parasitic infections: a population-based study in the southeast of Kerman province, southeastern Iran.

BACKGROUND: Determination of the prevalence and distribution pattern of intestinal parasites is a fundamental step to set up an effective control program to improve the health status. This study aimed to determine the prevalence of intestinal parasitic infections and associated risk factors among inhabitants of Rudbar-e Jonub county, southeast of Kerman province, southeastern Iran. METHODS: In this cross-sectional study, 861 stool specimens were collected from inhabitants of Rudbar-e Jonub county through a multistage cluster sampling method in 2018. The collected specimens were examined by parasitological methods including, direct wet-mounting (for the fresh specimens with a watery consistency), formalin-ethyl acetate sedimentation and agar plate culture. RESULTS: The prevalence of intestinal parasites was 34.2% (95% CI 30.1 to 38.2). The prevalence of protozoan parasites 32.3% (95% CI 28.4 to 36.5) was significantly higher than helminthic parasites 3.2% (95% CI 2.1 to 4.7). Blastocystis sp. (13.3%), Entamoeba coli (11.4%) and Giardia lamblia (10.6%) as protozoan parasite and Hymenolepis nana (2.4%) as helminthic parasite were the most common detected intestinal parasites in the study. Entamoeba histolytica/dispar (1.5%), Iodamoeba bütschlii (1.0%), Chilomastix mesnili (0.5%), Entamoeba hartmanni (0.4%), Enterobius vermicularis (0.3%) and Ascaris lambercoides (0.3%) were other detected parasites. Multiple logistic regression revealed a significant association of intestinal parasitic infections with source of drinking water and residency status (rural/urban). Multiple infections with 2 or 3 parasitic agents constituted 22.7% of 295 infected cases. CONCLUSIONS: This study revealed a high prevalence of intestinal protozoan infections among inhabitants of Rudbar-e Jonub county. Intestinal parasites especially protozoans remain a challenging public health problem wherever sanitation and health measures are limited in Iran.

A comparative study of parasites in three latrines from Medieval and Renaissance Brussels, Belgium (14th-17th centuries).

The aim of this study is to determine the species of parasite that infected the population of Brussels during the Medieval and Renaissance periods, and determine if there was notable variation between different households within the city. We compared multiple sediment layers from cesspits beneath three different latrines dating from the 14th-17th centuries. Helminths and protozoa were detected using microscopy and enzyme-linked immunosorbent assay (ELISA). We identified Ascaris sp., Capillaria sp., Dicrocoelium dendriticum, Entamoeba histolytica, Fasciola hepatica, Giardia duodenalis, Taenia sp. and Trichuris sp. in Medieval samples, and continuing presence of Ascaris sp., D. dendriticum, F. hepatica, G. duodenalis and Trichuris sp. into the Renaissance. While some variation existed between households, there was a broadly consistent pattern with the domination of species spread by fecal contamination of food and drink (whipworm, roundworm and protozoa that cause dysentery). These data allow us to explore diet and hygiene, together with routes for the spread of fecal-oral parasites. Key factors explaining our findings are manuring practices with human excrement in market gardens, and flooding of the polluted River Senne during the 14th-17th centuries.

Sympatric western lowland gorillas, central chimpanzees and humans are infected with different trichomonads.

We investigated intestinal trichomonads in western lowland gorillas, central chimpanzees and humans cohabiting the forest ecosystem of Dzanga-Sangha Protected Area in Central African Republic, using the internal transcribed spacer (ITS) region and SSU rRNA gene sequences. Trichomonads belonging to the genus Tetratrichomonas were detected in 23% of the faecal samples and in all host species. Different hosts were infected with different genotypes of Tetratrichomonas. In chimpanzees, we detected tetratrichomonads from 'novel lineage 2', which was previously reported mostly in captive and wild chimpanzees. In gorillas, we found two different genotypes of Tetratrichomonas. The ITS region sequences of the more frequent genotype were identical to the sequence found in a faecal sample of a wild western lowland gorilla from Cameroon. Sequences of the second genotype from gorillas were almost identical to sequences previously obtained from an anorexic French woman. We provide the first report of the presence of intestinal tetratrichomonads in asymptomatic, apparently healthy humans. Human tetratrichomonads belonged to the lineage 7, which was previously reported in domestic and wild pigs and a domestic horse. Our findings suggest that the ecology and spatial overlap among hominids in the tropical forest ecosystem has not resulted in exchange of intestinal trichomonads among these hosts.

