Synthesis of nonspecific esterase in small intestine enterocytes during experimental coccidiosis in suckling pigs]

The activity of nonspecific esterase (EC. 3.1.1.1.) was evaluated in the small intestine mucosa of 21 conventional piglets infected on day 5 after parturition (DAP) with oocysts of the Eimeria debliecki coccidium (infection dose of 200,000 oocysts) for this evaluation a microdensitometric analysis at the level of enterocytes was used. The same examination was also performed in the small intestine mucosa of four control conventional piglets at the age of 2-5 days (Tab. I). The synthesis of nonspecific esterase in the experimentally infected piglets was followed on day 1 to day 10 after infection (DAI). The activity of nonspecific esterase in the small intestine mucosa was found to decrease in a direction from duodenum absorption cells (D mean 34.15) to caudal ones (Fig. 1); ileum enterocytes have the optical density of the enzyme by 8.2% lower (D mean 31.38). The deposition of nonspecific esterase is localized mainly in the supranuclear zone of enterocytes while in the para- and infranuclear zones of absorption cells its concentration is only minute. In the experimentally infected piglets a marked increase in the optical density of nonspecific esterase of enterocytes was observed as soon as on day 1 after infection when the enzyme concentration increased by 19.4% (Tab. II). The maximum increase in the activity of nonspecific esterase of absorption cells was recorded on DAI 9 when the enzyme D mean value was higher by 165% in comparison with the activity of nonspecific esterase demonstrated in the control piglets (Fig. 2, 3, 4). But at the end of experimental infection (DAI 10) the total density of nonspecific esterase of enterocytes decreased by 38.2%.(ABSTRACT TRUNCATED AT 250 WORDS)     

Histochemistry of acid phosphatase in small intestine mucosa in experimental coccidiosis in suckling piglets]

The activity of acid phosphatase (phosphohydrolase of orthophosphate monoesters; EC. 3.1.3.2) was evaluated densitometrically in the mucosa of duodenum, jejunum and ileum of 22 conventional piglets which were experimentally infected by oocysts of the coccidiae Isospora suis (infection dose of 200,000 oocysts) on day one after parturition (DAP). The activity of the studied hydrolase was investigated in the infected piglets during days two to ten after infection (DAI) in the intestinal mucosa (enterocytes) and in goblet cells. The density of the reaction product of acid phosphatase was simultaneously determined in the same mucosal cells of different sections of the small intestine in five control conventional piglets at the age of 2-14 days. In the small intestine mucosa of control piglets the activity of acid phosphatase was demonstrated to be located especially in the supranuclear zone of enterocytes. As for goblet cells, the reaction product of acid phosphatase is distributed in all zones (supra-, para-, infranuclear zones); the lowest density of this enzyme was found in the infranuclear zone. The activity of acid phosphatase is also localized in intestinal crypts: in their cells the enzyme concentration is decreasing from duodenum to caudal sections. Important changes were revealed, in comparison with the control data, in the development of the activity of acid phosphatase in the intestinal mucosa cells in the experimentally infected piglets. In the period of investigation (DAI 2-10) there were two stages of the development of the density of the enzyme reaction product. The first stage can be characterized by an increase, the other by a decrease in the level of acid phosphatase activity. Enterocytes are influenced in both stages, but the decrease in the density of the reaction product of acid phosphatase was observed only in absorption cells, and not in goblet cells. The increase in the activity of acid phosphatase occurs in the periods of DAI 4 and 9-10. Enzymatic deviations occur mainly in the absorption cells of the mucosa of duodenum and middle jejunum; in the cells of posterior jejunum and ileum an increase in the density of the reaction product of acid phosphatase was also demonstrated, but at the lower quantitative level (especially on DAI 4). The decrease in the activity of acid phosphatase has a protracted development and it takes place on DAI 5 to 8.(ABSTRACT TRUNCATED AT 400 WORDS)     

Synthesis of glucose-6-phosphatase in small intestine mucosa during experimental coccidiosis in suckling piglets]

