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Background: Trypanosoma cruzi is transmitted to humans by hematophagous bugs belonging to the Triatominae subfamily. Its intra-vectorial cycle is complex and occurs exclusively in the insect's midgut. Dissecting the elements involved in the cross-talk between the parasite and its vector within the digestive tract should provide novel targets for interrupting the parasitic life cycle and affecting vectorial competence. These interactions are shaped by the strategies that parasites use to infect and exploit their hosts, and the host's responses that are designed to detect and eliminate parasites. The objective of the current study is to characterize the impact of T. cruzi establishment within its vector on the dynamics of its midgut. Methods: In this study, we evaluated the impact of T. cruzi infection on protein expression within the anterior midgut of the model insect Rhodnius prolixus at 6 and 24 h post-infection (hpi) using high-throughput quantitative proteomics. Results: Shortly after its ingestion, the parasite modulates the proteome of the digestive epithelium by upregulating 218 proteins and negatively affecting the expression of 11 proteins involved in a wide array of cellular functions, many of which are pivotal due to their instrumental roles in cellular metabolism and homeostasis. This swift response underscores the intricate manipulation of the vector's cellular machinery by the parasite. Moreover, a more in-depth analysis of proteins immediately induced by the parasite reveals a pronounced predominance of mitochondrial proteins, thereby altering the sub-proteomic landscape of this organelle. This includes various complexes of the respiratory chain involved in ATP generation. In addition to mitochondrial metabolic dysregulation, a significant number of detoxifying proteins, such as antioxidant enzymes and P450 cytochromes, were immediately induced by the parasite, highlighting a stress response. Conclusions: This study is the first to illustrate the response of the digestive epithelium upon contact with T. cruzi, as well as the alteration of mitochondrial sub-proteome by the parasite. This manipulation of the vector's physiology is attributable to the cascade activation of a signaling pathway by the parasite. Understanding the elements of this response, as well as its triggers, could be the foundation for innovative strategies to control the transmission of American trypanosomiasis, such as the development of targeted interventions aimed at disrupting parasite proliferation and transmission within the triatomine vector. [ABSTRACT FROM AUTHOR]

Background: Toxoplasma gondii (T. gondii) is a widespread, zoonotic protozoan intracellular parasite with a complex life cycle, which can cause toxoplasmosis, a potentially serious disease. During the invasion process, T. gondii proteins first bind to the relevant host cell receptors, such as glycosaminoglycan molecule (GAG-binding motif), which is one of the main receptors for parasites or virus to infect host cells. However, research on TGME49_216510 (T. gondii Trx21), a protein from Toxoplasma gondii, is limited. Methods: Bioinformatics analysis of the Trx21 protein was performed firstly. And specific primers were then designed using the conserved domain and GAG-binding motif to amplify, express, and purify a fragment of the Trx21 protein. The purified Trx21-GST protein was used for antioxidant and cell adhesion experiments. Simultaneously, mice were immunized with Trx21-His to generate specific polyclonal antibodies for subcellular localization analysis. Results: The Trx21 protein, consisting of 774 amino acids, included a transmembrane region, three GAG-binding motifs, and a Thioredoxin-like domain. The recombinant Trx21-His protein had a molecular mass of about 31 kDa, while the Trx21-GST protein had a molecular mass of about 55 kDa, which was analyzed by SDS-PAGE and Western blot. Subcellular localization analysis by IFA revealed that Trx21 is predominantly distributed in the cytoplasm of T. gondii. Furthermore, Trx21 exhibited a protective effect on supercoiled DNA against metal-catalyzed oxidation (MCO) and demonstrated adhesion abilities to Vero cells. Conclusions: These results indicate that Trx21 plays an important role in host cell interaction and oxidative damage. [ABSTRACT FROM AUTHOR]

Entamoeba histolytica is a protozoan parasite belonging to the phylum Amoebozoa that causes amebiasis, a global public health problem. E. histolytica alternates its form between a proliferative trophozoite and a dormant cyst. Trophozoite proliferation is closely associated with amebiasis symptoms and pathogenesis whereas cysts transmit the disease. Drugs are available for clinical use; however, they have issues of adverse effects and dual targeting of disease symptoms and transmission remains to be improved. Development of new drugs is therefore urgently needed. An untargeted lipidomics analysis recently revealed structural uniqueness of the Entamoeba lipidome at different stages of the parasite's life cycle involving very long (26–30 carbons) and/or medium (8–12 carbons) acyl chains linked to glycerophospholipids and sphingolipids. Here, we investigated the physiology of this unique acyl chain diversity in Entamoeba, a non-photosynthetic protist. We characterized E. histolytica fatty acid elongases (EhFAEs), which are typically components of the fatty acid elongation cycle of photosynthetic protists and plants. An approach combining genetics and lipidomics revealed that EhFAEs are involved in the production of medium and very long acyl chains in E. histolytica. This approach also showed that the K3 group herbicides, flufenacet, cafenstrole, and fenoxasulfone, inhibited the production of very long acyl chains, thereby impairing Entamoeba trophozoite proliferation and cyst formation. Importantly, none of these three compounds showed toxicity to a human cell line; therefore, EhFAEs are reasonable targets for developing new anti-amebiasis drugs and these compounds are promising leads for such drugs. Interestingly, in the Amoebazoan lineage, gain and loss of the genes encoding two different types of fatty acid elongase have occurred during evolution, which may be relevant to parasite adaptation. Acyl chain diversity in lipids is therefore a unique and indispensable feature for parasitic adaptation of Entamoeba. Author summary: Entamoeba, a protozoan parasite, thrives in its niche by alternating its form between a proliferative trophozoite and a dormant cyst. Entamoeba parasites were recently shown to generate lipid acyl chain diversity throughout their life cycle. Here, we provide insight into the mechanism generating this acyl chain diversity and its importance in maintaining the parasite's life cycle. We also show that the E. histolytica isozymes responsible for generating this diversity are reasonable targets for developing new anti-amebiasis drugs, and that the herbicides, flufenacet, cafenstrole, and fenoxasulfone, are promising leads for such drugs. Interestingly, gain and loss of the two genes encoding the isozymes controlling acyl chain diversity is plausibly related to E. histolytica host adaptation. [ABSTRACT FROM AUTHOR]

Background: Toxoplasma gondii is a zoonotic protozoan parasite with a heteroxenus life cycle that involves felids as the definitive hosts and any warm-blooded animal, including humans, as intermediate hosts. Cats are key players in parasite transmission as they are capable of shedding high numbers of oocysts in their feces that contaminate the environment. Methods: The study was performed on 31 domestic cats (31.23 ± 27.18 months old) originating from rural and urban areas (5.17:1) in the center and north-west Romania. Feces (n = 31), blood (n = 28), and heart samples (n = 27) were collected. Fecal samples were analyzed by flotation technique, and PCR (529 bp repetitive element). Fecal samples with T. gondii oocysts were bioassayed in mice. Serum samples were analyzed by modified agglutination test and ImmunoComb for the detection of specific anti-T. gondii IgG antibodies. Heart samples were bioassayed in mice, and analyzed by PCR. Toxoplasma gondii positive samples were genotyped by nPCR-RFLP targeting eleven genetic loci (SAG1, SAG2, alt-SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). Results: Toxoplasma gondii oocysts were found in 2 out of 31 fecal samples collected from a 3-months old stray kitten, and a 4-years old female. In total, 17 out of 27 sera were positive for T. gondii IgG antibodies. The antibody titers in MAT ranged from 1:6 to 1:384. Toxoplasma gondii DNA was detected in 7 out of 27 heart samples, and four of them were positive also by bioassay. Six T. gondii DNA samples from bioassayed mice could be assigned to ToxoDB PCR-RFLP genotype #1 or #3 (Type II) and one T. gondii DNA from heart digest to genotype #2 (Type III). Both of these genotypes are common in Europe. Conclusions: Our results revealed that the infection with T. gondii is still high in cats from Romania. The oocysts shedded by these cats represent an important source of infection for intermediate hosts, including humans. Further studies on a wider range of cases are necessary for a more exhaustive definition of the T. gondii genotypes circulating in Romania. [ABSTRACT FROM AUTHOR]

