Production and characterization of antibody against Opisthorchis viverrini via phage display and molecular simulation.

In this study, a key issue to be addressed is the safe disposal of hybridoma instability. Hybridoma technology was used to produce anti-O. viverrini monoclonal antibody. Previous studies have shown that antibody production via antibody phage display can sustain the hybridoma technique. This paper presents the utility of antibody phage display technology for producing the phage displayed KKU505 Fab fragment and using experiments in concomitant with molecular simulation for characterization. The phage displayed KKU505 Fab fragment and characterization were successfully carried out. The KKU505 hybridoma cell line producing anti-O. viverrini antibody predicted to bind to myosin was used to synthesize cDNA so as to amplify the heavy chain and the light chain sequences. The KKU505 displayed phage was constructed and characterized by a molecular modeling in which the KKU505 Fab fragment and -O. viverrini myosin head were docked computationally and it is assumed that the Fab fragment was specific to -O. viverrini on the basis of mass spectrometry and Western blot. This complex interaction was confirmed by molecular simulation. Furthermore, the KKU505 displayed phage was validated using indirect enzyme-linked immunosorbent assays (ELISA) and immunohistochemistry. It is worthy to note that ELISA and immunohistochemistry results confirmed that the Fab fragment was specific to the -O. viverrini antigen. Results indicated that the approach presented herein can generate anti-O. viverrini antibody via the phage display technology. This study integrates the use of phage display technology together with molecular simulation for further development of monoclonal antibody production. Furthermore, the presented work has profound implications for antibody production, particularly by solving the problem of hybridoma stability issues.

Expanded complement of Niemann-Pick type C2-like protein genes in Clonorchis sinensis suggests functions beyond sterol binding and transport.

BACKGROUND: The parasitic flatworm Clonorchis sinensis inhabits the biliary tree of humans and other piscivorous mammals. This parasite can survive and thrive in the bile duct, despite exposure to bile constituents and host immune attack. Although the precise biological mechanisms underlying this adaptation are unknown, previous work indicated that Niemann-pick type C2 (NPC2)-like sterol-binding proteins might be integral in the host-parasite interplay. Expansions of this family in some invertebrates, such as arthropods, have shown functional diversification, including novel forms of chemoreception. Thus, here we curated the NPC2-like protein gene complement in C. sinensis, and predicted their conserved and/or divergent functional roles. METHODS: We used an established comparative genomic-bioinformatic approach to curate NPC2-like proteins encoded in published genomes of Korean and Chinese isolates of C. sinensis. Protein sequence and structural homology, presence of conserved domains and phylogeny were used to group and functionally classify NPC2-like proteins. Furthermore, transcription levels of NPC2-like protein-encoding genes were explored in different developmental stages and tissues. RESULTS: Totals of 35 and 32 C. sinensis NPC2-like proteins were predicted to be encoded in the genomes of the Korean and Chinese isolates, respectively. Overall, these proteins had low sequence homology and high variability of sequence alignment coverage when compared with curated NPC2s. Most C. sinensis proteins were predicted to retain a conserved ML domain and a conserved fold conformation, with a large cavity within the protein. Only one protein sequence retained the conserved amino acid residues required in bovine NPC2 to bind cholesterol. Non-canonical C. sinensis NPC2-like protein-coding domains clustered into four distinct phylogenetic groups with members of a group frequently encoded on the same genome scaffolds. Interestingly, NPC2-like protein-encoding genes were predicted to be variably transcribed in different developmental stages and adult tissues, with most being transcribed in the metacercarial stage. CONCLUSIONS: The results of the present investigation confirms an expansion of NPC2-like proteins in C. sinensis, suggesting a diverse array of functions beyond sterol binding and transport. Functional explorations of this protein family should elucidate the mechanisms enabling the establishment and survival of C. sinensis and related flukes in the biliary systems of mammalian hosts.

Differential responses of epithelial cells from urinary and biliary tract to eggs of Schistosoma haematobium and S. mansoni.