Glucoso-6-phosphatase activity (EC. 3.1.3.9.) was evaluated microdensitometrically in the small intestine mucosa of 19 conventional piglets infected with Eimeria debliecki coccidium oocysts (infection dose of 200,000 oocysts) on day 1 after delivery (DAD). The synthesis of the observed hydrolase was followed within days 1 to 10 after infection (DAI). The activity of the same enzyme was also determined in four control conventional piglets at the age of two to five days. The small intestine mucosa of healthy piglets was found to have relatively balanced glucoso-6-phosphatase concentrations in all observed sections (duodenum, middle and posterior jejunum and ileum). The topographic gradient failed to be demonstrated in control piglets. In piglets experimentally infected with E. debliecki coccidium the glucoso-6-phosphatase activity was decreasing during infection already since DAI 1. The enzyme hypoactivity was still lower in the following days and reached the minimum value at the end of the target period (DAI 10), when its value made only 33% of the concentration recorded in the intestine mucosa of control piglets. The topographic predisposition to the lower activity of glucoso-6-phosphatase was not found to be high. The only exception are DAI 6 and 7, when the lowest enzyme concentration was observed in the duodenum mucosa (52% on DAI 6; 54% on DAI 7). The activity of the observed enzyme was higher in the other sections of small intestine (57% on DAI 6; 62% on DAI 7).     

Dehydrogenase activity in small intestine mucosa in experimental coccidiosis in suckling pigs]

The activities of 3-beta-hydroxybutyrate dehydrogenase (EC. 1.1.1.30.; HBD) and isocitrate dehydrogenase (EC. 1.1.1.41; ICD) were evaluated microdensitometrically in the mucosa of duodenum, jejunum and ileum of 19 conventional piglets infected on the first day after parturition (DAP) with oocysts of Eimeria debliecki coccidium (infection dose of 200,000 oocysts). The two investigated enzymes are deposited in mitochondria which are dispersed in the supra-, para- and infranuclear region of absorption cells (Fig. 1). The synthesis of the two dehydrogenases was investigated in the small intestine mucosa in the period of 1st to 10th day after infection (DAI). The HBD and ICD activities were also followed in the small intestine of four control conventional piglets at the age of 2-5 days (Tab. I). The two dehydrogenases could be characterized by a topographic gradient; it means that their activity was increasing in the small intestine mucosa through duodenum in an aboral direction. The ICD activity is higher in the intestinal mucosa of healthy piglets (Figs. 2 and 3), where its topic concentration was more marked while the HBD activity is dispersed in enterocytes (Fig. 4). In infected piglets the density of the two enzymes was demonstrated to decrease already in the starting period of experimental infection, and it reaches the lowest values for the first time on DAI 5-6 (Fig. 5, Tab. II), then on DAIs 9 (HBD; Fig. 6, graph 11)) or 8 (ICD, Fig. 7 and 10). In the period of experimental infection no statistically significant predisposition to the hypoactivity of target dehydrogenases nor its marked shift were observed. Somewhat rapid resumption of synthesis was demonstrated as soon as on DAI 8 in ICD (Fig. 8); its activity on DAI 10 in the intestinal mucosa corresponded to the 93% activity of this dehydrogenase recorded in the small intestine of control piglets. The density of HBD to the same day (DAI 10) reached in the intestinal mucosa of infected piglets the values making only 44.7% of those demonstrated in the intestinal mucosa of the control group of animals.     

Histochemistry of nucleoside phosphatases and phosphoglucomutase in the small intestine during coccidiosis in piglets]

The first day after birth, 22 conventional piglets were experimentally infected with the oocysts of the coccidia of I. suis (infection dose 200,000 oocysts). The activity of 5-nucleotidase (5-ribonucleotide phosphohydrolase, EC.3.1.3.5) and phosphoglucomutase (alpha-D-glucoso-1-phosphate phosphotransferase, EC.5.4.2.2) was densitometrically assessed in the mucosa of the small intestines of these piglets. Enzyme activities were studied in the infected piglets during the 2nd to 10th day after infection. The same histochemical examination was simultaneously performed in the intestinal mucosa of five control conventional piglets at an age of 2-14 days. 5-nucleotidase and phosphoglucomutase were found to have a high density in the mucosa of the small intestine of the control piglets: the high-density locations of these enzymes include, first of all, the supranuclear area of the absorption cells, the microvillous zone of enterocytes and the smooth muscle elements of lamina muscularis mucosae. The experimentally infected piglets showed a marked decline of the density of both enzymes during the infection. The deficit affected, for a transient period, the microvillous zone and the supranuclear region of enterocytes; the musculature of the mucous layer was affected permanently. The inactivity was more protracted in the case phosphoglutamase (especially 5 to 9 days after infection). The density of 5-nucleotidase showed a partial return to the normal already the 7th day after infection, with an interruption of resumption of activity on the 10th day. Resumption of enzyme activity in the lamina muscularis mucosae was not recorded during the infection. In the three locations under study, the density of none of the enzymes did reach parameters comparable with the controls at the end of the trial (10 days after infection).     