[Display omitted] • Microbe surface molecules can upregulate a mucin-like gene in Lutzomyia longipalpis. • Expression of the attacin gene is triggered as a generic immune response in sand flies. • Sand fly defensin gene expression is tuned, depending on the ingested microbe molecule. The interaction between pathogens and vectors' physiology can impact parasite transmission. Studying this interaction at the molecular level can help in developing control strategies. We study leishmaniases, diseases caused by Leishmania parasites transmitted by sand fly vectors, posing a significant global public health concern. Lipophosphoglycan (LPG), the major surface glycoconjugate of Leishmania , has been described to have several roles throughout the parasite's life cycle, both in the insect and vertebrate hosts. In addition, the sand fly midgut possesses a rich microbiota expressing lipopolysaccharides (LPS). However, the effect of LPG and LPS on the gene expression of sand fly midgut proteins or immunity effectors has not yet been documented. We experimentally fed Lutzomyia longipalpis and Phlebotomus papatasi sand flies with blood containing purified LPG from Leishmania infantum , Leishmania major , or LPS from Escherichia coli. The effect on the expression of genes encoding gut proteins galectin and mucin, digestive enzymes trypsin and chymotrypsin, and antimicrobial peptides (AMPs) attacin and defensins was assessed by quantitative PCR (qPCR). The gene expression of a mucin-like protein in L. longipalpis was increased by L. infantum LPG and E. coli LPS. The gene expression of a galectin was increased in L. longipalpis by L. major LPG, and in P. papatasi by E. coli LPS. Nevertheless, the gene expression of trypsins and chymotrypsins did not significantly change. On the other hand, both L. infantum and L. major LPG significantly enhanced expression of the AMP attacin in both sand fly species and defensin in L. longipalpis. In addition, E. coli LPS increased the expression of attacin and defensin in L. longipalpis. Our study showed that Leishmania LPG and E. coli LPS differentially modulate the expression of sand fly genes involved in gut maintenance and defence. This suggests that the glycoconjugates from microbiota or Leishmania may increase the vector's immune response and the gene expression of a gut coating protein in a permissive vector. [ABSTRACT FROM AUTHOR]

Echinococcus multilocularis is a cestode that causes human alveolar echinococcosis, a lethal zoonotic disease distributed in the northern hemisphere. The life cycle of this parasite is maintained in nature by voles as intermediate hosts and foxes as definitive hosts in Hokkaido, Japan. Although dogs are also susceptible to the parasite, the infection has been considered typically asymptomatic. We report the detection of E. multilocularis eggs in the diarrheal feces of a dog with chronic gastrointestinal signs, which disappeared after anthelmintic treatment. The mitochondrial genome sequence constructed by sequencing of the overlapping PCRs using DNA from the eggs was identical to the most predominant haplotype previously reported in red foxes in Hokkaido. This case highlights that Echinococcus infection should be considered as a differential diagnosis for diarrheal dogs in the disease endemic areas. Further efforts are needed to accumulate parasite genotypes in domestic dogs as well as humans to assess the risk of human infection from dogs. [ABSTRACT FROM AUTHOR]

Background: There are ten genotypes of Echinococcus granulosus with different intermediate and final hosts affecting the parasite's life cycle and its transmission to humans. Therefore, this study was conducted to determine the genotype of isolated hydatid cysts using the simple and fast high-resolution melting point analysis (HRM) method. Methods: The paraffin tissue samples of patients who underwent surgery were obtained from the pathology sample bank of Vasei and Emdad Hospitals in Sabzevar, Iran during 2010-2020. The DNA content of the samples was extracted after collecting and determining the characteristics using the DNA extraction kit. PCR was performed on the samples and the presence of the hydatid cyst genome was confirmed using the special Master Kit. Mix PCR of Solis Biodyne Company and Real-Time device (Bio-Rad) were used, and the genetic identity of hydatid cysts were determined. Results: Out of 33 paraffin samples, 21 samples contained hydatid cyst DNA, two of which were from the brain and 19 from the liver tissues; 12 samples did not contain hydatid cyst DNAs. All liver samples were from sheep species (G1), and the brain samples were from buffalo species (G3). Therefore, 9.53% of the Echinococcus species collected were buffalo (G3), and 90.47% were sheep (G1) strain. Conclusion: Based on previous patterns, HRM methods can be used for easy and quick identification of Echinococcus strains. The G1 strain was the dominant strain causing hydatid cyst in different human organs, including the liver and brain. [ABSTRACT FROM AUTHOR]

Orthonectida is an enigmatic group of animals with still uncertain phylogenetic position. Orthonectids parasitize various marine invertebrates. Their life cycle comprises a parasitic plasmodium and free‐living males and females. Sexual individuals develop inside the plasmodium; after egress from the host they copulate in the external environment, and the larva, which has developed inside the female infects a new host. In a series of studied orthonectid species simplification of free‐living sexual individuals can be clearly traced. The number of longitudinal and transverse muscle fibers is gradually reduced. In the nervous system, simplification is even more pronounced. The number of neurons constituting the ganglion is dramatically reduced from 200 in Rhopalura ophiocomae to 4–6 in Intoshia variabili. The peripheral nervous system undergoes gradual simplification as well. The morphological simplification is accompanied with genome reduction. However, not only genes are lost from the genome, it also undergoes compactization ensured by extreme reduction of intergenic distances, short intron sizes, and elimination of repetitive elements. The main trend in orthonectid evolution is simplification and miniaturization of free‐living sexual individuals coupled with reduction and compactization of the genome. Highlights: The main evolutionary trend in orthonectids is simplification and miniaturization of free‐living sexual individuals and reduction and compactization of the genome inextricably linked with these processes. [ABSTRACT FROM AUTHOR]

The trematodes, specifically the genus Schistosoma, are responsible for causing the parasitic disease known as schistosomiasis. In this research, the aim was to utilize the extract of Saussurea costus root as a molluscicidal substance to target the snails, which act as the intermediary host for the parasite species and disrupt the life cycle of the parasite. During the investigation, Biomphalaria alexandrina snails were treated with S. costus root extract for twenty four hours, before undergoing a 24-hour recuperation phase. According to the results, the extract was found to have a molluscicidal potential, with a lethal concentration (LC90) of 113.70ppm. Hemocyte abnormalities in B. alexandrina snails were brought about by exposure to ethanolic extract's LC10 and LC25. Furthermore, histopathological changes were recognized in the hermaphrodite and digestive glands which were confirmed through transmission electron microscope imaging. The genotoxic effects of the extract were also investigated using the RAPD-PCR technique, revealing a decrease in genomic stability (GTS) to 25% at the LC25 concentration. Moreover, the root extract of S. costus has negative effects on the viability and infection of Schistosoma mansoni miracidia and cercariae. The study examined the effects of exposing the infected B. alexandrina snails with S. mansoni miracidia to the LC10 and LC25 concentrations of the root extract after 3 and 21 days. The analysis using GC/MS identified seven compounds in the extract. Therefore, the root extract of S. costus as a bio control agent could replace the chemical molluscicides in reducing schistosomiasis. [ABSTRACT FROM AUTHOR]

Cephalopods are widely distributed in the world's oceans and are among the most important species in the marine ecosystem due to their short life cycle, fast growth rate and extremely rich biomass. Like other marine organisms, cephalopods are at risk of being infected by parasites, often acting as either intermediate or final hosts for different species of parasites. Due to the unique life cycle of parasites, their life cycle can run through the entire or partial life cycle of the host, greatly affecting the growth and survival of cephalopods. Currently, research on parasites in cephalopods mainly focuses on species composition, life history, infection status, and other aspects. This article summarizes the current research status of parasitic infections in cephalopod marine animals. The results show that the main parasites infecting cephalopods are Dicyemida, Cestoda and Nematoda. Dicyemida parasitize cephalopods characteristically, with maturation and reproduction taking place exclusively in the kidneys of cephalopods. Available statistics show that there are 149 species of Dicyemida infecting cephalopods, mainly found within the Sea of Japan, with fewer species found at lower latitudes. Cephalopods, as intermediate hosts, are also susceptible to infection by Cestoda larvae and have a high infection rate. Cestoda mainly parasitize the digestive tract of cephalopods. Due to the large size, stretching, and mobility of Cestoda larvae, they are prone to cause mechanical damage to the host. Nematodes are another common parasites infecting cephalopods, and are primarily ingested as third-stage larvae of Nematodes, which mostly parasitize the stomach and cecum. Nematodes are among the most abundant and common species of cephalopod parasitic infections, and can have significant pathological effects on the host. Other types of parasites can also affect the physiological characteristics of cephalopods to a certain extent. Research on parasites in cephalopods is still in its early stages. Therefore, future research should focus on accelerating the establishment of an open database of parasites in cephalopods, expanding and sharing information on parasite species. It is also important to deeply explore the life processes of parasitic species in cephalopods to further understand their impact on host growth. Additionally, using parasites in cephalopods to divide population composition can provide new ideas for the sustainable development and utilization of cephalopod fisheries. [ABSTRACT FROM AUTHOR]

Broomrape (Phelipanche ramosa) is an obligate root parasitic plant that is known to infest many important crops. It is emerging as a serious biotic threat for Brassica juncea cultivation in India. There is very limited information available about broomrape species and its infestation in B. juncea. Hence, the present study encompassed a comprehensive five-year field survey spanning from 2018 to 2023 of 47 different B. juncea fields in the states of Rajasthan and Haryana in India. The level of broomrape infestation varied from season to season and field to field. In the B. juncea fields, broomrape with blue/violet color corolla, along with occasionally white color type were also noticed. The hairy anther, corolla shape and use of mitochondrial DNA-specific molecular markers revealed that both these broomrape types belong to Phelipanche ramosa. Scanning electron microscopy (SEM) analysis showed that P. ramosa seeds have varying shapes and seed coat have fibrillar periclinal cell wall. In addition to all the known host species for P. ramosa till date, we also identified a new host, a wild Brassicaceae weed species; Lepidium didymum in Haryana. The presence of an alternative weed host may help P. ramosa survive even in the absence of major crop host species. Overall, the present study revealed some insights into the P. ramosa-B. juncea host-parasite complex and its effect on B. juncea which could be further investigated in more detail. [ABSTRACT FROM AUTHOR]

