Leukocyte numbers and intestinal mucosal morphometrics in horses with no clinical intestinal disease

Healthy horses and other animals have large numbers of resident leukocytes in the intestinal wall, but there is scant information regarding which and how many leukocytes are normally present in the equine intestinal wall. Our aim was to provide a reference range of leukocytes in the intestinal mucosal and submucosal propria of normal horses. We included in our study intestinal tissues from 22 Thoroughbred racehorses with no clinical intestinal disease, which had been euthanized because of catastrophic musculoskeletal injuries. Neutrophils, lymphocytes, eosinophils, macrophages, and plasma cells were counted in 5 random 17,600-µm² areas of villus lamina propria of the duodenum, jejunum, and ileum, and deep lamina propria of the duodenum, jejunum, ileum, right ventral colon, left ventral colon, left dorsal colon, right dorsal colon, and small colon. Other features investigated in the same intestinal segments included villus height and width (small intestine), presence of ciliated protozoa, Paneth cells number, subcryptal leukocyte layers (number of leukocyte layers between the bottom of the crypts and the muscularis mucosae), and submucosal leukocytes. Lymphocytes were the most numerous cells in all segments analyzed, followed by plasma cells, eosinophils, macrophages, and neutrophils. Eosinophil numbers were significantly higher in both lamina propria and submucosa of the large intestine than in the small intestine. The duodenum had shorter and thinner villi than either jejunum or ileum. The data provided from our study will be useful for diagnosticians examining inflammatory processes in the intestinal tract of horses.     

The development of the lesions caused by second generation schizonts of Eimeria necatrix.

First generation merozoites of Eimeria necatrix were found in epithelial cells of the crypts of Lieberkühn of chicks cells appeared to migrate into the lamina propria, forming nests of second generation schizonts that extended from the external muscularis to the lamina propria of the mid-villus. On the fourth and fifth days the mature schizonts, usually still within the infected cells were dishcarged into adjacent crypts. The muscularis mucosa appeared to be repaired by the formation of new, smooth muscle cells that spliced the broken ends of the ruptured muscle. 'Ghosts' of schizonts were found in the lamina propria on the sixth day after infection.     

贝氏莫尼茨绦虫感染绵羊对小肠局部黏膜淋巴组织的影响

为了研究贝氏莫尼茨绦虫自然感染绵羊对小肠黏膜免疫组织的影响,分别从宏观、微观及亚微观水平对自然感染贝氏莫尼茨绦虫的成年绵羊(感染组)肠道进行了细致地观察,并与正常成年绵羊(正常组)进行了比较.结果显示,感染组肠道所见虫体平均长度为1.5m,头节主要吸附在空肠淋巴集结分布丰富的部位,一般寄生数量为1～2条.眼观,虫体寄生部位黏膜增厚,表面有大量灰白色黏液附着,其间可见点状出血.镜下,局部黏膜上皮脱落,而在完整的黏膜上皮处,其上皮细胞、上皮内淋巴细胞、杯状细胞的数量都明显增多;固有层内毛细血管充血,淋巴细胞、浆细胞、弥散淋巴组织以及肠腺杯状细胞均有不同程度的增生,头节寄生处部分肠腺坏死;黏膜下层淋巴小结、淋巴集结显著增生,部分增生凸入固有层形成新的圆顶区;固有层与黏膜下层以及黏膜肌层可见大量嗜酸性粒细胞浸润.扫描电镜下,感染组肠黏膜上皮脱落;贝氏莫尼茨绦虫头节呈椭球状,有4个吸盘,无顶突,小沟,表面覆盖一层致密的微绒毛.研究结果表明,肠黏膜增厚,主要是局部黏膜免疫相关细胞在寄生虫虫体表面覆盖的微绒毛的不断刺激下,机体抗感染自身组织增生所致.成年绵羊对抗贝氏莫尼茨绦虫的感染可能是通过黏膜免疫相关组织增生来加强局部免疫力而实现的.     

