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We describe a new species of the genus Alippistrongylus (Nematoda: Trichostrongylina) in the small intestine of Delomys dorsalis (Rodentia: Sigmodontinae) found in Itatiaia National Park (Parque Nacional do Itatiaia, PNI), state of Rio de Janeiro, Brazil. The new species can be distinguished between the other two species previously described by the following morphological characteristics: pattern of the rays from the caudal bursa rays in males and the spherical-shaped appendage in the female. Additionally, we provide molecular genetic data from the new species. The discovery of Alippistrongylus itatiaiaensis sp. n. expands our understanding of nematode diversity and underscores the importance of parasite studies in biodiversity conservation. Its occurrence in a preserved area like the PNI emphasises the role of such habitats in maintaining unique ecological interactions.

Ascaris is one of the most widespread helminth infections, leading to chronic morbidity in humans and considerable economic losses in pig farming. In addition, pigs are an important reservoir for the zoonotic salmonellosis , where pigs can serve as asymptomatic carriers. Here, we investigated the impact of an ongoing Ascaris infection on the immune response to Salmonella in pigs. We observed higher bacterial burdens in experimentally coinfected pigs compared to pigs infected with Salmonella alone. The impaired control of Salmonella in the coinfected pigs was associated with repressed interferon gamma responses in the small intestine and with the alternative activation of gut macrophages evident in elevated CD206 expression. Ascaris single and coinfection were associated with a rise of CD4 - CD8α + FoxP3 + Treg in the lymph nodes draining the small intestine and liver. In addition, macrophages from coinfected pigs showed enhanced susceptibility to Salmonella infection in vitro and the Salmonella- induced monocytosis and tumor necrosis factor alpha production by myeloid cells was repressed in pigs coinfected with Ascaris . Hence, our data indicate that acute Ascaris infection modulates different immune effector functions with important consequences for the control of tissue-invasive coinfecting pathogens.IMPORTANCEIn experimentally infected pigs, we show that an ongoing infection with the parasitic worm Ascaris suum modulates host immunity, and coinfected pigs have higher Salmonella burdens compared to pigs infected with Salmonella alone. Both infections are widespread in pig production and the prevalence of Salmonella is high in endemic regions of human Ascariasis, indicating that this is a clinically meaningful coinfection. We observed the type 2/regulatory immune response to be induced during an Ascaris infection correlates with increased susceptibility of pigs to the concurrent bacterial infection., Competing Interests: The authors declare no conflict of interest.

Background: Macracanthorhynchus hirudinaceus (Pallas, 1781) is a zoonotic acanthocephalan that parasitizes the small intestine of wild boars. It is a pathogenic that causes economic losses, and poses a public health threat due to increased emergence., Purpose: The aims of this study is describes histopathologically the damage caused by M. hirudinaceus in the small intestine of wild boar Sus scrofa Linnaeus, 1758, and molecularly characterize this parasite (sequences, haplotypes, phylogeny) for the first time in Elazig city, Türkiye., Results: A high prevalence of infection was obtained. Upon separating the worms, it was discovered that there were ulcers resembling craters in the center, of the small intestine mucosa, surrounded by edema. The intestine wall where the parasite attached was damaged, with the villi epithelium and lamina propria in the mucosa being destroyed. The genomic DNA was isolated from all M. hirudinaceus samples, and PCR amplified the 489 bp gene fragments were sequenced and confirmed that all 21 sequences were M. hirudinaceus. The haplotype analysis of the sequences revealed the presence of a central star-shaped haplotype, in addition to four other haplotypes., Conclusion: After conducting sequence analysis, the genetic differences between the M. hirudinaceus sequences obtained in this study and those reported from Europe and Japan suggest that this parasite is endemic to Türkiye's local wild boar population. Also, four haplotypes were identified, distinguishing it from other haplotypes by 1-5 mutation steps. It is essential to consider the worm's sequences and the formation of haplotypes, since these intrinsic characteristics may impact in the epidemiology and pathology of the worm in the future., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Infection with the apicomplexan parasite Cryptosporidium is a leading cause of diarrheal disease. Cryptosporidiosis is of particular importance in infants and shows a strong association with malnutrition, both as a risk factor and as a consequence. Cryptosporidium invades and replicates within the small intestine epithelial cells. This is a highly dynamic tissue that is developmentally stratified along the villus axis. New cells emerge from a stem cell niche in the crypt and differentiate into mature epithelial cells while moving toward the villus tip, where they are ultimately shed. Here, we studied the impact of Cryptosporidium infection on this dynamic architecture. Tracing DNA synthesis in pulse-chase experiments in vivo, we quantified the genesis and migration of epithelial cells along the villus. We found proliferation and epithelial migration to be elevated in response to Cryptosporidium infection. Infection also resulted in significant cell loss documented by imaging and molecular assays. Consistent with these observations, single-cell RNA sequencing of infected intestines showed a gain of young and a loss of mature cells. Interestingly, enhanced epithelial cell loss was not a function of enhanced apoptosis of infected cells. To the contrary, Cryptosporidium -infected cells were less likely to be apoptotic than bystanders, and experiments in tissue culture demonstrated that infection provided enhanced resistance to chemically induced apoptosis to the host but not bystander cells. Overall, this study suggests that Cryptosporidium may modulate cell apoptosis and documents pronounced changes in tissue homeostasis due to parasite infection, which may contribute to its long-term impact on the developmental and nutritional state of children., Importance: The intestine must balance its roles in digestion and nutrient absorption with the maintenance of an effective barrier to colonization and breach by numerous potential pathogens. An important component of this balance is its constant turnover, which is modulated by a gain of cells due to proliferation and loss due to death or extrusion. Here, we report that Cryptosporidium infection changes the dynamics of this process increasing both gain and loss of enterocytes speeding up the villus elevator. This leads to a much more immature epithelium and a reduction of the number of those cells typically found toward the villus apex best equipped to take up key nutrients including carbohydrates and lipids. These changes in the cellular architecture and physiology of the small intestine may be linked to the profound association between cryptosporidiosis and malnutrition., Competing Interests: The authors declare no conflict of interest.