Echinococcus granulosus is a zoonotic parasite responsible for causing cystic echinococcosis in humans and animals. Cystic echinococcosis is recognized as a major public health problem in Iran, with numerous endemic areas spread throughout the country. Wild dogs (Canis familiaris) have been identified as the primary definitive hosts for E.granulosus and are known to play a vital role in the transmission and sustainability of the parasite's life cycle. Understanding the genetic diversity and distribution of E. granulosus genotypes in these wild dogs is important for effective control and prevention strategies. Between 2019 and 2022, a total of 68 peri-urban wild dogs, consisting of 47 males and 21 females, were captured, with unfortunate deaths due to car accidents or disease. Morphological and molecular investigation was performed to determine the presence of E. granulosus. The identification of E. granulosus genotypes was carried out by sequencing the COX1 and NADH1 genes. Of the 68 peri-urban wild dogs examined, 8 (11.7%) were positive for E. granulosus by morphological and molecular analysis. By performing PCR it was determined that the peri-urban wild dogs infected with E. granulosus carried the sheep strain (G1) genotype. This study successfully identified the presence of E. granulosus in peri-urban wild dogs, specifically with the G1 genotype. This finding highlights the potential risk that these dogs pose as carriers of this zoonotic parasite, which can be transmitted to humans and other animals. Further research and surveillance are essential to better understand the epidemiology of E. granulosus and to develop effective strategies for its control and eradication. [ABSTRACT FROM AUTHOR]

Background: In Europe, avian schistosomes of the genus Trichobilharzia are the most common etiological agents involved in human cercarial dermatitis (swimmer's itch). Manifested by a skin rash, the condition is caused by an allergic reaction to cercariae of nonhuman schistosomes. Humans are an accidental host in this parasite's life cycle, while water snails are the intermediate, and waterfowl are the final hosts. The study aimed to conduct a molecular and phylogenetic analysis of Trichobilharzia species occurring in recreational waters in North-Eastern Poland. Methodology: The study area covered three water bodies (Lake Skanda, Lake Ukiel, and Lake Tyrsko) over the summer of 2021. In total, 747 pulmonate freshwater snails (Radix spp., Lymnaea stagnalis) were collected. Each snail was subjected to 1–2 h of light stimulation to induce cercarial expulsion. The phylogenetic analyses of furcocercariae were based on the partial sequence of the ITS region (ITS1, 5.8S rDNA, ITS2 and 28SrDNA). For Radix spp. phylogenetic analyses were based on the ITS-2 region. Results: The prevalence of the Trichobilharzia species infection in snails was 0.5%. Two out of 478 (0.4%) L. stagnaliswere found to be infected with Trichobilharzia szidati. Moreover, two out of 269 (0.7%) snails of the genus Radix were positive for schistosome cercariae. Both snails were identified as Radix auricularia. One of them was infected with Trichobilharzia franki and the other with Trichobilharzia sp. Conclusions: Molecular identification of avian schistosome species, both at the intermediate and definitive hosts level, constitutes an important source of information on a potential threat and prognosis of local swimmer's itch occurrence, and helps to determine species diversity in a particular area. [ABSTRACT FROM AUTHOR]

In the Americas, P. vivax is the predominant causative species of malaria, a debilitating and economically significant disease. Due to the complexity of the malaria parasite life cycle, a vaccine formulation with multiple antigens expressed in various parasite stages may represent an effective approach. Based on this, we previously designed and constructed a chimeric recombinant protein, PvRMC-1, composed by PvCyRPA, PvCelTOS, and Pvs25 epitopes. This chimeric protein was strongly recognized by naturally acquired antibodies from exposed population in the Brazilian Amazon. However, there was no investigation about the induced immune response of PvRMC-1. Therefore, in this work, we evaluated the immunogenicity of this chimeric antigen formulated in three distinct adjuvants: Stimune, AddaVax or Aluminum hydroxide (Al(OH)3) in BALB/c mice. Our results suggested that the chimeric protein PvRMC-1 were capable to generate humoral and cellular responses across all three formulations. Antibodies recognized full-length PvRMC-1 and linear B-cell epitopes from PvCyRPA, PvCelTOS, and Pvs25 individually. Moreover, mice's splenocytes were activated, producing IFN-g in response to PvCelTOS and PvCyRPA peptide epitopes, affirming T-cell epitopes in the antigen. While aluminum hydroxide showed notable cellular response, Stimune and Addavax induced a more comprehensive immune response, encompassing both cellular and humoral components. Thus, our findings indicate that PvRMC-1 would be a promising multistage vaccine candidate that could advance to further preclinical studies. [ABSTRACT FROM AUTHOR]

The cGMP-dependent protein kinase (PKG) is the sole cGMP sensor in malaria parasites, acting as an essential signalling hub to govern key developmental processes throughout the parasite life cycle. Despite the importance of PKG in the clinically relevant asexual blood stages, many aspects of malarial PKG regulation, including the importance of phosphorylation, remain poorly understood. Here we use genetic and biochemical approaches to show that reduced cGMP binding to cyclic nucleotide binding domain B does not affect in vitro kinase activity but prevents parasite egress. Similarly, we show that phosphorylation of a key threonine residue (T695) in the activation loop is dispensable for kinase activity in vitro but is essential for in vivo PKG function, with loss of T695 phosphorylation leading to aberrant phosphorylation events across the parasite proteome and changes to the substrate specificity of PKG. Our findings indicate that Plasmodium PKG is uniquely regulated to transduce signals crucial for malaria parasite development. Author summary: Despite all efforts to control and eradicate malaria, the disease still poses a huge burden on human health. Almost half of the world's population lives in high malaria transmission areas with over half a million deaths occurring annually due to the disease, which is caused by a single-celled parasite called Plasmodium. Replication of the parasite inside red blood cells is responsible for all the clinical manifestations of the disease. At the end of each replicative cycle, parasites rupture the red blood cell in a process known as egress in order to invade new red blood cells. Previous studies have shown that an essential kinase termed PKG is a master regulator of egress. However, many aspects of PKG regulation are still unknown. In this work we examined the importance of phosphorylation on PKG function by replacing wild type PKG with mutant forms refractory to phosphorylation. We found that phosphorylation in a specific region of the protein is essential for parasite survival. Excitingly though, phosphorylation is not essential for kinase activity, as has been shown for mammalian PKG proteins but regulates its substrate specificity. Our results suggest that Plasmodium PKG is uniquely regulated compared to its mammalian counterparts, to facilitate parasite proliferation. [ABSTRACT FROM AUTHOR]

Background: Sarcocystis includes a global group of apicomplexan parasites with two-host life cycle frequently circulating in wildlife and domestic hosts, including humans. Two of the most important wild terrestrial carnivores acting as definitive hosts are the red fox and raccoon dog, due to their wide distribution in Europe and usage of wild and farmed animals as prey. This study was conducted to determine the prevalence of Sarcocystis in hunted red foxes and raccoon dogs from nine regions of the Czech Republic and to identify isolated sporocysts by molecular techniques. Methods: Approximately 5 g of the contents of large intestine from 200 animals (197 red foxes and three raccoon dogs) were examined by flotation centrifugation coprological method. Only samples of 50 red foxes and one raccoon dog positive to Sarcocystis spp. were used for the nested PCR (nPCR) method to amplify a fragment or partial sequence on the cox1 gene. Ten species-specific primer pairs for detection of Sarcocystis spp. using farm animals as intermediate hosts were utilized. Results: In total, 38.1% of the red foxes and 66.7% of the raccoon dogs were positive to Sarcocystis by light microscopy. The molecular characterization resulted in the identification of five species in the red fox: S. arieticanis, S. capracanis, S. cruzi, S. miescheriana, and S. tenella, while the PCR was negative for the sole raccoon dog. The highest intraspecific variation was found for S. miescheriana, while S. tenella was the most prevalent. Co-infections occurred in the large intestine of the red fox. No zoonotic species were found in our samples. Conclusion: This is the first study where the potential role of the red fox and raccoon dogs as spreaders of Sarcocystis to farm animals in the Czech Republic is shown. The use of species-specific primers provides a fast and easy method for screening multiple samples for a particular Sarcocystis species. [ABSTRACT FROM AUTHOR]

Climate change may exacerbate the impact of invasive parasites from warmer climates through pre‐existing temperature adaptations. We investigated temperature impacts on two closely related marine parasitic copepod species that share the blue mussel (Mytilus edulis) as host: Mytilicola orientalis has invaded the system from a warmer climate <20 years ago, whereas its established congener Mytilicola intestinalis has had >90 years to adapt. In laboratory experiments with temperatures 10–26°C, covering current and future temperatures as well as heat waves, the development of both life cycle stages of both species accelerated with increasing temperature. In the parasitic stages, the growth of the established invader increased evenly from 10°C to 22°C, whereas the recent invader barely grew at all at 10°C and grew faster already at 18°C. In contrast, temperature had little effect on the transition success between life cycle stages. However, the highest temperature (26°C) limited the egg development success of the established invader and the host entry success of both species, whereas the infection success of the established invader increased at 18°C and 22°C. In general, our experiments indicate that the main effect of temperature on both species is through development speed and not life cycle stage transition success. Based on regional long‐term temperature data and predictions, the numbers of completed life cycles per year will increase for both parasites. The established invader seems better adapted for low current temperatures (around 10°C), whereas the more recent invader barely develops at these temperatures but can cope in high temperatures (around 26°C). Hence, pre‐existing temperature adaptations of the recent invader may allow the species to better cope with heat waves. [ABSTRACT FROM AUTHOR]

