Pathogenesis of intestinal cryptosporidiosis in conventional and gnotobiotic piglets.

The pathogenesis of intestinal cryptosporidiosis was studied in 52 conventionally reared and 20 gnotobiotically reared piglets by inoculation with different doses of Cryptosporidium parvum oocysts. The prepatent period of C. parvum in both groups of animals were variable, depending on the number of oocysts administered. The patent period of C. parvum in conventionally reared piglets was 8 or 9 days; in gnotobiotic piglets cryptosporidia were found in feces until Day post infection (DPI) 16, when the last piglet was necropsied. Cryptosporidiosis in conventionally reared piglets is a self-limited diarrheal disease associated with morphological changes within the intestine. The most severe lesion was seen in the posterior jejunum and ileum from DPI 3 to DPI 7, and consisted of villous atrophy, crypt hyperplasia and inflammatory infiltration in the lamina propria. In gnotobiotic piglets cryptosporidia induced severe enterocolitis which occurred at least until DPI 16. The characteristics of enteric lesions were similar to those found in conventionally reared piglets. Intestinal cryptosporidiosis in both groups of animals shifted in the course of infection in the caudal direction and terminated in the large intestine. Examination by scanning electron microscope showed that infected absorptive cells had thicker and longer microvilli than those on non-infected cells; neighboring non-infected cells were hypertrophic, bulbously protuberant with minute microvilli with no distinct intercellular borders. Numerous cryptosporidia in the heterotopic glandular epithelium in the submucosa of cecum and colon on DPI 9 and 10 were found. No differences in the location and degree of cryptosporidial infection between colostrum-fed and colostrum-deprived conventionally reared piglets were found. Sow's colostrum does not appear to protect piglets from C. parvum infection. The role of intestinal microflora in the pathogenesis of cryptosporidiosis in piglets is discussed.     

Experimental Klebsiella and Salmonella infection in neonatal swine.

Twenty colostrum-fed piglets from three sows were separated from the sows 24 hours after birth and were randomly divided into five groups of four piglets each. Every piglet in each of four test groups was orally inoculated with about 10(10) colony forming units of Salmonella typhimurium, Salmonella choleraesuis var Kunzendorf or one of two isolates of Klebsiella pneumoniae. One group served as uninoculated controls. Piglets infected with K. pneumoniae developed severe diarrhea beginning about 12 hours after inoculation. They became dehydrated and weak but continued to drink. There were no morphological alterations in intestinal mucosa when piglets were killed and necropsied 48 or 72 hours after inoculation. Klebseilla pneumoniae was isolated from intestine and feces but not from liver or spleen. Piglets inoculated with S. choleraesuis became lethargic and disinterested in food by 24 hours after inoculation. Diarrhea developed by 48 hours after inoculation. Lesions at necropsy 60 or 72 hours postinoculation were subcutaneous edema, mesenteric lymphadenitis, diffuse intestinal superficial mucosal necrosis with villous atrophy, and focal deep ulceration in the ileum. Salmonella choleraesuis was isolated from all segments of intestine and from feces, liver and spleen. Piglets inoculated with S. typhimurium developed a relatively mild diarrheal disease with lesions similar to those with S. choleraesuis infection but less severe. The inoculated organism was recovered from all areas of intestine and from feces, liver and spleen. Serum from infected and control piglets had high (greater than 1:256) agglutinating titres against S. typhimurium but low titres (0 to 1:8) against S. choleraesuis. The agglutinins were assumed to originate from colostral antibodies.     

