Mechanisms of the host immune response and helminth-induced pathology during Trichobilharzia regenti (Schistosomatidae) neuroinvasion in mice.

Helminth neuroinfections represent serious medical conditions, but the diversity of the host-parasite interplay within the nervous tissue often remains poorly understood, partially due to the lack of laboratory models. Here, we investigated the neuroinvasion of the mouse spinal cord by Trichobilharzia regenti (Schistosomatidae). Active migration of T. regenti schistosomula through the mouse spinal cord induced motor deficits in hindlimbs but did not affect the general locomotion or working memory. Histological examination of the infected spinal cord revealed eosinophilic meningomyelitis with eosinophil-rich infiltrates entrapping the schistosomula. Flow cytometry and transcriptomic analysis of the spinal cord confirmed massive activation of the host immune response. Of note, we recorded striking upregulation of the major histocompatibility complex II pathway and M2-associated markers, such as arginase or chitinase-like 3. Arginase also dominated the proteins found in the microdissected tissue from the close vicinity of the migrating schistosomula, which unselectively fed on the host nervous tissue. Next, we evaluated the pathological sequelae of T. regenti neuroinvasion. While no demyelination or blood-brain barrier alterations were noticed, our transcriptomic data revealed a remarkable disruption of neurophysiological functions not yet recorded in helminth neuroinfections. We also detected DNA fragmentation at the host-schistosomulum interface, but schistosomula antigens did not affect the viability of neurons and glial cells in vitro. Collectively, altered locomotion, significant disruption of neurophysiological functions, and strong M2 polarization were the most prominent features of T. regenti neuroinvasion, making it a promising candidate for further neuroinfection research. Indeed, understanding the diversity of pathogen-related neuroinflammatory processes is a prerequisite for developing better protective measures, treatment strategies, and diagnostic tools.

Contribution of ultrasound examination in diagnosing platynosomiasis and correlation with macro and microscopic findings in callitrichids kept under human care.

BACKGROUND: Platynosomiasis in non-human primates kept under human care causes chronic disease of the bile ducts and liver, which initially presents with nonspecific signs and can culminate in the death of the animal. Diagnosing this disease is a challenge, and an ultrasound examination can be an excellent tool when it is suspected. METHODS: This study describes the ultrasound findings from 57 marmosets with suspected infection by Platynosomum sp., the correlated hepatobiliary changes, and the anatomopathological findings that confirmed the occurrence of platynosomiasis. RESULTS: In six marmosets (one C. aurita, two C. jacchus, and three Callithrix sp.), Platynosomum infection was confirmed macroscopically (presence of adult trematodes in the gallbladder) and microscopically (adults, larvae, and eggs in histological examinations and eggs in bile and feces). These findings were compatible with the hepatobiliary changes and with images suggestive of parasitic structures in ante-mortem assessments. CONCLUSION: Ultrasound examination demonstrated its usefulness within the clinical routine for investigating this parasitosis.

Anatomopathological lesions of infection caused by Platynosomum illiciens (Braun, 1901) in Neotropical primates kept in captivity.

This study described the hepatobiliary anatomopathological lesions associated with trematode Platynosomum illiciens parasitism in Neotropical primates kept in captivity. In the evaluated organs, we observed portal fibrosis, biliary epithelial hyperplasia, and inflammatory reaction with a predominance of lymphocytes and plasmocytes, and in some cases infiltration of eosinophils and neutrophils.

Characterization of digenetic trematodes infecting African catfish (Clarias gariepinus) based on integrated morphological, molecular, histopathological, and immunological examination.