Adenovirus inclusions in the ileum in coccidiosis in suckling piglets

Numerous intranuclear inclusion bodies in enterocytes were detected exclusively in the ileum of two nine-day piglets coming from a litter infected with diarrhea. The inclusion bodies were homogeneous in hematoxylin and eosine (HE), their staining was not clear enough, was amphophilic and they filled nearly the whole nucleus. They were eosinophil less often and had a halo on the periphery. Their staining was clearly orange-red in Feulgen's nuclear reaction after re-staining with G orange and bright green. Intranuclear inclusions were located exclusively on shortened villi and pseudovilli of ileum above the follicles of activated Peyer's patches. The findings of intranuclear inclusions in ileum demonstrate adenovirus enteral infections in suckling piglets.     

Lysosomal activity in enterocytes in the small intestine in piglets experimentally infected with Isospora suis

The lysosomal activity of enterocytes of the small intestine mucosa was investigated in gnotobiotic and conventional piglets experimentally infected on the first day after birth (DAB) by the oocysts of the coccidia Isospora suis. A method of the proof of beta-D-glucuronidase (EC.3.2.1.31.) activity was used to demonstrate lysosomes. The piglets were infected by different infection doses of oocysts (100,000 oocysts in gnotobiotic piglets and 200,000 oocysts in conventional piglets). In the gnotobiotic infected piglets the activity of beta-D-glucuronidase in enterocyte lysosomes was investigated in the period from day 3 to day 11 after infection (DAI) and in the infected conventional piglets in the period from day 2 to day 10 after infection. Comparing the control piglets, the group of gnotobiotic piglets at the age of 2-5 days and the group of conventional piglets at the age of 4-7 days, the higher activity of beta-D-glucuronidase was demonstrated in the lysosomes of intestinal mucosa enterocytes in the gnotobiotic control piglets (+5.30 of the average density value, Dx). In the infected gnotobiotic and conventional piglets the pattern of beta-D-glucuronidase activity was found to have three stages in the course of this infection. Two stages can be characterized by a great increase in the enzyme activity (DAI 3-9 in gnotobiotic piglets, DAI 2-3 and 7-9 in conventional piglets. The third stage, which is manifest mainly in the conventional infected piglets, is characterized by a marked decrease in the activity of beta-D-glucuronidase, reaching the level of control findings (DAI 10 and mainly 11 in gnotobiotic piglets. DAI 4-6 and 10 in conventional piglets). A topographical picture shows that the two stages of increase and the stage of beta-D-glucuronidase activity decrease occur in the whole small intestine without any predisposition defect of the enzyme in the different sections of the small intestine.     

The ratio of enterocytes and goblet cells in the mucosa of the small intestine in experimental infection of piglets with the coccidium Isospora suis

The proportions of cup-shaped cells and enterocytes were studied in piglets infected the first and fifth day after birth with 200,000 oocysts of Isospora suis. The greatest imbalance was found in the ratio of the cells of the intestinal mucous membrane in the region of middle and lower jejunum (ratio of 10:1.75)--this was recorded the third to fourth day after infection when the ratio made 10:0.62, whereas in the control animals it was 10:5.7. On the eighth to ninth day after infection the number of cup-shaped cells began to increase, the ratio reaching the values of 10:2.62. Even the 13th day after infection the ratio remained far from normal (10:2.50).     

