Emigration of polymorphonuclear leucocytes into the intestinal lumen of the neonatal piglet in response to challenge with K88-positive Escherichia coli

The emigration of neutrophils from the blood of neonatal piglets into the intestinal lumen in response to a K88-positive strain of Escherichia coli was investigated. The pig herd used was of known genetic susceptibility to K88-positive E coli and had recently experienced an outbreak of neonatal diarrhoea. Neutrophil emigration depended on certain factors. Neutrophils emigrated into ligated loops in susceptible piglets sucking immune colostrum from susceptible dams but not into loops in colostrum deprived resistant piglets or piglets sucking non-immune colostrum from resistant dams. In susceptible, colostrum deprived piglets neutrophils in intestinal contents were only associated with severe lesions. Large numbers of neutrophils which appeared at several foci on the villi were observed in three of six resistant piglets that sucked immune colostrum from susceptible dams. It was concluded that neutrophil emigration into the intestinal lumen of piglets could occur in response to K88-positive E coli and resulted not from the presence or absence of the intestinal K88-receptor but from the ingestion of immune colostrum.     

Review papers

Summary

The literature on the biology and pathology of mammalian small intestinal permeability to macromolecules is reviewed.

In mammals, macromolecules may penetrate the epithelial layer of the small intestinal mucosa, especially in the neonatal period. The neonatal uptake and transport of immunoglobulins is important in the acquisition of passive immunity in the newborn. In the mature small intestine the uptake of macromolecules almost ceases, except in M‐cells.

Excessive uptake and transport of macromolecules has been demonstrated in several experimental and spontaneous gastrointestinal diseases, in which increased concentration of macromolecules in the small intestinal lumen and/or damage of one or more components of the small intestinal mucosal barrier is present. Finally, methods to study macromolecular permeation of the small intestine are discussed.     

Effects of immune-mediated enteroluminal neltrophil emigration on intestinal function in pigs.

Intestinal function was assessed in cannulated loops of porcine proximal jejunum during immune-mediated emigration of neutrophils into the intestine. Net water, net sodium, net chloride, undirectional sodium and unidirectional chloride fluxes were measured before and after intestinal exposure to an antigen in both sensitized and nonsensitized pigs. Neutrophil emigration was assessed histologically. The results indicate that fluid and electrolyte movements in the intestine are not significantly altered during immune-mediated enteroluminal neutrophil emigration suggesting that the neutrophil response does not interfere with intestinal function.     

肠道菌群对动物肠黏膜免疫的调控作用

动物肠道共生着数量庞大、结构复杂的菌群,而肠壁内存在着为数众多、功能强大的黏膜淋巴细胞。肠道菌群具有促进肠黏膜免疫系统生长与发育和调控肠黏膜屏障与免疫功能的双重作用。本文主要从动物肠道菌群的定植与特性、肠黏膜免疫细胞的功能以及肠道菌群对肠黏膜屏障与免疫功能的调控作用进行综述。     

Intestinal crypt lesions associated with protein-losing enteropathy in the dog

Six dogs were diagnosed with protein losing enteropathy (PLE). There was no evidence of inappropriate inflammatory infiltrates or lymphangiectasia in multiple mucosal biopsies of the small intestine of 4 of the dogs. The 5th and 6th dogs had obvious lymphangiectasia and a moderate infiltrate of inflammatory cells in the intestinal mucosa. All 6 dogs had a large number of dilated intestinal crypts that were filled with mucus, sloughed epithelial cells, and/or inflammatory cells. Whether PLE occurs in these dogs because of protein lost from the dilated crypts into the intestinal lumen or whether the dilated crypts are a mucosal reaction due to another undetermined lesion that is responsible for alimentary tract protein loss is unknown. However, when large numbers of dilated intestinal crypts are present, they appear to be associated with PLE even if there are no other remarkable lesions in the intestinal mucosa.     