Recognizing that enteric tuft cells can signal the presence of nematode parasites, we investigated whether tuft cells are required for the expulsion of the cestode, Hymenolepis diminuta, from the non-permissive mouse host, and in concomitant anti-helminthic responses. BALB/c and C57BL/6 mice infected with H. diminuta expelled the worms by 11 days post-infection (dpi) and displayed DCLK1+ (doublecortin-like kinase 1) tuft cell hyperplasia in the small intestine (not the colon) at 11 dpi. This tuft cell hyperplasia was dependent on IL-4Rα signalling and adaptive immunity, but not the microbiota. Expulsion of H. diminuta was slowed until at least 14 dpi, but not negated, in tuft cell-deficient Pou2f3-/- mice and was accompanied by delayed goblet cell hyperplasia and slowed small bowel transit. Worm antigen and mitogen evoked production of IL-4 and IL-10 by splenocytes from wild-type and Pou2f3-/- mice was not appreciably different, suggesting similar systemic immune reactivity to infection with H. diminuta. Wild-type and Pou2f3-/- mice infected with H. diminuta displayed partial protection against subsequent infection with the nematode Heligmosomoides bakeri. We speculate that, with respect to H. diminuta, enteric tuft cells are important for local immune events driving the rapidity of H. diminuta expulsion but are not critical in initiating or sustaining systemic Th2 responses that provide concomitant immunity against secondary infection with H. bakeri., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Rajeev et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Epithelial cells secrete chloride to regulate water release at mucosal barriers, supporting both homeostatic hydration and the "weep" response that is critical for type 2 immune defense against parasitic worms (helminths). Epithelial tuft cells in the small intestine sense helminths and release cytokines and lipids to activate type 2 immune cells, but whether they regulate epithelial secretion is unknown. Here, we found that tuft cell activation rapidly induced epithelial chloride secretion in the small intestine. This response required tuft cell sensory functions and tuft cell-derived acetylcholine (ACh), which acted directly on neighboring epithelial cells to stimulate chloride secretion, independent of neurons. Maximal tuft cell-induced chloride secretion coincided with immune restriction of helminths, and clearance was delayed in mice lacking tuft cell-derived ACh, despite normal type 2 inflammation. Thus, we have uncovered an epithelium-intrinsic response unit that uses ACh to couple tuft cell sensing to the secretory defenses of neighboring epithelial cells., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The concentration of immunoglobulin (Ig) E is the lowest among serum Igs, but it can induces type I hypersensitivity and plays an important role in anti-parasitic infection. The present study aimed to explore the residence characteristics of IgE + cells in the sheep small intestine and the impact of Moniezia benedeni infection on them. The recombinant plasmids pET-28a-IgE were constructed and induced and expressed in Escherichia coli. BL21 (DE3). The rabbit anti-sheep IgE polyclonal antibody was prepared using the obtained recombinant protein as antigen. Finally, the levels of IgE + cells in the small intestine of healthy (Control group) and naturally M. benedeni-infected (Infected group) sheep were detected analyzed. The results showed that the rabbit anti-sheep IgE polyclonal antibody with good immunogenicity (titer = 1: 128000) could specifically bind to the heavy chain of natural sheep IgE. In the Control group, the IgE + cells were mainly distributed in lamina propria of the small intestine, and the densities were significantly decreased from duodenum to ileum (P<0.05), with respective values of (4.28 cells / 10 4 μm 2 , 1.80 cells / 10 4 μm 2 , and 1.44 cells / 10 4 μm 2 in duodenum, jejunum, and ileum. In the Infected group, IgE + cells density were 6.26 cells / 10 4 μm 2 , 3.01 cells / 10 4 μm 2 , and 2.09 cells / 10 4 μm 2 in duodenum, jejunum and ileum respectively, which were significantly higher in all segments compared to the Control group (P<0.05), increasing by 46.26%, 67.22% and 45.14%, respectively. In addition, compared with the Control group, the IgE protein levels were significantly increased in all intestinal segments of the Infected group (P<0.01), however, there was no significant differences among the different intestinal segments within the same group (P>0.05). The results demonstrated that M. benedeni infection could significantly increase the content of IgE and the distribution density of its secreting cells in sheep small intestine. The intestinal mucosal immune system of sheep presented obvious specificity against M. benedeni infection. This lays a good foundation for further exploring molecular mechanisms of the intestinal mucosal immune system monitoring and responding to M. benedeni infection., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Coccidiosis is a rapidly spreading and acute parasitic disease that seriously threatening the intestinal health of poultry. Matrine from leguminous plants has anthelmintic and anti-inflammatory properties., Purpose: This assay was conducted to explore the protective effects of Matrine and the AntiC (a Matrine compound) on Eimeria necatrix (EN)-infected chick small intestines and to provide a nutritional intervention strategy for EN injury., Study Design: The in vivo (chick) experiment: A total of 392 one-day-old yellow-feathered broilers were randomly assigned to six groups in a 21-day study: control group, 350 mg/kg Matrine group, 500 mg/kg AntiC group, EN group, and EN + 350 mg/kg Matrine group, EN + 500 mg/kg AntiC group. The in vitro (chick intestinal organoids, IOs): The IOs were treated with PBS, Matrine, AntiC, 3 μM CHIR99021, EN (15,000 EN sporozoites), EN + Matrine, EN + AntiC, EN + Matrine + CHIR99021, EN + AntiC + CHIR99021., Methods: The structural integrity of chicks jejunal crypt-villus axis was evaluated by hematoxylin and eosin (H&E) staining and transmission electron microscopy (TEM). And the activity of intestinal stem cells (ISCs) located in crypts was assessed by in vitro expansion advantages of a primary in IOs model. Then, the changes of Wnt/β-catenin signaling in jejunal tissues and IOs were detected by Real-Time qPCR,Western blotting and immunohistochemistry., Results: The results showed that dietary supplementation with Matrine or AntiC rescued the jejunal injury caused by EN, as indicated by increased villus height, reduced crypt hyperplasia, and enhanced expression of tight junction proteins. Moreover, there was less budding efficiency of the IOs expanded from jejunal crypts of chicks in the EN group than that in the Matrine and AntiC group, respectively. Further investigation showed that AntiC and Matrine inhibited EN-stimulated Wnt/β-catenin signaling. The fact that Wnt/β-catenin activation via CHIR99021 led to the failure of Matrine and AntiC to rescue damaged ISCs confirmed the dominance of this signaling., Conclusion: Our results suggest that Matrine and AntiC inhibit ISC proliferation and promote ISC differentiation into absorptive cells by preventing the hyperactivation of Wnt/β-catenin signaling, thereby standardizing the function of ISC proliferation and differentiation, which provides new insights into mitigating EN injury by Matrine and AntiC., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Increased permeability of the intestinal epithelial layer is linked to the pathogenesis and perpetuation of a wide range of intestinal and extra-intestinal diseases. Infecting humans with controlled doses of helminths, such as human hookworm (termed hookworm therapy), is proposed as a treatment for many of the same diseases. Helminths induce immunoregulatory changes in their host which could decrease epithelial permeability, which is highlighted as a potential mechanism through which helminths treat disease. Despite this, the influence of a chronic helminth infection on epithelial permeability remains unclear. This study uses the chronically infecting intestinal helminth Heligmosomoides polygyrus to reveal alterations in the expression of intestinal tight junction proteins and epithelial permeability during the infection course. In the acute infection phase (1 week postinfection), an increase in intestinal epithelial permeability is observed. Consistent with this finding, jejunal claudin-2 is upregulated and tricellulin is downregulated. By contrast, in the chronic infection phase (6 weeks postinfection), colonic claudin-1 is upregulated and epithelial permeability decreases. Importantly, this study also investigates changes in epithelial permeability in a small human cohort experimentally challenged with the human hookworm, Necator americanus. It demonstrates a trend toward small intestinal permeability increasing in the acute infection phase (8 weeks postinfection), and colonic and whole gut permeability decreasing in the chronic infection phase (24 weeks postinfection), suggesting a conserved epithelial response between humans and mice. In summary, our findings demonstrate dynamic changes in epithelial permeability during a chronic helminth infection and provide another plausible mechanism by which chronic helminth infections could be utilized to treat disease., (© 2024 The Authors. Immunology & Cell Biology published by John Wiley & Sons Australia, Ltd on behalf of the Australian and New Zealand Society for Immunology, Inc.)

Purpose: Using molecular techniques, we have previously shown that carnivorous mammals of the family Mustelidae might be common definitive hosts for various protozoan Sarcocystis species. In the present study we aimed to unravel whether Sarcocystis species using ungulates as intermediate hosts and canids or felids as definitive hosts can be found in intestine of mustelids., Methods: Small intestine samples of 93 individual mustelids of five different species from Lithuania were examined. Sarcocystis species were identified based on species-specific PCR and subsequent cox1 sequencing., Results: Six Sarcocystis species (S. arieticanis, S. bertrami, S. capracanis, S. capreolicanis, S. linearis and S. morae) defined by ungulate-canid life cycle were detected for the first time in small intestines of mustelids. By contrast, the prevalence of Sarcocystis characterised by ungulate-felid life cycle was low (3.2%). Overall, 76% of the examined animals were positive for at least one of the studied Sarcocystis species. Four species, S. arieticanis, S. bertrami, S. capracanis and S. morae were most commonly found, with the detection rate of about 40%., Conclusions: The current finding, in addition to our previous studies, suggests that mustelids play an important role in the spread of various Sarcocystis species., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Teniarynchosis (taeniasis) is a helminthiasis with chronic course, predominant lesion of the upper gastrointestinal tract and active release of segments. The causative agent of taeniasis is Taenia saginata. This disease is widespread, although endemic countries are Africa, Australia, South America and Asia. Cases of teniarynchosis are rare in surgical practice. We present a patient who admitted with blunt abdominal injury and isolated wound of small. After laparotomy, we found alive bovine tapeworm that entered into abdominal cavity through the wound in small intestine. The parasite was removed with subsequent intestinal wall repair. The patient was discharged. Thus, teniarynchosis can cause urgent surgical diseases or their complications, especially in endemic areas.

