Canine viral enteritis. II. Morphologic lesions in naturally occurring parvovirus infection.

The morphologic lesions associated with naturally occurring canine parvovirus infection are described. The disease is characterized by necrosis of crypt epithelium in the small intestine and of lymphoid tissue in Peyers patches, lymph nodes, spleen and thymus.     

Influence of a probiotic Enterococcus faecium strain on development of the immune system of sows and piglets

The influence of the probiotic bacterium Enterococcus faecium SF68 on the immune system and the intestinal colonization of pigs were determined in a feeding experiment with sows and piglets. Mucosal immunity of the developing piglets was monitored by isolation and detection of intestinal lymphocyte cell populations from the proximal jejunal epithelium and the continuous Peyers patches by the use of flow cytometry. The levels of intestinal IgA in both groups of piglets were compared, as well as total IgG in the serum of sows and piglets. Feces of the sows and intestinal contents of the piglets were taken for determination of total anaerobe and coliform bacterial counts in both probiotic and control groups. Villus length and depth of the crypts were measured in the jejunum of sacrificed piglets to monitor the development of the intestinal mucosal surface amplification. Total serum IgG of the sows appeared to be unaffected. Piglets of both groups showed similar IgG levels up to 5 weeks after birth with a slight tendency toward lower values in the probiotic group. At an age of 8 weeks the total IgG levels of the probiotic animals were significantly lower (p<0.01). No differences were observed in the populations of CD4+ and CD8+ T cells in the Peyers patches. However, the levels of cytotoxic T cells (CD8+) in the jejunal epithelium of piglets of the probiotic group were significantly reduced. The depth of the jejunal crypts and length of the villi were similar in both groups, suggesting the relative T-cell population differences were not due to alterations in the epithelial cell numbers. The total anaerobe and coliform bacterial populations were not significantly affected by the probiotic treatment, either in sows or in the piglets. However, a remarkable decline in the frequency of beta-haemolytic and O141 serovars of Escherichia coli was observed in the intestinal contents of probiotic piglets, suggesting an explanation for the reduction in cytotoxic T-cell populations.     

Mycobacterium avium subsp. paratuberculosis enters the small intestinal mucosa of goat kids in areas with and without Peyer's patches as demonstrated with the everted sleeve method

The main lesions of paratuberculosis in ruminants are in the small intestine. Previous studies have shown that the bacterium enters the small intestine through M cells found in the follicle-associated epithelium lining the domes of the Peyer's patches. The everted sleeve method, devised for the in vitro study of intestinal absorption, was used in this study to investigate the uptake of Mycobacterium avium subsp. paratuberculosis in goat intestine. Everted small intestinal sleeves of goat kids, prepared from areas with and without Peyer's patches, were incubated for 60 min in 3H-labeled bacterial solution. The results of this study imply that the bacteria can enter the intestinal mucosa of the jejunum, both in areas with and without Peyer's patches. These findings indicate, therefore, that M. avium subsp. paratuberculosis bacteria not only enter through M cells but also through enterocytes.     

Age-related changes in the anatomical characteristics of Peyer's patches in small intestine of Bactrian camels (Camelus bactrianus)

The distribution, size, and appearance of Peyer's patches vary according to species. In order to determine the anatomical characteristics of Peyer's patches in small intestine of Bactrian camel, and age-related changes in the number of Peyer's patches, 40 Bactrian camels of the following four age groups were studied: young (0.5–2 years), pubertal (3–5 years), middle-aged (6–16 years), and old (17–20 years). The exact number of Peyer's patches was recorded, and the appearance of Peyer's patches was described in detail. The results indicated that Peyer's patches of Bactrian camels not only have a particular anatomical location and distinct appearance but also change with age. They were distributed in the whole small intestine and there were four distinct types of Peyer's patches: nodular, faviform, cup-shaped, and cystic form Peyer's patches. However, the nodular and cystic form Peyer’s patches are specific to Bactrian camel, which have not been found in other animals including Dromedary camel. In addition, the distribution density of Peyer's patches in ileum was the maximum, then was jejunum and duodenum. Further statistical analysis showed that the number of Peyer's patches was altered with age. The number peaked in 5-year-old camels and declined subsequently with age. However, there was little change in the size of Peyer's patches in different age groups; no age-related macroscopic variations in the shape or size of the Peyer's patches were found. Results obtained from this study provide the basic information to further study on the gastrointestinal mucosal immunity of Bactrian camel.     