An immunohistochemical study of S-100 protein in the intestinal tract of Chinese soft-shelled turtle, Pelodiscus sinensis

The present work describes the distribution of S-100 protein in the intestinal tract of a Chinese soft-shelled turtle specimen (Pelodiscus sinensis). S-100 protein positive cells were located in the intestinal tract, from the proximal small to distal large intestine. S-100 protein positive dendritic cells had irregular shape and were positive in both cytoplasm and nucleus. Most of them were located both lamina propria and submucosa in the small intestine, while few were found in the large intestine. S-100 protein, C-kit positive ICCs and Silver staining glial cells were predominantly observed in three locations: (1) in the interspace between the submucosa and circular muscle layer; (2) in the circular muscle layer; and (3) between the circular and longitudinal muscle layers of the intestine. Fewer were found in the large intestinal lamina propria and submucosa. Three types of positive cells (S-100 protein positive cells, C-kit positive ICCs and Silver staining glial cells) with 1–2 long or 2–3 short processes were distributed as lace-like or surrounding blood vessels in the different locations mentioned above. In the lamina propria, all the positive cells with irregular processes were connected with each other and formed a network. In the submucosa, all the positive cells were found surrounding the blood vessels.     

猪A群轮状病毒SCJY-13株的分离鉴定及致病性分析

【目的】 在从腹泻仔猪粪便中分离鉴定猪A群轮状病毒(Rotavirus group A, RVA)并了解其致病性。【方法】 应用MA-104细胞从仔猪腹泻粪便中分离鉴定RVA, 将其经口感染健康初生仔猪, 观察其临床症状, 并利用实时荧光定量PCR检测排毒、HE染色观察病理变化、免疫组织化学(immunohistochemistry, IHC)试验了解病毒分布。【结果】 分离到1株可引起MA-104细胞明显细胞病变的G9P[23] RVA, 命名为RVA/Pig-tc/CHN/SCJY-13/2017/G9P[23](简称SCJY-13株), 病毒滴度为10^5.5 TCID₅₀/100 μL。经口感染SCJY-13株的仔猪在感染后11 h出现腹泻, 持续到感染后165 h完全恢复, 其发病率为100%(7/7), 病死率为28.57%(2/7)。感染后8 h可从仔猪肛拭子检测到病毒核酸, 直至感染后192 h, 峰值出现在感染后24 h。感染后54 h各段小肠病毒载量达到最高, 其中回肠中病毒载量最高, 显著高于十二指肠、空肠(P<0.05);HE染色和IHC检测结果显示, SCJY-13在感染仔猪小肠的绒毛及隐窝中大量聚集, 尤其是回肠段; SCJY-13株感染引起十二指肠、空肠和回肠绒毛固有层大量淋巴细胞浸润、黏膜上皮细胞和肠绒毛尖端空泡变性、柱状细胞增多、绒毛断裂脱落等, 以回肠段较严重。【结论】 SCJY-13株能引起新生仔猪100%发病, 其主要靶位区在回肠, 感染后排毒时间长。本试验结果为猪RVA的致病性研究提供了一定的参考依据。     

An immunohistochemical study of S-100 protein in the intestinal tract of Chinese soft-shelled turtle,Pelodiscus sinensis

The present work describes the distribution of S-100 protein in the intestinal tract of a Chinese soft-shelled turtle specimen (Pelodiscus sinensis). S-100 protein positive cells were located in the intestinal tract, from the proximal small to distal large intestine. S-100 protein positive dendritic cells had irregular shape and were positive in both cytoplasm and nucleus. Most of them were located both lamina propria and submucosa in the small intestine, while few were found in the large intestine. S-100 protein, C-kit positive ICCs and Silver staining glial cells were predominantly observed in three locations: (1) in the interspace between the submucosa and circular muscle layer; (2) in the circular muscle layer; and (3) between the circular and longitudinal muscle layers of the intestine. Fewer were found in the large intestinal lamina propria and submucosa. Three types of positive cells (S-100 protein positive cells, C-kit positive ICCs and Silver staining glial cells) with 1–2 long or 2–3 short processes were distributed as lace-like or surrounding blood vessels in the different locations mentioned above. In the lamina propria, all the positive cells with irregular processes were connected with each other and formed a network. In the submucosa, all the positive cells were found surrounding the blood vessels.     

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.     