Infection with the protozoan parasite Giardia duodenalis (syn. intestinalis , lamblia ) has been associated with intestinal mucus disruptions and microbiota dysbiosis. The mechanisms remain incompletely understood. Mucus consists primarily of densely glycosylated mucin glycoproteins. Mucin O -glycans influence mucus barrier properties and mucin-microbe interactions and are frequently altered during disease. In this study, we observed time-dependent and regiospecific alterations to intestinal mucin glycosylation patterns and the expression of mucin-associated glycosyltransferase genes during Giardia infection. Glycosylation alterations were observed in Giardia -infected mice in the upper small intestine, the site of parasite colonization, and in the distal colon, where active trophozoites were absent. Alterations occurred as early as day 2 post-infection and persisted in mice after parasite clearance. We also observed small intestinal goblet cell hyperplasia and thinning of the distal colon mucus barrier during early infection, and microbiota alterations and altered production of cecal SCFAs. Giardia -induced alterations to mucin glycosylation were at least in part dependent on microbiota dysbiosis, as transplantation of a dysbiotic mucosal microbiota collected from Giardia -infected mice recapitulated some alterations. This study describes a novel mechanism by which Giardia alters intestinal mucin glycosylation, and implicates the small intestinal microbiota in regulation of mucin glycosylation patterns throughout the gastrointestinal tract.

Helminth parasites are adept manipulators of the immune system, using multiple strategies to evade the host type 2 response. In the intestinal niche, the epithelium is crucial for initiating type 2 immunity via tuft cells, which together with goblet cells expand dramatically in response to the type 2 cytokines IL-4 and IL-13. However, it is not known whether helminths modulate these epithelial cell populations. In vitro, using small intestinal organoids, we found that excretory/secretory products (HpES) from Heligmosomoides polygyrus blocked the effects of IL-4/13, inhibiting tuft and goblet cell gene expression and expansion, and inducing spheroid growth characteristic of fetal epithelium and homeostatic repair. Similar outcomes were seen in organoids exposed to parasite larvae. In vivo, H. polygyrus infection inhibited tuft cell responses to heterologous Nippostrongylus brasiliensis infection or succinate, and HpES also reduced succinate-stimulated tuft cell expansion. Our results demonstrate that helminth parasites reshape their intestinal environment in a novel strategy for undermining the host protective response., Competing Interests: Disclosures: The authors declare no competing interests exist., (© 2021 Drurey et al.)

The nematophagous fungus Duddingtonia flagrans is used in integrated management of gastrointestinal nematodes in ruminants. The chlamydospores of the fungus, orally administered, pass through the segments of the ruminant digestive tract and, in the feces, capture the nematodes preventing their migration to grasslands. The drastic conditions of the gastrointestinal segments can negatively affect the fungus' biocontrol activity. The aim of this study was to assess the effect of in vitro conditions of the sheep's main gastrointestinal segments on the concentration, viability and nematode predatory ability of D. flagrans chlamydospores. The segments evaluated separately in vitro were the oral cavity, rumen, abomasum, and small intestine. The results showed that chlamydospores concentration was not affected by exposure to the different segments. The viability of the chlamydospores after exposure to the oral cavity (2.53 × 10 6  CFU/mL) and small intestine (1.24 × 10 5  CFU/mL) was significantly lower than its control treatment, with values of 6.67 × 10 6  CFU/mL and 2.31 × 10 5  CFU/mL respectively. Nematode predatory ability after rumen exposure was reduced by 7% compared to the control treatment, by 25% after abomasum exposure and by 17% after small intestine. This study revealed the individual in vitro effect of each segment of ovine gastrointestinal tract on the integrity of this strain of the fungus D. flagrans affecting its viability and nematode predatory ability under the evaluated conditions. Delivery systems could be designed to protect chlamydospores considering the impact of each gastrointestinal segment., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Paneth cells constitutively produce antimicrobial peptides and growth factors that allow for intestinal homeostasis, host protection, and intestinal stem cell replication. Paneth cells rely heavily on the glycolytic metabolic program, which is in part controlled by the kinase complex Mechanistic target of rapamycin (mTORC1). Yet, little is known about mTOR importance in Paneth cell integrity under steady-state and inflammatory conditions. Our results demonstrate that IFN-γ, a crucial mediator of the intestinal inflammation, acts directly on murine Paneth cells to alter their mitochondrial integrity and membrane potential, resulting in an TORC1-dependent cell death mechanism distinct from canonical cell death pathways including apoptosis, necroptosis, and pyroptosis. These results were established with the purified cytokine and a physiologically relevant common Th1-inducing human parasite Toxoplasma gondii . Given the crucial role for IFN-γ, which is a cytokine frequently associated with the development of inflammatory bowel disease and compromised Paneth cell functions, the identified mechanisms underlying mTORC1-dependent Paneth cell death downstream of IFN-γ may provide promising novel approaches for treating intestinal inflammation., Competing Interests: AA, AS, EB, AL, SG, EC, AM, SG, FY No competing interests declared, (© 2021, Araujo et al.)

Giardia duodenalis is a common cause of infection in children and travelers. The most frequent symptom is diarrhea in these patients. G. duodenalis trophozoites use a highly specialized adhesive disc to attach the host intestinal epithelium to induce intestinal damages. Pathological features of the small intestine following giardiasis include villous atrophy; infiltration of granulocytes, lymphocytes, and plasma cells into the lamina propria; and nodular lymphoid hyperplasia. The disturbed intestinal microbiota has been observed in patients with giardiasis. Therefore, a growing body of evidence has emphasized restoring the gut microbiome by probiotics in giardiasis. This study aimed to review the literature to find the pathologic features of giardiasis and its relationship with imbalanced microbiota. Then, benefits of probiotics in giardiasis and their potential molecular mechanisms were discussed. It has been illustrated that using probiotics (e.g., Lactobacillus and Saccharomyces) can reduce the time of gastrointestinal symptoms and repair the damages, particularly in giardiasis. Probiotics' capability in restoring the composition of commensal microbiota may lead to therapeutic outcomes. According to preclinical and clinical studies, probiotics can protect against parasite-induced mucosal damages via increasing the antioxidant capacity, suppressing oxidative products, and regulating the systemic and mucosal immune responses. In addition, they can reduce the proportion of G. duodenalis load by directly targeting the parasite. They can destroy the cellular architecture of parasites and suppress the proliferation and growth of trophozoites via the production of some factors with anti-giardial features. Further researches are required to find suitable probiotics for the prevention and treatment of giardiasis., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Allintoshius Chitwood, 1937 is the only genus of the family Ornithostrongylidae (Travassos, 1937) Durette-Desset and Chabaud, 1981 that parasitizes bats. Currently, there are 10 valid species in the genus, of which 3 were described from Brazil. This study describes a new species of Allintoshius and records the first occurrence of a nematode of this genus parasitizing Artibeus lituratus (Olfers). Allintoshius gomesae n. sp. is characterized by having anterior region coiled, cephalic vesicle with cuticular dilation striated transversely, and claviform esophagus. Synlophe in females consists of 16 cuticular ridges at the mid-body. Males have large caudal bursa, and conic and small spicules, and the gubernaculum is absent. Females have uterus didelphic, amphidelphic, tail tip tapered, and ovijector divided into 2 divergent branches, subequal in length. The new species differs from its congeners especially by the shape of the tail tip, vulvar opening, and size of spicules. Allintoshius gomesae is the fourth species of Allintoshius from Brazil and the first report in Ar. lituratus, increasing the number of species recognized of the genus., (© American Society of Parasitologists 2021.)

Group 2 innate lymphoid cells (ILC2s) are early effectors of mucosal type 2 immunity, producing cytokines such as interleukin (IL)-13 to mediate responses to helminth infection and allergen-induced inflammation. ILC2s are also present in lymph nodes (LNs) and can express molecules required for antigen presentation, but to date there are limited data on their dynamic behaviour. We used a CD2/IL-13 dual fluorescent reporter mouse for in vivo imaging of ILC2s and Th2 T cells in real time following a type 2 priming helminth infection or egg injection. After helminth challenge, we found that ILC2s were the main source of IL-13 in lymphoid organs (Peyer's patches and peripheral LNs), and were located in T cell areas. Intravital imaging demonstrated an increase in IL-13 + ILC2 size and movement following helminth infection, but reduced duration of interactions with T cells compared with those in homeostasis. In contrast, in the intestinal mucosa, we observed an increase in ILC2-T cell interactions post-infection, including some of prolonged duration, as well as increased IL-13 + ILC2 movement. These data suggest that ILC2 activation enhances cell motility, with the potential to increase the area of distribution of cytokines to optimise the early generation of type 2 responses. The prolonged ILC2 interactions with T cells within the intestinal mucosa are consistent with the conclusion that contact-based T cell activation may occur within inflamed tissues rather than lymphoid organs. Our findings have important implications for our understanding of the in vivo biology of ILC2s and the way in which these cells facilitate adaptive immune responses., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Lok, Walker, Jolin, Scanlon, Ishii, Fallon, McKenzie and Clatworthy.)

