Developmental Stages Of Parasite Research Articles

Arginase activity is associated with fibrosis in experimental infection with Taenia crassiceps, but does not play a major role in resistance to infection

Murine infection with Taenia crassiceps cysticerci is used as an experimental model for human and animal cysticercosis. In this infection parasites can be found associated with an inflammatory infiltrate enriched with macrophages. Experimental evidence exists supporting a role for either NO-producing classically activated (CAMΦ) or arginase- and CD301-expressing alternatively activated macrophages (AAMΦ) in T. crassiceps resistance. In both cell types, arginine is utilized as an important mediator in macrophage effector functions. To investigate whether there is an association between arginine availability, susceptibility to T. crassiceps and other parameters such as fibrosis, BALB/c mice were infected intraperitoneally with cysticerci and treated daily with the arginase inhibitor nor-NOHA or supplemented with l-arginine and followed for eight weeks. The numbers and developmental stages of parasites were evaluated as well as the presence of CD301+ AAMΦ, arginase activity and collagen deposition in the peritoneal membrane. Treatment with the arginase inhibitor or supplementation with l-arginine did not change the parasitic load or profile of the infection. However, the arginase inhibitor significantly decreased the deposition of collagen. These results suggest that arginase activity does not interfere with parasite control during experimental infection with T. crassiceps, but it is important for fibrosis in cysticercosis.

Dynamics of the major histocompatibility complex class I processing and presentation pathway in the course of malaria parasite development in human hepatocytes: implications for vaccine development.

Control of parasite replication exerted by MHC class I restricted CD8+ T-cells in the liver is critical for vaccination-induced protection against malaria. While many intracellular pathogens subvert the MHC class I presentation machinery, its functionality in the course of malaria replication in hepatocytes has not been characterized. Using experimental systems based on specific identification, isolation and analysis of human hepatocytes infected with P. berghei ANKA GFP or P. falciparum 3D7 GFP sporozoites we demonstrated that molecular components of the MHC class I pathway exhibit largely unaltered expression in malaria-infected hepatocytes until very late stages of parasite development. Furthermore, infected cells showed no obvious defects in their capacity to upregulate expression of different molecular components of the MHC class I machinery in response to pro-inflammatory lymphokines or trigger direct activation of allo-specific or peptide-specific human CD8+ T-cells. We further demonstrate that ectopic expression of circumsporozoite protein does not alter expression of critical genes of the MHC class I pathway and its response to pro-inflammatory cytokines. In addition, we identified supra-cellular structures, which arose at late stages of parasite replication, possessed the characteristic morphology of merosomes and exhibited nearly complete loss of surface MHC class I expression. These data have multiple implications for our understanding of natural T-cell immunity against malaria and may promote development of novel, efficient anti-malaria vaccines overcoming immune escape of the parasite in the liver.

Using Genetic Methods To Define the Targets of Compounds with Antimalarial Activity

Although phenotypic cellular screening has been used to drive antimalarial drug discovery in recent years, in some cases target-based drug discovery remains more attractive. This is especially true when appropriate high-throughput cellular assays are lacking, as is the case for drug discovery efforts that aim to provide a replacement for primaquine (4-N-(6-methoxyquinolin-8-yl)pentane-1,4-diamine), the only drug that can block Plasmodium transmission to Anopheles mosquitoes and eliminate liver-stage hypnozoites. At present, however, there are no known chemically validated parasite protein targets that are important in all Plasmodium parasite developmental stages and that can be used in traditional biochemical compound screens. We propose that a plethora of novel, chemically validated, cross-stage antimalarial targets still remain to be discovered from the ~5,500 proteins encoded by the Plasmodium genomes. Here we discuss how in vitro evolution of drug-resistant strains of Plasmodium falciparum and subsequent whole-genome analysis can be used to find the targets of some of the many compounds discovered in whole-cell phenotypic screens.

Cytometry in malaria--a practical replacement for microscopy?

Malaria, caused by protozoan Plasmodium parasites, kills ~800,000 people each year. Exact figures are uncertain because presumptive diagnoses are often made without identifying parasites in patients' blood either by microscopy, using Giemsa's century-old stain, or by simpler tests that are ultimately dependent on microscopy for quality control. Microscopy itself relies on trained observers' ability to detect subtle morphological features of parasitized red blood cells, only a few of which may be present on a slide. Quantitative and objective flow cytometric measurements of cellular constituents such as DNA, RNA, and the malaria pigment hemozoin are now useful in research in malaria biology and pharmacology, and can provide more reliable identification of parasite species and developmental stages and better detection of low-density parasitemia than could microscopy. The same measurements can now be implemented in much smaller, simpler, cheaper imaging cytometers, potentially providing a more accurate and precise diagnostic modality.