Follicle associated epithelium of the gut associated lymphoid tissue of cattle

The morphology of the gut-associated lymphoid tissue of the small and large intestine in three gnotobiotic calves was examined by scanning and transmission electron microscopy, and the distribution of specialized membranous cells present in the follicle associated epithelium was defined. Isolated follicles remaining in the ileum of a cow after involution of the continuous Peyer's patch were examined by scanning electron microscopy. The presence of membrane-bound particles, reported to be exclusively associated with the continuous Peyer's patch, was investigated in other gut-associated tissue of the small and large intestine of the calf. The presence of two types of follicle associated epithelium in the small intestine of the calf was confirmed, and the follicle associated epithelium of the large intestine proved to be a homogeneous population of specialized membranous cells, similar to that of the continuous Peyer's patch of the small intestine. In the discrete Peyer's patches, some specialized membranous cells were completely hidden by adjacent enterocytes and could only be identified by cytoplasmic extensions into the intestinal lumen. In the proximal part of the continuous Peyer's patch, a transitional zone was detected where the follicle associated epithelium of some doomed villi was composed of a homogeneous population of specialized membranous cells, while the epithelium covering other doomed villi consisted of a mixture of absorptive and specialized membranous cells, usually only found in the discrete Peyer's patches. Membrane-bound particles were observed associated with gut-associated lymphoid tissue in the small and large intestine.     

CD8+T细胞在不同年龄家兔胃肠道的分布规律

探讨不同年龄组家兔胃肠道各段CD8⁺T细胞的分布特征,为进一步研究CD8⁺T细胞的作用奠定形态学基础。采用免疫组织化学技术对不同年龄组(幼年组、青年组、老年组)家兔胃肠道内CD8⁺T细胞进行染色、观察及统计分析。结果显示,家兔的CD8⁺T细胞多呈圆形、椭圆形或呈不规则形。在不同年龄组家兔,胃肠不同部位的分布有如下特点:家兔的CD8⁺T细胞主要分布于胃肠道的固有层中;青年组CD8⁺T细胞的分布数量显著高于幼年组和老年组;各肠段之间CD8⁺T细胞分布的数量差异不显著,但其分布的总量按照小肠、大肠、胃的顺序依次降低。研究结果显示,CD8⁺T细胞在家兔小肠的分布最多,并且其数量随着年龄的增长呈现先上升后下降的趋势,青年家兔胃肠的分布最为丰富。     

Pathogenesis of canine parvovirus-2 in dogs: histopathology and antigen identification in tissues

The pathogenesis of canine parvovirus-2 was studied in orally inoculated conventional dogs using histopathological and peroxidase anti-peroxidase staining techniques. Lymphoid necrosis and depletion of lymphocytes from lymphoid tissues were most notable on days 5 and 6 after exposure. Lymphocyte hyperplasia occurred following day 7. Epithelial cell changes in segments of the small intestine were more severe on days 6 to 9 after exposure in areas associated with Peyer's patches and in the upper segments of the small intestine. The lymphocyte was the primary infected cell. Virus infected cryptal epithelial cells were not detected until 24 hours after the identification of infected cells in lymphoid tissues on day 4 after exposure. The majority of virus infected epithelial cells were found in crypts intimately associated with or adjacent to Peyer's patches in the upper segments of the small intestine.     

长春封闭群日本大耳白兔内脏器官组织结构研究报告

利用奥林帕斯光学显微镜,对长春封闭群日本大耳白兔胃、小肠、大肠、卵巢、睾丸的组织结构分别进行了研究。     

Ultrastructure and alkaline phosphatase activity of the dome epithelium of canine Peyer's patches

The dome epithelium of Peyer's patches from different parts of the intestine of four dogs was examined by scanning and transmission electron-microscopy and by alkaline phosphatase histochemistry on glycol-methacrylate embedded sections. M cells were scattered among more numerous enterocytes in duodenal and jejunal Peyer's patches, but constituted the major cellular component of dome epithelia of the ileal Peyer's patches. Alkaline phosphatase histochemistry demonstrated low enzyme activity in the brush border of M cells, as compared to enterocytes, in the duodenum and jejunum, allowing identification of M cells at the light microscopic level. Alkaline phosphatase activity was too low in ileal enterocytes to permit visualization of M cells. The presence of intrafollicular invaginations of dome epithelium is a consistent finding in duodenal Peyer's patches of the dog and these invaginations were characterized by few M cells, many intraepithelial lymphocytes and strong alkaline phosphatase activity.     