"Taste-like" tuft cells in the intestine trigger type 2 immunity in response to worm infection. The secretion of interleukin-13 (IL-13) from type 2 innate lymphoid cells (ILC2) represents a key step in the tuft cell-ILC2 cell-intestinal epithelial cell circuit that drives the clearance of worms from the gut via type 2 immune responses. Hallmark features of type 2 responses include tissue remodeling, such as tuft and goblet cell expansion, and villus atrophy, yet it remains unclear if additional molecular changes in the gut epithelium facilitate the clearance of worms from the gut. Using gut organoids, we demonstrated that IL-4 and IL-13, two type 2 cytokines with similar functions, not only induced the classical type 2 responses (e.g., tuft cell expansion) but also drastically up-regulated the expression of gasdermin C genes ( Gsdmc s). Using an in vivo worm-induced type 2 immunity model, we confirmed the up-regulation of Gsdmc s in Nippostrongylus brasiliensis -infected wild-type C57BL/6 mice. Consistent with gasdermin family members being principal effectors of pyroptosis, overexpression of Gsdmc2 in human embryonic kidney 293 (HEK293) cells triggered pyroptosis and lytic cell death. Moreover, in intestinal organoids treated with IL-4 or IL-13, or in wild-type mice infected with N. brasiliensis , lytic cell death increased, which may account for villus atrophy observed in worm-infected mice. Thus, we propose that the up-regulated Gsdmc family may be major effectors for type 2 responses in the gut and that Gsdmc-mediated pyroptosis may provide a conduit for the release of antiparasitic factors from enterocytes to facilitate the clearance of worms., Competing Interests: The authors declare no competing interest.

The role of p53 in tumor suppression has been extensively studied and well-established. However, the role of p53 in parasitic infections and the intestinal type 2 immunity is unclear. Here, we report that p53 is crucial for intestinal type 2 immunity in response to the infection of parasites, such as Tritrichomonas muris and Nippostrongylus brasiliensis. Mechanistically, p53 plays a critical role in the activation of the tuft cell-IL-25-type 2 innate lymphoid cell circuit, partly via transcriptional regulation of Lrmp in tuft cells. Lrmp modulates Ca 2+ influx and IL-25 release, which are critical triggers of type 2 innate lymphoid cell response. Our results thus reveal a previously unrecognized function of p53 in regulating intestinal type 2 immunity to protect against parasitic infections, highlighting the role of p53 as a guardian of immune integrity.

Co-infections of mammalian hosts with intestinal helminths and bacterial pathogens are common, especially in areas with inadequate sanitation. Interactions between co-infecting species and host microbiota can cause significant changes in host immunity, disease severity, and pathogen transmission, requiring unique treatment for each case. A greater understanding of the influences of parasite-bacteria co-infections will improve diagnosis and therapeutic approaches to control infectious diseases. To study the influence of the trematode parasite Echinostoma caproni on commensal and pathogenic bacteria in the mouse gut, we examined the abundance of intestinal lactic acid bacteria and Salmonella enterica serovar Typhimurium in control mice not exposed to E. caproni (P-) or S. Typhimurium (S-), E. caproni-infected (P+S-), S. Typhimurium-infected (P-S+), and E. caproni-S. Typhimurium co-infected (P+S+) mice, and determined bacterial burdens in the livers and spleens of the P-S+ and P+S+ mice. We also examined a subset of P+S- and P+S+ mice for survival and the relative location of E. caproni in the small intestine. The numbers of presumptive lactic acid bacteria were significantly higher in the P+S+ and P-S+ mice compared to the uninfected mice, and S. Typhimurium colonization in the liver and spleen was significantly reduced in the P+S+ mice compared to the P-S+ mice. Echinostoma caproni were located anteriorly in the intestine of P+S- mice, while in the P+S+ mice, the parasites were distributed more posteriorly. Survival of E. caproni was unaffected in either group. The results of our study suggest that E. caproni facilitates a higher abundance of presumptive lactic acid bacteria in the mouse intestine and reduces colonization of S. Typhimurium in the liver and spleen of the co-infected host., (© American Society of Parasitologists 2021.)

Background: Toxoplasma gondii is a protozoan with worldwide distribution and triggers a strong Th1 immune response in infected susceptible hosts. On the contrary, most helminth infections are characterized by Th2 immune response and the use of helminth-derived antigens to regulate immune response in inflammatory disorders has been broadly investigated., Objectives: The aim of this study was to investigate whether treatment with Strongyloides venezuelensis antigen extract (SvAg) would alter immune response against T gondii., Methods: C57BL/6 mice were orally infected with T gondii and treated with SvAg, and parasitological, histological and immunological parameters were investigated., Results: It was observed that SvAg treatment improved survival rates of T gondii-infected mice. At day 7 post-infection, the parasite load was lower in the lung and small intestine of infected SvAg-treated mice than untreated infected mice. Remarkably, SvAg-treated mice infected with T gondii presented reduced inflammatory lesions in the small intestine than infected untreated mice and decreased intestinal and systemic levels of IFN-γ, TNF-α and IL-6. In contrast, SvAg treatment increased T gondii-specific IgA serum levels in infected mice., Conclusions: S venezuelensis antigen extract has anti-parasitic and anti-inflammatory properties during T gondii infection suggesting as a possible alternative to parasite and inflammation control., (© 2020 John Wiley & Sons Ltd.)

Raillietina echinobothrida (R. echinobothrida) is one of the most pathogenic and prevalent tapeworms threat to the commercial chickens in China. However, there is a lack of research on their molecular identification and morphological characteristics. This study explored the molecular identification markers for R. echinobothrida in North China based on 18s ribosomal RNA (18s rRNA) gene and the ribosomal DNA second internal transcribed spacer (ITS-2) gene. The BLAST results of 18s rRNA (1643 bp) and ITS-2 (564 bp) gene sequences showed that the isolated intestinal tapeworms were R. echinobothrida. Phylogenetic trees obtained by maximum likelihood (ML) or neighbor-joining (NJ) method revealed that the R. echinobothrida in North China had the closest evolutionary relationship with the species found on the Qinghai-Tibet plateau, China. Morphological observations by hematoxylin staining and scanning electron microscope showed four round suckers and a retractable rostellum on the spherical scolex of R. echinobothrida. Two rows of alternately arranged hooks distributed around the rostellum. There were 30-40 testes in each mature segment. A well-developed cirrus pouch lied outside the excretory duct of mature segment. The gravid segment contained 200-400 eggs and there was a well-developed oncosphere in each egg. In addition, abundant ultrastructural features in mature proglottid of R. echinobothrida in North China were identified by transmission electron microscopy. In conclusion, the present study established ways of molecular phylogenetic identification for R. echinobothrida based on 18s rRNA and ITS-2 gene, and identified the morphological and ultrastructural characteristics of R. echinobothrida in North China.

Background: Toxoplasma gondii is a parasite that primarily infects through the oral route. Nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs) play crucial roles in the immune responses generated during parasitic infection and also drive the inflammatory response against invading parasites. However, little is known about the regulation of NLRs and inflammasome activation in T. gondii-infected human small intestinal epithelial (FHs 74 Int) cells., Methods: FHs 74 Int cells infected with T. gondii were subsequently evaluated for morphological changes, cytotoxicity, expression profiles of NLRs, inflammasome components, caspase-cleaved interleukins (ILs), and the mechanisms of NLRP3 and NLRP6 inflammasome activation. Immunocytochemistry, lactate dehydrogenase assay, reverse transcription polymerase chain reaction (RT-PCR), real-time quantitative RT-PCR, and western blotting techniques were utilized for analysis., Results: Under normal and T. gondii-infected conditions, members of the NLRs, inflammasome components and caspase-cleaved ILs were expressed in the FHs Int 74 cells, except for NLRC3, NLRP5, and NLRP9. Among the NLRs, mRNA expression of NOD2, NLRP3, NLRP6, and NAIP1 was significantly increased in T. gondii-infected cells, whereas that of NLRP2, NLRP7, and CIITA mRNAs decreased significantly in a time-dependent manner. In addition, T. gondii infection induced NLRP3, NLRP6 and NLRC4 inflammasome activation and production of IL-1β, IL-18, and IL-33 in FHs 74 Int cells. T. gondii-induced NLRP3 inflammasome activation was strongly associated with the phosphorylation of p38 MAPK; however, JNK1/2 had a weak effect. NLRP6 inflammasome activation was not related to the MAPK pathway in FHs 74 Int cells., Conclusions: This study highlighted the expression profiles of NLRs and unraveled the underlying mechanisms of NLRP3 inflammasome activation in T. gondii-infected FHs 74 Int cells. These findings may contribute to understanding of the mucosal and innate immune responses induced by the NLRs and inflammasomes during T. gondii infection in FHs 74 Int cells.