Spray-dried porcine plasma affects intestinal morphology and immune cell subsets of weaned pigs

The aim of this study was to evaluate the effects of dietary spray-dried porcine plasma (SDPP) on the productive performance, intestinal morphology and leukocyte cell subsets of piglets. Sixteen early-weaned piglets (20 ± 2 d) were distributed into two dietary treatments: 1) free access to control diet or 2) 6% SDPP in the control diet instead of soy-protein concentrate. Intestinal morphometry of the small and large intestine, haematology and immune cell flow cytometric analysis of blood, ileo-colic lymph node and ileal Peyer's patches were performed in all pigs. Although SDPP treatment did not increase growth performance, it improved feed efficiency. We observed that SDPP diminishes blood monocytes, and macrophages (SWC3+), B lymphocytes (CD21+) and γδ T cells (γδTCR+) in gut lymphoid tissues. SDPP-treated piglets also showed lower intraepithelial lymphocyte numbers and lamina propria cell density in the small and large intestine. All these results suggest lower activation of the immune system of the SDPP-piglets during the post-weaning period. Moreover, the use of SDPP can be considered a valid alternative to antibiotic growth promoters.     

益气健脾中药对腹泻仔猪小肠黏膜组织结构的影响

为了研究益气健脾中药对腹泻仔猪小肠黏膜结构的影响,试验用抗生素及中药复方颗粒剂治疗腹泻仔猪,颈静脉放血处死后,腹腔解剖,无菌采集小肠黏膜,通过制作石蜡切片、HE染色观察小肠黏膜病理组织学变化;电镜包埋切片观察小肠黏膜细胞结构的变化。结果显示,中药复方颗粒剂治疗组小肠黏膜除有少量出血外,未发现明显病理变化。抗生素治疗组则小肠黏膜脱落,有广泛充血、出血;黏膜上皮细胞变性坏死;嗜酸性粒细胞浸润,有大量以淋巴细胞(LELs)、杯状细胞(GSs)为主的炎性细胞浸润;微绒毛排列紊乱、脱落;细胞受损严重,线粒体等细胞器结构异常。结果表明,抗生素治疗仔猪腹泻造成小肠黏膜明显的病理变化和功能障碍,而中药复方颗粒剂治疗有利于仔猪小肠黏膜结构的修复,为防制仔猪腹泻、提高仔猪的生长性能提供理论依据。     

中药复方颗粒剂对断奶腹泻仔猪小肠黏膜超微结构的影响

为研究中药颗粒剂对断奶腹泻仔猪小肠黏膜超微结构的影响,选择（28±1）d、体重10.04 kg左右的杜长大三元杂种断奶腹泻仔猪18头,随机分为抗生素治疗组和中药复方颗粒剂治疗组,每组3个重复,每个重复3头猪。两组分别于用药后14 d每组随机抽取3头猪颈动脉放血处死,腹腔解剖观察病理变化,无菌采集小肠黏膜进行细胞学观察。结果显示,中药复方颗粒剂治疗组小肠黏膜除有少量出血外,未发现明显病理变化。抗生素治疗组小肠黏膜脱落,有广泛充血、出血;黏膜上皮细胞变性坏死;嗜酸性粒细胞浸润,有大量以淋巴细胞（LELs）、杯状细胞（GSs）为主的炎性细胞浸润;微绒毛排列紊乱,脱落;细胞受损严重,线粒体等细胞器结构异常。表明抗生素治疗仔猪腹泻造成小肠黏膜显著的病理变化和功能障碍,而中药复方颗粒剂治疗有利于仔猪小肠黏膜结构的修复。     

Paucibacillary paratuberculosis in a goat

A 5-year-old Saanen doe was presented with emaciation, good appetite, normal feces, and a dry flaky hair coat. Johne's disease was tentatively diagnosed. Gross postmortem findings were a thickened, corrugated ileum and enlarged edematous mesenteric lymph nodes. Histologically, there was extensive lymphocytic infiltration of the ileal lamina propria. Mycobacterium paratuberculosis could not be identified or cultured.     