Parasitic infection may cause massive losses in Clarias gariepinus fries and fingerlings. Therefore, this study aimed to characterize the digenetic trematodes species (two adults' flukes and one metacercariae) infecting African catfish Clarias gariepinus, as well as their histopathological impacts on infected fish. The intestinal flukes were identified as Orientocreadium batrachoides and Masenia bangweulensis based on their morphological and molecular characteristics. Sequencing of their 28S (LSU rRNA) and 18S rRNA (SSU rRNA) genes confirmed that these trematodes belong to the families Orientocreadiidae and Cephalogonimidae, respectively. The metacercariae trematode infecting skin and muscles were only morphologically identified as Cyanodiplostomum sp. The gene expression levels of MHC II increased in naturally infected fish either with O. batrachoides or Cyanodiplostomum sp. alone, compared with uninfected catfish. In addition, lysozyme levels in individual fish serum increased in catfish infected either with O. batrachoides or Cyanodiplostomum sp. alone. Histopathological examination of the skin revealed embedded parasitic cysts that displaced tissue in the dermis. Surrounding tissues were infiltrated with melanomacrophages and displayed dermal edema. Histopathological analysis showed O. batrachoides or M. bangweulensis between the gastric folds of the stomach of infected catfish, causing infiltration of mononuclear inflammatory cells in the lamina propria.

Renicolidae infection in Manx shearwater (Puffinus puffinus): is parasitism implicated on renal lesions?

Renicolid digeneans are frequently observed in the renal tubules and ureters of seabirds, such Puffinus puffinus, a migratory species distributed along the Brazilian coast. However, few studies have focused on the relationship between renicolid infection and health status in P. puffinus. Thus, the aim of this study was to describe (i) renal and systemic alterations, (ii) the renicolids and (iii) the biological aspects associated with the presence of renicolids in P. puffinus. Gross and histological assays were performed in 93 P. puffinus stranded on the Paraná coast, southern Brazil, and renicolids were submitted to morphological and molecular assays. A high prevalence of renicolids in P. puffinus (71/93) was observed. In the kidney, the main microscopic findings were lymphocytic interstitial infiltrate, ductal ectasia and tubular necrosis. The renal lesions were significantly associated with the parasite infection. The morphological (n = 84) and molecular analyses (n = 2) confirmed the species as Renicola sloanei (100% and 95.9% of nucleotide identity with R. sloanei strains from P. puffinus and from Spheniscus demersus, respectively). In both parasitized and non-parasitized animals, cardiac and skeletal muscle degeneration and necrosis were the most frequent systemic changes. Therefore, the results suggest renicolids being a possible cause for the demonstrated renal alterations. A contribution of this parasite to a decreased health status of Puffinus puffinus along their migratory route is possible.

Pathological effects of Cichlidogyrus philander Douëllou, 1993 (Monogenea, Ancyrocephalidae) on the gills of Pseudocrenilabrus philander (Weber, 1897) (Cichlidae).

Histopathological changes of Cichlidogyrus philander Douëllou, 1993 on the gills of Pseudocrenilabrus philander (Weber, 1897) were studied using light and scanning electron microscopy. Observations revealed that C. philander attaches to its host by alternating the prohaptor (for temporary attachment or feeding) or haptor (using haptoral parts for firm and secured attachment). The sharp terminal ends of the anchors are inserted basally into the gill lamella, between two adjacent secondary gill lamellae and the marginal hooklets assist by superficially penetrating, holding and lifting epithelial tissue in the proximal region of the secondary gill lamella. The attachment of C. philander resulted in compression, rupturing of the interlamellar epithelium, change in the organization of epithelial cells in both primary and secondary gill lamella, displacement of the extracellular cartilaginous matrix, occasional rupturing of blood vessels and erythrocytes and some cells becoming ill-defined. At the site of attachment, the host response comprises of hyperplasia, increase in the number of mucous cells and infiltration with neutrophils. It was concluded that the effect of C. philander is mild in natural conditions.

Pathology associated with three new Clinostomum metacercariae from Argentina with morphological and DNA barcode identification.