The hawksbill turtle Eretmochelys imbricata is a critically endangered species with a worldwide distribution. Limited information is available about the naturally occurring intestinal parasites of this species and what impact these parasites may have on the health of the hawksbill turtle. Diaschistorchis pandus was identified postmortem in 5 hawksbill turtles from Grenada, West Indies, using morphologic characterization. Sanger sequencing was performed for conserved ribosomal regions (5.8S, ITS2, 28S) and the mitochondrial cytochrome c oxidase subunit 1 gene (COI). Phylogenetic analysis of the 28S rRNA gene sequence data shows D. pandus clustering with other trematodes in the family Pronocephalidae, corroborating morphological classification. No genetic sequences have been previously reported for this trematode species, which has limited the collection of objective epidemiological data about this parasite of marine turtles., (© American Society of Parasitologists 2021.)

Alveolar echinococcosis and cystic echinococcosis are severe zoonotic diseases caused by Echinococcus multilocularis and Echinococcus granulosus s.l. in Europe. To present knowledge, in the European continent, the most important definitive hosts of these parasites belong to the Canidae family. The golden jackal as an opportunistic mesopredator frequently preys on rodents including arvicolids and other easily available food resources, such as viscera and other carrion. By these reasons, the golden jackal can promote the maintenance of both Echinococcus multilocularis and Echinococcus granulosus s.l. Our investigation was conducted in the southwestern part of Hungary where one of the densest golden jackal populations exists. We examined altogether 173 golden jackal small intestines to determine the presence of Echinococcus multilocularis and Echinococcus granulosus s.l. After the molecular diagnostic procedure, we found 27 Echinococcus multilocularis-positive (prevalence: 15.6%; mean intensity: 664 worms) and three Echinococcus granulosus s.l. infected hosts (prevalence: 1.7%; mean intensity: 554.3 worms). We suggest the invasion of the golden jackal in Europe can enhance the spread of both Echinococcus multilocularis and Echinococcus granulosus s.l. This novel epidemiological situation can influence the geographical distribution of these helminths and the characteristics of their endemic in different host species, as well as in humans., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

The aim of this research was to determine the effect of gut health parameters on the flock's final weight of broilers and to calculate an accurate equation to estimate this weight with information available at 7, 14, and 21 days, in field conditions. Gut health parameters (gizzard erosion, coccidiosis, feed passage, and redness, gut tone, consistency of content, and presence of mucus for each part of the small intestine [duodenum, jejunum, and ileum], and color, consistency, and presence of gas for caeca content) were evaluated at 7 and 14 days. Other parameters evaluated for impact on flock final weight were body weight and mortality, both at 7, 14, and 21 days; stocking density; litter reuse; and downtime period. Structural equation model evaluation of the data showed that stocking density and litter reuse did not affect (P > 0.05) flock final weight, while downtime period, body weight (14 and 21 days), and mortality (14 and 21 days) directly affected (P ≤ 0.05) the flock final weight. Gut health parameters did not directly affect the flock's final weight; however, they affected body weight and mortality at 14 days, thus showing an indirect effect on the flock's final weight. It was also possible to determine two accurate equations to estimate the flock's final weight using information available at both 14 (R 2  = 0.56) and 21 (R 2  = 0.77) days.

Falcaustra vietnamensis n. sp. is described from the small intestine of Physignathus cocincinus from north-central Vietnam. The new species is characterized by the large male worms (20.2-28.8 mm in length and 557-724 μm in width) relative to known members of the genus, 2 sharply pointed alate spicules of equal length (1,128-1,256 μm in length), gubernaculum including 2 separate pieces, 1 ventral with a pointed distal end and 1 dorsal with a blunt distal end (164-192 μm and 155-172 μm in length, respectively), and 12 pairs of caudal papillae. Female worms are larger than male worms (24.2-34.1 mm in length and 532-735 μm in width), with the vulva situated in the posterior half of body, and elliptical eggs, 60-70 μm long by 42-47 μm wide. Falcaustra vietnamensis n. sp. represents the 38th species assigned to the genus and the third species recorded from a lizard host in the Oriental biogeographical region. Partial sequences of the 18S ribosomal RNA gene (rDNA), internal transcribed spacer regions (ITS), and cytochrome c oxidase subunit 1 (COI) are provided for the new species. The molecular phylogenetic position of the genus Falcaustra is briefly discussed., (© American Society of Parasitologists 2021.)

Objective: To evaluate the dynamics of S. venezuelensis infection in Wistar rats of different age ranges., Design: Thirty-five (n = 35, 7 per group) male Wistar rats were distributed according to age into five groups: 2, 3, 6, 12 and 18 months old (mo). The rats were infected by S. venezuelensis and eggs per gram of feces (EPG) were measured at 3, 9, 15 and 21 days post-infection (dpi). All animals were killed at 21 dpi, thymus, lungs and small intestines were removed, and relative weight calculated. The adult worms recovered from the small intestines and blood cells were counted., Results: Rats in advanced age presented higher parasite oviposition at 9 dpi and posterior reduction of EPG, while young rats still showed higher oviposition at 15 dpi and 21 dpi. At 12 and 18 mo, the rats had greater number of adult worms, which with low fecundity, eosinophilia and least concentration of monocytes. The fecundity of worms was more expressive in young rats. A strong correlation was observed between age and EPG at 9 dpi (R = 0.72, p < 0.0001), at 15 (R = -0.66, p < 0.0001) and at 21 dpi (R = -0.65, p < 0.0001), as well as age and numbers of worms at 21 dpi (R = 0.74, p < 0.0001). The relative weight of the thymus, lungs and small intestines were higher in rats at 2 and 3 mo in comparison to the older groups of rats., Conclusions: Aging process interfered on host-parasite relationship and changed the dynamics of infection of S. venezuelensis in Wistar rats., (Published by Elsevier Inc.)

The prevalence of ectoparasites and intestinal helminths of different pigeon taxa in Medina, Saudi Arabia, with special emphasis on the feral pigeon, Columba livia domestica (Columbiformes: Columbidae), was evaluated. Fifty-four pigeons were examined externally for ectoparasites and 28 feral pigeons were examined for helminths. Two ectoparasites were recorded on feral C. l. domestica (Harami) pigeons, including the shaft louse Menopon gallinae (Phthiraptera: Menoponidae), and the pigeon fly, Pseudolynchia canariensis (Diptera: Hippoboscidae), with 100 and 88.90% prevalence, respectively. Ectoparasites were also collected from 5 other breeds of C. l. domestica (Pakistani, Farensi, Turki, Kori, and Qatifi). Menopon gallinae infected Pakistani, Farensi, and Turki pigeons with 100% prevalence. A third ectoparasite, the brown poultry louse, Goniodes dissimilis (Psocodea: Philopteridae), infected Farensi, Turki, and Kori pigeons at rates of 100, 50, and 50%, respectively. Qatifi pigeons were not infected with any ectoparasites. Two types of intestinal helminths were recovered from feral pigeons: cestodes of Raillietina spp. (Cyclophyllidea: Davaineidae) and nematodes of Ascaridia sp. (Ascaridida: Ascaridiidae) (with 10.71 and 3.57% prevalences, respectively). To the best of our knowledge, this is the first study to shed light on the parasites of pigeons in Medina, Saudi Arabia., (© American Society of Parasitologists 2020.)