Expression of different classes of immunoglobulin in intraepithelial plasma cells of the Harderian gland of domestic ducks Anas platyrhynchos

The Harderian gland of chickens contains numerous plasma cells and is considered as a peripheral lymphoid organ. Data about this gland in other avian species are scarce or inexistent. Considering that ducks show some unique characteristics regarding the immune system, which are important in evolutionary context, and that unusual location of plasma cells into the epithelium was recently described in primitive avian species, here we investigated the occurrence and characterized intraepithelial plasma cells in the Harderian gland of ducks, according to the immunoglobulin produced. Numerous intraepithelial plasma cells were found confined to the Harderian gland ducts. Plasma cells were also found in the ducts lamina propria. IgM-positive cells were the most abundant into the epithelium. In contrast, IgY- or IgA-positive cells were predominant in the lamina propria. The constancy of intraepithelial plasma cells in all specimens examined indicates that they may be essential mediator for an effective immunesurvaillance of the ocular mucosa.     

Hexamita meleagridis (Spironucleus meleagridis) infection in chukar partridges associated with high mortality and intracellular trophozoites

An outbreak of infectious catarrhal enteritis, associated with the flagellated protozoan Spironucleus meleagridis (syn. Hexamita meleagridis), is reported from a commercial flock of chukar partridges in California. The disease affected birds between the ages of 4 and 6 wk and resulted in diarrhea, listlessness, depression, and high mortality. Concurrent infection with other intestinal pathogens, including Cryptosporidia, group E Salmonella, long-segmented filamentous microorganisms (LSFMOs), and Rotavirus-like virus particles, was found in some but not all affected birds. Dermatitis of the face, shanks, and feet, suggestive of B-complex vitamin deficiency, was present in most affected birds as well. Flagellated protozoan parasites could be found in the lumen of the duodenum and jejunum and in the intestinal crypts. In some cases the flagellates were wedged between epithelial cells or were located intracellularly within cells of the mucosal epithelium and the intestinal lamina propria.     

Immune development in jejunal mucosa after colonization with selected commensal gut bacteria: a study in germ-free pigs

The immunological structure of the porcine jejunal lamina propria in germ-free piglets was compared with that of their counterparts associated with two strains of commensal Escherichia coli, A0 34/86 serotype O83:K24:H31 and the O86 E. coli strain, up to 20 days post-colonization. In the antigen-presenting compartment, both dendritic cells (DC) and cells expressing CD163, probably macrophages were investigated. In addition we also assessed the number of CD2+/CD3+ (T) cells. In contrast to some previous reports, we show a total lack of both DC and T cells for germ-free animals in the diffuse lymphoid tissue of villi and crypts of the jejunum. Association with either strain of commensal E. coli had a profound effect on the immune structure and resulted in extensive recruitment of DC to the lamina propria and of T cells to epithelium and lamina propria. The data suggest that the earliest immigrant cells were monocytes, which soon acquired the phenotype of mucosal DC. T cells migrated in at a slightly slower rate. Nevertheless, the response could be extremely rapid: within 3 days of colonization with O83, the magnitude of this response was comparable to that observed 20 days post-colonization.     

Eimeria organisms develop in the epithelial cells of equine small intestine

Histopathologic and immunohistochemical examinations were performed to determine the origin of host cells parasitized by Eimeria in the small intestines collected from five foals. Eimeria organisms at various stages (mainly microgametes and macrogametes) were frequently found in the cytoplasm of hypertrophied host cells in the lamina propria at the tips of villi of the jejunum and ileum. The cytoplasm of the host cell was immunohistochemically positive for cytokeratin AE1/AE3 and cytokeratin 13 and was negative for vimentin, desmin, alpha-smooth muscle actin, chromogranin A, neuron-specific enolase, and factor VIII. The host cells parasitized by Eimeria species had the immunostaining characteristics of epithelial cells but not of mesenchymal cells, endothelial cells of lacteals or capillaries, smooth muscle cells or neuroendocrine cells. These results suggest that the host cell of Eimeria species is possibly derived from intestinal epithelial cells and then displaced into the lamina propria of the small intestine.     