Investigation of bovine gut associated lymphoid tissue (GALT) using monoclonal antibodies against bovine lymphocytes

Gut associated lymphoid tissue of the small and large intestine of calves and cows has been compared morphologically and quantitatively using monoclonal antibodies to bovine lymphocytes. B cells were significantly decreased in the ileum of the cow compared to the calf. Significantly increased numbers of T cells were present in cell suspensions of all lymphoid areas of the cow compared to the calf. T lymphocyte subsets were quantified into cryostat sections of lymphoid tissues expressing BoT4, and BoT8 antigens demonstrated increased numbers in follicular and dome areas of the discrete Peyer's patches of the small and large intestine of the cow. BoT4+, BoT8+, and the non-BoT4/BoT8+ T cell subsets were increased in the mucosa of the cow as compared to the calf. Similarities in structure and lymphocyte composition of the discrete Peyer's patches of the small intestine, cecum and colon and isolated single follicles in the large intestine suggest similar functional properties.     

年龄差异对家兔CD4+T细胞在胃肠分布的影响

为深入了解家兔消化道中CD4⁺T细胞的组织学定位以及在消化道不同解剖学部位的数量变化,本试验运用免疫组织化学方法,探讨了家兔胃(胃底腺区、幽门腺区)和小肠及大肠不同解剖学部位中的CD4⁺T细胞定位特点和数量变化规律。结果表明,在胃肠中,CD4⁺T细胞主要分布于固有层。胃底腺区阳性细胞数目要明显比幽门腺区少。小肠、大肠各段的阳性细胞分布数量未见明显差异,由十二指肠到回肠表现为小幅度递增,由盲肠到直肠表现为小幅度递减。胃肠中CD4⁺T细胞在各部位的数量随年龄增长表现出先增后减的变化趋势,老年组和幼年组CD4⁺T细胞数量在大肠中未见明显差异。结果提示,CD4⁺T细胞在青年家兔胃肠中的分布最为丰富。     

Mycotoxicosis caused by aerosolized T-2 toxin administered to female mice

Thymus, spleen, adrenal glands, and small intestine of female mice exposed to aerosolized T-2 mycotoxin were examined at postexposure hours (PEH) 0.25, 1, 2, 4, 6, 9, 12, and 24. Lymphocyte necrosis was observed at PEH 1 in the thymus, spleen, and lamina propria and Peyer patches of the small intestine. Necrosis of small intestinal crypt epithelial cells was observed at PEH 2, and necrosis of parenchymal cells and increased number of neutrophils were seen in sinusoids of the adrenal cortex at PEH 4. These results indicated that the earliest microscopic evidence of T-2 mycotoxicosis after aerosol exposure was necrosis of lymphocytes in the thymus, spleen, and lamina propria and Peyer patches of the small intestine.     

Distribution of the pores of epithelial basement membrane in the rat small intestine

The distribution and diameter of the pores of epithelial basement membrane in the intestinal villi and the lymph nodules of ileal Peyer's patches were investigated in the rat small intestine by scanning electron microscopy after the removal of the overlying epithelial cells with OsO(4) maceration. In the duodenum, jejunum and ileum, the pores were mainly distributed at the upper three fourths of the villi, but were scarce around the top of the villi. The diameter of some of the pores in the upper three fourths of the villi was larger than that of those in the lower portion. The protrusion of lymphocytes and the cytoplasmic processes of macrophages were also seen at the orifices of the pores. In ileal Peyer's patches, in contrast, pores were densely distributed in the lower one third of the follicle-associated epithelium (FAE) where M cells were mainly seen. Furthermore, these pores were larger than those found in the upper two thirds. Lymphocytes or cytoplasmic processes of macrophages were frequently seen in the lower one third of FAE. These results suggest that the pores at the basement membrane correspond to the passage of the immunocompetent cells which are in contact with M cells or villous columnar epithelial cells and that the abundance of pores is a sign of aggressive interaction between the particular epithelial cells and the immunocompetent cells at the upper three fourths of intestinal villi and the lower one third of FAE in the rat small intestine.     

Glycoconjugate histochemistry in the small and large intestine of normal and Solanum glaucophyllum-intoxicated rabbits

Vitamin D participates in mineral homeostasis, immunomodulation, cell growth and differentiation. The leaves of Solanum glaucophyllum contain high levels of 1,25-dihydroxyvitamin D3 as glycoside derivatives and their chronic ingestion generates a hypervitaminosis D-like state. We analyzed changes on carbohydrate expression as a cell differentiation indicator on samples of the small and large intestine of S. glaucophyllum-intoxicated rabbits, using conventional and lectin histochemistry. Male New Zealand white rabbits were intoxicated with S. glaucophyllum during two or four weeks and killed the day after. A group of animals (“possibly recovered group”) were intoxicated during 15 days and killed at day 45 of the beginning of the experiment. We found changes in the lectin binding pattern in the small and large intestine of the intoxicated rabbits. Some of these changes were reverted in the possibly recovered group. Vitamin D could be a new regulator factor of the intestinal glycosylation process.     