Sea turtles are difficult to sample because of their protected status; however, museum collections and sea turtle stranding networks provide unique opportunities for parasitological research. Four gastrointestinal tracts from stranded, endangered green turtles, Chelonia mydas, were collected between 1993 and 1995 from the upper Texas coast and opportunistically sampled for parasite fauna. Two new species of Telorchis, a common freshwater amphibian and reptilian intestinal parasite genus, were found and described. Telorchis marinus n. sp. differs from Telorchis mydas n. sp. by its short body length, lack of pharyngeal glands, long esophagus relative to total body length, short and straight cirrus sac, short ventral sucker to ovary length relative to total body length, and an ovary located in the anterior one-third of body; it differs from its congeners in the number of ovary lengths between the ventral sucker and ovary, the number of ventral sucker lengths the cirrus sac extends beyond the posterior margin of the ventral sucker, and the vitelline field extent. Telorchis mydas differs from its congeners in the number of ovary lengths between the ventral sucker and ovary, the number of ventral sucker lengths the cirrus sac extends beyond the posterior margin of the ventral sucker, and the combination of having its ovary position near the midbody and a long, sinuous cirrus sac that is 35-44% of the total body length. Given the taxonomic complexities within Telorchis, a revised key to North American species is provided using morphological characteristics to assist future researchers in delineating true species and appropriate synonymies with molecular explorations. We reject the majority of synonymies in the genus until molecular data are available; we accept the synonymies of Telorchis necturi as Telorchis stunkardi and Telorchis gutturosi as Telorchis chelopi. Both Telorchis linstowi and Telorchis stossichi should be considered as species inquirenda. This is the first confirmed report of Telorchis from a marine host and the first report on parasites of cheloniid sea turtles in Texas, and this study adds to the ever-growing evidence that collections are essential to understanding biodiversity., (© American Society of Parasitologists 2020.)

Background: Trichostrongylus is one of the most important zoonotic trichostrongylid nematodes, infecting mostly livestock. Data on its genetic characteristics are lacking in Iran., Methods: We determined the phylogenetic relationships of Trichostrongylus species in three counties of Kohgiloyeh and Boyerahmad (K-B) province, southwest Iran. Small intestine and abomasum of 70 sheep and goats were investigated., Results: A total of 35 isolates of Trichostrongylus worms were detected and all were genetically identified as Trichostrongylus vitrinus. Analysis of 321 bp of the internal transcribed spacer 2 (ITS2) of ribosomal DNA revealed 16 genotypes. All genotypes were single nucleotide polymorphisms, including some hypervariable points. All sequences were trimmed to 170 bp, compared with sequences on GenBank including short sequences from other endemic foci of Iran and other countries and all isolates were used to generate a maximum likelihood phylogenetic tree, which consisted of two clades A and B. Clade A included isolates from Iran, Russia, New Zealand, Australia and the UK; clade B only contained South African isolates. Most clade A isolates (north, southwest and west Iran, Russia, New Zealand, Australia and UK) were in a similar phylogenetic position. One subclade was detected in clade A (isolates from Southwest Iran, New Zealand and UK)., Conclusions: We hypothesize that drug resistant T. vitrinus may account for its exclusive detection in our samples. The high similarity of genotypes from Iran, New Zealand and UK may be due to their close political relationships during the colonial era. More research is needed to understand better the phylogeny of T. vitrinus and its relationship with drug resistance and human transmission.

Toxoplasma gondii is a protozoan parasite capable of infecting a large number of warm-blooded animals and causes serious health complications in immunocompromised patients. T. gondii infection of the feline small intestine is critical for the completion of the life cycle and transmission of T. gondii. Protein acetylation is an important posttranslational modification, which plays roles in the regulation of various cellular processes. Therefore, understanding of how T. gondii reprograms the protein acetylation status of feline definitive host can help to thwart the production and spread of T. gondii. Here, we used affinity enrichment and high-resolution liquid chromatography with tandem mass spectrometry to profile the alterations of the acetylome in cat small intestine 10 days after infection by T. gondii Prugniuad (Pru) strain. Our analysis showed that T. gondii induced significant changes in the acetylation of proteins in the cat intestine. We identified 2606 unique lysine acetylation sites in 1357 acetylated proteins. The levels of 334 acetylated peptides were downregulated, while the levels of 82 acetylated peptides were increased in the infected small intestine. The proteins with differentially acetylated peptides were particularly enriched in the bioenergetics-related processes, such as tricarboxylic acid cycle, oxidative phosphorylation, and oxidation-reduction. These results provide the first baseline of the global acetylome of feline small intestine following T. gondii infection and should facilitate further analysis of the role of acetylated protein in the pathogenesis of T. gondii infection in its definitive host.

In Argentina, trichinellosis is an endemic disease acquired mainly through consumption of raw pork infected with nematodes larvae from the Trichinella genus. For years, the only species involved in outbreaks in humans and pig foci in Argentina was Trichinella spiralis. In 2008 the presence of a new Trichinella taxon from a cougar (Puma concolor) was detected and recorded in the province of Rio Negro, Argentina, and the finding was established as a new species in 2012: Trichinella patagoniensis. To the best of our knowledge, there is no information available on the intestinal phase and antibody response in a susceptible host during T. patagoniensis infection. Therefore, our research has been designed to study experimental infection with T. patagoniensis compared to infection with T. spiralis in BALB/c mice. One hundred and twenty eight BALB/c mice were divided into two groups and individuals in each group were infected per os with 500 larvae of T. patagoniensis or 500 larvae of T. spiralis, respectively. After that, they were euthanized on different days. Adult worm recovery from small intestines and artificial digestion of each carcass was performed. Histopathology of small intestines was performed using hematoxylin-eosin staining. Systemic cytokines and antibody kinetics were evaluated. Intestinal adult worm recovery of T. patagoniensis and T. spiralis took place until day 17 and 25, respectively. Systemic IFN-γ, IL-10, and TNF showed significant variations in T. patagoniensis infected mice. Seroconversion was detected in animals as from 15 days post-infection (pi) for both T. patagoniensis and T. spiralis, reaching the highest OD value at 42 days pi. Similar microscopic lesions were observed in the small intestine from mice infected with the same dose of T. spiralis and T. patagoniensis. Our findings contribute new information regarding the intestinal phase and the antibody kinetics of T. patagoniensis in BALB/c mice., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Canine schistosomiasis, caused by the trematode Heterobilharzia americana, can pose a diagnostic challenge due to nonspecific symptoms. The aim of this multicenter, retrospective, descriptive study was to compare the prevalence and extent of sonographic changes associated with schistosomiasis between affected and infection-free dogs. Medical records of two referral centers were searched for dogs with confirmed schistosomiasis that had undergone an abdominal ultrasound. Fifty-five cases fulfilled the inclusion criteria, and a contemporaneous control group was derived from dogs that tested negative for H. americana. Two blinded reviewers evaluated the images. The majority of Heterobilharzia-infected (further termed H-pos) dogs (82%) had ultrasonographic abnormalities in the small intestine ± liver. Abnormal layering of the small intestine was noted in 38 of 54 H-pos dogs, compared to six of 54 control dogs (P < .0001). Pinpoint hyperechoic foci were noted in the small intestinal submucosa or muscularis layers in 25 of 54 H-pos dogs, but only three controls (P < .0001). Heterogeneity of the hepatic parenchyma and pinpoint hyperechoic foci were more prevalent in H-pos dogs (65% vs 40%; P = .0213 and 44% vs 18%; P = .0068, respectively). Pinpoint hyperechoic foci within mesenteric lymph nodes were noted in seven H-pos dogs and none of the controls (P = .0128). The combination of heterogeneous small intestine wall layering and pinpoint hyperechoic foci in small intestine, liver, or mesenteric lymph nodes was the most reliable indication of infection (P = .0001; odds ratio = 36.87), with positive predictive value of 94%, yet modest sensitivity for the detection of infection (58%). Observing these sonographic features suggests schistosomiasis and should prompt further testing., (© 2020 American College of Veterinary Radiology.)