Identification of porcine circovirus type 2 in retrospective cases of pigs naturally infected with porcine epidemic diarrhoea virus

The identification of porcine circovirus type 2 (PCV2) was studied in fresh intestinal tissues by polymerase chain reaction (PCR) and in formalin-fixed, paraffin-wax-embedded intestinal tissues by in situ hybridisation. The tissues came from pigs naturally infected with porcine epidemic diarrhoea virus (PEDV). A total of 35 (32.7%) of 107 small intestinal samples from pigs naturally infected with PEDV were found to be positive using PCR. Positive signals for PCV2 were detected in 32 (29.9%) of 107 small intestinal samples from pigs naturally infected with PEDV by in situ hybridisation. The distribution of positive cells in the jejunum and ileum was multifocal or patchy. Distinct positive labelling was found throughout the lamina propria in the small intestines. The results of this study indicate that PCV2 is highly prevalent in pigs naturally infected with PEDV.     

Reduced differentiation of intestinal epithelial cells in wasting marmoset syndrome

Wasting marmoset syndrome (WMS) is a serious disease in captive common marmoset (Callithrix jacchus) colonies. Because of the high mortality rates, elucidation of the underlying mechanisms is essential. In this study, we compared the histopathology, the number of each epithelial cell in the jejunum and colon, and the expression patterns of some molecular markers between healthy and WMS-affected marmosets. Atrophy of villi in the jejunum and mononuclear cell infiltration in the lamina propria were observed in the intestinal tract of WMS-affected marmosets. Although the numbers of transient amplifying cells and tuft cells were increased, the number of goblet cells was obviously decreased in the jejunum and colon of WMS-affected marmosets compared to healthy marmosets. In addition, the number of enterocytes in the jejunum was decreased in WMS animals. There was no apparent difference in the numbers of stem cells, enteroendocrine cells, or Paneth cells. The expression of β-catenin and Tcf7l2 was increased in WMS, and the co-existence of β-catenin and Tcf7l2/Cyclin D1 was observed around the crypts in WMS-affected marmosets. These findings suggest that cell proliferation continues, but cell differentiation is halted in the intestinal tract due to the enhanced β-catenin/Tcf7l2/Cyclin D1signaling pathway in WMS, which results in malfunction of the villus and mucosa.     

Intestinal lesions in specific-pathogen-free lambs associated with a cryptosporidium from calves with diarrhea

Small and large intestines of seven specific pathogen-free lambs infected with cryptosporidia from calves with diarrhea were examined by scanning and transmission electron microscopy and by light microscopy. The small intestine was infected in all the lambs, and the cecum and colon in three. Small intestinal alterations were severe villous atrophy and dilatation of the crypts of Lieberkühn. Epithelial cross-bridging between contiguous villi caused much villous fusion. Epithelial cells constituting the bridges were connected by desmosomal junctions, and were continuous with the epithelial coverings of the associated villi. The lamina propria was heavily infiltrated with neutrophil leukocytes. Infected crypts in cecum and colon were dilated and devoid of mucus-secreting cells, while the ridges between crypts were hypertrophied, and the lamina propria was infiltrated by neutrophils. Cell vegetations with adherent bacteria were present in the surface intestinal epithelium of two lambs infected for 11 and 14 days, respectively. No adherent bacteria were seen in any site in lambs killed up to six days post-inoculation.     

A light microscopical study of the intestinal tract of the Nile crocodile (Crocodylus niloticus, Laurenti 1768).

Although the histology of the intestinal tract of Crocodylus niloticus is touched on in overall studies on reptilian intestinal tract, a more comprehensive light microscopical study on this area is lacking. Specimens for histological examination were taken from the duodenum, the jejunum, the ileum and the rectum. The data obtained revealed that the mucosa is thrown into folds and simple, slightly branched tubular intestinal glands. The mucosal folds diminished in height and eventually disappeared upon reaching the ileorectal junction. The epithelium covering the folds and crypts was of the simple columnar type. Clear marginal cells, goblet cells and argentaffin cells were observed throughout the intestinal tract. No Paneth cells were seen in this study. The lamina propria was rich in lymphocytic infiltrations while the muscularis mucosa consisted mainly of an outer longitudinal layer, the inner circular layer being rudimentary or absent. The submucosa was extremely narrow, and the circular and longitudinal layers of the tunica muscularis contained distinct layers of dense fibrous connective tissue. The histology of the intestinal tract of C. niloticus is shown to be in line with the situation in crocodilians and also exhibits a resemblance to that of carnivorous mammals.     