Organic acid profiles in feces of pigs with pathogenic or non-pathogenic diarrhea.

A chemical characteristic of the feces of diarrheal piglets permits differentiation among piglets receiving antibiotic treatment and those with colibacillosis or dyspepsia. A high concentration of lactic or succinic acid was observed in the diarrheic feces of piglets receiving antibiotic treatments and those with dyspepsia; however, no lactic or succinic acids were detected in piglets with colibacillosis. There was, however, little difference in the total concentration of organic acids among the three types of diarrheal illnesses. A quantitative analysis of lactic and succinic acids in diarrheic feces might provide a means for rapidly differentiating between colibacillosis and non-pathogenic diarrheas in piglets.     

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.     

The effect of Lactobacillus acidophilus and Bifidobacterium spp. administration on the morphology of the gastrointestinal tract, liver and pancreas in piglets

The aim of the study was to investigate the influence of administration of probiotic bacteria on morphology of the gastrointestinal tract, liver and pancreas. The experiment was performed on 15 piglets at the age from 3 to 35 days, intragastrically administered with Bifidobacterium breve, B. animalis, and Lactobacillus acidophilus bacteria. In all piglets examined, the development of the gastrointestinal tract, liver and pancreas was observed to progress normally. After microflora administration to the piglets, an increase in the number of fibrocytes and fibroblasts was observed in the mucosa lamina propria of stomach and intestines. An increase was also reported in the number and activation of endocrine cells in the stomach and small intestines. The activity of alkaline and acid phosphatases, as well as succinic (SDH) and lactic (LDH) dehydrogenases, was found to be higher after the administration of probiotics. The administration of bacteria, especially of Lactobacillus acidophilus, caused an increase in the number of lymphocytes and lymphoid cells in lamina propria and intraepithelial lymphocytes in the small intestine. Enhanced proliferation of crypt cells was observed in the crypts of intestinal glands; however, there were no statistically significant differences in the PCNA index between the control and probiotic-administered groups. The performed study showed that the administration of probiotic bacteria has no negative impact on the morphology of the gastrointestinal tract, liver and pancreas and is found beneficial to its functioning and immune processes.     

Canine coccidiosis attributed to an Isospora ohioensis-like organism: a case report

Coccidiosis due to an Isospora ohioensis-like organism was diagnosed in a 10-week-old pup. The pup had diarrhea, dehydration, and weight loss before it died 7 days after onset of clinical signs. Lesions were limited to the intestinal tract. Beginning in the distal end of the small intestine and extending through cecum and colon were mild histiocytic proliferation in the lamina propria and multifocal cryptitis. Numerous meronts and gamonts of an unidentified coccidium were in or near the lesions. Parasites were in the villous epithelium, lamina propria, and intestinal glands of the distal one-half of the small intestine, cecum, and colon. It was concluded that infection was due to an unidentified coccidium with oocysts structurally similar to those of I ohioensis.     

An epidemiological survey on pigs showing symptoms of infectious enteric diseases and dyspepsia in Japan

Diarrhea in pigs has the potential to have a serious economic impact on the swine industry. Previously, we suggested that the likely cause of the presence of non-infectious diarrhea in pigs characterized by lactate accumulation was dyspepsia. In this experiment, the prevalence of enteropathogens and hyper-lactate accumulation in feces of piglets in 4 distinct growth stages was examined. The feces were collected when veterinarian experts recognized abnormalities in sporadic outbreaks. Prevalence of enteropathogens in diarrheal feces was 100% in fattening pigs (FP), 75% in weaning pigs (WP), 50% in suckling pigs (SP), and 42% in growing pigs (GP). Prevalence of enteropathogens in loose feces was 53% in WP, 50% in SP, 40% in FP, and 28% in GP. Prevalence of hyper-lactate accumulation in diarrheal feces was 33% in GP, 33% in SP, 25% in WP, and 25% in FP. Prevalence of hyper-lactate accumulation in loose feces was 40% in GP, 0% in SP, 7% in WP, and 5% in FP. Accordingly, non-infectious dyspepsia is frequent in growing pigs. In this period, pigs are potentially exposed to needless antimicrobial therapeutic treatments in sporadic cases.     