Plankton parasites such as viruses, bacteria, fungi, and unicellular eukaryotes are associated symbionts colonizing algal groups in aquatic ecosystems. They occur within a network of microbe–microbe interactions in which they parasitize phytoplankton and seaweeds, i.e., primary producers generating organic carbon and forming the basis of marine food webs. These parasites use algae as a source of nutrients and reproduce at the expense of their host, causing infectious symptoms leading to disease and death. Plankton parasites can reduce the algal populations, and the infection of one specific species may, in turn, favor the development of another, influencing the seasonal succession of algal blooms in oceans, seas, and lakes. Many parasites have free-living stages that zooplankton graze upon, representing a significant trophic link in food webs. The biology and life cycle of plankton parasites are well investigated in marine and freshwater algal hosts. Still, the chemical signaling mediating these microbial interactions and the effect of co-occurring symbionts remain underexplored. This review focuses on the diversity of plankton parasites infecting algae, with a particular emphasis on unicellular eukaryotes. The ecological role of plankton parasites, the mechanisms regulating cellular infection and host resistance, and the interplay of alga-parasite interactions with coexisting microorganisms are discussed. [ABSTRACT FROM AUTHOR]

Introduction: Human cysticercosis occurs when humans ingest Taenia solium eggs from the fecal-oral route. The infection in the central nervous system is called neurocysticercosis and is considered the principal cause of late-onset epilepsy in endemic countries in Latin America, Asia, and Africa. Case report: 71-year-old female in a state of neglect with a medical history of untreated rheumatoid arthritis. She presented her first seizure episode. Cerebral magnetic resonance imaging with contrast reported parenchymal and subarachnoid neurocysticercosis in different stages. Discussion: The high incidence of neurocysticercosis in endemic countries is associated with poverty conditions such as less hygienic conditions and permanent contact with domestic animals. The four stages of classification of neurocysticercosis is useful for pathologists and radiologists to identify the life cycle of the parasite within the body and the specific imaging findings of each phase. After diagnosis, treatment includes the use of niclosamide or praziquantel. Conclusions: neurocysticercosis is a public health problem in endemic countries that requires further attention. Clinical manifestations are variable and neuroimaging findings are essential to making a correct diagnosis. [ABSTRACT FROM AUTHOR]

Post-translational modifications (PTMs) are essential for regulating protein functions, influencing various fundamental processes in eukaryotes. These include, but are not limited to, cell signaling, protein trafficking, the epigenetic control of gene expression, and control of the cell cycle, as well as cell proliferation, differentiation, and interactions between cells. In this review, we discuss protein PTMs that play a key role in the malaria parasite biology and its pathogenesis. Phosphorylation, acetylation, methylation, lipidation and lipoxidation, glycosylation, ubiquitination and sumoylation, nitrosylation and glutathionylation, all of which occur in malarial parasites, are reviewed. We provide information regarding the biological significance of these modifications along all phases of the complex life cycle of Plasmodium spp. Importantly, not only the parasite, but also the host and vector protein PTMs are often crucial for parasite growth and development. In addition to metabolic regulations, protein PTMs can result in epitopes that are able to elicit both innate and adaptive immune responses of the host or vector. We discuss some existing and prospective results from antimalarial drug discovery trials that target various PTM-related processes in the parasite or host. [ABSTRACT FROM AUTHOR]

The eggs of the blood fluke Schistosoma mansoni are the main cause of the clinical manifestations of chronic schistosomiasis. After laying, the egg "winners" attach to the endothelium of the mesenteric vein and, after a period of development, induce the growth of a small granuloma, which facilitates their passage to the intestinal lumen. Egg "losers" carried by the bloodstream to non-specific tissues also undergo full development and induce large granuloma formation, but their life ends there. Although these trapped eggs represent a dead end in the parasite life cycle, the vast majority of studies attempting to describe the biology of the S. mansoni eggs have studied these liver-trapped "losers" instead of migrating intestinal "winners". This raises the fundamental question of how these eggs differ. With robust comparative transcriptomic analysis performed on S. mansoni eggs isolated 7 weeks post infection, we show that gene expression is critically dependent on tissue localization, both in the early and late stages of development. While mitochondrial genes and venom allergen-like proteins are significantly upregulated in mature intestinal eggs, well-described egg immunomodulators IPSE/alpha-1 and omega-1, together with micro-exon genes, are predominantly expressed in liver eggs. In addition, several proteases and protease inhibitors previously implicated in egg-host interactions display clear tissue-specific gene expression patterns. These major differences in gene expression could be then reflected in the observed different ability of liver and intestinal soluble egg antigens to elicit host immune responses and in the shorter viability of miracidia hatched from liver eggs. Our comparative analysis provides a new perspective on the biology of parasite's eggs in the context of their development and tissue localization. These findings could contribute to a broader and more accurate understanding of parasite eggs interactions with the host, which have historically been often restricted to liver eggs and sometimes inaccurately generalized. Author summary: The egg of Schistosoma mansoni is the main causative agent of pathologies of chronic schistosomiasis, a neglected tropical disease with a significant global health burden. Pathology is primarily caused by a high number of eggs being lodged in host organs over time, causing inflammation and tissue damage. While trapped "loser" eggs causing the disease have been extensively studied, little attention has been paid to the successful "winner" eggs that pass through the intestinal wall and are responsible for the continuation of the life cycle. Here we provide a comprehensive analysis of the gene expression profiles of the parasite's eggs based on their maturity and, most importantly, their tissue localization. We revealed that eggs differ substantially based on their tissue localization from an early development. In addition to genes associated with development and metabolism, we have described differential expression of known immunomodulators, which may be reflected by the differential capacity of eggs to elicit a host immune response. Furthermore, we demonstrate that genes encoding proteins potentially involved in host-parasite interactions exhibit clear tissue-dependent expression patterns. Our research provides novel insights into the biology of S. mansoni eggs and provides a robust data source for new avenues of investigation related to this important pathogen. [ABSTRACT FROM AUTHOR]

Toxoplasma, an important intracellular parasite of humans and animals, causes life-threatening toxoplasmosis in immunocompromised individuals. Although Toxoplasma secretory proteins during acute infection (tachyzoite, which divides rapidly and causes inflammation) have been extensively characterized, those involved in chronic infection (bradyzoite, which divides slowly and is surrounded by a cyst wall) remain uncertain. Regulation of the cyst wall is essential to the parasite life cycle, and polysaccharides, such as chitin, in the cyst wall are necessary to sustain latent infection. Toxoplasma secretory proteins during the bradyzoite stage may have important roles in regulating the cyst wall via polysaccharides. Here, we focused on characterizing the hypothetical T. gondii chitinase, chitinase-like protein 1 (TgCLP1). We found that the chitinaselike domain containing TgCLP1 is partially present in the bradyzoite microneme and confirmed, albeit partially, its previous identification in the tachyzoite microneme. Furthermore, although parasites lacking TgCLP1 could convert from tachyzoites to bradyzoites and make an intact cyst wall, they failed to convert from bradyzoites to tachyzoites, indicating that TgCLP1 is necessary for bradyzoite reactivation. Taken together, our findings deepen our understanding of the molecular basis of recrudescence and could contribute to the development of novel strategies for the control of toxoplasmosis. [ABSTRACT FROM AUTHOR]