Chronic urogenital schistosomiasis can lead to squamous cell carcinoma of the bladder. The International Agency for Research on Cancer classifies the infection with S. haematobium as a group 1 carcinogen, a definitive cause of cancer. By contrast, hepatointestinal schistosomiasis due to the chronic infection with S. mansoni or S. japonicum associated with liver periportal fibrosis, does not apparently lead to malignancy. The effects of culturing human epithelial cells, HCV29, established from normal urothelium, and H69, established from cholangiocytes, in the presence of S. haematobium or S. mansoni eggs were investigated. Cell growth of cells co-cultured with schistosome eggs was monitored in real time, and gene expression analysis of oncogenesis, epithelial to mesenchymal transition and apoptosis pathways was undertaken. Schistosome eggs promoted proliferation of the urothelial cells but inhibited growth of cholangiocytes. In addition, the tumor suppressor P53 pathway was significantly downregulated when exposed to schistosome eggs, and downregulation of estrogen receptor was predicted in urothelial cells exposed only to S. haematobium eggs. Overall, cell proliferative responses were influenced by both the tissue origin of the epithelial cells and the schistosome species.

Endogenous development of Cystoisospora belli in intestinal and biliary epithelium of humans.

Cystoisospora (Isospora) belli is a coccidian parasite of humans. It can cause serious digestive disorders involving infection of intestines, biliary tract and gallbladder, especially in those with depressed immunity. It has a direct fecal-oral transmission cycle. After ingestion of sporulated oocysts, the parasite multiplies asexually and sexually within host epithelial cells, resulting in unsporulated oocysts that are excreted in feces. The details of asexual and sexual stages are not known and certain inclusions in epithelial cells in biopsy samples have been erroneously identified recently as C. belli. Here, we provide details of developmental stages of C. belli in two patients, in duodenal biopsy of one and biliary epithelium of the other. Immature and mature asexual stages (schizonts/meronts) were seen in epithelial cells. The merozoites were seen singly, in pairs and in groups in single parasitophorous vacuole (pv) in host cytoplasm. Immature and mature meronts were seen together in the same pv; up to eight nuclei were seen in meronts that retained elongated crescent shape; round multinucleated schizonts, seen in other coccidians, were not found. Meronts were up to 25 µm long and contained up to ten merozoites that were 8-11 µm long. The merozoites and meronts contained PAS-positive granules. Microgamonts (male) contained up to 30 nuclei that were arranged at the periphery and had condensed chromatin; 1-3 PAS-positive, eosinophilic, residual bodies were left when microgametes were formed. The microgametes were 4 µm long and PAS-negative. All stages of macrogamonts, including oocysts were PAS-positive. The detailed description of the life cycle stages of C. belli reported here should facilitate in histopathologic diagnosis of this parasite.

Misleading findings of liver-specific MR contrast agent for radiological diagnosis of cysto-biliary communication in hydatid cysts.

OBJECTIVE: To investigate the effectiveness of gadoxetic acid-enhanced magnetic resonance cholangiopancreatography (CE-MRCP) and T2-weighted half-Fourier acquisition single-shot turbo spin-echo (T2W HASTE) sequences for diagnosis of cysto-biliary communication in hydatid cysts compared to surgical results. METHODS: Preoperative abdominal magnetic resonance imaging examinations of patients who underwent surgery for hepatic hydatid cysts were reviewed by two radiologists retrospectively. A total of 45 patients with hydatid cysts were included. Of 45, 27 also had CE-MRCPs. T2W HASTE sequences and CE-MRCPs were investigated separately for cysto-biliary communication. The relationship between radiological and surgical results was analyzed. The interobserver agreement was evaluated. RESULTS: Of 45 hydatid cysts, there were surgically proven 21 cysts without biliary communications and 24 cysts with biliary communications. All cysts with biliary communications were shown on T2W HASTE sequences. There was no leakage of gadoxetic acid into these cysts (n = 24). Sensitivity, specificity, negative predictive value, positive predictive value (PPV), and accuracy of diagnosis of cyst with biliary communication on T2W HASTE sequences was 100%, 63.64%, 100%, 66.67%, and 78.95%, respectively. Specificity (77.78%), PPV (87.50%), and accuracy (91.30%) were increased in ≥ 10 cm cysts. There was almost perfect interobserver agreement (K = 0.81-1.00). CONCLUSION: Leakage of gadoxetic acid inside the cyst indicates biliary communication. However, the lack of leakage does not rule out cysto-biliary communication. When biliary communication is clearly shown on T2W HASTE sequences, it should be reported as cysto-biliary communication even if there is no leakage of gadoxetic acid into the cyst on CE-MRCP.

Clonorchiasis sinensis detected by laparoscopic exploration of biliary tracts in two patients with obstructive jaundice.