Variation in the immuno-pathological responses of lambs after experimental infection with different strains of Mycobacterium avium subsp. paratuberculosis

Ruminant infection by Mycobacterium avium subsp. paratuberculosis (MAP) causes a granulomatous inflammatory response in the intestine and associated lymph nodes. Differences either in the affected organs or in the inflammatory infiltrate were observed between species and individuals. Such differences are usually attributed to variations in host immune responses or to inconsistent effects among different MAP strains. To evaluate if different MAP strains induce different immuno-pathological responses in lambs, 28 one-month-old individuals were divided into six groups and inoculated with different MAP strains. Groups 1 and 2 were inoculated with two bovine strains isolated in Argentina that showed different genetic patterns after BstEII-IS900-RFLP (hereafter strains E and A respectively). Group 3 was inoculated with a bovine strain isolated in Spain obtained after a previous step of culture (patterns C1). Group 4 was inoculated with a homogenate of intestinal mucosa of a clinical case affected by the same bovine strain as that of group 3. Group 5 was inoculated with an ovine strain that was directly purified from the intestinal mucosa of a clinical case, and group 6 was kept as control (i.e. no inoculation). Peripheral immune responses were assessed until 150 days post-infection (dpi), when lambs were humanely killed. Pathological studies were performed in tissues from the intestine and lymph nodes. Lesion types and inflammatory infiltrates were examined as indicators of pathogenicity. All the lambs infected with bovine MAP strains showed a common lesion pattern regardless of the strain type. Such pattern was characterized by focal lesions mainly in the mesenteric lymph nodes, the presence of fibrous tissue, and, occasionally, necrosis in the granulomas as well as the presence of numerous giant cells. Differences in lesion severity were observed among groups: lambs from groups 1 and 2 had the highest number of granulomas and the largest lymph node area affected. Lesions in animals from group 5 (infected with an ovine strain) were more severe and occurred mostly in the intestinal lymphoid tissue; necrosis, fibrosis or giant cells were never detected in this group. These results indicate that the MAP strain type induces different pathological responses in lambs.     

In vitro Studies of Group E Streptococci in Swine Leukocytes I. Phagocytic and Bactericidal Properties of Polymorphonuclear Leukocytes from Swine

Serum from both immune and nonimmune ten-week-old swine contained factors which promoted phagocytosis of group E Streptococci (GES). The factors in nonimmune serum, which were heat labile at 70°C for ten minutes, were less efficient than the factors present in immune serum.

Bactericidal activity of the polymorphonuclear (PMN) leukocytes against GES was observed with serum from both immune and nonimmune ten-week-old swine, as well as with serum from normal sows and piglets. However, the bactericidal activity of PMN leukocytes in serum from either normal sows or immune ten-week-old swine was greater than the bactericidal activity of PMN leukocytes in either piglet serum or serum from nonimmune ten-week-old swine. When the serum was either heated to 70°C for ten minutes or treated with 2-mercaptoethanol, bactericidal activity of PMN leukocytes against GES was only observed in the presence of immune serum.

     

Modulation by colostrum-acquired maternal antibodies of systemic and mucosal antibody responses to rotavirus in calves experimentally challenged with bovine rotavirus

The effect of colostral maternal antibodies (Abs), acquired via colostrum, on passive protection and development of systemic and mucosal immune responses against rotavirus was evaluated in neonatal calves. Colostrum-deprived (CD) calves, or calves receiving one dose of pooled control colostrum (CC) or immune colostrum (IC), containing an IgG1 titer to bovine rotavirus (BRV) of 1:16,384 or 1:262,144, respectively, were orally inoculated with 105.5 FFU of IND (P[5]G6) BRV at 2 days of age. Calves were monitored daily for diarrhea, virus shedding and anti-BRV Abs in feces by ELISA. Anti-rotavirus Ab titers in serum were evaluated weekly by isotype-specific ELISA and virus neutralization (VN). At 21 days post-inoculation (dpi), all animals were euthanized and the number of anti-BRV antibody secreting cells (ASC) in intestinal and systemic lymphoid tissues were evaluated by ELISPOT. After colostrum intake, IC calves had significantly higher IgG1 serum titers (GMT=28,526) than CC (GMT=1195) or CD calves (GMT<4). After BRV inoculation, all animals became infected with a mean duration of virus shedding between 6 and 10 days. However, IC calves had significantly fewer days of diarrhea (0.8 days) compared to CD and CC calves (11 and 7 days, respectively). In both groups receiving colostrum there was a delay in the onset of diarrhea and virus shedding associated with IgG1 in feces. In serum and feces, CD and CC calves had peak anti-BRV IgM titers at 7 dpi, but IgA and IgG1 responses were significantly lower in CC calves. Antibody titers detected in serum and feces were associated with circulation of ASC of the same isotype in blood. The IC calves had only an IgM response in feces. At 21 dpi, anti-BRV ASC responses were observed in all analyzed tissues of the three groups, except bone marrow. The intestine was the main site of ASC response against BRV and highest IgA ASC numbers. There was an inverse relationship between passive IgG1 titers and magnitude of ASC responses, with fewer IgG1 ASC in CC calves and significantly lower ASC numbers of all isotypes in IC calves. Thus, passive anti-BRV IgG1 negatively affects active immune responses in a dose-dependent manner. In ileal Peyer's patches, IgM ASC predominated in calves receiving colostrum; IgG1 ASC predominated in CD calves. The presence in IC calves of IgG1 in feces in the absence of an IgG1 ASC response is consistent with the transfer of serum IgG1 back into the gut contributing to the protection of the intestinal mucosa.     