Mast cells in the bovine lower respiratory tract: morphology, density and distribution

The lower respiratory tract of six 8-day-old calves and six adult cows was fixed in either isotonic formal-acetic-acid or neutral buffered formalin in order to study the morphology, density and distribution of mast cells. They were found at all levels of the tract with the highest density in the major bronchi. Tissues from cows had significantly more mast cells than those from calves. There were significantly more mast cells detected in calf airways fixed with isotonic formal-acetic-acid compared with those seen in the same tissues fixed with neutral buffered formalin. Regardless of the age and fixation, mast cells were located predominantly in the alveolar septa and in the lamina propria of airways. They were also commonly encountered within the mucosal epithelium of the trachea. Ultrastructurally, mast cells in bovine airways and lung contained two types of intracytoplasmic granules as described in other species.     

Ontogeny of gut-associated lymphoid tissue (GALT) in the dogfish Scyliorhinus canicula L

Cartilaginous fish occupy a fundamental position in vertebrate phylogeny and it is likely that this group has retained some of the ancestral immune mechanisms. The ontogeny of GALT has received little attention in elasmobranchs and this study correlates this development with morphological differentiation, development of other lymphoid organs, exposure to seawater and transition from yolk dependence to exogenous food as a source of nutrient. GALT was first represented by individual lymphocyte-like and macrophage-like cells in the lamina propria. In later stages accumulations and intraepithelial leucocytes were recorded prior to hatching. The size of accumulations and the number of lymphocyte and macrophage-like cells infiltrating the lamina propria and epithelium increased in fish as they became dependent upon an exogenous diet. Although GALT developed after the thymus and lymphoid-like tissue in the kidney and at approximately the same time as the epigonal, Leydig and spleen, the source of cells populating the gut is unknown. Plasma cells and granulocytes were not observed in the developing fish until 6 months post-hatch after which the fish had a similar GALT distribution and content to the adult fish.     

Intestinal trichomonads (Tritrichomonas mobilensis) in the natural host Saimiri sciureus and Saimiri boliviensis

A retrospective study of cecal and colonic tissues from 28 squirrel monkeys (Saimiri sciureus and Saimiri boliviensis) demonstrated enteric trichomonads within luminal crypts. Twenty-one of 28 (75%) had trichomonads in the mucosal epithelium either in cup-like depressions or intraepithelial vacuoles. Organisms were also beneath the superficial luminal mucosal epithelium and between the basement membrane and crypt epithelial cells. Immunoperoxidase staining also identified organisms within the lamina propria and submucosa. Additional histologic changes included mucosal ulceration, multifocal cryptitis, and focal epithelial necrosis. Most areas containing trichomonads did not have an associated inflammatory response.     

Gut-associated lymphoid tissue in the large intestine of calves. II. Electron microscopy

Scanning electron microscopy of lymphoid tissue in the large intestine of three germfree calves (age 3, 6, and 7 days) revealed two different units: propria nodules and lymphoglandular complexes (LGC). Propria nodules had lymphoid tissue predominantly in lamina propria and were covered by distinct follicle-associated epithelium which lacked goblet cells; nodules were surrounded by wide crypts, which were also lined by follicle-associated epithelium towards the luminal side. Lymphoglandular complexes had lymphoid follicles in the tunica submucosa; epithelial diverticulae extended through the muscularis mucosae branching into the lymphoid nodule. In centers of lymphoglandular complexes, protrusions of lymphoid tissue were covered with distinct follicle-associated epithelium. By transmission electron microscopy cells compatible with M cells in the small intestine of calves and cells with characteristics of both enteroabsorptive and M cells were found. Follicle-associated epithelium of propria nodules and lymphoglandular complexes differed only in the relative frequency of cell types.     