Ultrastructural Study of Host-Parasite Relationship and Pathogenicity of Eimeria Sp. Infecting Libyan Jird ( Meriones Libycus ) ( Lichtenstein , 1828 )

The relationship of Eimeria sp. and its host fat LibyanJird (Meriones libycus) was studied on an ultrastructural level. The host cellular organelles (nucleus, mitochondria, endoplasmic reticulum and golgi apparatus) and the changes of its infected intestinal epithelial cells during the development of parasitic stages (schizogony and gamogony) were studied and compared with those non-infected cells. The ultrastructures of intravacuolar tubules and folds in the parasitophorous vacuole (P.V.) were described. These fine structures may involve in the transporttation of materials from the host cell to across the parasitovorous vacuole (P.V.).

Parasite-induced and parasite development-dependent alteration of the swimming behavior of fish hosts

Parasites with complex life cycles have the ability to change the behavior of their intermediate host in a way that increases their transmission rate to the next host. However, the level of behavioral changes can vary considerably, depending on the stage of parasite development and parasite intensity. To investigate the influence of such parameters, we evaluated the locomotory activity of the fish Poecilia vivipara prior to experimental infections, 7 days post-infection (dpi) and 14dpi with cercariae of the digenean Ascocotyle (Phagicola) pindoramensis. The locomotory activity was monitored using an image system, Videomex®, linked to with a video camera able to record the swimming behavior of the fishes. At the end of the experiments, fishes were dissected and all metacercariae from the gills and mesenteries, the specific sites utilized by A. (P.) pindoramensis, were recovered and counted. There was a significant decrease in the swimming behavior of fishes after 14dpi. Similarly, we found a significant correlation between the swimming behavior of the fishes and parasite intensity in both sites of infection. It is surmised that the decrease in locomotory activity of P. vivipara caused by A. (P.) pindoramensis can disturb its predator–prey relationship in natural environment.

Plasmodium berghei MAPK1 displays differential and dynamic subcellular localizations during liver stage development.

Mitogen-activated protein kinases (MAPKs) regulate key signaling events in eukaryotic cells. In the genomes of protozoan Plasmodium parasites, the causative agents of malaria, two genes encoding kinases with significant homology to other eukaryotic MAPKs have been identified (mapk1, mapk2). In this work, we show that both genes are transcribed during Plasmodium berghei liver stage development, and analyze expression and subcellular localization of the PbMAPK1 protein in liver stage parasites. Live cell imaging of transgenic parasites expressing GFP-tagged PbMAPK1 revealed a nuclear localization of PbMAPK1 in the early schizont stage mediated by nuclear localization signals in the C-terminal domain. In contrast, a distinct localization of PbMAPK1 in comma/ring-shaped structures in proximity to the parasite’s nuclei and the invaginating parasite membrane was observed during the cytomere stage of parasite development as well as in immature blood stage schizonts. The PbMAPK1 localization was found to be independent of integrity of a motif putatively involved in ATP binding, integrity of the putative activation motif and the presence of a predicted coiled-coil domain in the C-terminal domain. Although PbMAPK1 knock out parasites showed normal liver stage development, the kinase may still fulfill a dual function in both schizogony and merogony of liver stage parasites regulated by its dynamic and stage-dependent subcellular localization.

Macrophage migration inhibitory factor (MIF) of the protozoan parasite Eimeria influences the components of the immune system of its host, the chicken

Macrophage migration inhibitory factor (MIF) is a soluble factor produced by sensitized T lymphocytes that inhibits the random migration of macrophages. Homologues of MIF from invertebrates have been identified, making it an interesting molecule from a functional perspective. In the present study, the localization of a parasite MIF protein as well as its effect on the host was characterized. Western blot analysis shows that Eimeria MIF (EMIF) is found during all parasite developmental stages tested. Transmission electron microscopy shows that MIF is distributed throughout cytosol and nucleus of Eimeria acervulina merozoites. Immunohistochemical analysis suggests that EMIF may be released into the surrounding tissues as early as 24h after infection, while later during oocyst formation, MIF expression is localized to areas immediately surrounding the oocysts, as well as in wall-forming bodies. The chemotaxis assay revealed an inhibitory function of EMIF on chicken monocyte migration. Quantitative real-time PCR was performed to examine the effect of EMIF on host immune system by measuring the transcripts of inflammatory mediators. An ex vivo stimulation study showed that E. acervulina MIF (EaMIF) enhanced expression of pro-inflammatory cytokines and chemokines in the presence of lipopolysaccharide (LPS). Furthermore, sequential treatment of adherent peripheral blood mononuclear cells with EaMIF, chicken MIF, and LPS in 2-h intervals led to the highest levels of interleukin (IL)-1B, chemokine CCLi3, IL-18, and interferon-gamma mRNA expression. This study shows that parasite MIF is widely expressed and may have potential effects on the immune system of the host.