低聚糖对断奶羔羊小肠黏膜免疫相关细胞的影响

选用陶赛特F2代断奶公羔40只,随机分成4组,研究低聚糖对断奶羔羊小肠黏膜免疫相关细胞的影响。结果表明:日粮中添加低聚果糖FOS(0.3%)、低聚异麦芽糖IMO(0.3%)和复合低聚糖(0.1%FOS+0.2%IMO)均可提高断奶羔羊胸腺指数和脾脏指数(P>0.05)及小肠黏膜上皮内sIEL数量(P<0.05);FOS可提高小肠黏膜上皮GC的数量I,MO和复合低聚糖可降低GC数量(P>0.05)I;MO和复合低聚糖可提高断奶羔羊小肠黏膜中MC细胞的数量(P>0.05)。提示断奶羔羊日粮中添加低聚糖可增加其小肠黏膜免疫相关细胞的数量,增强断奶羔羊小肠黏膜免疫功能,提高其免疫力。     

Pregnancy-associated glycoprotein, chymosin and pepsinogen immunoreactivity of proteins extracted from fetal gastric tissue in bovine species

The objective of this work was to investigate the expression of gastric aspartic proteinases in fundic and pyloric mucosa removed from bovine fetuses. For this purpose, fractions issued from classical biochemical protocols were analyzed by proteolytic method, by PAG-RIA and by Western blot with the use of antisera raised against both pepsinogens and PAG. A strong reaction of proteins extracted from the fundic mucosa collected at the beginning of pregnancy was revealed with both anti-bPAG-I and anti-bPAG-II antisera, suggesting the expression of pepsinogen F in bovine species. Concerning pyloric mucosa, the analysis by Western blot highlighted a very strong immunoreaction with the anti-bovine chymosin serum. Amino-terminal sequencing allowed to identify bovine fetuin and albumin in fundic extracts, chymosin in the pyloric mucosa extracts, as well as some unknown proteins in both mucosa. Despite no N-terminal microsequence corresponding to the hypothetical pepsinogen F could be identified, it cannot be excluded that an existing bovine pepsinogen F-like molecule could be degraded during the purification procedure or that co-purified proteins could be responsible for masking its N-terminal microsequence.     

玉屏风多糖对小鼠派氏结形态结构及其T细胞亚群的影响

通过研究玉屏风多糖（YPF-P）对小鼠派氏结（Peyer's patches,PPs）形态结构及其T细胞亚群的影响,探讨其对小鼠肠黏膜的免疫调节作用。选取96只SPF小鼠随机分为6组,分别为空白对照组（0.2 mL生理盐水）、YPF-P阳性对照组（YPF-P 200 mg/kg）、环磷酰胺（cyclophosvnamide,Cy）免疫抑制组（80 mg/kg Cy）及YPF-P低、中、高剂量组（100、200、400 mg/kg YPF-P）,饲喂1周,取小肠PPs,常规切片HE染色后应用图像分析技术检测PPs形态结构的变化;体外培养小鼠PPs淋巴细胞,采用流式细胞术研究PPs中T淋巴细胞亚群的变化。结果表明,YPF-P对小鼠小肠PPs的生长发育具有促进作用,Cy可极显著降低小鼠小肠PPs面积、小肠纵切面面积及PPs面积和小肠纵切面面积的比值（P < 0.01）,低、中、高剂量YPF-P可在一定程度上缓解Cy对小鼠小肠的损伤作用。同时,YPF-P可显著或极显著提高PPs中CD3⁺、CD4⁺ T淋巴细胞及CD4⁺/CD8⁺（P < 0.05; P < 0.01）。结果显示,YPF-P能提高Cy诱导的免疫抑制小鼠的肠黏膜免疫功能,并能促进PPs中相关T淋巴细胞增殖,对小鼠肠黏膜功能具有增强作用。     