In the Laboratory of Parasites of Fishes, Crustaceans and Mollusks (CEPAVE), we undertook a parasitological study on three species of fish from the Espinal and Esteros del Iberá ecoregions of Argentina. Clinostomid metacercariae were found parasitizing Characidium rachovii, Crenicichla vittata and Gymnogeophagus balzanii. In this study, we analysed the damage that these parasites inflict on their hosts through the evaluation of histological sections. In addition, Clinostomum metacercariae were identified using morphological characters and DNA barcoding. In the pathological analysis, we observed that muscle tissue was the most affected. The inflammatory response showed vascular congestion areas and infiltration of numerous inflammatory cells, mainly lymphocytes. The molecular and morphological approach supports the presence of three new lineages of clinostomid metacercariae in Argentina. This could lead to the discovery of a high number of lineages or species of Clinostomum from South America.

Multiple food-borne trematodiases with profound systemic involvement: a case report and literature review.

BACKGROUND: Food-borne trematodiases are an important group of neglected global diseases. Affected patients in regions with low prevalence usually experience delayed diagnosis, especially when presenting with atypical clinical symptoms. Here, we presented a rare case of a Chinese patient infected with three food-borne trematodiases. CASE PRESENTATION: A 42-year-old man presented with diarrhea, lower extremity edema, and symptoms of cardiac dysfunction. He had a history of intermittent consumption of raw freshwater fishes for 6-7 years. Upon evaluation, he had eosinophilia, anemia, intrahepatic bile duct dilatation and a growing space-occupying lesion in the left atrium. The patient underwent a cardiac surgery which revealed an endocardial hematoma due to mechanical injuries. Imaging investigations also revealed intracranial and pulmonary lesions. A total of three trematodiases were diagnosed based upon microscopic stool examination, from which eggs of Clonorchis sinensis, Heterophyidae and Echinostomatidae were identified. Deposition of Clonorchis sinensis eggs was also observed from ileocecal squash slides. The patient was successfully treated with three cycles of praziquantel. CONCLUSIONS: Food-borne trematodiases may present with systemic involvement. Patients with dietary history of high risk or atypical ingestions should be evaluated for parasitic infection, even in non-endemic areas.

Fascioloides magna in Moose ( Alces alces) From Elk Island National Park, Alberta.

Thirty-seven adult female moose ( Alces alces) from 2 distinct but adjacent populations in Elk Island National Park (EINP), Alberta, Canada (19 in north EINP and 18 in south EINP), were fitted with mortality-sensing VHF radio-collars, and radio signals were acquired daily to ascertain mortality status. At capture, serum, whole blood, and feces were collected; pregnancy was determined; teeth were aged by visual inspection; and a portion of liver was assessed by ultrasound examination. Postmortem examination was conducted on 20 suitable carcasses. Clinical pathological abnormalities, including eosinophilia, polycythemia, elevated levels of liver enzymes in serum, hemoglobin, hematocrit, and red blood cell distribution, and liver damage as seen in ultrasound images occurred only in moose from north EINP. Infected moose had 4.7 ± 4.8 Fascioloides magna flukes per liver (mean ± SD). The proportion of moose pregnant at capture was similar in both populations (74% in north EINP, 61% in south EINP). Proportional mortality was significantly higher in moose from the north (68%) than the south (32%). Fascioloides magna was associated as a cause of death in 7 of 14 (50%) moose in the north where cause of death was determined, while predation ( n = 1), acute toxemic syndrome ( n = 3), dystocia ( n = 1), and roadkill and undetermined causes ( n = 3) were additional causes of mortality. F. magna was associated with poor body condition and was a major cause of mortality in north EINP but not south EINP, despite very similar habitat and proximity, suggesting a significant role for these flukes in affecting health and viability of naturally infected moose populations.

Histopathological changes induced by the digenean intestinal parasite Masenia nkomatiensis Dumbo, Dos Santos, & Avenant-Oldewage, 2019 of the catfish Clarias gariepinus (Burchell) from Incomati Basin, Mozambique.