Enteric parasitic infections are among the most prevalent infections in lower- and middle-income countries (LMICs) and have a profound impact on global public health. While the microbiome is increasingly recognized as a key determinant of gut health and human development, the impact of naturally acquired parasite infections on microbial community structure in the gut, and the extent to which parasite-induced changes in the microbiome may contribute to gastrointestinal symptoms, is poorly understood. Enteric parasites are routinely identified in companion animals in the United States, presenting a unique opportunity to leverage this animal model to investigate the impact of naturally acquired parasite infections on the microbiome. Clinical, parasitological, and microbiome profiling of a cohort of 258 dogs revealed a significant correlation between parasite infection and composition of the bacterial community in the gut. Relative to other enteric parasites, Giardia was associated with a more pronounced perturbation of the microbiome. To compare our findings to large-scale epidemiological studies of enteric diseases in humans, a database mining approach was employed to integrate clinical and microbiome data. Substantial and consistent alterations to microbiome structure were observed in Giardia -infected children. Importantly, infection was associated with a reduction in the relative abundance of potential pathobionts, including Gammaproteobacteria , and an increase in Prevotella -a profile often associated with gut health. Taken together, these data show that widespread Giardia infection in young animals and humans is associated with significant remodeling of the gut microbiome and provide a possible explanation for the high prevalence of asymptomatic Giardia infections observed across host species. IMPORTANCE While enteric parasitic infections are among the most important infections in lower- and middle-income countries, their impact on gut microbiota is poorly understood. We reasoned that clinical symptoms associated with these infections may be influenced by alterations of the microbiome that occur during infection. To explore this notion, we took a two-pronged approach. First, we studied a cohort of dogs naturally infected with various enteric parasites and found a strong association between parasite infection and altered gut microbiota composition. Giardia , one of the most prevalent parasite infections globally, had a particularly large impact on the microbiome. Second, we took a database-driven strategy to integrate microbiome data with clinical data from large human field studies and found that Giardia infection is also associated with marked alteration of the gut microbiome of children, suggesting a possible explanation for why Giardia has been reported to be associated with protection from moderate to severe diarrhea., (Copyright © 2020 Berry et al.)

Thirty white-spotted geckos, Tarentola annularis, from the South Sinai desert in Egypt, were examined for helminth parasites. Spauligodon aspiculus was observed to infect 19 geckos with 63.33% as a prevalence of parasitic infection. The present nematode species is separated from congeners by morphological and metrical characteristics such as lateral alae, aspinose filamentous tail, and no spicule, and three pairs of caudal papillae with posterior pair excluded from envelopment by the caudal alae in the male worms, and knobbed eggs, and postbulbar vulva in females. It compared morphometrically with other Spauligodon species described previously and showed few differences in measurements. Molecular characterization based on the partial 28S rRNA nuclear ribosomal gene sequence showed that there was a close identity, up to 72%, with other sequences retrieved from GenBank. Phylogenetic analysis showed that the parasite sequence in conjunction with existing data facilitates the investigation of the placement of this pharyngodonid species within Oxyuridae. The present species is deeply embedded in the genus Spauligodon with close relationships to previously described Spauligodon nicolauensis (gb| JN619349.1, and JF829243.1) as more related sister taxa. This study highlights the importance of combining genetic and morphological data with taxonomy in pharyngodonid species., (© 2020 Wiley Periodicals, Inc.)

Entamoeba histolytica is a protozoan parasite responsible for amoebiasis, a disease with a high prevalence in developing countries. Establishing an amoebic infection involves interplay between pathogenic factors for invasion and tissue damage, and immune responses for protecting the host. Here, we review the pathogenicity of E. histolytica and summarize the latest knowledge on immune response and immune evasion mechanisms during amoebiasis.

Hymenolepis diminuta is a zoonotic cestode parasitizing the small intestine of rodents (definitive hosts). Humans can accidentally enter into the life cycle of this tapeworm via the ingestion of infected insects (intermediate hosts) containing cestode cysticercoids in their body cavity. More than two centuries after the first record in humans, there are no accurate estimates of the number of human cases around the world. In order to have a more precise idea about the number of human cases with H. diminuta and the current status of the disease (hymenolepiasis) worldwide, we conducted a literature review of published records on human infection with H. diminuta. One thousand five hundred and sixty-one published records of infection with H. diminuta from 80 countries were identified. This review presents an estimate of the number of human cases with H. diminuta and a current overview of the prevalence, geographic distribution, symptoms, diagnosis, exposure to infective stages, and therapeutic approaches for this underestimated zoonotic tapeworm.

Coccidiosis is caused by protozoa of the genus Eimeria. These are intracellular parasites of enterocytes that rupture the host cell, causing lesions in the intestinal mucosa. The lesions caused by Eimeria reduce nutrient absorption capacity, negatively affecting productive gains in broilers, and representing a gateway for other enteropathogens. The objective of this study was to analyze the correlation between lesions caused by Eimeria and the prevalence of coccidiosis and other alterations found in the gastrointestinal tracts of broilers produced in Brazil from 2017 to 2018. Intestinal health evaluations were performed in 32 integrations (farm) of broilers in Brazil, totaling 726 birds analyzed between the ages of 22 and 40 days. Necropsied chickens were collected at three different points, with at least three birds per shed. We analyzed the following changes in the gastrointestinal tract: presence of cellular desquamation, fluid and mucus excess, ingestion of bedding, small and large intestine lesion, food passage, altered tone, "Turkish towel" lesions, worm infection, enteritis and gizzard erosion. The definition of macroscopic lesion scores caused by Eimeria acervulina, Eimeria maxima, Eimeria tenella followed a specific methodology. Mucosal oocyst counts for E. maxima (E. maxima micro) was performed using an optical microscope with a magnification of 100×. We found that the species E. acervulina had the highest prevalence (5.5%). With respect to E. acervulina, a positive correlation was observed with cellular desquamation, bedding ingestion and passage of food. The second highest prevalence was E. maxima (average of 4%), showing positive correlations with cellular desquamation, fluid excess, bed ingestion, feed passage and E. acervulina. E. tenella represented the lowest prevalence (0.8%) among the species of Eimeria analyzed, showing a positive correlation with altered intestinal tone. On microscopic evaluation, E. maxima was present in 45% of mucosa scrapings, representing subclinical coccidiosis of 1125% (11.25-fold) greater than the rate of clinical coccidiosis. Regarding other alterations that were visualized in the gastrointestinal tract, we have recorded the incidence of altered intestinal tone (0.1%), worm infection (0.4%), small intestine (0.8%), enteritis (1%), duodenitis (1.5%), "Turkish towel" lesions (3.3%), excess fluid (4.5%), bed ingestion (6.9%), excess mucus (8.4%), food passage (10.3%), cellular desquamation (11%) and gizzard erosion (13.4%). We conclude that monitoring is of paramount importance to understand the intestinal health status of poultry lots. Microscopic E. maxima is present in 45%. We identified factors that correlate with reduction in intestinal health, impairing zoo-economic performance., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Introduction: A new coccidian species of the genus Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae), is reported from the bat host Myotis riparius Handley from Ilha Grande, a large island off the coast of the State of Rio de Janeiro, in southeastern Brazil., Methods: Bats were captured in 13 mist nets (10 × 3 m), which were set within the experimental plots, and through active searches of the daytime roosts of Molossus molossus Pallas found in Vila Dois Rios. Containment was made in bags for the collection of feces and identification of coccidia. A survey was conducted on the coccidia species described so far (Table 2)., Results: The oöcysts of Eimeria riparii n. sp. are ellipsoidal to cylindroidal with an extremely thin, bi-layered wall, slightly rough. Two polar granules are present, micropyle and oöcyst residuum are both absent. The sporocysts are ellipsoidal, the sporocyst residuum is formed by sparse, rounded granules of varying sizes; the Stieda body is trapezoidal and a sub-Stieda body is absent. Sporozoites are banana shaped. With the new species described here, a total of 40 Eimeria spp. have been described infecting bat hosts, belonging to 30 species of 18 genera and 5 families., Conclusion: The subsequent increase in the known diversity of bats has been derived from the ongoing expansion of research in a number of different areas of taxonomy and ecology although the number of studies of the associated coccidian parasites of the family Eimeriidae has increased more slowly.