Background: Plasmodium vivax represents the most geographically widespread human malaria parasite affecting civilian and military populations in endemic areas. Targeting the pre-erythrocytic (PE) stage of the parasite life cycle is especially appealing for developing P. vivax vaccines as it would prevent disease and transmission. Here, naturally acquired immunity to a panel of P. vivax PE antigens was explored, which may facilitate vaccine development and lead to a better understanding of naturally acquired PE immunity. Methods: Twelve P. vivax PE antigens orthologous to a panel of P. falciparum antigens previously identified as highly immunogenic in protected subjects after immunization with radiation attenuated sporozoites (RAS) were used for evaluation of humoral and cellular immunity by ELISA and IFN-γ ELISpot. Samples from P. vivax infected individuals (n = 76) from a low endemic malaria region in the Peruvian Amazon Basin were used. Results: In those clinical samples, all PE antigens evaluated showed positive IgG antibody reactivity with a variable prevalence of 58–99% in recently P. vivax diagnosed patients. The magnitude of the IgG antibody response against PE antigens was lower compared with blood stage antigens MSP1 and DBP-II, although antibody levels persisted better for PE antigens (average decrease of 6% for PE antigens and 43% for MSP1, p < 0.05). Higher IgG antibodies was associated with one or more previous malaria episodes only for blood stage antigens (p < 0.001). High IgG responders across PE and blood stage antigens showed significantly lower parasitaemia compared to low IgG responders (median 1,921 vs 4,663 par/µl, p < 0.05). In a subgroup of volunteers (n = 17),positive IFN-γ T cell response by ELISPOT was observed in 35% vs 9–35% against blood stage MSP1 and PE antigens, respectively, but no correlation with IgG responses. Conclusions: These results demonstrate clear humoral and T cell responses against P. vivax PE antigens in individuals naturally infected with P. vivax. These data identify novel attractive PE antigens suitable for use in the potential development and selection of new malaria vaccine candidates which can be used as a part of malaria prevention strategies in civilian and military populations living in P. vivax endemic areas. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii is an intracellular parasite that is important in medicine and veterinary science and undergoes distinct developmental transitions in its intermediate and definitive hosts. The switch between stages of T. gondii is meticulously regulated by a variety of factors. Previous studies have explored the role of the microrchidia (MORC) protein complex as a transcriptional suppressor of sexual commitment. By utilizing immunoprecipitation and mass spectrometry, constituents of this protein complex have been identified, including MORC, Histone Deacetylase 3 (HDAC3), and several ApiAP2 transcription factors. Conditional knockout of MORC or inhibition of HDAC3 results in upregulation of a set of genes associated with schizogony and sexual stages in T. gondii tachyzoites. Here, our focus extends to two primary ApiAP2s (AP2XII-1 and AP2XI-2), demonstrating their significant impact on the fitness of asexual tachyzoites and their target genes. Notably, the targeted disruption of AP2XII-1 and AP2XI-2 resulted in a profound alteration in merozoite-specific genes targeted by the MORC–HDAC3 complex. Additionally, considerable overlap was observed in downstream gene profiles between AP2XII-1 and AP2XI-2, with AP2XII-1 specifically binding to a subset of ApiAP2 transcription factors, including AP2XI-2. These findings reveal an intricate cascade of ApiAP2 regulatory networks involved in T. gondii schizogony development, orchestrated by AP2XII-1 and AP2XI-2. This study provides valuable insights into the transcriptional regulation of T. gondii growth and development, shedding light on the intricate life cycle of this parasitic pathogen. [ABSTRACT FROM AUTHOR]

The 6-cysteine protein family is one of the most abundant surface antigens that are expressed throughout the Plasmodium falciparum life cycle. Many members of the 6- cysteine family have critical roles in parasite development across the life cycle in parasite transmission, evasion of the host immune response and host cell invasion. The common feature of the family is the 6-cysteine domain, also referred to as s48/45 domain, which is conserved across Aconoidasida. This review summarizes the current approaches for recombinant expression for 6-cysteine proteins, monoclonal antibodies against 6- cysteine proteins that block transmission and the growing collection of crystal structures that provide insights into the functional domains of this protein family. [ABSTRACT FROM AUTHOR]

The WHO reported an estimated 249 million malaria cases and 608,000 malaria deaths in 85 countries in 2022. A total of 94% of malaria deaths occurred in Africa, 80% of which were children under 5. In other words, one child dies every minute from malaria. The RTS,S/AS01 malaria vaccine, which uses the Plasmodium falciparum circumsporozoite protein (CSP) to target sporozoite infection of the liver, achieved modest efficacy. The Malaria Vaccine Implementation Program (MVIP), coordinated by the WHO and completed at the end of 2023, found that immunization reduced mortality by only 13%. To further reduce malaria death, the development of a more effective malaria vaccine is a high priority. Three malaria vaccine targets being considered are the sporozoite liver infection (pre-erythrocytic stage), the merozoite red blood cell infection (asexual erythrocytic stage), and the gamete/zygote mosquito infection (sexual/transmission stage). These targets involve specific ligand-receptor interactions. However, most current malaria vaccine candidates that target two major parasite population bottlenecks, liver infection, and mosquito midgut infection, do not focus on such parasite ligands. Here, we evaluate the potential of newly identified parasite ligands with a phage peptide-display technique as novel malaria vaccine antigens. [ABSTRACT FROM AUTHOR]

Background: Pathogenic parasites are responsible for multiple diseases, such as malaria and Chagas disease, in humans and livestock. Traditionally, pathogenic parasites have been largely an evasive topic for vaccine design, with most successful vaccines only emerging recently. To aid vaccine design, the VIOLIN vaccine knowledgebase has collected vaccines from all sources to serve as a comprehensive vaccine knowledgebase. VIOLIN utilizes the Vaccine Ontology (VO) to standardize the modeling of vaccine data. VO did not model complex life cycles as seen in parasites. With the inclusion of successful parasite vaccines, an update in parasite vaccine modeling was needed. Results: VIOLIN was expanded to include 258 parasite vaccines against 23 protozoan species, and 607 new parasite vaccine-related terms were added to VO since 2022. The updated VO design for parasite vaccines accounts for parasite life stages and for transmission-blocking vaccines. A total of 356 terms from the Ontology of Parasite Lifecycle (OPL) were imported to VO to help represent the effect of different parasite life stages. A new VO class term, 'transmission-blocking vaccine,' was added to represent vaccines able to block infectious transmission, and one new VO object property, 'blocks transmission of pathogen via vaccine,' was added to link vaccine and pathogen in which the vaccine blocks the transmission of the pathogen. Additionally, our Gene Set Enrichment Analysis (GSEA) of 140 parasite antigens used in the parasitic vaccines identified enriched features. For example, significant patterns, such as signal, plasma membrane, and entry into host, were found in the antigens of the vaccines against two parasite species: Plasmodium falciparum and Toxoplasma gondii. The analysis found 18 out of the 140 parasite antigens involved with the malaria disease process. Moreover, a majority (15 out of 54) of P. falciparum parasite antigens are localized in the cell membrane. T. gondii antigens, in contrast, have a majority (19/24) of their proteins related to signaling pathways. The antigen-enriched patterns align with the life cycle stage patterns identified in our ontological parasite vaccine modeling. Conclusions: The updated VO modeling and GSEA analysis capture the influence of the complex parasite life cycles and their associated antigens on vaccine development. [ABSTRACT FROM AUTHOR]

Malaria continues to pose a serious global health threat, and artemisinin remains the core drug for global malaria control. However, the situation of malaria resistance has become increasingly severe due to the emergence and spread of artemisinin resistance. In recent years, significant progress has been made in understanding the mechanism of action (MoA) of artemisinin. Prior research on the MoA of artemisinin mainly focused on covalently bound targets that are alkylated by artemisinin‐free radicals. However, less attention has been given to the reversible noncovalent binding targets, and there is a paucity of information regarding artemisinin targets at different life cycle stages of the parasite. In this study, we identified the protein targets of artemisinin at different stages of the parasite's intraerythrocytic developmental cycle using a photoaffinity probe. Our findings demonstrate that artemisinin interacts with parasite proteins in vivo through both covalent and noncovalent modes. Extensive mechanistic studies were then conducted by integrating target validation, phenotypic studies, and untargeted metabolomics. The results suggest that protein synthesis, glycolysis, and oxidative homeostasis are critically involved in the antimalarial activities of artemisinin. In summary, this study provides fresh insights into the mechanisms underlying artemisinin's antimalarial effects and its protein targets. Highlights: We identified the targets of artemisinin at different stages of the intraerythrocytic cycle of Plasmodium falciparum using a photoaffinity probe of artemisinin.Artemisinin can interact with parasite proteins in vivo through both covalent and noncovalent mechanisms.Artemisinin may exert its antimalarial effects by interfering with the parasite's protein synthesis, glycolysis, and oxidative homeostasis pathways.This study provides fresh insights into the mechanisms underlying artemisinin's antimalarial effects and its protein targets. [ABSTRACT FROM AUTHOR]

The molecular revolution in parasitology has led to extraordinary achievements, but also to an excessive focus on technology. Molecular tools are not perfect; to use them effectively, we need to understand their limitations and weaknesses. 'Classical' (non-molecular) parasitology should not be underestimated and neglected, as this could have negative consequences, for example, misidentification of parasites. The opportunity to use modern methods to better understand parasite life cycles remains untapped, and long-term and risky ecological research is not supported. The integrative approach, that is, a combination of 'classical' and modern methods, is the only way to better understand the complexity of parasites. Research should focus on the complexity of the interactions of parasites with their hosts and the environment (One Health concept) in today's rapidly changing world. We live in the age of molecular biology and '-omics', and molecular methods have opened up unimagined possibilities for biological research, including parasitology. However, too one-sided a focus on new approaches can lead to major gaps as less 'cool' topics are neglected. Selected areas of research are briefly discussed to highlight the gaps caused by the current excessive focus on molecular and '-omics' methods. It is crucial to combine both 'classical' and modern methods without neglecting the complexity of the interactions of parasites with their hosts and the environment (One Health concept), which is even more urgent in today's rapidly changing world. Parasitologists should be more involved in field studies and multidisciplinary assessment of parasites. [ABSTRACT FROM AUTHOR]

The article discusses the role of parasite sex in the transmission and adaptation of Cryptosporidium, a leading cause of severe diarrheal disease worldwide. The research highlights the importance of parasite sex in fueling the spread of the disease and its ability to adapt to different hosts. The article also discusses the rise of cryptosporidiosis cases in the United States and the evolution of hypertransmissible strains. Laboratory studies have provided insights into the molecular details of the parasite's life cycle and the high frequency of mating, which contributes to the rapid spread of genes that confer parasite benefit. The article concludes by emphasizing the potential of understanding parasite sex as a powerful tool for discovering new insights into Cryptosporidium and its interaction with hosts. [Extracted from the article]