First detection of Alaria alata mesocercariae in wild boars (Sus scrofa Linnaeus, 1758) from Bulgaria

The trematode Alaria alata, an intestinal parasite of different carnivore species is widely distributed throughout Europe. The mesocercarial stages of Alaria spp. may infect almost all vertebrate species, including humans, and, in particular, omnivorous scavengers such as wild boars serve as paratenic hosts for the parasite. The introduction of the A. alata mesocercariae migration technique (AMT) opened the way to a reliable detection of Alaria spp. mesocercariae in different body tissues of their paratenic hosts. For the first time, it was possible to detect vital A. alata mesocercariae from two Bulgarian wild boars by means of this new method. In addition, polymerase chain reaction (PCR) examination of the respective parasitic DNA allowed the unequivocal species identification of the parasites as A. alata. Isolation and molecular biological identification of the parasite's developmental stages make significant contributions to completion of data on both the distribution of Alaria spp. in stocks of European game and the relationship between different Eurasian Alaria spp. isolates.

Explaining variability in parasite aggregation levels among host samples

Aggregated distributions among individual hosts are a defining feature of metazoan parasite populations. Heterogeneity among host individuals in exposure to parasites or in susceptibility to infection is thought to be the main factor generating aggregation, with properties of parasites themselves explaining some of the variability in aggregation levels observed among species. Here, using data from 410 samples of helminth parasites on fish hosts, I tested the contribution of (i) within-sample variation in host body size, taken as a proxy for variability in host susceptibility, and (ii) parasite taxon and developmental stage, to the aggregated distribution of parasites. Log-transformed variance in numbers of parasites per host was regressed against log mean number across all samples; the strong relationship (r²= 0.88) indicated that aggregation levels are tightly constrained by mean infection levels, and that only a small proportion of the observed variability in parasite aggregation levels remains to be accounted for by other factors. Using the residuals of this regression as measures of 'unexplained' aggregation, a mixed effects model revealed no significant effect of within-sample variation in host body size or of parasite taxon or stage (i.e. juvenile versus adult) on parasite aggregation level within a sample. However, much of the remaining variability in parasite aggregation levels among samples was accounted for by the number of individual hosts examined per sample, and species-specific and study-specific effects reflecting idiosyncrasies of particular systems. This suggests that with most differences in aggregation among samples already explained, there may be little point in seeking universal causes for the remaining variation.

A new microsporidium, Triwangia caridinae gen. nov., sp. nov. parasitizing fresh water shrimp, Caridina formosae (Decapoda: Atyidae) in Taiwan

A new microsporidium was isolated from the endemic, Taiwanese shrimp, Caridina formosae (Decapoda, Atyidae) from northern Taiwan. A conspicuous symptom of infection was presence of opaque white xenomas located in the proximity of the alimentary tract, the surface of the hepatopancreas, and the gills. A fully developed xenoma consisted of a hard, thick capsule filled with sporophorous vesicles containing multiple spores. Microsporidia developed synchronously within the same sporophorous vesicle, although the stage of parasite development differed among the vesicles. Fresh spores were pyriform, mononucleated and measured 6.53×4.38μm. The polar filament was anisofilar with 9–11 coils. Phylogenetic analysis based on the small subunit ribosomal DNA sequence showed that the isolate is most similar to the fish microsporidian clade containing the genera Kabatana, Microgemma, Potaspora, Spraguea, and Teramicra. The highest sequence identity, 80%, was with Spraguea spp. Based on pathogenesis, life cycle and phylogenetic analysis, we erect a new genus and species, Triwangia caridinae for the novel microsporidium.