Fimbriae in Escherichia coli isolated from the small intestine of piglets

Ninety E. coli strains, isolated from piglets which had died from neonatal diarrhea, were tested for the presence of K88, K99, 987P and type 1 fimbriae. Two or more types of fimbriae were demonstrated in 14 of the strains, a single fimbria! type in 44 strains while in 32 strains no fimbriae were detected. Of the 14 E. coli strains with more than 1 type of fimbriae, 12;, 10, 8 and 4 strains showed K88, K99, 987P and type 1, respectively.Twelve E. coli strains were isolated from piglets which had died in the neonatal period without showing signs of neonatal diarrhea at necropsy. One strain showed 987P and 3 strains showed type 1 fimbriae, while the remaining 8 strains were unfimbriated.Sixteen fimbriated E. coli strains were subcultured in order to examine the stability of fimbrial expression in the strains. The K88 and the type 1 fimbriae were regularly expressed, while the K99 and 987P were inconsistently demonstrated.     

Mucus synthesis in the goblet cells of the small intestine in experimental infection with the coccidium Isospora suis in piglets

The state of mucus synthesis in the goblet cells of the small intestine was studied in conventional piglets infected with a dose of 200,000 oocytes of the coccidium Isospora suis the first and fifth day after parturition. The synthesis of mucus and its chemical characteristics undergo significant changes during the third and fourth day after infection. The activity of acid and neutral mucous substances declines; their level and the physiological synthetic function of goblet cells begin to return to the normal during the period starting on the eight to tenth day after infection. However, there were no fully functioning goblet cells in the broken numerical ratio even at the end of the period of investigation, i.e. the 13th day after infection. The thin surface layer of mucus remained almost unchanged within the whole extent of the small intestine parts studied.     

仔猪小肠肌间神经丛中神经元数量的定量测定

自1921年Langley提出“肠道神经系统”(enteric nervous system，ENS)这一概念以来，肠道神经系统对肠道运动的控制和功能调节的重要性已日益引起人们的注意。国内外学者对ENS的形态结构、神经元的性质等做了广泛而深入的研究。不过从现有资料看，有关猪的ENS资料较少。     

日粮添加谷氨酰胺对断奶仔猪小肠形态结构的影响

试验选用60头21d断奶、体重均一的杜长大仔猪。按单因素完全随机化设计随机分为2组,一组为对照组,饲喂基础日粮;一组为试验组,基础日粮添加0.5%的谷氨酰胺,研究谷氨酰胺处理对断奶仔猪小肠黏膜形态变化的影响。在断奶当天及断奶后7d和14d每个处理随机选取3头仔猪屠宰,取小肠组织,测定肠黏膜的形态结构变化。结果表明:日粮中添加谷氨酰胺提高了小肠绒毛高度和固有层厚度及皱褶深度。     

Developmental changes in intercellular junctions and Kv channels in the intestine of piglets during the suckling and post-weaning periods

Background: The intestinal epithelium is an important barrier that depends on a complex mixture of proteins and these proteins comprise different intercellular junctions. The purpose of this study was to investigate the postnatal and developmental changes in morphology, intercellular junctions and voltage-gated potassium(Kv) channels in the intestine of piglets during the suckling and post-weaning periods.Results: Samples of the small intestine were obtained from 1-, 7-, 14-, and 21-d-old suckling piglets and piglets on d 1, 3, 5, and 7 after weaning at 14 d of age. The results showed that the percentage of proliferating cell nuclear antigen(PCNA)-positive cells and alkaline phosphatase(AKP) activity, as well as the abundances of E-cadherin,occludin, and Kv1.5 m RNA and claudin-1, claudin-3, and occludin protein in the jejunum were increased from d 1to d 21 during the suckling period(P 0.05). Weaning induced decreases in the percentage of PCNA-positive cells,AKP activity and the abundances of E-cadherin, occludin and zonula occludens(ZO)-1 m RNA or protein in the jejunum on d 1, 3 and 5 post-weaning(P 0.05). There were lower abundances of E-cadherin, occludin and ZO-1m RNA as well as claudin-1, claudin-3 and ZO-1 protein in the jejunum of weanling piglets than in 21-d-old suckling piglets(P 0.05). The abundances of E-cadherin, occludin, ZO-1 and integrin m RNA were positively related to the percentage of PCNA-positive cells.Conclusion: Weaning at 14 d of age induced damage to the intestinal morphology and barrier. While there was an adaptive restoration on d 7 post-weaning, the measured values did not return to the pre-weaning levels, which reflected the impairment of intercellular junctions and Kv channels.     