Inhibition of passive systemic anaphylaxis in guinea pigs by Trichinella spiralis infection

Thirty of 33 guinea pigs experimentally infected with Trichinella spiralis survived passive systemic anaphylactic reactions which were fatal for 32 of 33 uninfected controls. Guinea pigs were infected with viable excysted trichinella larvae 20 to 32 days before they and uninfected controls were sensitized by intracardiac injection of rabbit anti-bovine serum albumin. Animals were challenged 24 hours later by intracardiac injection of bovine serum albumin. Respiratory anaphylaxis was observed in all challenged animals. Protection from passive anaphylaxis in parasitized guinea pigs was most likely due to preemption of mast cell receptors by parasite-induced antibody with consequent blockage of passive sensitization.     

Haemorrhagic Enteropathies in Red Deer (Cervus Elaphus)

Sir, — I would like to draw your readers’ attention to the occurrence of post-stress haemorrhagic enteropathies in red deer (Cervus elaphus). Haemorrhagic enteropathy has been observed in approximately 75 percent of red deer necropsied at the Invermay Animal Health Laboratory and has also been seen in wapiti (Cervus canadensis). Deer dying with post-stress haemorrhagic enteropathy commonly have splash haemorrhages on the serosal surfaces of the abomasum and intestine and to a lesser extent on the rumen and omasum. The intestinal mucosa is heavily congested and haemorrhagic with blood present in the lumen. In many cases mucosal haemorrhage may be so severe the intestinal contents appear similar to blood. The degree of severity of the condition varies from slight to very severe and the length of intestine involved also varies from a few centimetres to the entire gastro-intestinal tract from the rumen to the rectum.     

乳酸杆菌在调节肠道屏障功能中的作用

乳酸杆菌不仅是动物肠道内存在的正常微生物,也是益生菌的重要组成部分。肠道屏障主要是由肠道黏膜免疫系统组成的免疫屏障、肠黏液层和肠上皮细胞层组成的肠黏膜物理屏障及肠道微生物构成的生物屏障构成。作者综述了近年来国内外有关乳酸杆菌制剂对肠道屏障功能调控作用的研究进展。     

The interaction between plant lectins and the small intestinal epithelium: A primary cause of intestinal disturbance

Summary

The literature concerning the effects of plant lectins on the small intestinal epithelium is reviewed. It appears that after oral intake, intact plant lectins can reach the small intestinal lumen. Their binding to the mucosal surface evokes an increased synthesis of glycoproteins and a degeneration of the intestinal epithelium. The epithelial alterations may result in hyperregenerative villus atrophy and endogenous nitrogen loss. These changes ultimately can lead to less efficient feed conversion, diminished growth, scouring, wasting and death. The possible significance of plant lectins in digestive disturbances in farm animals is suggested.     

In vivo investigation of the efficacy of a customized solution to attenuate injury following low-flow ischemia and reperfusion injury in the jejunum of horses

OBJECTIVE: To evaluate the efficacy of a customized solution to attenuate intestinal injury following 20% low-flow ischemia and reperfusion in the jejunum of horses. ANIMALS: 10 healthy adult horses. PROCEDURE: Two 30.5-cm-long segments of jejunum were exteriorized through a ventral midline incision and the mesenteric artery and vein supplying that portion of the intestine were instrumented with flow probes. Blood flow was decreased to 20% of baseline for 90 minutes followed by 90 minutes of reperfusion. In 5 horses, 60 mL of the customized solution was placed in the lumen of each segment (treatment-group horses), and 60 mL of lactated Ringer's solution was placed in the lumen of 5 additional horses (control-group horses). Biopsy specimens were obtained from 1 segment in both groups for histologic evaluation. Aliquots of luminal fluid were obtained from the other segment in both groups for determination of albumin concentrations as an index of mucosal permeability. RESULTS: Compared with control-group horses, treatment-group horses had a significant decrease in luminal albumin concentration following reperfusion. Although differences in mucosal grades were not significantly different between control- and treatment-group horses, treatment-group horses had significantly greater jejunal villous length and area, compared with that of control-group horses. CONCLUSIONS AND CLINICAL RELEVANCE: Intraluminal administration of the customized solution in the jejunum, compared with lactated Ringer's solution, results in an improvement in histologic findings and mucosal translocation of albumin in horses with mild intestinal injury.     