Eimeria necatrix is a high pathogenic pathogen second to Eimeria tenella causing chicken coccidiosis. However, the precise underlying molecular mechanisms of interaction between E. necatrix and chickens are not fully understood. Accumulating evidences suggest that micro-RNAs (miRNAs) play pivotal regulatory roles in various diseases, including parasitic diseases. In the present study, the expression profile of miRNAs in Hy-line variety white chicken small intestines infected with E. necatrix was studied by using deep sequencing. A total of 35 miRNAs (including 16 significantly upregulated and 19 significantly downregulated miRNAs) were significantly differentially expressed (DE) in infected tissues at 108 h post-infection (pi). Real-time polymerase chain of 10 miRNAs (including 5 upregulated and 5 downregulated) randomly selected successfully confirmed the effectiveness of deep sequencing. Target prediction showed that 4,568 mRNAs could be regulated by 21 (including 12 upregulated and 9 downregulated) of 35 differentially expressed miRNAs. Functional analysis indicated that target genes of these differentially expressed miRNAs would be involved in pathways related to infection of E. necatrix, including cell differentiation, adhesion, proliferation, and apoptosis (e.g., MAPK signaling pathway and PPAR signaling pathway). To our best knowledge, this is the first study on the miRNA expression profile of small intestines during E. necatrix infection, and the findings in the present study suggested that these DE miRNAs would play important regulatory role in the interaction between E. necatrix and chicken intestines., (Copyright © 2020. Published by Elsevier Inc.)

Giardiasis and other protozoan infections are major worldwide causes of morbidity and mortality, yet development of new antimicrobial agents with improved efficacy and ability to override increasingly common drug resistance remains a major challenge. Antimicrobial drug development typically proceeds by broad functional screens of large chemical libraries or hypothesis-driven exploration of single microbial targets, but both strategies have challenges that have limited the introduction of new antimicrobials. Here, we describe an alternative drug development strategy that identifies a sufficient but manageable number of promising targets, while reducing the risk of pursuing targets of unproven value. The strategy is based on defining and exploiting the incompletely understood adduction targets of 5-nitroimidazoles, which are proven antimicrobials against a wide range of anaerobic protozoan and bacterial pathogens. Comprehensive adductome analysis by modified click chemistry and multi-dimensional proteomics were applied to the model pathogen Giardia lamblia to identify dozens of adducted protein targets common to both 5'-nitroimidazole-sensitive and -resistant cells. The list was highly enriched for known targets in G. lamblia, including arginine deiminase, α-tubulin, carbamate kinase, and heat shock protein 90, demonstrating the utility of the approach. Importantly, over twenty potential novel drug targets were identified. Inhibitors of two representative new targets, NADP-specific glutamate dehydrogenase and peroxiredoxin, were found to have significant antigiardial activity. Furthermore, all the identified targets remained available in resistant cells, since giardicidal activity of the respective inhibitors was not impacted by resistance to 5'-nitroimidazoles. These results demonstrate that the combined use of click chemistry and proteomics has the potential to reveal alternative drug targets for overcoming antimicrobial drug resistance in protozoan parasites., Competing Interests: The authors have declared that no competing interests exist.

Trichinellosis is a serious food-borne zoonotic infection of cosmopolitan distribution. Currently, treatment for trichinellosis is far from ideal. Given the important role of oxidative stress and immune-mediated inflammation in the pathogenesis of trichinellosis, this study was designed to evaluate the possible protective effects of resveratrol (RSV) during the intestinal and muscular phases of Trichinella spiralis infection in mice. The oral administration of RSV at a dose of 20 mg/kg once daily for two weeks resulted in significant reductions in both adult and larval counts; significant improvements in the redox status of the small intestine and muscles; a significant reduction in interleukin 4, pentraxin 3 and vascular endothelial growth factor expression; and the mitigation of intestinal and muscular inflammation. In conclusion, this study identifies RSV as a promising agent for the treatment of experimental trichinellosis, and more studies in experimental animals and humans are worth consideration.

There is limited information regarding responses by slow cycling stem cells during T. spiralis-induced T-cell mediated intestinal inflammation and how such responses may relate to those of Paneth cells. Transgenic mice, in which doxycycline induces expression of histone 2B (H2B)-green fluorescent protein (GFP), were used. Following discontinuation of doxycycline ("chase" period), retention of H2B-GFP enabled the identification of slow cycling stem cells and long-lived Paneth cells. Inflammation in the small intestine (SI) was induced by oral administration of T. spiralis muscle larvae. Epithelial retention of H2B-GFP per crypt cell position (cp) was studied following immunohistochemistry and using the Score and Wincrypts program. Compared to non-infected controls, there was significant reduction in the number of H2B-GFP-retaining stem cells in T. spiralis-infected small intestines. H2B-GFP-retaining stem cells peaked at around cp 4 in control sections, but smaller peaks at higher cell positions (>10) were seen in sections of inflamed small intestines. In the latter, there was a significant increase in the total number of Paneth cells, with significant reduction in H2B-GFP-retaining Paneth cells, but a marked increase in unlabelled (H2B-GFP-negative) Paneth cells. In conclusion, following T. spiralis-infection, putative slow cycling stem cell numbers were reduced. A marked increase in newly generated Paneth cells at the crypt base led to higher cell positions of the remaining slow cycling stem cells., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The use of helminth infections as tools to understand the type 2 immune response is a well-established technique and important to many areas of immunological research. The phenotype and function of immune cell populations at the site of infection is a key determinant of pathogen clearance. However, infections with helminths such as the murine nematode Heligomosmoides polygryrus cause increased mucus production and thickening of the intestinal wall, which can result in extensive cell death when isolating and analysing cells from the lamina propria (LP). Populations of larger immune cells such as macrophages and dendritic cells are often trapped within mucus or dying tissues. Here we describe an optimised protocol for isolating LP leukocytes from the small intestine of H.polygyrus -infected mice, and we demonstrate phenotypic and functional identification of myeloid and CD4 + T cell subsets using cytokine staining and flow cytometry. Our protocol may provide a useful experimental method for the immunological analysis of the affected tissue site during helminth infections., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Fasciola hepatica is the causative agent of fasciolosis, a parasitic zoonosis of global distribution causing significant economic losses in animal production and a human public health problem in low-income countries. Hosts are infected by ingestion of aquatic plants carrying metacercariae. Once ingested, the juvenile parasites excyst in the small intestine and, after crossing it, they follow a complex migratory route that lead the parasites to their definitive location in the bile ducts. Despite being a critical event in the progression of the infection, the available data on the cross-talk relationships between the parasite and the host at an early stage of the infection are scarce. The objective of the present work is to characterize the proteomic changes occurring in both the parasite and the host, through the development of a novel in vitro model, to shed light on the molecular pathways of communication between the newly excysted juveniles (NEJ) from F. hepatica and the host's intestinal epithelium. For this, in vitro excystation of F. hepatica metacercariae was carried out and NEJ were obtained. Additionally, optimal conditions of growth and expansion of mouse primary small intestinal epithelial cells (MPSIEC) in culture were fine-tuned. Tegumentary and somatic parasite antigens (NEJ-Teg and NEJ-Som), as well as host cell protein lysate (MPSIEC-Lys) were obtained before and after 24 h co-culture of NEJ with MPSIEC. We used an isobaric tags for relative and absolute quantitation (iTRAQ)-based strategy to detect 191 and 62 up-regulated, and 112 and 57 down-regulated proteins in the NEJ-Teg and NEJ-Som extracts, respectively. Similarly, 87 up-regulated and 73 down-regulated proteins in the MPSIEC-Lys extract were identified. Taking into account the biological processes in which these proteins were involved, interesting mechanisms related to parasite development, invasion and evasion, as well as manipulation of the host intestinal epithelial cell adhesion, immunity and apoptosis pathways, among others, could be inferred, taking place at the host-parasite interface. The further understanding of these processes could constitute promising therapeutic targets in the future against fasciolosis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The aim of this work was to identify the molecular characteristics of a chymotrypsin-like enzyme from Trichinella spiralis (Tschy) and its facilitation of larval penetration into enteral epithelial cells (EECs). The complete Tschy cDNA sequence was cloned and expressed in Escherichia coli BL21. RT-PCR, IIFA and western blotting showed that Tschy was expressed at the T. spiralis muscle larvae (ML), intestinal infective L1 larvae (IL1), adult worms (AW) and embryo stages and was primarily located in the stichosome of this parasite. The results of ELISA, IIFA and Far-western assays showed that there was a specific binding between rTschy and EECs, and the binding was dependent on the dose of both rTschy and EEC proteins. Confocal microscopy demonstrated that the binding was located in the EEC cytoplasm. rTschy facilitated T. spiralis larval penetration of EECs, and anti-rTschy antibodies impeded the larval intrusion of EECs. These results demonstrate that Tschy facilitated the larval intrusion of the host's enteral epithelium and could be a candidate molecular target for vaccine against the enteral invasive phase of T. spiralis., (Copyright © 2019 Elsevier Ltd. All rights reserved.)