Differences in the development of the small intestine between gnotobiotic and conventionally bred piglets

The effects of age, weaning and breeding conditions on the small intestinal morphology and the distribution of immunocompetent cells were investigated. The villus height and numbers of CD3+ T-lymphocytes, measured in the duodenum, jejunum and ileum, were determined in both the gnotobiotic and conventionally bred piglets. The diet of gnotobiotic piglets was composed of milk-replacement and feed mixtures. The application of milk replacement was finished on day 28. Conventional piglets were fed on sow's milk and feed mixtures. The animals were weaned on day 28. Small intestines were collected from 12 conventional and 12 gnotobiotic piglets slaughtered at the age of 2, 7, 14, 21, 28 and 35 days. The morphology results demonstrated that duodenal and ileal villi were significantly higher (p < 0.001) in gnotobiotic piglets during the entire period of the experiment. However, the weight of conventional piglets was higher during the experiment, in some cases significantly (p < 0.05). A marked reduction, in some cases significant (p < 0.001, p < 0.01 respectively), of the villi height on day 7 after the diet change for both groups of animals was recorded. The results demonstrate the differences in the gut development between both groups and their relationship to the breeding conditions. Nevertheless, the effect of crucial diet changes was observed independently of them. The immunohistochemistry results showed significantly (p < 0.001, p < 0.01 respectively) higher numbers of CD3+ T-lymphocytes in the jejunal villi of conventionally bred piglets. Similar results, in some cases significant (p < 0.001, p < 0.01 respectively), were also obtained from the other parts of the small intestine. These observations confirm reduced microorganism exposure under the gnotobiotic conditions.     

Postnatal development of lymphocyte subsets in different compartments of the small intestine of piglets

In contrast to rodents, all compartments of the porcine small intestine contain lymphoid cells at birth. During the first few days of life maternal antibodies and leukocytes are taken up by the intestinal epithelium. The number of intraepithelial lymphocytes (IEL) increases by a factor of 12 from Day 1 to Day 60, but in germfree pigs only a minor increase is observed. Immediately after birth, low numbers of T cells are present in the small intestinal lamina propria (LP), and many of these express neither CD4 nor CD8 on their surface. This type of subset composition is still present in germfree pigs at an age of 1.5 months. The appearance of IgA+ and IgM+ lymphocytes in the LP differs. Many more of these B cells are found in the LP of the crypts than of the villi. The development of the discrete jejunal Peyer's patches (PP) differs from that of the continuous PP in the terminal ileum. The lymphocyte subset composition shows the most obvious differences between conventional and germfree piglets and between the jejunal and ileal PP at an age of 1.5 months. Several markers, e.g. for antigen-presenting cells, memory T cells, M cells in the domes of PP, have recently become available for the pig. These should now be applied in experiments on pigs in the early postnatal period to study the mechanisms of the development of tolerance and protective immune reactions.     

Use of the anticoccidial agent, tortrazuril (Baycox, Bayer) in coccidiosis in suckling pigs]

At present neonatal coccidiosis caused by Isospora suis is known to be an important disease in piglets. The coccidium I. suis was diagnosed as the causative agent of diarrhoeic disease in piglets on a large pig farm with continuous farrowing operation. The diagnosis of coccidiosis was based on clinical history, gross lesions, histopathology, stained impression smears and aided by detection of oocysts in the faeces. Coccidiosis was associated with diarrhoea in piglets at the age of five to fourteen days. The disease was characterized by variable morbidity and only a portion of the litters was usually affected at one time. The piglets appeared listless, and suffered from yellow watery scours progressing to yellow pasty scours over a three- to five-day period. The lack of response to common antimicrobial therapy, and/or vaccination of sows for E. coli was observed. The prophylactic and therapeutic efficacy of tortrazuril (Baycox, Bayer) against coccidiosis was evaluated in naturally infected piglets. Tortrazuril at a dose of 20 mg/kg, given perorally to each piglet on days 6 and 8 of age, proved to be efficacious in preventing clinical coccidiosis in piglets. Tortrazuril fully controlled the oocyst output, prevented the development of diarrhoea but not improved the weight gains in three-week-old piglets if compared to the untreated controls. The prophylactic and therapeutic efficacy of tortrazuril was compared with another drug--amprolium (Amprovin, MSD). Amprolium at a dose of 100 mg/kg, given perorally to piglets from day 6 to 8 of age, was not efficacious in preventing clinical porcine neonatal coccidiosis. Amprolium reduced the oocyst output, but not prevented the development of diarrhoea. Tortrazuril (Baycox, Bayer) is clearly effective against porcine neonatal coccidiosis caused by I. suis.     

