beta-D-glucosidase in the microvillous zone of small intestine enterocytes in experimental coccidiosis in suckling piglets]

In the small intestine mucosa of 24 gnotobiotic farrows experimentally infected with the oocysts of coccidiosis of Isospora suis (infection administration--100,000 oocysts) on the first day after the delivery, we carried out the microdensitometric evaluation of the activity of beta-D-glucosidase (phlorizin-hydrolase; hetero-beta-galactosidase; lactase-beta-glucosidase complex; EC. 3.2.1.21). Great attention was paid to the topochemistry of enzyme, deposited in a microvillous zone of enterocytes. We studied likewise the activity of beta-D-glucosidase in the striped fringe of enterocytes of the four control gnotobiotic farrows, in the age from 2 to 5 days. We found out that in healthy farrows the reaction product of studied disaccharidase is located in high concentrations in the microvillous zone of absorptive cells of the whole small intestine. We proved a topographic gradient at which the beta-D-glucosidase activity decreases in control farrows the duodenum mucosa in the aboral direction. When using the choice substrate for beta-D-glucosidase (5-Br-4-Cl-beta-indolyl-3-D-glucoside) we did not prove the enzyme deposition in the small intestine wall. The negative enteral effect of coccidiosis I. suis was provable in the farrows experimentally infected already on the first day after the infection (DPI) when the beta-D-glucosidase activity decreased within the whole small intestine by 15% (ileum) and even by 23% (middle jejunum). The activity reduction had been deepening since the first after the infection and it reached its maximum on the 9th day after the infection when the enzyme concentration in the microvillous zone of absorptive cells reached only 11% of the activity level found in control farrows. On the 10th and 11th day after the infection we registered the increase of the density of beta-D-glucosidase reaction product, however the microvillous zone was even in that final stage of experimental infection significantly deficient (31% of intestine mucosa activity of control farrows).     

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)     

Densitometric analysis of aminopeptidase M activity in small intestine mucosa in piglets experimentally infected with Isospora suis

In gnotobiotical and conventional piglets infected a day post partum (DPP) with oocysts of the coccidium Isospora suis, densitometrical analysis of the activity of aminopeptidase M (EC.3.4.11.2; APM) was performed in the area of microvillous zone of the small intestine. Piglets were infected with different infection doses of oocysts (100,000 oocysts) in gnotobiotical piglets and 200,000 oocysts in conventional piglets). In infected gnotobiotical piglets, the APM activity was studied in the period from the 3rd to 11th day after infection (DAI) and in infected conventional piglets in the period of to the 2nd to 10th day after infection (DAI). Control piglets, in the group of the gnotobiotical animals at the age of 2 to 5 days in the group of the conventional animals at the age of 4 to 7 days, had different APM activity in the microvillous zone of the intestinal mucosa. It was stated that the microvillous zone of the intestinal mucosa gained higher values in control conventional piglets (+7.01 mean values of density). In infected gnotobiotical piglets the density fall of the reaction product of APM was demonstrated already on the third day with further marked reduction of APM density on the 4th day after infection in the whole small intestine with predominance of the persisting APM activity in ileum. Even despite the slight increase in the density of the reaction product of APM in the period from the 5th to 7th DAI (the highest increase in APM density on the 6th DAI), a further decrease of the activity was recorded again on the 8th and namely the 9th DAI in the whole small intestine (the lowest value of density was found in the rear jejunum), the ileum mucosa being affected, too. A slightly higher density of the reaction product of APM was found in the duodenum. On the 10th DAI the APM density started to change and on the 11th DAI in the duodenum and in the middle jejunum it even reached higher values in comparison with the control data. Some differences were proved in the infected conventional piglets in comparison with the development of the APM activity in the small intestine mucosa in the infected gnotobiotical piglets. On the 3rd and 4th DAI APM defect occurred in the whole small intestine, with APM density prevailing in the ileum mucosa (like in the group of infected gnotobiotical piglets). The second period of decrease in APM activity lasted for almost four days (6th to 9th DAI).(ABSTRACT TRUNCATED AT 400 WORDS)     

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.     

