Goblet Cell Types in Intestine of Tiger Barb and Black Tetra (Cyprinidae,Characidae: Teleostei)

Histochemical properties of goblet cells in intestine of a stomach‐less teleost, tiger barb (Puntius tetrazona), and a stomach‐containing teleost, black tetra (Gymnocorymbus ternetzi), are described and compared. The intestine goblet cells were mostly wide in both species, but in tiger barb, some of them were markedly thinner. In black tetra, all the intestine goblet cells displayed magenta colour after PAS, whereas in the tiger barb, only the thinner goblet cells displayed such affinity. The latter cell type was coloured strongly magenta when the tissue was treated with alcian blue (pH 2.5) followed by PAS, whereas the wide goblet cells in tiger barb and all goblet cells in black tetra displayed mainly a blue colour after such treatment. Further, the goblet cells in both species were coloured cleanly blue after high iron diamine followed by alcian blue (pH 2.5). The intestine goblet cells in both species displayed a moderate affinity to WGA and concanavalin A lectins and no affinity to DBA. Most of the goblet cells displayed no affinity to PNA, but some of them in the tiger barb displayed a moderate or strong affinity to this lectin. The affinity to WGA was somewhat strengthened after pre‐treatment with neuraminidase. These results suggest that tiger barb contains two types or variants of intestinal goblet cells: high numbers of wide cells filled by acidic, non‐sulphated mucin and some thinner cells filled by neutral mucin. The intestine goblet cells in black tetra were filled by variable amounts of neutral and acidic mucin, but the total number of such cells is much less than in tiger barb. The present lectin and neuraminidase results suggest that the intestinal mucins in both species contain significant amounts of N‐acetylglucosamine, sialic acid and glucose/mannose, but seem to lack N‐acetylgalactosamine. However, some of these cells in tiger barb contain moderate to large amounts of galactose. Together, these results suggest significant species‐specific features of the intestine goblet cells and mucin types in tiger barb and black tetra. In conclusion, the present results suggest that the diet and feeding habits in stomach‐less teleosts compared with stomach‐containing teleosts, greatly influence the number of intestine goblet cells and type of mucin in these cells.     

Acidification of the colonic mucins following polyvalent colonization of the germ-free rat.

Colonic mucins of germ-free (GF) and conventional rats (CV) were compared. After isolation by gel filtration on Sepharose CL-4B and purification by density gradient centrifugation, the content of isolated colonic mucins was estimated by determination of PAS positive carbohydrates. Purified mucins were subjected to carbohydrate and amino acid analysis and separated into mucin subclasses by ion exchange chromatography. While the total amount of colonic mucins was not statistically different in GF and CV animals, analysis of carbohydrate composition demonstrated an increased amount of sialic acid in CV rat mucin. This was in accordance with results of ion exchange chromatography, revealing a significant higher amount of negative charged mucin subclasses in CV mucin, compared to the germ-free counterpart. The results of amino acid analysis were similar in both groups. The compositional differences in carbohydrate moieties are attributed to modulations by the intestinal flora. A selective bacterial degradation of the neutral mucin subclasses and modifications in the mucin composition due to a stimulated synthesis are discussed.     

Myxoid leiomyosarcoma of the gizzard in a broiler chicken

A leiomyosarcoma was found in the gizzard of a 57-day-old female broiler chicken weighing 1.8 kg. Grossly, the tumor mass, 13.0 x 8.5 x 10.0 cm, enveloped the gizzard and had a gelatinous appearance due to the rich production of mucin. Miliary metastatic tumors were noted in the liver. Histopathologically, there was marked production of mucus throughout the tumor tissue, and densely or loosely arranged long spindle-shaped leiomyosarcoma cells proliferated. The tumor cells had a low rate of mitosis, showed slight cellular atypia, and, immunohistochemically, were positive for actin, alpha-smooth muscle actin, and desmin. Electron microscopically, various amounts of microfibrils with focal densities, dense patches, and basal plates were observed.     

Binding of E. coli isolates from pigs with postweaning diarrhea or edema disease to crude intestinal mucin of a weaned pig.