Cystic Echinococcosis (CE) as a prevalent tapeworm infection of human and herbivorous animals worldwide, is caused by accidental ingestion of Echinococcus granulosus eggs excreted from infected dogs. CE is endemic in the Middle East and North Africa, and is considered as an important parasitic zoonosis in Iran. It is transmitted between dogs as the primary definitive host and different livestock species as the intermediate hosts. One of the most important measures for CE control is dog deworming with praziquantel. Due to the frequent reinfection of dogs, intensive deworming campaigns are critical for breaking CE transmission. Dog reinfection rate could be used as an indicator of the intensity of local CE transmission in endemic areas. However, our knowledge on the extent of reinfection in the endemic regions is poor. The purpose of the present study was to determine E. granulosus reinfection rate after praziquantel administration in a population of owned dogs in Kerman, Iran. A cohort of 150 owned dogs was recruited, with stool samples collected before praziquantel administration as a single oral dose of 5 mg/kg. The re-samplings of the owned dogs were performed at 2, 5 and 12 months following initial praziquantel administration. Stool samples were examined microscopically using Willis flotation method. Genomic DNA was extracted, and E. granulosus sensu lato-specific primers were used to PCR-amplify a 133-bp fragment of a repeat unit of the parasite genome. Survival analysis was performed using Kaplan-Meier method to calculate cumulative survival rates, which is used here to capture reinfection dynamics, and monthly incidence of infection, capturing also the spatial distribution of disease risk. Results of survival analysis showed 8, 12 and 17% total reinfection rates in 2, 5 and 12 months following initial praziquantel administration, respectively, indicating that 92, 88 and 83% of the dogs had no detectable infection in that same time periods. The monthly incidence of reinfection in total owned dog population was estimated at 1.5% (95% CI 1.0–2.1). The results showed that the prevalence of echinococcosis in owned dogs, using copro-PCR assay was 42.6%. However, using conventional microscopy, 8% of fecal samples were positive for taeniid eggs. Our results suggest that regular treatment of the dog population with praziquantel every 60 days is ideal, however the frequency of dog dosing faces major logistics and cost challenges, threatening the sustainability of control programs. Understanding the nature and extent of dog reinfection in the endemic areas is essential for successful implementation of control programs and understanding patterns of CE transmission. Author summary: Cystic echinococcosis (CE), caused by the small tapeworm of dogs, Echinococcus granulosus, is considered as a prevalent zoonotic infection of human and livestock worldwide. Dogs play a crucial role in the parasite life cycle, serving as definitive hosts for the tapeworm and contributing to the transmission of the disease to humans and other livestock. Praziquantel (PZQ) dosing of dogs is proposed as one of the main elements of CE control programs. Frequent PZQ dosing is required because of reinfection of farm dogs following feeding offal to them and this presents major logistic and financial problems. The frequency of PZQ dosing is dependent on the rate of dog reinfection in each endemic region. We explored the significance of farm dogs reinfection in an endemic area in southeastern Iran, to provide better understanding of CE transmission and developing effective control programs. We monitored a cohort of 150 dogs previously treated for cystic echinococcosis by praziquantel. We showed that the prevalence of echinococcosis in the farm dogs, using PCR assay was 42.6% before praziquantel administration. To our surprise, a significant proportion of these dogs, approximately 17%, experienced reinfection with E. granulosus, 12 months following initial praziquantel administration. This finding was both alarming and informative. Our study emphasized the importance of responsible dog ownership behavior and proper disposal of livestock offal in endemic areas. [ABSTRACT FROM AUTHOR]

Host diversity has been shown to play a major role in shaping parasite diversity. However, it is less clear if this relationship holds across various hosts in complex life cycles, and what aspects of host diversity are regulating parasite diversity. This study tests whether host diversity at lower trophic levels (i.e., invertebrate hosts) is linked to parasite diversity at higher trophic levels (i.e., vertebrate hosts). This study then compares whether invertebrate host taxonomic vs. functional diversities have a greater influence on this relationship. To test this, fish parasites and macroinvertebrates were sampled from multiple sites along rivers in the New Jersey Pine Barrens. Upon examining diversity, macroinvertebrate taxonomic diversity was not significantly correlated with parasite diversity in vertebrate hosts. On the contrary, there was a significant positive correlation between macroinvertebrate functional feeding group diversity and parasite diversity in vertebrate hosts. These results suggest that intermediate host feeding strategies may serve as a means of structuring parasite communities in subsequent fish hosts. However, a better understanding of parasite life cycles in the N.J. Pine Barrens is necessary to support this hypothesis. [ABSTRACT FROM AUTHOR]

Various types of RNA modification in parasites affect parasite virulence, viability, differentiation and reproduction. Epitranscriptomics represents a novel regulatory layer and a new hallmark in parasites. Prediction of parasite homologs of the regulators of RNA modifications reported in humans facilitates prediction of the types of RNA modification and related regulators in parasites and elucidates the evolution of the parasitic epitranscriptional machinery. Multiple forms of crosstalk affect epitranscriptome variations and may be an important mechanism for the regulation of RNA modifications in parasites. RNA modifications of host cells are also an important component of the host–parasite interface, which is involved in the upregulation of host antiparasite immunity and influences the transmission and progression of parasitic diseases. RNA modifications (epitranscriptome) – such as N6-methyladenosine (m6A), 5-methylcytosine (m5C), and pseudouridine (Ψ) – modulate RNA processing, stability, interaction, and translation, thereby playing critical roles in the development, replication, virulence, metabolism, and life cycle adaptations of parasitic protozoa. Here, we summarize potential homologs of the major human RNA modification regulatory factors in parasites, outline current knowledge on how RNA modifications affect parasitic protozoa, highlight the regulation of RNA modifications and their crosstalk, and discuss current progress in exploring RNA modifications as potential drug targets. This review contributes to our understanding of epitranscriptomic regulation of parasitic protozoa biology and pathogenesis and provides new perspectives for the treatment of parasitic diseases. [ABSTRACT FROM AUTHOR]

Plasmodium sporozoite development in and egress from oocysts in the Anopheles mosquito remains largely enigmatic. In a previously performed high‐throughput knockout screen, the putative subunit 5 of the prefoldin complex (PbPCS5, PBANKA_0920100) was identified as essential for parasite development during mosquito and liver stage development. Here we generated and analyzed a PbPCS5 knockout parasite line during its development in the mosquito. Interestingly, PbPCS5 deletion does not significantly affect oocyst formation but leads to a growth defect resulting in aberrantly shaped sporozoites. Sporozoites produced in the absence of PbPCS5 were thinner, markedly elongated, and did, in most cases, not contain a nucleus. Sporozoites contained fewer subpellicular microtubules, which reached deep into the sporoblast during sporogony where they contacted and indented nuclei. These aberrantly shaped sporozoites did not reach the salivary glands, and we, therefore, conclude that PbPCS5 is essential for sporogony and the life cycle progression of the parasite during its mosquito stage. [ABSTRACT FROM AUTHOR]

An essential step in the life cycle of malaria parasites is their egress from hepatocytes, which enables the transition from the asymptomatic liver stage to the pathogenic blood stage of infection. To exit the liver, Plasmodium parasites first disrupt the parasitophorous vacuole membrane that surrounds them during their intracellular replication. Subsequently, parasite‐filled structures called merosomes emerge from the infected cell. Shrouded by host plasma membrane, like in a Trojan horse, parasites enter the vasculature undetected by the host immune system and travel to the lung where merosomes rupture, parasites are released, and the blood infection stage begins. This complex, multi‐step process must be carefully orchestrated by the parasite and requires extensive manipulation of the infected host cell. This review aims to outline the known signaling pathways that trigger exit, highlight Plasmodium proteins that contribute to the release of liver‐stage merozoites, and summarize the accompanying changes to the hepatic host cell. [ABSTRACT FROM AUTHOR]

Ion channels serve many cellular functions including ion homeostasis, volume regulation, signaling, nutrient acquisition, and developmental progression. Although the complex life cycles of malaria parasites necessitate ion and solute flux across membranes, the whole-genome sequencing of the human pathogen Plasmodium falciparum revealed remarkably few orthologs of known ion channel genes. Contrasting with this, biochemical studies have implicated the channel-mediated flux of ions and nutritive solutes across several membranes in infected erythrocytes. Here, I review advances in the cellular and molecular biology of ion channels in malaria parasites. These studies have implicated novel parasite genes in the formation of at least two ion channels, with additional ion channels likely present in various membranes and parasite stages. Computational approaches that rely on homology to known channel genes from higher organisms will not be very helpful in identifying the molecular determinants of these activities. Given their unusual properties, novel molecular and structural features, and essential roles in pathogen survival and development, parasite channels should be promising targets for therapy development. [ABSTRACT FROM AUTHOR]