Submicroscopic structure of small intestine cells of lambs during a course of experimental inflammation]

The changes observed in the submicroscopic structure of the enterocytes of the jejunum of lambs in which inflammation has been chemically evoked justify the assumption that 12 hours after application of the chemical agent some differences exist, particularly in cellular reaction in the submucosa, in comparison with inflammations evoked by the infective agent. The submicroscopic changes in the enterocytes correspond to acute inflammation and indicate alterations on cell components which considerably reduce the absorptive area of the intestinal epithelium, and on organellae essential for the function of the enterocyte. In this way the natural barriers in the intestinal wall are damaged, leading to derangements in the absorption and passage of the nutrients taken in.     

牛至油对吮乳仔猪生长性能影响

仔猪腹泻是集约化养猪生产条件下的一种典型的多因素性疾病,该病是目前最严重的仔猪疾病之一,也是引起仔猪死亡的重要原因.据调查,仔猪因腹泻死亡占仔猪死亡总数的39.8%.而腹泻又主要集中在仔猪的三个年龄段,即出生后1～3日龄、7～14日龄和刚断奶后的仔猪.据报道,即使在养猪业较发达的国家,如美国仔猪断奶前死亡率也占15.5%,荷兰占11.5%～14.2%.故在吮乳仔猪早期补料的日粮中,通过添加某些天然植物抗生素抑制吮乳仔猪的肠道有害菌群,提高其自身免疫能力,已成为养猪业生产的一项重要课题.     

Effect of dietary phytic acid on performance and nutrient uptake in the small intestine of piglets

An experiment was conducted with piglets to determine the effect of dietary phytic acid supplementation on performance, electrophysiological properties of jejunum mounted in Ussing chambers, sodium-dependent glucose transporter 1 (SGLT1) protein expression in jejunum, and plasma glucose and Na concentrations. Sixteen piglets with an average initial BW of 7.40 ± 0.36 kg were randomly assigned to 2 experimental diets with 8 piglets per diet. The diets were casein-cornstarch-based and were either unsupplemented or supplemented with 2% phytic acid (as Na phytate). The basal diet was formulated to meet the recommendation of NRC (1998) for energy, AA, minerals, and vitamins for piglets. The experiment lasted for 21 d, and at the end, BW gain and feed consumption were determined, and blood samples were collected for determination of plasma glucose and Na concentrations. The piglets were then euthanized to determine jejunal electrophysiological properties (transmural potential difference and short-circuit current) and SGLT1 protein expression. Phytic acid supplementation reduced ADG (P = 0.002), ADFI (P = 0.017), and G:F (P = 0.001) from 316.1 to 198.2 g, 437.4 to 360.3 g, and 0.721 to 0.539 g/g, respectively. Phytic acid supplementation also tended to reduce (P = 0.088) potential difference (-3.80 vs. -2.23 mV) and reduced (P = 0.023) short-circuit current from 8.07 to 0.1 μA/cm(2). However, phytic acid supplementation did not affect SGLT1 protein, and blood plasma glucose and Na concentrations. In conclusion, dietary phytic acid reduced growth performance and transmural short-circuit current in the jejunum of piglets. The reduced transmural short-circuit current in the jejunum by phytic acid implies reduced active Na transport in the jejunum by the phytic acid. Therefore, it seems that dietary phytic acid reduces growth performance of pigs partly through reduced capacity of the small intestine to absorb Na.