The presence of the fedA (gene coding F18 fimbriae) and genes coding STa and LTI enterotoxins and verotoxin Stx2v was determined in 30 E. coli strains isolated from weaned pigs with postweaning diarrhea (PWD) and edema disease (ED). The fedA gene was detected in 22 strains (73.3%). It was mostly associated with the presence of ST gene determinant (14 from 22 fedA positive strains, 63.6%). Two strains possessed ST/Stx2v or LTI/Stx2v combination of genes for both toxins and two strains were negative for investigated toxin determinants. Among 8 fedA-negative strains, five strains without gene determinants for toxins were detected. All 30 E. coli strains were investigated for their binding to crude intestinal mucin of a weaned pig fixed in wells of microtitre plates. Positive mucin binding was observed in most of strains, however, great differences were shown between individual strains. Nineteen strains were classified as strongly adherent, 10 strains as weakly adherent, and only one nonadherent strain was found. Three E. coli strains, selected among the best mucin binders, bound to mucin in a concentration-dependent manner. A high mucin binding by E. coli strains was observed only after their cultivation on blood agar plates. Their cultivation in LB broth or on McConkey agar plates had negative effect on the mucin binding by these strains. The mucin binding is not restricted by the presence of fedA gene because the strains displaying very good binding are found either among fedA positive (1, 602/2, 4/3, 576/6) or fedA negative (DK 6, DK 8) E. coli strains. E. coli strains with the highest mucin binding ability belong to potential ST producents (strains 1, 602/2, 4/3, 6/2, 602/4) while the strains without genes coding toxin production displayed lower binding to mucin substratum with exception of the strains ZV5 and 13.     

Inositol hexaphosphate increases mucin loss from the digestive tract of ducks

The effects of different forms of inositol hexaphosphate (IP6, phytate) on mucin excretion from the digestive tract of ducks were investigated. Forty-eight ten-wk-old male ducks were grouped by weight into eight blocks of six cages with one bird per cage. On the first day of experimentation, birds received by intubation six dextrose-based diets arranged in a 2 × 3 factorial consisting of phytase (0 or 1000 units) and a form of IP6 (no IP6, free phytic acid or magnesium-potassium phytate). During the 54 h following feeding, excreta were continuously collected and frozen until analysed. The amount of mucin and amino acids, threonine, valine and tyrosine, in excreta increased in ducks fed with either magnesium-potassium phytate or free phytic acid. The increase in mucin excretion was more in birds fed with magnesium-potassium phytate than those fed with free phytic acid. The loss of arginine, isoleucine, leucine, lysine and phenylyalanine in excreta was reduced by the presence of microbial phytase. It is concluded that the form of IP6 fed to ducks affects the extent of mucin excretion and also the extent and nature of endogenous amino acid losses in the excreta. Supplementation with exogenous microbial phytase reduced some of the IP6 feeding-induced endogenous intestinal amino acid losses.     

Coating of Objects Introduced into the Oviduct of Pseudopregnant Rabbit Does

This investigation addresses the possibility of providing mouse embryos or other foreign objects with a protective mucin coat by transferring them into the oviduct of a life rabbit doe. Mouse embryos at the 8 or 16-cell stage, rabbit oocytes and latex spheres resembling mouse embryos in size were transferred to the ligated oviducts of ovulation-induced rabbit does. The does were killed 24 h later to have their oviducts flushed. A large proportion of the latex spheres (89%) and of the ovulated oocytes of the recipient does (92%) was recovered. The recovery rates for transferred rabbit oocytes, either intact or with the zona pellucida removed, were 61% and 51%, respectively, whereas that for mouse embryos was extremely poor (20%). Rabbit oocytes with or without zona were enveloped in a thick mucin coat regardless whether they had been transferred or ovulated by the recipients. The same applied to empty rabbit zonae. Mouse embryos and latex spheres were also covered by a mucin coat, but it was four times thinner. While residing in the rabbit oviduct, the mouse embryos continued developing to a stage comparable to what would have been expected in situ . During the subsequent in vitro culture, mouse embryos continued developing to the expanded blastocyst stage. They did, yet, not hatch from the zona. It may be concluded that particles of various origins, when placed into the oviduct of ovulated rabbit does, will be provided with a mucin covering which is, however, considerably thinner than that surrounding oocytes or zonae pellucidae originating from rabbits.     