Free Ca2+ Initiates and Regulates Malaria Parasite Egress Program in Infected Erythrocytes

Malaria parasite egress from infected erythrocytes finalizes the asexual cycle of this organism and leading to parasite dissemination and disease progression. The egress mechanism is not fully understood, but several tightly coordinated steps and pathways were described recently. Mature parasites breach two sets of membranes to escape an infected host cell. We investigated the role of free calcium in the initiation and control of individual steps in the egress program of Plasmodium falciparum. The entire egress pathway (tens of minutes before release) is independent of extracellular free calcium, using rather intracellular calcium accumulated within the parasite during the trophozoite-schizont stages of parasite development. Based on experiments with calcium chelators and ionophore, inhibitors of the parasite endoplasmic reticulum, fluorescence microscopy and morphological analysis of mature parasites we propose that the schizont endoplasmic reticulum is the calcium source for egress initiation and that several post-initiation steps in the egress program are affected by increased intracellular Ca2+. We also reveal a new calcium-dependent step in the parasite egress mechanism: swelling of the parasitophorous vacuole (PV). This major event in parasite egress leads to the rupture of critically swelled vacuoles and mature parasite extrusion from the host cell. We found that this vacuolar swelling is exaggeratedly slow in dehydrated sickle cells, leading to a severe defect in parasite egress, probably due to low levels of ions conducting osmotic driven water translocation from erythrocyte cytoplasm to PV. Parasite egress can be accelerated in mature schizonts by pharmacological intervention; the mechanism of this phenomenon involves PV swelling. However, the same treatment of immature schizonts leads to immediate parasite death within the host cell due to erythrocyte hemolysis of a critically expanded PV. Exploring this stage-dependent parasite killing may be useful for development of anti-malarials.

Histopathological and Histochemical Study of Intestinal Cryptosporidiosis in Mice

This study was conducted to investigate the histopathological and histochemical alteration of intestinal mice infected with C. parvum isolated from calf feces. The result elucidate that histopathologcal changes represented by hypertrophy and hyperplasia of epithelium villi in mucosa and submucosal gland, presence of different developmental stages of parasite attached to brush border of epithelium and in crypt in addition to proliferation of mononuclear inflammatory cell and blunts of villi have been observed as compared with control group. Histochemcially, histological sections of infected mice revealed strong positive reaction with AB pH 1.0. In addition to positive reaction with PAS – AB pH 2.5 as compared with control group.Conclusion: This study investigated that infection with C. parvum cause changes in mucosubstances secreted from goblet cells of mucosal villi and submucosal glands.

The nature of changes of some immunophysiological characteristics in bream (Abramis brama) infected with plerocercoids (Ligula intestinalis) at various stages of parasite development

The data from studies of the antimicrobial properties of blood serum, the content of total lipids, and antioxidant activity of immunocompetent tissues and organs of breams Abramis brama infected with plerocercoids Ligula intestinalis depending on the phase of development of the parasite are presented. The quantitative characteristics of the studied parameters are determined.

Effect of heterozygous beta thalassemia on the phosphorylative response to Plasmodium falciparum infection

Malaria parasites interact with the host cell membrane inserting new proteins and inducing oxidative and phosphorylative changes of erythrocyte proteins. In the present report we monitored the time dependent oxidative and phosphorylative modifications induced by parasites in heterozygous beta thalassemia (Het-βThal). Het-βThal causes mild anemia and is known to determine a pro-oxidant milieu and a protective effect against severe malaria. In malaria cultures Het-βThal has been reported to induce accumulation of hemoglobin denaturation products. At early parasite development stages (rings), tyrosine hyper-phosphorylation of band 3 was the most notable modification, and at later development stages (trophozoites), additional membrane proteins displayed significant hyper-phosphorylation of their serine and tyrosine residues (adducins, ankyrin, catalase). Het-βThal also caused membrane destabilization. Free radical scavengers effectively inhibited the phosphorylative response and membrane destabilization. Kinase inhibitors exerted similar effects suggesting a causal relationship between oxidative stress, membrane protein hyper-phosphorylation and increased membrane damage exacerbated by Het-βThal. In conclusion, different lines of evidence suggest that Het-βThal enhances the redox stress caused by malaria parasites inducing its protective effect destabilizing the host cell membrane. This article is part of a Special Issue entitled: Integrated omics.

Therapeutic and Residual Efficacy of a Pour-On Formulation of Novaluron against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) on Infested Cattle

The effectiveness of a pour-on formulation of novaluron (Novatack Pour-on 5% Al, Clarion Biociencias LTDA, Goiania, Brazil), a benzoylphenyl urea acaricide, was evaluated by applying a low and high concentrations (1 mg/20 kg body weight and 1 mg/10 kg body weight) to cattle infested with all parasitic developmental stages (adults, nymphs, and larvae) of Rhipicephalus (Boophilus) microplus (Canestrini). The therapeutic percentage reduction in the number of females that reached repletion following treatment was 24.8 and 24.6% for the low and high rates, respectively. No difference in the weight of engorged females recovered from treated calves or in the index of fecundity (IF) was observed for either rate in comparison with the untreated control, and no significant difference in the overall therapeutic level of control was observed between the low and the high rates. Results obtained from the analysis of persistent (residual) efficacy of both rates showed no significant differences between the treatments at weekly intervals; however, the percent control of the IF observed against larvae infested 1 and 2 wks after treatment of both rates was moderate indicating that novaluron appears to be more effective against larvae that come into contact with cattle after treatment than against adults or nymphs already infesting the animal at the time of treatment.