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 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).     

Activity of mucosubstances and the goblet cell count in the large intestine of piglets infected with the coccidium, Isospora suis

In a group of conventional and gnotobiotic piglets experimentally infected with the Isospora suis coccidia the quantitative presence of acid and neutral mucous substances in the large intestine and the counts of goblet cells in the surface mucosa and in Lieberkühnis crypts (in the following text called just the crypts) were investigated. In conventional piglets infected with the dose of 200,000 oocysts of I. suis coccidia the lowest content of acid mucous substances was recorded from the eighth to tenth day after infection (DAI). A decrease in the activity of neutral mucous substances was somewhat slower. The lowest count of goblet cells was found on DAI 9, especially on the surface mucosa (4.89 to 4.91 goblet cells per 10 enterocytes). There was observed no difference in the piglets infected the first and fifth day after parturition (DAP). Gnotobiotic piglets infected with the dose of 100,000 oocysts of I. suis coccidia on DAP 1 showed the lowest content of mucous substances in the large intestine from the ninth to tenth day after infection. Unlike the conventional piglets, in gnotobiotic piglets there was recorded a decrease in the content of acid and neutral mucous substances. The gnotobiotic piglets had the lowest counts of goblet cells in the surface mucosa (10:4.57) and in the crypts (10:7.71) on DAI 9. As to the quantitative proportions, in the conventional and gnotobiotic piglets neutral mucous substances prevailed on the other days (DAI 3-7 and DAI 11), similarly like on DAI 8. The results of this investigation revealed a functional disease of the large intestine in conventional and gnotobiotic piglets infected experimentally with the Isospora suis coccidia.     

Density of selected enzymes in the goblet cells of the small intestine in piglets experimentally infected with the coccidium Isospora suis

The density of selected enzymes in the goblet cells of the mucous membrane of the small intestine was studied in a group of 12 gnotobiotic piglets experimentally infected with the coccidium Isospora suis one day after parturition (DPP), using the Vickers M-786 scanning and integrating microdensity meter. At an infecting dose of 100,000 oocysts of I. suis, the histochemistry of the goblet cells of the mucous membrane of the piglets changed significantly in the period of 4 to 10 days after infection (DPI). Increases occur in the density of non-specific esterase (EC. 3.1.1.1.) and acid phosphatase (EC. 3.13.2.). The density of acid and neutral muco-substances declines and the densities of alkaline phosphatase (EC. 3.1.3.1.) and aminopeptidase M (EC. 3.4.11.2) are significantly high. The goblet cells of the mid and posterior parts of jejunum are very similar in their histochemistry in the experimentally infected gnotobiotic piglets. In the duodenum and ileum the histochemical picture of the goblet cells shows no substantial difference from the data recorded in the goblet cells of the mucous membrane of the small intestine of the four control piglets at an age of two to seven days.     

Morphometric analysis of the activity of alkaline phosphatase in the small intestine of piglets infected with the coccidium Isospora suis

Gnotobiotical one-day-old piglets were infected with 100,000 Isospora suis coccidia oocysts, and were immediately killed. In piglets killed on the 3rd to 11th day after infection (DAI), the morphometric analysis of alkaline phosphatase activity was performed in the area of the microvillus zone of small intestine. In the control 2-7 days old animals, the small intestine was not equally supplied with alkaline phosphatase. In duodenum the activity reached 88.37 per cent of the active length of absorbent surface (% LAac), in the middle jejunum 95.98 per cent LAac, in the dorsal jejunum 78.63% LAac and the ileum 90.55 % LAac. The width of the active area was more balanced and ranged from 5.003 microM in the ileum to 6.129 microM in the dorsal jejunum. In infected gnotobiotical piglets the lowest activity was found out on the 3rd to 4th DAI, with a greater decline on the 9th day after infection. The range from 25.99 to 40.50 per cent LAac with minimum in the duodenum and maximum in the ileum was observed on the 3rd DAI. In the middle and dorsal ileum the activity was nearly equal (28.34 and 27.69 per cent LAac). nI the dorsal jejunum a moderate increasing was up to 47.13% LAac on the 4th DAI, with the exception of the ileum, where the activity of alkaline phosphatase decreased to 24.96% LAac. On the 9th DAI the activity of alkaline phosphatase was nearly equal in the whole small intestine (from 55.70 to 60.01% LAac) with the maximum in the middle jejunum. In the width of the reaction product a direct dependence on the total activity of alkaline phosphatase was evident only in the segment of the middle and dorsal jejunum and ileum, but merely on the period of the 3rd to 4th DAI. The lowest values were measured in the middle jejunum (0.982 micron on the 3rd DAI and 0.709 micron on the 4th DAI). No dependence was observed between the total activity and the reaction product in the middle jejunum (0.982 micron on the 3rd DAI and 0.709 micron on the 4th DAI), there was no general stabilisation of the activity of alkaline phosphatase.     