BACKGROUND: Hepatic clonorchiasis is one of the most prevalent food-borne parasitic diseases worldwide. Clonorchis sinensis, the pathogen, is the major parasitic trigger contributing to cholangitis, cholelithiasis, and even cholangiocarcinoma. Unfortunately, unspecific clinical manifestations of patients with hepatic clonorchiasis tend to mislead clinicians to neglect or misdiagnose them, following ignorance of appropriate therapy. Our case report may shed light on definite diagnosis of clonorchiasis with concomitant cholelithiasis, methodology for surgical drainage of the parasites, and postoperative anthelmintic therapy. CASE PRESENTATION: Two patients with habit of eating infected raw or undercooked freshwater fish were hospitalized due to right upper quadrant pain and jaundice. Magnetic resonance cholangiopancreatography (MRCP)/computed tomography (CT) detection indicated cholangiolithiasis and cholangiolithiasis with concurrent cholecystolithiasis, respectively. Fecal examinations were both negative for adult worms or eggs of parasites. However, adults of Clonrochis sinensis were detected within hepatobiliary tracts during laparoscopic cholecystectomy. Postoperative drainage and anthelmintic therapy contributed to complete recovery with good prognosis. CONCLUSIONS: Clonorchiasis provokes cholangiolithiasis and cholecystolithiasis. Standardized treatments for these gallstone patients with concomitant clonorchiasis include surgical removal of the calculus, postoperative T tubule drainage and anthelmintic therapy. Serological test or polymerase chain reaction (PCR)-based approaches might be helpful for diagnosis of clonorchiasis when no eggs are found by stool microscopy. Public health promotion on ceasing to eat raw freshwater fish is essential for prevention and control of clonorchiasis.

Biliary Taeniasis with Cholecystitis: An Unusual Case of Taenia solium Infection with a Literature Review.

Taeniasis is a cosmopolitan helminthic disease caused by Taenia species, which included Taenia solium, Taenia saginata, and Taenia asiatica. These parasites typically infect the small intestine, but cases of aberrant migration have been reported. We treated a 70-year-old man who presented with vomiting and colicky abdominal pain. On physical examination, Murphy's sign was positive, and laboratory findings indicated severe inflammation. Computed tomography and magnetic resonance cholangiopancreatography revealed typical features of cholecystitis. An 82-cm-long, slender and degenerated, parasite-like organism was aspirated through a percutaneous transhepatic gallbladder drainage tube. After extensive washing of the organism, we detected yellowish-brown colored, spherical 37.9 × 33.8-µm-sized taenid eggs with thick transverse striations. Hematoxylin-eosin-stained worm sections also contained Taeniidae eggs. Polymerase chain reaction amplification of DNA extracted from the worm with species-specific cytochrome c1 (cox1) primer sets detected a T. solium-specific fragment. Because of sustained high fever combined with inflammatory signs, the patient underwent laparoscopic cholecystectomy and inflamed gallbladder removal. A histopathologic specimen demonstrated chronic reactive cholecystitis. The patient's fever and leukocytosis rapidly resolved after surgery. We experienced an uncommon case of biliary taeniasis representing cholecystitis caused by adult worm of T. solium.

Co-occurrence of opisthorchiasis and diabetes exacerbates morbidity of the hepatobiliary tract disease.

Complications arising from infection with the carcinogenic liver fluke Opisthorchis viverrini cause substantial morbidity and mortality in Thailand and adjacent lower Mekong countries. In parallel, the incidence rate of diabetes mellitus (DM) is increasing in this same region, and indeed worldwide. Many residents in opisthorchiasis-endemic regions also exhibit DM, but the hepatobiliary disease arising during the co-occurrence of these two conditions remains to be characterized. Here, the histopathological profile during co-occurrence of opisthorchiasis and DM was investigated in a rodent model of human opisthorchiasis in which diabetes was induced with streptozotocin. The effects of excretory/secretory products from the liver fluke, O. viverrini (OVES) on hepatocyte and cholangiocyte responses during hyperglycemic conditions also were monitored. Both the liver fluke-infected hamsters (OV group) and hamsters with DM lost weight compared to control hamsters. Weight loss was even more marked in the hamsters with both opisthorchiasis and DM (OD group). Hypertrophy of hepatocytes, altered biliary canaliculi, and biliary hyperplasia were more prominent in the OD group, compared with OV and DM groups. Profound oxidative DNA damage, evidenced by 8-oxo-2'-deoxyguanosine, proliferating cell nuclear antigen, and periductal fibrosis characterized the OD compared to OV and DM hamsters. Upregulation of expression of cytokines in response to infection and impairment of the pathway for insulin receptor substrate (IRS)/phosphatidylinositol-3-kinases (PI3K)/protein kinase B (AKT) signaling attended these changes. In vitro, OVES and glucose provoked time- and dose-dependent effects on the proliferation of both hepatocytes and cholangiocytes. In overview, the co-occurrence of opisthorchiasis and diabetes exacerbated pathophysiological damage to the hepatobiliary tract. We speculate that opisthorchiasis and diabetes together aggravate hepatobiliary pathogenesis through an IRS/PI3K/AKT-independent pathway.