Structure and function of Peyer's patches in the intestines of different animal species

Structure and function of Peyer's patches in man and animals are compared reviewing reports of recent years. Differences exist regarding number and distribution of Peyer's patches along the small intestine and regarding distribution of m-cells of dome epithelium. M-cells trigger the contact between infectious organisms and intestinal epithelium. Several different ways of interaction are possible: 1. Selective uptake of organisms by m-cells. 2. Paracellular transport of organisms between m-cells. 3. Multiplication of organisms above dome epithelial areas. 4. Restriction of interaction between organism and mucosa to absorptive enterocytes. 5. Specific damage of organisms to lymphatic areas of Peyer's patches.     

Studies on the evolution, pathology, and immunity of commercial fattening rabbits affected with epizootic outbreaks of diarrhoeas in Mexico: a case report

Epizootic outbreaks of diarrhoeas have emerged and disseminated in different rabbit farms in Mexico causing great economical losses, during the past years. Seven, 5-weeks-old New Zealand White (NZW) rabbits chosen at random from 35 ill animals that were remitted for postmortem, histopathology, and ultrastructural examinations were studied. Bacteriological and parasitological studies were carried out in three additional ill rabbits of same age. In a field trail 45, 5-weeks-old apparently healthy NZW rabbits were observed daily for sanitary status for a 5-week period. Some of the rabbits did not response to the preventive drug treatment and were therefore, used to study the development of the disease. Clinical signs, gross lesions, and mortality throughout the fattening period were recorded. Eight, 8-weeks-old NZW rabbits who survived an outbreak were assessed for gamma-globulins in serum of the total protein fraction during a 3-week period. Gamma-globulins were also measured in eight free-disease healthy rabbits of same breed and age. Lesions of the small intestine consisted of mucoid enteropathy, lymphocytic plasmocytic enteritis with atrophy and fusion of villi, and hyperplasia of globet cells. Serosal edema was present. Ultrastructural examinations of jejunum and ileum from 3/7 diseased rabbits, revealed enterocytes in apoptosis, mixed with degenerative and/or necrotic changes together with infiltration of lymphocytes, macrophages, neutrophils, and loss of microvillus. There were electron dense structures suggestive of virus particles inside the nuclei and cytoplasm of some enterocytes. There was lymphoid spleen atrophy and proliferation of reticuloendothelial cells in 7/7 rabbits. Interstitial pneumonia in 4/7 rabbits was found. Encephalitozoon cuniculi was detected in the brain of 1/7 rabbits. Escherichia coli were detected in 3/3 cases and Eimeria spp. in 2/3 cases. Mortality rate in the field study was 51.1% and the spread of the disease occur in 9/9 cages. The proportion of gamma-globulins in rabbits who survive an outbreak was much lower (P=0.0001) than free-disease healthy rabbits (8.1+/-1.0 and 14.0+/-1.0, respectively). The disease was multifactorial and consisted of sub-acute mucoid enteropathy probably induced by viral infection and aggravated by the proliferation of opportunistic pathogens common to rabbits. This may explain the severe degenerative and necrotic changes observed in the small intestine of diarrhoeic rabbits.     

Penetration in vitro of newly excysted juvenile flukes of Japanese Fasciola sp. through ligated intestines of rabbits, mice, rats and chickens.

Ligated intestines of rabbits, mice, rats and chickens were used to examine the penetration of newly excysted juvenile flukes of Japanese Fasciola sp. in vitro. In rabbit intestines, the penetration rate was relatively high in the rectum and duodenum. Penetration rates in the jejunum, ileum, cecum and colon were comparable to those in the rectum and duodenum, although it was lower in the appendix. In the case of mouse, juvenile flukes penetrated the duodenum, jejunum, cecum, and rectum at considerably high rates. In rat intestine, penetration by flukes was less in the duodenum and rectum, although flukes were detected in the jejunum. In chicken intestine, flukes barely penetrated the duodenum, jejunum and rectum. Consequently, newly excysted flukes of Fasciola sp. seem to penetrate any region of the intestine in rabbits and mice. In rats, the middle small intestine may be the site suitable for flukes to penetrate. In chickens, the difficulty in penetration of the intestinal wall may be one of the reasons why chickens are scarcely infected with Fasciola sp.     