Morphological,morphometric and histochemical characterization of the gastric mucosa of the camel (Camelus dromedarius)

Mucosal morphology, morphometry and mucin histochemistry of the stomach were studied in the one-humped camel. The lining of the stomach was divided into eight grossly identifiable regions. The first region was non-glandular, occupied the body of the first compartment of the stomach and constituted 53.2% of the gastric mucosa. The other seven regions were lined by a glandular mucosa. Histological, histochemical and morphometric investigations have shown that glandular mucosa comprises pseudo-cardiac, cardiac, fundic and pyloric regions. The pseudo-cardiac region was characterized by widely separated short tubular serous glands. It constituted 36.2% of the gastric mucosa; it extended over the entire lining of the second compartment and parts of the first and third compartments. The cardiac region was confined to the initial zone of the third compartment amidst the psuedo-cardiac region but contiguous with the distal end of the gastric groove. It constituted 3.4% of the gastric mucosa and was characterized by neutral and acid mucin positive glands. The fundic and pyloric regions occupied the distal distended part of the third compartment. The fundic region constituted 4.3% of the gastric mucosa. It was packed with typical fundic glands characterized by chief cells, parietal cells and acid mucin positive neck cells. The pyloric region constituted 2.9% of the gastric mucosa. Its glands were positive to acid mucins except for their bases that were positive to neutral mucins. Differences in volume densities of the mucosal components and reactivity of the surface epithelium and gastric pits to mucin stains were noted in the different regions of the stomach.     

A mucinous histochemical study on malignancy of aberrant crypt foci (ACF) in rat colon

The relationship between malignancy and number of crypts (crypt multiplicity) comprising aberrant crypt foci (ACF) was investigated, by studying changes in the mucous nature of ACF with 5 crypts or less, ACF with 6-13 crypts, adenomas and invasive adenocarcinomas induced by 1,2-dimethylhydrazine in distal colon of rats. A paradoxical Con A-staining was performed for goblet cell mucins. Of the sulfomucin-dominant ACF with 1-3 crypts, 82.6% had labile class III mucin, similar to the distal colon in the normal rats. However, in most of the goblet cell mucin produced by the ACF with 4-5 crypts with an indicated relation to colorectal carcinoma or the sialomucin (SiM) -dominant ACF with 1-3 crypts, mucin types other than class I were rarely present. The incidence of class I mucin decreased with the increase in crypt multiplicity of ACF or in the degree of histological malignancy, with the lowest incidence of 40% in adenocarcinomas. In contrast, the incidence of class II mucin increased markedly with the increase in crypt multiplicity of ACF or in the degree of histological malignancy, with the highest incidence in adenocarcinomas (95%). The ACF with 6-13 crypts had a mucous profile similar to that of adenomas. These results suggested that malignancy of ACF related to the crypt multiplicity. In the ACF with 1-3 crypts, SiM-dominant ACF had the potential to progress to malignant lesions.     

Mammary mucinous carcinoma in the cat

Mucinous carcinoma of the mammary gland is a rare tumor characterized by excessive mucin production. In human and canine pathology, the diagnosis of mucinous carcinoma is based on the demonstration of an epithelial phenotype of mucus-producing cells and periodic acid-Schiff (PAS)-diastase positivity of the mucin. The histologic and immunohistologic characteristics of feline mucinous mammary carcinoma were examined. Of 656 cases of feline mammary neoplasms and dysplasias, 3.2% were found to be mucin-producing tumors. Cytokeratin 19 (16 cases positive, 4 heterogenous, and 1 negative) and vimentin (15 cases positive, 2 heterogenous, and 4 negative) expression were examined, and the mucin produced was alcian blue positive. PAS-diastase staining was variable (38.1%). Based on these findings, mucinous mammary carcinoma in the cat varies significantly from the human and canine varieties and alcian blue is the prominent stain in the diagnosis of feline mucinous carcinoma.     