Biliary Tract-Infecting Myxosporeans from Estuarine and Reef Stonefish (Scorpaeniformes: Synanceiidae) Off Eastern Australia, with Descriptions of Sphaeromyxa horrida n. sp. and Myxidium lapipiscis n. sp. (Myxosporea: Bivalvulida).

Two new species of myxosporeans are described from the gallbladders of estuarine stonefish, Synanceia horrida, and reef stonefish, Synanceia verrucosa, from localities off Cairns, in tropical north Queensland and in Moreton Bay in southern Queensland, Australia. Sphaeromyxa horrida n. sp. can be distinguished from congeners in the morphologically distinct "balbianii" species group within Sphaeromyxa on the basis of morphometric differences in length and width of mature spores, length and width of polar capsules, and unique small-subunit (SSU) ribosomal (rDNA) sequence composition relative to other taxa. Replicate SSU rDNA sequences generated from Sph. horrida n. sp. collected from Sy. horrida and Sy. verrucosa in tropical north Queensland and from Sy. horrida in Moreton Bay were identical, suggesting that this species is widely distributed along the east coast of Australia. Myxidium lapipiscis n. sp. can be distinguished from the majority of described Myxidium species on the basis of its relatively small mature spore size (6.1-7.9 µm long × 3.1-3.9 µm wide), and its unique SSU rDNA sequence. Specimens putatively identified as M. lapipiscis n. sp. were found in Sy. horrida from both tropical north Queensland and Moreton Bay, suggesting that this taxon is also widely distributed along the east coast of Australia. However, no molecular data were available for the specimens from tropical north Queensland for comparative genetic analyses. Bayesian inference and maximum likelihood analysis of the SSU rDNA sequences for these 2 new species revealed that Sph. horrida n. sp. formed a strongly supported clade with Sphaeromyxa zaharoni Diamant, Whipps, and Kent, 2004, which was described from the scorpaeniform, Pterois miles, from the Red Sea. This is the first report of myxozoans infecting stonefish (Synanceiidae).

Rejection of the synonymization of Pegosomum saginatum (Ratz, 1898) Ratz, 1903 with Pegosomum asperum (Wright, 1879) Ratz, 1903.

The Pegosomum Ratz, 1903 are digenean parasites of piscivorous birds. They exhibit few morphological autapomorphies and some of their identification features (number of collar spines) can be altered before or during fixation. Several re-classifications within the genus were suggested, but they have never been supported by molecular analyses. We addressed the synonymization of species within Pegosomum asperum/saginatum complex suggested by Dubinin, Dubinina and Saidov. We analyzed one nuclear (ITS2) and two mitochondrial (CO1, ND1) loci of two central European species of Pegosomum, namely Pegosomum asperum (Wright, 1879) Ratz, 1903 and Pegosomum saginatum (Ratz, 1898) Ratz, 1903. Our combined molecular and comparative morphological analyses confirmed the validity of the two Pegosomum spp. Both species had highly similar morphology and occurred sympatrically in the gall bladder and bile duct of Ardea alba (Linnaeus, 1758). P. saginatum occurred more frequently in hosts infected concurrently with P. asperum. We also provided host-, age- and sex-specific prevalence and intensity of infections, and comparative measurements of the two Pegosomum spp. based on an extensive dataset collected from 1962 to 2016. These species can be morphologically distinguished based on the extent of vitellarium, which reached anteriorly to the pharynx in P. asperum but extended anteriorly to the midline of the esophagus in P. saginatum. The species also differed in the cirrus diameter (P. asperum≥447, P. saginatum≤447).

Fasciola hepatica-a "Diver" in the Biliary Tree.

Fasciola hepatica is an endemic trematode that affects millions of people worldwide. The human being can be an accidental host through consumption of contaminated food or water. The authors present a case of hepatic fascioliasis in a 69-year-old Portuguese woman who recently traveled from Egypt, Brazil, and India.