Observations on avian intestinal spirochaetosis

Three SPF-laying hens were inoculated into the crop with avian intestinal spirochaetes which previously had been passaged in broiler chicks by oral inoculation (isolate 1380). Mild persisting gastrointestinal disorder developed; at nine months post inoculation spirochaetes were readily demonstrated in caecal faeces. Histologic examination of the caecal mucosa revealed many spirochaetes covering the mucosal surface and filling up the crypts lumina. Spirochaetes were found in intra- and subepithelial locations and in gaps running through the epithelium. These gaps often opened into subepithelial cavities crowded with spirochaetes ('gaplike lesions'). These lesions were seen mostly on the tips of the villi and in the deeper parts of the crypts. Massive erosion or desquamation of epithelium heavily infested by spirochaetes occurred. These findings indicate colonisation of the mucosal surface and of the crypts, penetration of the mucosa and colonisation of subepithelial compartments with spirochasetes in poultry suffering from intestinal spirochaetosis.     

Characteristics of lactoferrin receptor in bovine intestine: higher binding activity to the epithelium overlying Peyer's patches

Several lines of evidence have recently demonstrated the occurrence of specific lactoferrin (Lf) receptors in different cells. We report here, for the first time, the characteristics of binding, and distribution of Lf receptors in the bovine intestinal tract with special emphasis on the epithelium overlying Peyer's patches (EOPP). Brush-border membrane vesicles (BBMV) were prepared from the mucosa of duodenum, jejunum, ileum, colon, EOPP in jejunum and EOPP in ileum. Receptor binding assays were carried out using 125I-labelled bovine Lf. Specific and saturable Lf receptors were found in BBMV of all the intestinal segments examined. Non-linear regression and Scatchard plot analyses clearly revealed that EOPP had the highest binding maximal (Bmax), and lowest in colon. The maximum dissociation constant (Kd) 3.74 microm was in the ileum. We found that bovine transferrin competed with Lf for the same binding site of receptors. In contrast, no binding of bovine serum albumin occurred. It was concluded that Lf receptors in the mucosal lining are attributable to mediate multifunctional activities of Lf in the gut, especially in the EOPP.     

Intestinal mucosal immune response against virulent duck enteritis virus infection in ducklings

Duck virus enteritis (DVE) is an acute and contagious herpes virus infection of duck, geese and swans with high morbidity and mortality. The development of specific mucosal immune system against duck enteritis virus (DEV) infection for ducks has been hindered by a lack of knowledge concerning the purification of immunoglobulin A (IgA) of duck. In the present work, the method for purification of duck immunoglobulin A was developed, and the induction of intestinal mucosal immune responses against DEV was studied by orally infected ducklings with virulent DEV. The results showed that a continuous increased DEV DNA levels were observed in blood and various organs examined of orally infected ducklings throughout the infection, which was accompanied by the development of infection in ducklings from mild progressed to severe pathological lesions. Furthermore, a marked increased level of DEV-specific IgA and IgG antibodies in bile, serum and the intestinal tract, as well as the density of IgA⁺ cells in intestine were detected between 1 and 12 days p.i., followed by a drastic reduction of the antibody levels and the density of IgA⁺ cells at 15 days p.i. The results indicate that the DVE infection can stimulate both IgA-dominated antibody immune responses in the intestinal tract, and IgG-dominated antibody systemic immunity in the serum of ducklings orally inoculated with virulent DEV. The severe lesions of the villus epithelial cells and the lymphoid organs can suppress the intestinal mucosal immune responses.     

Report on a hogget ill-thrift trial

A method of cannulating the small intestine of the calf is described. Indwelling cannulas gave direct access to the intestinal lumen and mucosa, providing direct study of intestinal function and morphology.