Histochemical and morphologic changes of porcine airway epithelial cells in response to infection with Mycoplasma hyopneumoniae.

Mycoplasma hyopneumoniae causes pneumonia in pigs. The effect of infection by this organism on histochemical characteristics of airway mucin within epithelial cells was studied. Seven- to 10-week-old pigs were inoculated intratracheally with M hyopneumoniae or culture broth, and lung tissues were collected from inoculated and control pigs at 2, 4, and 6 weeks after inoculation. Tissue sections were stained with periodic acid-Schiff/Alcian blue, pH 2.5 or high iron diamine/Alcian blue. Histologic features of randomly selected bronchi, bronchioles, and submucosal glands were compared in sections stained with periodic acid-Schiff/Alcian blue. Bronchial goblet cell sulfomucin and sialomucin were quantitated by image analysis of sections stained with high iron diamine/Alcian blue. Bronchi and bronchioles of infected pigs contained proportionately fewer goblet cells with mucin at all stages of infection than age-matched control pigs. Goblet cells in bronchi of infected pigs contained significantly less total mucin and sialomucin, and significantly more sulfomucin than goblet cells of control pigs. Increased sulfated mucin in bronchial goblet cells may reflect altered glycoprotein production or secretion in response to infection with M hyopneumoniae.     

Structure of the Oesophagus and Morphometric, Histochemical–Immunohistochemical Profiles of the Oesophageal Gland During the Post-hatching Period of Japanese Quails (Coturnix coturnix japonica)

In the oesophagus, mucins, which originate from oesophageal submucosal glands, play an important role in the mucosal protection as a pre-epithelial barrier. In this study, the structure of cervical and thoracic parts of oesophagus of Japanese quail during the post-hatching period was compared, and the contents of carbohydrate and gastric mucin MUC5AC of the oesophageal glands in these parts were analysed at the light microscope levels by applying conventional histochemical and immunohistochemical methods. The oesophageal glands were present at hatching, located in the laminae propria. The numbers of glands were different in the cervical and thoracic parts, but the differences were found to be insignificant. The thoracic part has the oesophageal tonsils which are associated with the glands. Oesophageal tonsil was formed from day 5 after hatching. In quail of all ages, the secretory epithelium of glands contained neutral sialomucins and weakly sulphomucins. The cells in the neck region of secretory units contained sialomucins, while the cells of excretory ducts had strongly sulphomucins. Sialomucin containing cells in the secretory units increased with the advance of age and glandular development. But, in the secretory units, the sulphomucin content of glands was more in the thoracic part. The secretory epithelium of tonsil-associated glands contained mostly sulphomucins and a little sialomucin. From the hatching, MUC5AC mucin was detected in the cells of excretory ducts. Although the lymphoepithelium of the tonsil units exhibited negative reactions to all histochemical methods, it showed positive reaction to MUC5AC mucin antibody. In conclusion, the cervical and thoracic parts may be functionally different and the thoracic part of oesophagus was transformed into an immunological organ following day 5 after hatching.     

Influence of bacitracin on microbial functions in the gastrointestinal tract of horses

This study investigated the influence of zinc bacitracin on the intestinal flora of horses. The functionally active intestinal flora was examined in 6 horses during treatment with zinc bacitracin. Utilising gas chromatography, spectrophotometry, gel electrophoresis and paper chromatography, samples were analysed on biochemical markers reflecting the action of parts of the intestinal flora. The following 5 flora-related functions were studied in faecal samples and intestinal samples from different sections of the hindgut: conversion of cholesterol to coprostanol and of bilirubin to urobilinogens, degradation of mucin and of beta-aspartylglycine and inactivation of tryptic activity. Conversion to coprostanol, conversion to urobilinogens and degradation of mucin were affected by treatment of zinc bacitracin and conversion to coprostanol was most sensitive. All functions were normalised in a short time, in contrast to man and rats. Differences in environmental exposures are probably the reason for a more rapid normalisation of the intestinal flora functions in horses.     