Purpose: To study the morphology of all the life stages and the life cycle of the parasitic isopod Cymothoa indica Schioedte and Meinert, 1884, and its prevalence in different commercial fishes in the Chilika Lagoon, India. Methods: From April 2021 to December 2022, different life stages of the said parasites were collected from five different fish species from four localities of the Chilika Lagoon. The parasites were carefully removed from the host with a pair of forceps and then identified following standard literature and description. The morphology of all life stages, life cycle, and prevalence of the isopod C. indica were carried out as per the standard protocol. Results: A total of 1252 host fishes were analysed. The total prevalence was 44.5%. The host with the highest prevalence of C. indica was Glossogobius giuris, with 58.79% of the hosts parasitized by at least one parasite individual. Followed by the host Datnoides polota (50.98%), Johnius dussumieri (46.25%), Etroplus suratensis (28.45%) and the least prevalence was in Lates calcarifer (14.70%). The description of the life stages is based on the observation of 523 females, 116 males, 46 Juveniles, and 50 embryonic stages of the said parasites. Conclusion: The description of all the life stages and life cycles certainly helps to identify this species with clarity and ease in understanding the species' biology. In future, this study may be helpful in the effective management towards the improvement of aquaculture productivity. [ABSTRACT FROM AUTHOR]

Background: Babesia orientalis is an intra-erythrocytic protozoan parasite that causes babesiosis in water buffalo. The genome of B. orientalis has been reported and various genes have been accurately annotated, including heat shock proteins (HSP). Three B. orientalis HSPs (HSP90, HSP70 and HSP20) have been previously identified as potential antigenic targets. Here, a new validation strategy for the chaperone activities and cell protection characteristics of the three HSPs was developed in vitro. Methods: BoHSP20, BoHSP70 and BoHSP90B were amplified from cDNA, followed by cloning them into the pEGFP-N1 vector and transfecting the vector plasmid separately into 293T and Hela mammalian cells. Their expression and localization were determined by fluorescence microscopy. The biological functions and protein stability were testified through an analysis of the fluorescence intensity duration. Their role in the protection of cell viability from heat-shock treatments was examined by MTT assay (cell proliferation assay based on thiazolyl blue tetrazolium bromide). Results: Fusion proteins pEGFP-N1-BoHSP20, pEGFP-N1-BoHSP70, and pEGFP-N1-BoHSP90B (pBoHSPs: pBoHSP20; pBoHSP70 and pBoHSP90B) were identified as 47 kDa/97 kDa/118 kDa with a 27 kDa GFP tag, respectively. Prolonged fluorescent protein half-time was observed specifically in pBoHSPs under heat shock treatment at 55 °C, and BoHSP20 showed relatively better thermotolerance than BoHSP70 and BoHSP90B. Significant difference was found between pBoHSPs and controls in the cell survival curve after 2 h of 45 °C heat shock. Conclusion: Significant biological properties of heat stress-associated genes of B. orientalis were identified in eukaryote by a new strategy. Fusion proteins pBoHSP20, pBoHSP70 and pBoHSP90B showed good chaperone activity and thermo-stability in this study, implying that BoHSPs played a key role in protecting B. orientalis against heat-stress environment during parasite life cycle. In conclusion, the invitro model explored in this study provides a new way to investigate the biological functions of B. orientalis proteins during the host–parasite interaction. [ABSTRACT FROM AUTHOR]

Trypanosomes are single-celled extracellular parasites that infect mammals, including humans and livestock, causing global public health concerns and economic losses. These parasites cycle between insect vectors, such as tsetse flies and vertebrate hosts, undergoing morphological, cellular, and biochemical changes. They have remarkable immune evasion mechanisms to escape the host's innate and adaptive immune responses, such as surface coat antigenic variation and the induction of the loss of specificity and memory of antibody responses, enabling the prolongation of infection. Since trypanosomes circulate through the host body in blood and lymph fluid and invade various organs, understanding the interaction between trypanosomes and tissue niches is essential. Here, we present an up-to-date overview of host–parasite interactions and survival strategies for trypanosomes by introducing and discussing the latest studies investigating the transcriptomics of parasites according to life cycle stages, as well as host cells in various tissues and organs, using single-cell and spatial sequencing applications. In recent years, this information has improved our understanding of trypanosomosis by deciphering the diverse populations of parasites in the developmental process, as well as the highly heterogeneous immune and tissue-resident cells involved in anti-trypanosome responses. Ultimately, the goal of these approaches is to gain an in-depth understanding of parasite biology and host immunity, potentially leading to new vaccination and therapeutic strategies against trypanosomosis. [ABSTRACT FROM AUTHOR]

Opisthorchis viverrini is a parasitic liver fluke contracted by consumption of raw fish, which affects over 10 million people in Southeast Asia despite sustained control efforts. Chronic infections are a risk factor for the often fatal bile duct cancer, cholangiocarcinoma. Previous modeling predicted rapid elimination of O. viverrini following yearly mass drug administration (MDA) campaigns. However, field data collected in affected populations shows persistence of infection, including heavy worm burden, after many years of repeated interventions. A plausible explanation for this observation is systematic adherence of individuals in health campaigns, such as MDA and education, with some individuals consistently missing treatment. We developed an agent-based model of O. viverrini which allows us to introduce various heterogeneities including systematic adherence to MDA and education campaigns at the individual level. We validate the agent-based model by comparing it to a previously published population-based model. We estimate the degree of systematic adherence to MDA and education campaigns indirectly, using epidemiological data collected in Lao PDR before and after 5 years of repeated MDA, education and sanitation improvement campaigns. We predict the impact of interventions deployed singly and in combination, with and without the estimated systematic adherence. We show how systematic adherence can substantially increase the time required to achieve reductions in worm burden. However, we predict that yearly MDA campaigns alone can result in a strong reduction of moderate and heavy worm burden, even under systematic adherence. We predict latrines and education campaigns to be particularly important for the reduction in overall prevalence, and therefore, ultimately, elimination. Our findings show how systematic adherence can explain the observed persistence of worm burden; while emphasizing the benefit of interventions for the entire population, even under systematic adherence. At the same time, the results highlight the substantial opportunity to further reduce worm burden if patterns of systematic adherence can be overcome. Author summary: Opisthorchis viverrini is a liver parasite contracted by consumption of raw fish which affects over 10 million people in Southeast Asia despite sustained control efforts, with heavy infections being a major risk factor for often fatal bile duct cancer. Efforts to reduce the worm burden in populations include education of individuals on the safe consumption of fish meals, improvement of sanitation to interrupt the parasitic life cycle and mass drug administration campaigns, where ideally all targeted individuals of a population receive a drug against the parasite without a prior test of whether they are infected. We present a new agent-based mathematical model of Opisthorchis viverrini transmission and control. This model allows us to include systematic adherence to interventions, which describes the pattern of some individuals consistently being missed by interventions aimed at an entire (sub-)population. Our findings show how systematic adherence can explain the observed persistence of worm burden, but how interventions can be greatly beneficial even for individuals that systematically miss treatment. At the same time, we show the substantial opportunity to further reduce worm burden if patterns of systematic adherence can be overcome. [ABSTRACT FROM AUTHOR]

Malaria is a deadly disease caused by protozoan pathogens of the Plasmodium parasite. Transmission to humans occurs through the bite of an infected female Anopheles mosquito. According to the World Health Organization (WHO), an estimated 247 million cases of malaria were recorded worldwide in 2021, with approximately 619 000 malaria deaths. The initial signs of malaria can be mild and challenging to diagnose due to the signs and symptoms being similar to those of other illnesses. The malaria burden remains largely concentrated in the WHO sub-Saharan African region and has been recognised as a significant contributor to morbidity and mortality. This review aims to contribute to the existing knowledge on malaria in South Africa, a region within sub-Saharan Africa, focusing on the epidemiology and life cycle of the malaria parasite as well as diagnostic approaches for detecting malaria. In addition, nonpharmacological and pharmacological interventions for treating and preventing malaria infections will also be discussed herein. While there has been a significant reduction in the global burden of this disease, malaria remains a public health issue in South Africa. As such, the implementation of effective preventative measures and strategies, early diagnosis, and appropriate treatment regimens are crucial to reducing the malaria burden in South Africa. [ABSTRACT FROM AUTHOR]

Simple Summary: Hypersaline lakes provide services like nest sites and food for migratory waterbirds. The crustacean Artemia (brine shrimp) is an intermediate host for many cestode parasites completing their life cycle in multiple waterbirds (flamingos, grebes, waders, and ducks) associated with the hypersaline lakes. The molecular identification of larval stages of six cestodes in Artemia from three countries correlated well with the morphology of advanced larval stages. This study provides the first parasite molecular data on Artemia-cestode-bird relationships to facilitate our understanding of food web dynamics and service provisioning in hypersaline lakes, which are threatened by human-caused perturbations, including climate change. Determining molecular markers for parasites provides a useful tool for their identification, particularly for larval stages with few distinguishable diagnostic characters. Avian cestodes play a key role in the food webs and biodiversity of hypersaline wetlands, yet they remain understudied. Using naturally infected Artemia, we identified cestode larvae (cysticercoids), assessed their genetic diversity, and explored phylogenetic relationships in relation to larval morphology and waterbird final hosts. We obtained partial 18S rDNA sequences for 60 cysticercoids of the family Hymenolepidae infecting Artemia spp. from seven localities and three countries (Spain, the USA, and Chile). We present the first DNA sequences for six taxa: Confluaria podicipina, Fimbriarioides sp., Flamingolepis liguloides, Flamingolepis sp. 1, Flamingolepis sp. 2, and Hymenolepis californicus. Intraspecific sequence variation (0.00–0.19% diversity) was lower than intergroup genetic distance (0.7–14.75%). Phylogenetic analysis revealed three main clades: 1—Flamingolepis, 2—Fimbriarioides, 3—Confluaria and Hymenolepis, all of which separated from hymenolepidids from mammals and terrestrial birds. This clear separation among taxa is congruent with previous morphological identification, validating the 18S gene as a useful marker to discriminate at generic/species level. Working with intermediate hosts allows the expansion of knowledge of taxonomic and genetic diversity of cestodes in wildlife, as well as elucidation of their life cycles. [ABSTRACT FROM AUTHOR]