Modelling evolution of virulence in populations with a distributed parasite load.

Modelling evolution of virulence in host-parasite systems is an actively developing area of research with ever-growing literature. However, most of the existing studies overlook the fact that individuals within an infected population may have a variable infection load, i.e. infected populations are naturally structured with respect to the parasite burden. Empirical data suggests that the mortality and infectiousness of individuals can strongly depend on their infection load; moreover, the shape of distribution of infection load may vary on ecological and evolutionary time scales. Here we show that distributed infection load may have important consequences for the eventual evolution of virulence as compared to a similar model without structuring. Mathematically, we consider an SI model, where the dynamics of the infected subpopulation is described by a von Förster-type equation, in which the infection load plays the role of age. We implement the adaptive dynamics framework to predict evolutionary outcomes in this model. We demonstrate that for simple trade-off functions between virulence, disease transmission and parasite growth rates, multiple evolutionary attractors are possible. Interestingly, unlike in the case of unstructured models, achieving an evolutionary stable strategy becomes possible even for a variation of a single ecological parameter (the parasite growth rate) and keeping the other parameters constant. We conclude that evolution in disease-structured populations is strongly mediated by alterations in the overall shape of the parasite load distribution.

Dynamics of avian haemosporidian assemblages through millennial time scales inferred from insular biotas of the West Indies.

Although introduced hemosporidian (malaria) parasites (Apicomplexa: Haemosporida) have hastened the extinction of endemic bird species in the Hawaiian Islands and perhaps elsewhere, little is known about the temporal dynamics of endemic malaria parasite populations. Haemosporidian parasites do not leave informative fossils, and records of population change are lacking beyond a few decades. Here, we take advantage of the isolation of West Indian land-bridge islands by rising postglacial sea levels to estimate rates of change in hemosporidian parasite assemblages over a millennial time frame. Several pairs of West Indian islands have been connected and separated by falling and rising sea levels associated with the advance and retreat of Pleistocene continental glaciers. We use island isolation following postglacial sea-level rise, ca. 2.5 ka, to characterize long-term change in insular assemblages of hemosporidian parasites. We find that assemblages on formerly connected islands are as differentiated as assemblages on islands that have never been connected, and both are more differentiated than local assemblages sampled up to two decades apart. Differentiation of parasite assemblages between formerly connected islands reflects variation in the prevalence of shared hemosporidian lineages, whereas differentiation between islands isolated by millions of years reflects replacement of hemosporidian lineages infecting similar assemblages of avian host species.

Alsinol, an arylamino alcohol derivative active against Plasmodium, Babesia, Trypanosoma, and Leishmania: past and new outcomes.

Malaria, babesiosis, trypanosomosis, and leishmaniasis are some of the most life-threatening parasites, but the range of drugs to treat them is limited. An effective, safe, and low-cost drug with a large activity spectrum is urgently needed. For this purpose, an aryl amino alcohol derivative called Alsinol was resynthesized, screened in silico, and tested against Plasmodium, Babesia, Trypanosoma, and Leishmania. In silico Alsinol follows the Lipinski and Ghose rules. In vitro it had schizontocidal activity against Plasmodium falciparum and was able to inhibit gametocytogenesis; it was particularly active against late gametocytes. In malaria-infected mice, it showed a dose-dependent activity similar to chloroquine. It demonstrated a similar level of activity to reference compounds against Babesia divergens, and against promastigotes, and amastigotes stages of Leishmania in vitro. It inhibited the in vitro growth of two African animal strains of Trypanosoma but was ineffective in vivo in our experimental conditions. It showed moderate toxicity in J774A1 and Vero cell models. The study demonstrated that Alsinol has a large spectrum of activity and is potentially affordable to produce. Nevertheless, challenges remain in the process of scaling up synthesis, creating a suitable clinical formulation, and determining the safety margin in preclinical models.