The migration of larval Toxocara canis in mice. I. Migration through the intestine in primary infections

Toxocara canis second stage larvae (L2) hatched in the stomach of mice and within 2 h reached all parts of the small intestine, the posterior half being the preferred site for larval penetration. Following penetration at the base of crypts of leiberkuhn they followed tortuous routes in the lamina propria, and entered the tunica muscularis obliquely. Larvae only appeared to be arrested in their migration in the submucosa and lamina propria by an inflammatory reaction composed mainly of lymphocytes and eosinophils. Larvae were seen entering and within lymphatic vessels as well as the peritoneal cavity and invasion of the vascular system followed though actual penetration of intestinal blood vessels was not seen. Proteolytic enzyme activity was detected in culture media in which L2 larvae were maintained.     

A comparison of the effects of dietary spray-dried bovine colostrum and animal plasma on growth and intestinal histology in weaner pigs

An experiment was conducted to evaluate the effects of dietary spray-dried bovine and porcine plasma and spray-dried bovine colostrum on growth performance and intestinal histology in weaner pigs. Thirty-two 21-day-old piglets (6.65 ± 0.14 kg) were allocated to receive one of four dietary treatments: control, bovine plasma, porcine plasma, and bovine colostrum at weaning and another 8 piglets were killed at weaning to provide baseline data. The experimental diets were offered ad libitum for one week, after which animals were killed and post-mortem measurements obtained. No differences in average daily feed intake and growth rate were observed among dietary treatment groups (P > 0.05). Baseline piglets had taller villi and shallower crypts (P < 0.05) in the proximal jejunum, mid jejunum and distal ileum than those observed a week after weaning, irrespective of dietary treatment. Weaning-related expansion of intestinal lamina propria CD4+ and CD8+ T lymphocyte populations was observed (P < 0.05). Complex and variable treatment effects on villus height, crypt depth, villus and crypt goblet cell density, and lamina propria T cell density were observed, suggesting that the tested protein sources do not share a common or simple mode of action.     

Immunohistochemical and histoplanimetrical study on the endothelial receptor involved in transportation of minute chylomicrons into subepithelial portal blood in intestinal villi of the rat jejunum

A portion of the minute chylomicrons less than 75 nm in diameter are transcytosed from the extravascular tissue into the subepithelial blood capillaries (sBC) in the villous apices of the rat jejunum. However, the details of the transportation mechanism have not been clarified. In this study, the endothelial receptor involved in the transportation of minute chylomicrons into the sBC’s lumina was immunohistochemically and histoplanimetrically examined in intestinal villi of the rat jejunum. Immunopositivity for very low density lipoprotein (VLDL) receptor was detected on the luminal and basal surfaces of the endothelial cells of sBC in approximately 68% of those apices of jejunal villi that possessed numerous chylomicrons in the lamina propria, while VLDL receptor was detected on the endothelial cells of sBC in only approximately 8% of intestinal villi that possessed few or no chylomicrons in the lamina propria. No immunopositivity for LDL receptor was detected in the sBC of all intestinal villi. These findings suggest that VLDL receptor is expressed by the endothelial cells of the sBC in conjunction with the filling of the lamina propria of jejunal villi with many chylomicrons produced by the villous columnar epithelial cells and that the VLDL receptor mediates the transportation of minute chylomicrons, maybe VLDL, into the subepithelial portal blood from the extravascular tissue of the rat jejunal villi.     

Canine lymphocytic plasmocytic enteritis: an immunopathological investigation of intestinal plasma cells.

The numbers of IgA-, IgG- and IgM-containing plasma cells in the lamina propria of duodenum, jejunum, ileum, and coecum were quantitated in 3 patients suffering from lymphocytic plasmacytic enteritis and 3 normal control dogs. A great increase was found in all 3 types of plasma cells and at all levels of the intestinal tract. Especially the IgG-containing cells showed a remarkable increase particularly in 2 patients. The possible reasons for these heavy infiltrations of immunologically competent cells are discussed, and chronic antigenic stimulations of food origin is suggested.