The intestines of 154 Clarias gariepinus were examined of which 29 were naturally infected with Masenia nkomatiensis, and of these, seven (intensity ranging from 8 to 231) were examined for pathology. Destruction of the epithelium covering the villi, detachment of epithelial cells and parts of villi were observed. Excessive mucus secretion occurred in the vicinity of the worm and catarrh was observed, indicative of an inflammatory response. The number of mucous and mast cells was higher at the attachment site than at an area 5,000 µm away and in uninfected individuals, suggesting that the parasite triggered a localized innate immune response. The number of neutrophils, basophils and lymphocytes in infected tissue was not significantly different from uninfected tissue confirming that no acquired immune response was produced against the maseniid. The caecae in the anterior part of the parasites' intestine consisted of convoluted epithelium forming invaginations or "crypts." Contraction of the thick layer of circular muscle fibres of the caeca facilitates the movement of digested material. Observation of digested host cells and cell debris within the caecae provides further evidence that M. nkomatiensis is consuming host cells.

A Case of Unexpected Clinostomum complanatum Infection Initially Presenting as Foreign Body in Pharynx.

A 46-year old man visited our outpatient clinic with complaint of foreign body sensation in throat after consuming raw freshwater fish 5 days ago. Laryngoscopic examination revealed a motile worm attached on posterior pharyngeal wall. The worm was removed using biopsy forceps under transnasal endoscopy and evidently identified as Clinostomum complanatum after microscopic examination. Patient's subjective foreign body sensation of throat and hyperemia of laryngeal mucosa remained for approximately 2 weeks post-removal, which were eventually resolved after administration of non-steroidal anti-inflammatory drug and anti-refluxant drug for 2 weeks. Treatment was ended at three weeks since the first visit. C. complanatum infections in humans are rare, and only four cases have been reported in Korea. Symptoms resembling pharyngitis or laryngitis occurs by consumption of raw, infected freshwater fish and treatment is done by mechanically removing the parasite.

Necrotizing Enteritis Caused by Pharyngostomum cordatum Infection in a Stray Cat.

A stray female cat of unknown age, presenting bright red watery diarrhea, was submitted to the Animal and Plant Quarantine Agency for diagnosis. In the small intestines extracted from the necropsied cat, numerous white oval-shaped organisms were firmly embedded in the mucosa and there was thickening of intestinal wall. Histopathological analysis revealed severe necrotizing enteritis, together with atrophied intestinal villi, exfoliated enterocytes, and parasitic worms. Recovered worms were identified as Pharyngostomum cordatum by morphological observation and genetic analysis. Although P. cordatum is known to occur widely in Korea, this is the first clinical description of an infection by P. cordatum causing severe feline enteritis.

Prominent hepatic ductular reaction induced by Oschmarinella macrorchis in a Hubbs' beaked whale Mesoplodon carlhubbsi, with biological notes.

Beaked whales are among the least known group of cetaceans, and information regarding their pathology and parasitology is especially scarce. We describe a case of significant parasitism by a trematode found in the liver of an adult male Hubbs' beaked whale Mesoplodon carlhubbsi that stranded in Hokkaido, Japan. Post-mortem examinations revealed a localised area of discolouration restricted to the hilar region of the left hepatic lobe, where spindle-shaped trematodes occupied the dilated and hypertrophic bile ducts. Histologically, the intrahepatic bile ducts were characterised by adenomatous hyperplasia with goblet cell metaplasia of the biliary epithelium. Findings in the adjacent hepatic parenchyma included pseudocarcinomatous ductular reactions obliterating hepatocytes, a histomorphology not previously reported in marine mammals. Morphological identification of the trematode corresponded to Oschmarinella macrorchis, which has only been reported once in a Stejneger's beaked whale, M. stejnegeri. PCR amplification and sequencing analyses of the parasite's mtDNA ND3, 18S and 28S rRNA regions generated novel gene sequences. Environmental contaminant levels were measured to explore its potential relationship with the parasitism but there was no conclusive association. A high level of polychlorinated biphenyl (30000 ng g-1 lipid weight) was detected in the blubber of this individual, when compared to those of 3 other male Hubbs' beaked whales stranded in Japan. Stomach contents were also analysed, indicating the presence of various squid species and unidentified fish. Our results contribute to the knowledge of a little-known beaked whale and provide evidence for the first time of the pathobiological response caused by O. macrorchis.