Glycosylphosphatidylinositols (GPIs) are crucial for Plasmodium falciparum development and survival in both mosquitos and humans. The reaction steps of GPI precursor biosynthesis and GPI biosynthesis of malaria parasites are important targets for drug development. In P. falciparum , β- O -GlcNAcylation occurs in functionally important protein classes, such as transcription factors, energy metabolism pathway, stress response, cytoskeletal motor assembly and signaling, and thus it may have vital functional relevance. Fucosylation and tryptophan C -mannosylation of the TRAP family of proteins of P. falciparum are essential for the protein structural stability, gametocyte egress, ookinete and oocyst development, sporogenesis, and infection transmission. In-depth knowledge of structure and functions of parasite glycans may provide important targets for the development of effective vaccines and/or drugs for malaria. In malaria parasites, although post-translational modification of proteins with N -. O -, and C -glycosidic bond-linked glycans is limited, it is confined to relatively fewer proteins in which the glycans are present at significant levels and may have important functions. Furthermore, several proteins are modified with glycosylphosphatidylinositols (GPIs) which represent the predominant glycan synthesized by parasites. Modification of proteins with GPIs is obligatory for parasite survival as GPI-anchored proteins (GPI-APs) play essential roles in all life cycle stages of the parasites, including development, egress, gametogenesis, motility, and host cell adhesion and invasion. Here, we discuss the current knowledge on the structures and potential functions of the glycan moieties of parasite proteins. The knowledge has important implications for the development of drugs and vaccines for malaria. In malaria parasites, although post-translational modification of proteins with N -. O -, and C -glycosidic bond-linked glycans is limited, it is confined to relatively fewer proteins in which the glycans are present at significant levels and may have important functions. Furthermore, several proteins are modified with glycosylphosphatidylinositols (GPIs) which represent the predominant glycan synthesized by parasites. Modification of proteins with GPIs is obligatory for parasite survival as GPI-anchored proteins play essential roles in all life cycle stages of the parasites, including development, egress, gametogenesis, motility, and host cell adhesion and invasion. Here, we discuss the current knowledge on the structures and potential functions of the glycan moieties of parasite proteins. The knowledge has important implications for the development of drugs and vaccines for malaria. [ABSTRACT FROM AUTHOR]

The epidemiology of Strongyloides stercoralis is briefly reviewed with an emphasis on cross section and longitudinal studies of infection prevalence stratified by age, performance of different diagnostic tools, mass drug administration (MDA) impact and estimates of key population parameters within the complex life cycle of the parasite that determine transmission intensity and response to control measures. The paucity of studies is highlighted, and gaps in current knowledge identified about the population biology of this very prevalent infection in tropical and sub-tropical regions around the world. A stochastic individual based stochastic model is described in part to highlight gaps in knowledge. The impact of repeated MDA is simulated to illustrate some aspects of transmission dynamics of this helminth infection. Specifically, the impact and bounce back times in either the intervals between treatment rounds, or post the cessation of treatment, depend critically on the magnitude of two distinct components of the basic reproductive number R0. The absence of data on these key components is highlighted, as is the value of studies of longitudinal cohorts of people in regions of endemic infection post rounds of MDA to record how infection levels bounce back post treatment at individual and population levels of study. This article is part of the Theo Murphy meeting issue 'Strongyloides: omics to worm-free populations'. [ABSTRACT FROM AUTHOR]

Simple Summary: The neglected tropical disease schistosomiasis is a worm infection that is caused by parasitic blood flukes. The disease is found in sub-Saharan Africa, the Middle East, Southeast Asia, and the New World. Worldwide, 240 million people are infected, and 700 million people are at risk. Schistosomiasis is a debilitating, chronic disease, and the mortality is estimated at 200,000 deaths per year. Schistosomiasis control relies on the drug praziquantel, but it does not prevent reinfection after treatment. The development of new vaccines, drugs, and diagnostic methods and the investigation of the biological basis of infectivity are, therefore, of critical importance. The development of transgenesis systems, as have been used for other pathogens, has been hampered by the complexity of the parasite and its life cycle. Now, after 25 years, tools to genetically manipulate the parasite have developed to the point that we can analyse this parasite's biology in detail. In this review, we describe the progress that has been made in establishing gene manipulation in schistosomes and discuss numerous examples of studies that have led to a better understanding of molecules that are essential for host–parasite interplay. This insight will lead to new strategies and methods for combatting and managing schistosomiasis. As research on parasitic helminths has entered the post-genomic era, research efforts have turned to deciphering the function of genes in the public databases of genome sequences. It is hoped that, by understanding the role of parasite genes in maintaining their parasitic lifestyle, critical insights can be gained to develop new intervention and control strategies. Methods to manipulate and transform parasitic worms are now developed to a point where it has become possible to gain a comprehensive understanding of the molecular mechanisms underlying host–parasite interplay, and here, we summarise and discuss the advances that have been made in schistosome transgenesis over the past 25 years. The ability to genetically manipulate schistosomes holds promise in finding new ways to control schistosomiasis, which ultimately may lead to the eradication of this debilitating disease. [ABSTRACT FROM AUTHOR]

Schistosomes are blood flukes with specialised tissues and organs, each one playing a pivotal role in perpetuating the parasite life cycle. Herein, we describe a detailed methodology for preserving the proteome of adult Schistosoma mansoni worms during manual dissection for enrichment of tissues associated with the parasite's alimentary tract. We provide step‐by‐step directions for specimen storage and dissection while in preservative solution, tissue homogenisation, protein extraction and digestion using a methodology fully compatible with downstream quantitative liquid chromatography–mass spectrometry analysis. Our methodology uses label‐free and QconCAT‐based absolute quantification for detection of S. mansoni oesophageal gland products proposed as vaccine candidates. Through stabilisation of the proteome and minimising sample degradation during dissection our approach has allowed us to access the hidden proteome of target tissues not readily available from total lysates because of their small volume. This protocol can be replicated or adapted to other Schistosoma species lacking quantitative proteomics characterisation of specialised tissues for discovery of proteins with potential diagnostic and therapeutic utility. [ABSTRACT FROM AUTHOR]

Chagas disease is a neglected infectious disease caused by the protozoan Trypanosoma cruzi, primarily transmitted by triatomine vectors, and it threatens approximately seventy-five million people worldwide. This parasite undergoes a complex life cycle, transitioning between hosts and shifting from extracellular to intracellular stages. To ensure its survival in these diverse environments, T. cruzi undergoes extreme morphological and molecular changes. The metacyclic trypomastigote (MT) form, which arises from the metacyclogenesis (MTG) process in the triatomine hindgut, serves as a crucial link between the insect and human hosts and can be considered the starting point of Chagas disease. This review provides an overview of the current knowledge regarding the parasite's life cycle, molecular pathways, and mechanisms involved in metabolic and morphological adaptations during MTG, enabling the MT to evade the immune system and successfully infect human cells. [ABSTRACT FROM AUTHOR]

Background: Infections of cats with Echinococcus granulosus is uncommon because the cat is not part of the parasite life cycle that a carnivorous and another herbivore represent. Nevertheless, it occurs incidentally when eating food or drinking water contaminated with the worm's larva, especially with the presence of the definitive host (dogs), in this case, the infections are concentrated in stray or outside cats. For this reason, this study examined the possibility of cat infection with E. granulosus and diagnosed the common genotype of this infection. Objective: This study examined the possibility of cat infection with E. granulosus and diagnosed the common genotype of this infection. Methods: Four of the 37 cats that had died in different accidents developed cystic echinococcosis (CE). The cytochrome c oxidase subunit I (COX1) gene was initially amplified and sequenced to determine if these cysts belonged to E. granulosus, in beginning. The DNA fragments resulting from sequencing were then compared and aligned with other sequences using the Gene Bank database. Finally, a phylogenetic tree was drawn according to the sequence data obtained from cox1 genes sequencing, and the MEGA 7.0 phylogenetic analysis program was utilized. Results: Four different sequences were deposited in the Gen Bank with accession numbers (ON795961 to ON795964), all of which belong to the G1 genotype. Approximately 84% and 100% of these sequences aligned with G1 (AB622277.1) and G1 (MG722980.1), respectively. Conclusions: G1 is the dominant genotype that causes cat infections, even though the cat's EC infection was incidental. [ABSTRACT FROM AUTHOR]