[Trans-hepatico cystic coledochostomy for the opening hydatid cyst in the biliary tract: Indications and outcome evaluation of 25 cases]. / La cholédochostomie transhépaticokystique dans le traitement des kystes hydatiques ouverts dans les voies biliaires : revue des indications et analyse des résultats d'une série de 25 cas.

The treatment of hydatid cysts of the liver opened in the bile ducts is sometimes difficult and complex. The trans-hepaticocystic coledochostomy (CTHK) is one of these processes. We conducted a review of the indications for this surgical procedure and its perioperative outcomes through a series of 25 patients and then we analyzed predictor factors of complications. During the period's study, 909 patients with liver hydatid cyst went under the surgery. Ninety two (92, i.e. 10.1%) of them had an opening in the bile ducts through a large fistula. Twenty five (25, i.e. 27%) had a trans-hepaticocystic coledochostomy. The sex ratio was 0.6. The population was young with an average age of 47 years. The trans-hepaticocystic coledochostomy was performed because of a thick pericyst associated with a nonsutured fistula in 18 cases, a voluminous residual cavity with a soft pericyst but the fistula was unexposed in six cases and in a case where the pericyst was thick, fistula was exposed. Its suture was associated with a high risk of bile duct stenosis. Specific morbidity was 20% due to a length of the intraductal coledochostomy under 2 cm (P = 0.016), the absence of an epiploplasty (P = 0.004) and the existence of a leak of contrast material outside the fistula on the cholangiography (P = 0.005). The trans-hepaticocystic coledochostomy is a safe and reliable technique, often indicated when other conservative methods are failing. The application of its technical requirements and avoidance of mounting errors allow reducing its morbidity.

Carbonyl stress phenomena during chronic infection with Opisthorchis felineus.

Infection with the fish borne liver fluke Opisthorchis felineus is common in the Eastern Europe (Ukraine, European part of Russia), Northern Asia (Siberia) and Central Asia (Northern Kazakhstan). Better understanding of the molecular pathogenesis of the biliary tract and liver during chronic opisthorchiasis can be expected to improve protection against and management of complications of this disease. We hypothesize that infection with O. felineus associates with formation of methylglyoxal and carbonyl stress in the liver and hence here we investigated the glyoxalase system and the receptor for advanced glycated end products (RAGE) in the liver of hamsters infected with this liver fluke. Expression of mRNA encoding glyoxalase 1 decreased at 8weeks of the infection and catalytic activity as well decreased at 8 and 12weeks after infection, and the expression of the glyoxalase 2 decreased until 36week post-infection, which associated with the decreasing activity of the enzyme at 8 and 12weeks post-infection. Glutathione levels in infected livers had decreased at week 8, whereas up-regulation of RAGE at mRNA levels was seen for the extended duration of the experimental infection of the hamsters. This outcome supported the notion of hepatic dicarbonyl stress during chronic opisthorchiasis. The inhibition of the glyoxalase system and accumulation of methylglyoxal at the early stages of the infection may underpin development of insulin resistance during opisthorchiasis.

Levels of 8-OxodG Predict Hepatobiliary Pathology in Opisthorchis viverrini Endemic Settings in Thailand.

Opisthorchis viverrini is distinct among helminth infections as it drives a chronic inflammatory response in the intrahepatic bile duct that progresses from advanced periductal fibrosis (APF) to cholangiocarcinoma (CCA). Extensive research shows that oxidative stress (OS) plays a critical role in the transition from chronic O. viverrini infection to CCA. OS also results in the excision of a modified DNA lesion (8-oxodG) into urine, the levels of which can be detected by immunoassay. Herein, we measured concentrations of urine 8-oxodG by immunoassay from the following four groups in the Khon Kaen Cancer Cohort study: (1) O. viverrini negative individuals, (2) O. viverrini positive individuals with no APF as determined by abdominal ultrasound, (3) O. viverrini positive individuals with APF as determined by abdominal ultrasound, and (4) O. viverrini induced cases of CCA. A logistic regression model was used to evaluate the utility of creatinine-adjusted urinary 8-oxodG among these groups, along with demographic, behavioral, and immunological risk factors. Receiver operating characteristic (ROC) curve analysis was used to evaluate the predictive accuracy of urinary 8-oxodG for APF and CCA. Elevated concentrations of 8-oxodG in urine positively associated with APF and CCA in a strongly dose-dependent manner. Urinary 8-oxodG concentrations also accurately predicted whether an individual presented with APF or CCA compared to O. viverrini infected individuals without these pathologies. In conclusion, urinary 8-oxodG is a robust 'candidate' biomarker of the progression of APF and CCA from chronic opisthorchiasis, which is indicative of the critical role that OS plays in both of these advanced hepatobiliary pathologies. The findings also confirm our previous observations that severe liver pathology occurs early and asymptomatically in residents of O. viverrini endemic regions, where individuals are infected for years (often decades) with this food-borne pathogen. These findings also contribute to an expanding literature on 8-oxodG in an easily accessible bodily fluid (e.g., urine) as a biomarker in the multistage process of inflammation, fibrogenesis, and infection-induced cancer.