Histopathological and ultrastructural assessment of two mugilid species infected with myxozoans and helminths.

The histopathology and ultrastructure of the intestine of mullets, Liza ramada and Liza saliens, from Comacchio lagoons (northern Italy) naturally infected with myxozoans and helminths were investigated and described. Sixty-two (80.5%) of 77 mullets harboured one or more of the following parasites species: Myxobolus mugchelo (Myxozoa), Neoechinorhynchus agilis (Acanthocephala), Haplosplanchnus pachysomus and Dicrogaster contractus (Digenea). Co-occurrence of helminths with myxozoans was common. The main damage caused by digeneans was destruction of the mucosal epithelium of the villi, necrosis and degeneration of intestinal epithelial cells. More severe intestinal damage was caused by acanthocephalans which reach the submucosa layer with their proboscis. At the site of helminths infection, several mast cells (MCs), rodlet cells (RCs), mucous cells and few neutrophils and macrophages were observed in the epithelium. RCs and mucous cells exhibited discharge activity in close vicinity to the worm's tegument. M. mugchelo conspicuous plasmodia were encysted mainly in muscle and submucosa layers of the intestine. Indeed, spores of M. mugchelo were documented within the epithelial cells of host intestine and in proximity to MCs. Degranulation of the MCs near the myxozoans was very frequent.

Gastric lesions in dolphins stranded along the Eastern Adriatic coast.

Stranded cetaceans are often found with gastric lesions associated with the presence of parasites; most frequently, nematodes of the genus Anisakis and the heterophyd digenean trematode Pholeter gastrophilus. In this study, we present histopathology mainly (but not exclusively) related to these 2 parasite species. Macroscopically, lesions associated with the presence of Anisakis spp. were characterised by the presence of ulcers within the gastric mucosa, while the digenean P. gastrophilus was found within large submucosal fibrotic nodules in the gastric wall. Anisakis-induced alterations included severe ulcerative gastritis with mixed inflammatory infiltrate often associated with colonies of bacteria, and mild to moderate granulomatous gastritis with eosinophilic infiltrate. P. gastrophilus-associated lesions were characterised by fibrogranulomatous gastritis with mixed inflammatory infiltrate. Additionally, immunohistochemical (IHC) analysis of P. gastrophilus lesions was consistent with the histopathologic findings, revealing inflammation-mediated stimulation. IHC-positive localisation of CD3+, iNOS+ and caspase-3+ cells suggests intensive accumulation of cytotoxic T-cells, proinflammatory cytokines and execution-phase of cell apoptosis at the parasitized area. In contrast, mechanical damage, rather than visible inflammatory response could be observed at the site of attachment of Braunina cordiformis recorded in 4 animals. Lesions not associated with the presence of parasites were mostly characterised by focal loss of superficial epithelial cells and accumulation of brown hemosiderin-like pigment or fibrous gastritis with lymphoplasmacytic infiltrate. In light of these results, we argue that observed 'tolerant' host-parasite interactions that led toward gastric lesions do not represent the cause of death and stranding of cetaceans included in this study.

Moderate pathogenic effect of Ligophorus uruguayense (Monogenoidea, Ancyrocephalidae) in juvenile mullet Mugil liza (Actinopterygii, Mugilidae) from Brazil.

Monogenoidea pathogenic activity can elicit various histological responses in fish. Species of Ligophorus are specific parasites of mullets, and its relationship with host fish may result in a moderate pathogenic action. In order to ascertain this relationship, estuarine mullets (Mugil liza) were collected in an estuary, reared in laboratory, for three weeks, and forwarded for histological and parasitological analyses. Ligophorus uruguayense (Monogenoidea) infestation in the gills of the mullets was identified. The severe infestation by only one species of Monogenoidea may result from the specificity of these parasites to mullets. Mullets submitted to histological analysis exhibited respiratory epithelium detachment; mild, moderate and severe hyperplasia of the respiratory epithelium; atrophy; and telangiectasia of the gills. This is the first study reporting that mullets highly infested by Monogenoidea can show mild (100%) to severe (20%) gill changes with a distinct frequency of occurrence. Because of the high prevalence of mild alterations observed, it is possible to accept that L. uruguayense is moderately pathogenic to M. liza, even during high prevalence and intensity of infestation, as a result of its specificity.