The tear film and ocular mucins

Abstract The trilaminar tear film, composed of the lipid, aqueous and mucin layers, has many functions including defending the ocular surface. The aqueous layer has several soluble antimicrobial factors that protect the ocular surface. Ocular mucins have recently been studied with regard to their role in the defense of the eye as well as in dry eye syndromes. To date, 15 mucin genes have been identified, and six of these mucin genes are localized to or secreted by ocular glands or epithelia. Understanding the production, secretion and function of ocular mucins will aid in the treatment of dry eye syndromes and ocular surface microbial infections.     

Surface mucus in the non-glandular region of the equine stomach

In horses, ulceration of the non-glandular region of the stomach is common and has been attributed to the lack of a protective mucus covering. This study aimed to determine whether the non-glandular region is covered by a mucus layer. A mixture of antibodies raised against human gastric mucin (MUC 5 AC) showed a tissue distribution in the glandular region of the equine stomach similar to that seen in humans. Dot blots of mucus from the glandular and non-glandular regions showed cross-reactivity with these antibodies. Various histological fixation and processing techniques were compared for their ability to preserve mucus in the non-glandular region. Fixing frozen sections on-slide for 20 seconds in 20 per cent formalin/1 per cent cetylpyridinium chloride was considered the best method. In conclusion, the equine stomach expresses a gene homologous to human MUC 5 AC. Its product is expressed as a neutral mucin, which is present in the mucus that covers both the glandular and non-glandular regions. Future comparison of mucus composition in the healthy and ulcerated stomach will improve our understanding of gastric ulceration in the horse.     

Expression of secreted and membrane-bound mucins in the airways of piglets experimentally infected with Mycoplasma hyopneumoniae

Immunohistochemistry was used to demonstrate secreted mucins MUC2, MUC5AC and MUC5B and membrane-bound mucin MUC4 in the pulmonary bronchioles of piglets experimentally infected with Mycoplasma hyopneumoniae. Conventional status, Landrace-Duroc cross-bred piglets, 13 days of age, were randomised to two groups. One group (n=20) was infected by the intra-tracheal route with the SNU98703 strain of M. hyopneumoniae, and a group of 12 animals acted as uninfected controls. Five infected and three uninfected piglets were euthanased on the day of infection and at 7, 21, and 35 days post-inoculation (PI). Membrane-bound MUC4 and secreted MUC5AC were the predominant mucins produced in the bronchioles of the piglets in response to M. hyopneumoniae infection, but by day 35 PI, all labelled mucins had returned to pre-infection levels, contemporaneous with reduced pulmonary lesion scores. The increased mucin production may result from direct stimulation of the epithelium by mycoplasmal infection, or may arise indirectly following M. hyopneumoniae-induced ciliostasis.     

Screening of pigs resistant to F4 enterotoxigenic Escherichia coli (ETEC) infection

The present study analysed quantitatively the mucin 4 polymorphism for determining the F4ac/ab receptor status of a total of 63 pigs by comparing it with the in vitro villous adhesion assay. The probability of a susceptible genotype for the mucin 4 increases significantly with increasing F4ab or F4ac ETEC adhesion per 250 microm villi (P=0.029 for F4ab, P=0.030 for F4ac), with the odds ratio for each unit increase of F4ab or F4ac equal to, respectively, 1.036 (95% CI [1.004-1.069]) and 1.018 (95% CI [1.002-1.034]). In the phenotypic in vitro villous adhesion test, a cut-off value of 5 bacteria was chosen as a criteria for the distinction between an F4R positive and F4R negative pig. The sensitivity and specificity for the in vitro villous adhesion test, with the genotyping test for mucin 4 as golden standard, is 100% and 24%, respectively, for F4ab as well as F4ac. Absence of adhesion of F4ac and F4ab ETEC to the villous brush borders was not associated with genotypic resistance suggesting that there is at least one other receptor for F4ab/ac Escherichia coli. As a consequence, not only mucin 4 gene polymorphism but also expression of these other receptor(s) has to be included in a screening assay for F4ac/ab receptor negative pigs.     