Cryptosporidium sp.-associated enteritis without gastritis in rough green snakes (Opheodrys aestivus) and a common garter snake (Thamnophis sirtalis).

An epizootic of severe Cryptosporidium sp.-associated enteritis occurred in a group of 15 wild-caught juvenile rough green snakes (Opheodrys aestivus) at the Baltimore Zoo quarantine facility. All of the animals died with no premonitory signs. Histopathologic examination of the small and proximal large intestine of eight of the green snakes showed moderate to severe Cryptosporidium sp. infection and enteritis characterized by dense heterophilic and lymphocytic inflammatory infiltrates throughout the lamina propria with epithelial necrosis. Cryptosporidium sp. was also found in feces of an adult common garter snake (Thamnophis sirtalis) that was wild caught on zoo grounds and held in quarantine during the epizootic. After euthanasia, histologic examination of the garter snake showed a severe small intestinal Cryptosporidium sp. infection with only mild enteritis consisting of sparse heterophilic and lymphocytic infiltrates. There was no gross or histologic evidence of Cryptosporidium sp. gastritis in the nine snakes evaluated, and this is the first report of Cryptosporidium sp.-associated enteritis in snakes without gastric lesions.     

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.     

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.     

寡果糖对人源菌群仔猪肠道中IgA和IgG分泌细胞的影响

本文主要研究寡果糖对人源菌群仔猪肠道中IgA和IgG分泌细胞的影响。通过无菌剖腹产获取15头无特定病原菌(specific pathogen free,SPF)仔猪,随机分为三组。第一组为SPF组,经口灌服磷酸钠缓冲液(内含10%甘油)以示对照;第二组为人源菌群(human flora-associated,HFA)组,经口服途径接种人源菌群;第三组为寡果糖(fructo-oligosaccharides,FOS)组,口服途径接种人源菌群且灌喂寡果糖。仔猪饲养于屏障系统内,无菌条件下人工哺育45天。应用免疫组织化学方法进行研究。结果表明:(1)所有仔猪小肠和结肠的固有层中均分布有IgA和IgG分泌细胞。(2)IgA和IgG分泌细胞在十二指肠中分布最多,随着肠段的向后推移IgA和IgG分泌细胞数量有逐渐下降趋势。(3)HFA组和FOS组IgA分泌细胞数量在回肠显著高于SPF组(P<0.01);十二指肠中HFA组IgG分泌细胞数量显著高于SPF组(P<0.01)。(4)FOS组IgA分泌细胞数量在空肠显著高于HFA组外(P<0.05),其他肠段总体上低于HFA组,但差异不显著。本结果提示给新生仔猪接种人源菌群能促进仔猪肠道中IgA和IgG分泌细胞的发育,而寡果糖使肠道IgA和IgG分泌细胞数量呈现下降的趋势。     

Colitis associated with Clostridium difficile in specific-pathogen-free C3H-scid mice

Soft feces and a decreased delivery rate were observed in a specific-pathogen-free (SPF) C3H-scid mouse breeding colony. Grossly, the ceca were shrunken and edematous in the affected mice. Histopathologically, severe edema in the cecal submucosa as well as infiltration of inflammatory cells in the lamina propria and submucosa of the ceca and colon were observed. No pathogenic microorganisms were detected by the routine microbiological tests. By anaerobic bacterial-examination, Clostridium (C.) difficile with toxin A was isolated from the cecal contents of the affected mice. The mice were diagnosed with C. difficile-associated colitis. This case appears to be the first report of natural infection with C. difficile in SPF mice with clinical signs.