Enterocecocolitis associated with intraepithelial Campylobacter-like bacteria in rabbits (Oryctolagus cuniculus)

We examined 28 suckling, weanling, and young adult rabbits with lethargy, inappetence, and mucinous, semifluid feces. Sixteen of the rabbits had intestinal lesions. In eight of these rabbits, the primary changes were multifocal to diffuse epithelial proliferation and accumulation of lymphocytes, macrophages, or both in the lamina propria of the small intestine, cecum, and sacculated colon. In two of these rabbits, the accumulation of macrophages in the lamina propria was extensive. The other eight rabbits had erosive and suppurative cecocolitis, and four of the rabbits with proliferative lesions also had suppurative cecocolitis. In Warthin-Starry-stained sections of affected intestine, curved or spiral bacteria were visible within degenerated or hyperplastic epithelium, in luminal exudate, or in both. Such organisms were sparse or not found in the other 12 rabbits, which did not have intestinal lesions. The bacteria ultrastructurally resembled intraepithelial Campylobacter-like bacteria previously observed in proliferative enteritis in a variety of species and in acute typhlitis in young rabbits. In immunofluorescence tests, Campylobacter-like bacteria in epithelial cells, crypt lumina, and in luminal exudates in both proliferative and erosive lesions bound monoclonal antibodies and polyclonal antisera prepared against intracellular bacteria found in proliferative enteritis in pigs, hamsters, and ferrets. These observations indicate that a condition similar to proliferative enteritis of swine, hamsters, and other species also occurs in laboratory rabbits.     

谷氨酰胺对断奶肉兔肠道发育的影响

本试验旨在研究谷氨酰胺对肉兔肠道发育的影响。试验选择28日龄断奶新西兰白兔360只,随机分成4个组,每个组9个重复,每个重复10只(公母各占1/2)。4组试验兔分别饲喂在基础饲粮基础上添加0(对照)、0.4%、0.8%和1.2%谷氨酰胺的试验饲粮,试验期为4周。结果表明:饲粮添加谷氨酰胺对肉兔35、42和56日龄时的胃重、盲肠重和小肠长度以及56日龄时的小肠重无显著影响(P0.05),但饲粮添加0.8%谷氨酰胺可以显著提高肉兔35和42日龄时的小肠重(P0.05);饲粮添加0.8%和1.2%谷氨酰胺可以显著提高肉兔35和42日龄时的十二指肠和空肠绒毛高度(P0.05),同时饲粮添加0.8%谷氨酰胺还可以显著提高肉兔35和42日龄时的回肠绒毛高度以及56日龄时的十二指肠和空肠绒毛高度(P0.05);饲粮添加0.8%谷氨酰胺可以显著降低肉兔35和42日龄时的十二指肠和空肠隐窝深度以及35日龄时的回肠隐窝深度(P0.05)。由此可见,饲粮添加谷氨酰胺可以促进肉兔断奶后1~2周的肠道发育,本试验条件下,肉兔饲粮中谷氨酰胺适宜添加水平为0.8%。     

断奶日龄对肉兔肠道发育的影响

本试验旨在研究断奶日龄对肉兔肠道发育的影响。试验选择初生肉兔120窝,随机分成4组,每组30窝,分别在出生后21、25、28和35日龄断奶,试验期56 d。结果表明:1)断奶日龄对肉兔28和35日龄胃、小肠和盲肠重有显著或极显著影响(P0.05或P0.01),随着断奶日龄延后呈增加趋势,断奶日龄对49和56日龄各消化器官重和各日龄小肠长度无显著影响(P0.05);2)断奶日龄对十二指肠(28、35和49日龄)、空肠(28、35、42和49日龄)和回肠绒毛高度(28、35、42和49日龄)有显著或极显著影响(P0.05或P0.01),随着断奶日龄延后呈先增加后降低趋势,而对56日龄各肠段绒毛高度无显著影响(P0.05),肉兔断奶后绒毛高度需要2~3周才能恢复,21日龄断奶组恢复最慢,35日龄断奶组恢复最快;3)断奶日龄对十二指肠(28和35日龄)、空肠(28、35和42日龄)和回肠隐窝深度(35日龄)有显著影响(P0.05),随着断奶日龄延后呈降低趋势,而对49和56日龄各肠段隐窝深度无显著影响(P0.05);4)断奶日龄对28和35日龄十二指肠、空肠和回肠绒毛高度/隐窝深度有极显著影响(P0.01),随着断奶日龄延后呈增加趋势,而对56日龄各肠段空肠绒毛高度/隐窝深度无显著影响(P0.05)。由此可见,断奶日龄对肉兔肠道49日龄前发育影响较大,对49日龄后影响变小;早期断奶降低了小肠黏膜绒毛高度,增加了隐窝深度;随断奶日龄延后,断奶造成的肉兔小肠黏膜受损程度减轻,断奶后一般需要2~3周才能恢复小肠黏膜结构。