黏蛋白侧链岩藻糖在肠道细菌与宿主互作过程中的作用

消化道表面黏液层中含有大量黏蛋白,这些黏蛋白分子侧链往往被糖基化修饰,这些糖基在肠道菌的黏附、定植及免疫调节过程中发挥重要作用。岩藻糖是肠道黏蛋白分子侧链中的一种重要糖基,本文总结了岩藻糖作为肠道菌黏附靶位点和信号调节分子,以及调节肠道功能等方面的作用。     

Sheep intelectin-2 co-purifies with the mucin Muc5ac from gastric mucus

Secretion of gastric mucins plays an essential role in host protection, and modifications in mucus properties are characteristic of the protective immune responses to pathogens. This study describes the purification and characterisation of sheep gastric mucins, and identification of those proteins that co-purify with mucins, with the potential to modify mucus properties. Gastric mucus was collected and pooled from four abattoir sheep and separated by CsCl density gradient centrifugation. Proteomic analysis of the mucin-containing fraction indicated the presence of gastric mucin (Muc5ac) and several co-purifying proteins, including intelectin-2 (Itln2). Further experimentation indicated that a combination of denaturation and reduction was required to fully release Itln2 from gastric mucin. A putative correlation was found between mucin-bound intelectin concentration and rheological properties in further sheep gastric mucus samples. In conclusion, this study provides the first characterisation of sheep gastric mucins and their purification partners, revealing potentially important mucin-intelectin interactions.     

Quantification of mucin gene expression in tracheobronchial epithelium of healthy dogs and dogs with chronic bronchitis

OBJECTIVE: To develop a real-time PCR assay for the quantification of mucin gene expression in tracheobronchial brushing specimens from dogs and compare mucin gene expression in specimens from dogs with naturally occurring chronic bronchitis with that in specimens from healthy dogs. ANIMALS: 7 healthy dogs and 5 dogs with chronic bronchitis. PROCEDURES: Primers that were designed to span the predicted intron-exon boundaries of a canine MUC5AC-like gene were used to develop a real-time PCR assay for quantification of expression of that gene. Total mRNA was isolated from tracheobronchial brushing specimens obtained from dogs with and without bronchitis during anesthesia; MUC5AC-like gene expression in those samples was quantified by use of the real-time PCR assay. RESULTS: The PCR assay was sensitive and specific for the target sequence, the predicted amino acid sequence of which had greatest homology with human, porcine, and rat MUC5AC. The assay was able to quantify the target over a wide dynamic range. Dogs with chronic bronchitis had a 3.0-fold increase in the quantity of MUC5AC-like mRNA, compared with healthy dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The ability to measure mucin gene expression from tracheobronchial brushing specimens collected from client-owned dogs during routine bronchoscopy should prove to be a useful tool for the study of bronchitis in dogs and expand the usefulness of airway inflammation in dogs as a model for bronchitis in humans.     

Dietary fibre and indigestible protein increase ileal glycoprotein output without impacting colonic crypt goblet cells in weaned piglets

Mucus plays an important role in gut health by favouring colonisation resistance. The aim of this study was to quantify the impact of fibre and protein on mucin recovery in ileal digesta and on goblet cell histochemistry in the proximal colon. A control diet with highly digestible protein and low fibre and three complex diets containing indigestible protein associated with low, soluble or insoluble fibre sources were tested for 14 days in piglets weaned at 28 days of age. Mucin concentration was determined by ethanol precipitation. Goblet cell subtypes in colonic crypts were analysed by histochemistry. The ileal mucin output was higher with the complex diets than with the control diet (31.6 on average vs 21.7 g/kg DM intake). Increases observed with the low and soluble fibre diets were similar (34 g/kg DM intake). This suggests a limited effect of soluble fibre in presence of indigestible protein. Surprisingly, the observed increase was lower with insoluble fibre (27.2 g/kg DM intake). DM and N output and digestibility were not affected by the diets, but a linear relationship was found between DM and mucin output. In proximal colon, crypt depth, goblet cell numbers per crypt and glycosylation subtypes were not affected by the diet suggesting no change in the capacity of mucus to protect the gut. To conclude, introducing highly indigestible protein in the diet increased ileal mucin output, without effects of these factors on crypt goblet cell patterns in the proximal colon. Introducing fibre in a diet containing highly indigestible protein had a marginal effect.