A Study of Eimeria Species in Sheep in Mosul City

The study was conducted to diagnose and study species of Eimeria in sheep in Mosul city from beginning of September 2009 to end May 2010, as well as to determine the percentage and intensity of infection of Eimeria species. Five hundred fecal samples of sheep with different ages were collected from different areas of the Mosul city. The results showed that total percentage of Emeria infection was 63.6%. The variations in percentage of infection were recorded according to month of study. Highest percentage was recorded in March being 89.2% and the lowest in September 25.9%. The species E. ovina recorded the highest infection rate 86.7%, while the species E. granulosa represented lowest infection rate 10%. Moreover the intensity of infection was higher in young ages and lower in adult. The results were detected that indoor sheep infection with high parasitic infection 69.9% whereas outdoor animals have an infection rate 25.3%. The morphological characters of oocysts were varied according to species of Eimeria has been studied. Fifty of intestinal and abomasal samples from both slaughtered in shops butchery in Mosul city and dead animals were examined to detect Eimeria infection, and results show that infection percentage was 56.4% in intestine of slaughtered animals and 36.3% in dead animal. Moreover no infection of Eimeria were detected in abomasums in both slaughtered and dead animals. The oocysts of (E. parva, E.pallida and E. ovinoidalis) detected at more than 5000 oocysts per gram of intestinal contents. The intestinal secraping stained with Giemsa stain reveals the presence of different developmental stages of parasites in wall of intestine. The histopathological sections of intestine revealed the different pathological changes concerning of Eimeria infection.

Gene selective mRNA cleavage inhibits the development of Plasmodium falciparum

Unique peptide-morpholino oligomer (PMO) conjugates have been designed to bind and promote the cleavage of specific mRNA as a tool to inhibit gene function and parasite growth. The new conjugates were validated using the P. falciparum gyrase mRNA as a target (PfGyrA). Assays in vitro demonstrated a selective degradation of the PfGyrA mRNA directed by the external guide sequences, which are morpholino oligomers in the conjugates. Fluorescence microscopy revealed that labeled conjugates are delivered into Plasmodium-infected erythrocytes during all intraerythrocytic stages of parasite development. Consistent with the expression of PfGyrA in all stages of parasite development, proliferation assays showed that these conjugates have potent antimalarial activity, blocking early development, maturation, and replication of the parasite. The conjugates were equally effective against drug sensitive and resistant P. falciparum strains. The potency, selectivity, and predicted safety of PMO conjugates make this approach attractive for the development of a unique class of target-specific antimalarials and for large-scale functional analysis of the malarial genome.

Peroxiredoxins in parasites.

Parasite survival and virulence relies on effective defenses against reactive oxygen and nitrogen species produced by the host immune system. Peroxiredoxins (Prxs) are ubiquitous enzymes now thought to be central to such defenses and, as such, have potential value as drug targets and vaccine antigens. Plasmodial and kinetoplastid Prx systems are the most extensively studied, yet remain inadequately understood. For many other parasites our knowledge is even less well developed. Through parasite genome sequencing efforts, however, the key players are being discovered and characterized. Here we describe what is known about the biochemistry, regulation, and cell biology of Prxs in parasitic protozoa, helminths, and fungi. At least one Prx is found in each parasite with a sequenced genome, and a notable theme is the common patterns of expression, localization, and functionality among sequence-similar Prxs in related species. The nomenclature of Prxs from parasites is in a state of disarray, causing confusion and making comparative inferences difficult. Here we introduce a systematic Prx naming convention that is consistent between organisms and informative about structural and evolutionary relationships. The new nomenclature should stimulate the crossfertilization of ideas among parasitologists and with the broader redox research community. The diverse parasite developmental stages and host environments present complex systems in which to explore the variety of roles played by Prxs, with a view toward parlaying what is learned into novel therapies and vaccines that are urgently needed.

Delivering resistance to a major constraint for rain‐fed rice production

Delivering resistance to a major constraint for rain‐fed rice production