Localization of Tubulin from the Carcinogenic Human Liver Fluke, Opisthorchis viverrini.

BACKGROUND: Opisthorchiasis caused by Opisthorchis viverrini, is of considerable public health importance in Southeast Asia, particularly in Lao People's Democratic Republic and Thailand. The infection is associated with a number ofhepatobiliary diseases including cholangitis, obstructive jaundice, hepatomegaly, cholecystitis, cholelithiasis and cholangiocarcinoma. OBJECTIVE: This study aims to localize the expression sites of 0. viverrini tubulin using immunohistochemistry by monoclonal anti α-tubulin (MAαT) and anti ß-tubulin (MAßT). MATERIAL AND METHOD: The adult worms of O. viverrini, and the adult worm of O. viverrini in biliary system of hamsters were fixed, cryo-sectioned and then immunohistochemically stained. The sections were incubated with MAαTor MAßT. A positive test required the observation of brown-staining in the fluke's organs. RESULTS: The immunohistochemistry of MAαT and MAßT in adult worms of O. viverrini, and the adult worm of O. viverrini in biliary system of hamsters strongly expresses in the sperm and seminal vesicles of the worm. MAαT and MAßT expressed slightly in sub-tegumental tissue, stromalparenchyma, muscle fibers, and miracidium in the mature egg of the worm. No staining in the spermatogonia, gut epithelium, immature egg, tegument or vitelline glands of the worm nor in the hamster bile duct epithelium was seen. CONCLUSION: The present study indicates that O. viverrini tubulin is present in the reproductive organs and other important organs of the worm. Because it plays a key role in the biological processes of cellular motility and fertility it should be further studied in detail including the characterization, production of recombinant proteases, and their application in immunodiagnosis.

Schistosome infections induce significant changes in the host biliary proteome.

Schistosomiasis is a disease caused by blood trematodes affecting man and animals that represents an important human health and veterinary problem. Main damages caused by this infection are a consequence of the host inflammatory reaction against the parasite eggs trapped inside the liver. Despite that the hepatic pathology of schistosomiasis is very well known, there are no specific studies dealing with the schistosome infection effects on the biliary function. The purpose of this work was to analyse the changes induced by Schistosoma bovis infection in the biliary proteome. For this, whole gallbladders from S. bovis-infected and non-infected mice were dissected, homogenized and fractionated by differential centrifugation. The resulting protein fractions were resolved by SDS-PAGE, the gels were sliced, and the gel pieces analysed by LC-MS/MS. Altogether, we identified 1937 proteins, which were classified according to their "protein class" and "molecular function", and then subjected to an "Enrichment analysis". The differences found in gallbladder proteomes between S. bovis-infected and non-infected mice are analysed. We show that chronic schistosome infections cause significant changes in the biliary proteome that may produce physiological alterations and affect the therapeutic actions of drugs when administered to human patients and animals with schistosomiasis. BIOLOGICAL SIGNIFICANCE: To identify the changes induced by the schistosome infection in bile protein composition, and therefore in bile function, we compared the proteome of the gallbladders collected from non-infected healthy mice and from mice infected with S. bovis during 4months. For this, gallbladders from both groups of mice were homogenized and these homogenates were fractionated by serial centrifugation and acrylamide gel electrophoresis. The proteins were in gel digested and analysed by LC-MS/MS for identification. The present work reports the first data on the proteome of the mouse gallbladder and provides a comprehensive catalogue of biliary proteins that may be of great use in other studies addressing biliary physiology and pathology. We observed that there were significant differences in the biliary proteome of mice infected and non-infected with S. bovis and that chronic schistosome infections may produce important physiological alterations in the bile function.