Calicophoron daubneyi and Fasciola hepatica: characteristics of natural and experimental co-infections of these digeneans in the snail Lymnaea glabra.

A retrospective study on different Lymnaea glabra samples collected from central France between 1993 and 2010 was carried out to determine the prevalence of natural co-infections with Calicophoron daubneyi and Fasciola hepatica, and to specify the composition of redial burdens. Experimental infections of L. glabra performed during the same period of time were also analysed to study metacercarial production of each digenean in co-infected snails. Controls were naturally or experimentally co-infected Galba truncatula. In natural co-infections, prevalence was 0.7% in L. glabra (186/25,128) and 0.4% in G. truncatula (137/31,345). Low redial burdens were found in these snails, with F. hepatica rediae significantly more numerous in L. glabra than in G. truncatula (7.5 per snail instead of 5.2). In contrast, the total numbers of C. daubneyi rediae in both lymnaeids were close to each other (4.3 and 3.0 rediae, respectively). In experimentally co-infected groups, prevalence was greater in G. truncatula than in the other lymnaeid (6.3% instead of 3.0%). Significantly shorter patent periods and lower metacercarial production for each digenean were noted in L. glabra than in G. truncatula. However, in both lymnaeids, the two types of cercariae were released during the same shedding waves and several peaks during the patent period were synchronous. In spite of a greater shell height for L. glabra, metacercarial production of both digeneans in co-infected snails was lower than that in G. truncatula, thus indicating a still incomplete adaptation between these French L. glabra and both parasites.

Cytologic Diagnosis of Heterobilharzia americana Infection in a Liver Aspirate From a Dog.

Heterobilharzia americanais a trematode of the Schistosomatidae family that infects dogs, raccoons, and other mammals as definitive hosts. This parasite is considered endemic in the southern Atlantic and Gulf coasts; however, only a few cases are reported. A 7-year-old dog from Louisiana was referred for persistent hypercalcemia, hyperglobulinemia, and weight loss. Abdominal ultrasound revealed diffuse hyperechogenicity of the liver with several hyperechoic nodules of varying size. Cytologic examination of fine-needle aspirates of the liver revealed few ovoid to round basophilic thin-walled eggshell fragments and rare ciliated miracidia.H. americanaeggs were identified on fecal sedimentation.

Malformations of the gill filaments of the ruffe Gymnocephalus cernuus (L.) (Pisces) caused by echinostomatid metacercariae.

In parasite surveys of fishes from Lake Balaton and its tributaries in Hungary, infections with metacercariae of a species of the digenean genus Echinochasmus (Trematoda: Echinostomatidae) were found in seven species of fish. In ruffe, Gymnocephalus cernuus, malformations of the gill filaments apparently caused by these infections were observed. These malformations were in the form of bifurcations of the filaments at about their mid-length. At the point where the filaments bifurcate, an Echinochasmus metacercaria was always embedded in the cartilaginous ray of the gill filament. All specimens of the ruffe were found to be infected by these metacercariae, and each ruffe specimen was infected by 30-300 metacercariae. Such a bifurcation was found in all of the ruffe specimens, but, apart from these gill malformations, the metacercariae produced only local changes in the cartilage. In the other six infected fish species, only local signs were observed in the cartilage. Experimental infections of chicks with metacercariae resulted in the finding of the sexual adult (marita) of an unidentified species of Echinochasmus. ITS sequences of the adult and metacercaria corresponded with each other, and also with a cercaria isolated from a gravel snail (Lithoglyphus naticoides), with a 99.5-100% similarity.