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).     

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.     

Impact of Yersinia enterocolitica enteritis on disaccharidase activity and small intestinal morphology in colostrum-deprived newborn piglets

The effect of enteritis on the development of the small intestine was examined in newborn, colostrum-deprived piglets infected with a human isolate of Y. enterocolitica (serotype 0:3, biotype 4) soon after birth. The piglets were killed 3 days (n = 6) or 5 days (n = 8) after infection, or antibiotic therapy was commenced on day 5 and the animals killed on day 14 (n = 5). Compared with the non-infected controls, infected animals had reduced mucosal lactase and sucrase, but not maltase activity, while after antibiotic therapy, previously infected piglets had a lower lactase and a higher maltase and sucrase activity. Lactase activity was significantly reduced in the duodenum and jejunum, and mean values were lower in the ileum, but the difference did not reach significance; maltase activity was greater at all ages from the distal jejunum to the mid-ileum; sucrase activity was reduced in all segments up to day 5 but after antibiotic therapy was increased in the jejunum and appeared early in the ileum. Enzyme profiles were more mature along the crypt-villus axis in some segments of the intestine in previously infected piglets. Sodium-potassium-ATPase activity was unchanged. There was a reduced villus height:crypt depth ratio, crypt hyperplasia and increased crypt cell proliferation. Morphological maturation, indicated by loss of vacuoles and location of the nucleus at the base of the enterocyte, proceeded distally from the duodenum to ileum from 3 to 14 days of age when only the ileum remained immature. In infected piglets, there was reduced vacuolation and earlier location of the nucleus at the base of the cell in the distal intestine. Accelerated maturity of specific disaccharidases and enterocyte morphology in infected piglets appears to be due to physical damage to the mucosa resulting in faster proliferation of crypt cells and migration of enterocytes. It is suggested that this may reduce macromolecular internalisation and impair the ability to utilise dietary carbohydrate and may have long-term effects on growth and immunological responses of the gut.     

Alkaline phosphatase activity in goblet cells in the small intestine of piglets experimentally infected with the coccidium Isospora suis

Alkaline phosphatase activity (EC. 3.1.3.1.) in goblet cells was investigated in the small intestine of 16 gnotobiotic piglets infected one day after delivery (DAD) by different rates of oocysts of Isospora suis coccidia. At a high infection rate of I. suis (750,000) the goblet cells were found to be highly positive to alkaline phosphatase on day 3 to day 4 after infection (DAI). In piglets infected by a low infection rate of I. suis oocysts (100,000) the activity of alkaline phosphatase activity in goblet cells was proved on days 4 to 10 after infection. In the first group of piglets, the positive goblet cells prevailed in the middle region of jejunum, with the peak on 4th DAI. It the second group of piglets a marked increase in the alkaline phosphatase activity was recorded in the goblet cells in the posterior part of jejunum on days 4 to 5 after infection and on 10th DAI. No alkaline phosphatase activity in the goblet cells was demonstrated in the control gnotobiotic piglets at the age of two to seven days.     

Concurrent infection of enterocytes with Eimeria scabra and other enteropathogens in swine

Bacteria were detected in the enterocytes of the distal jejunum in weaned pigs on Days 7 and 9 post-infection (DPI) with Eimeria scabra in addition to the developmental stages of the coccidia. Short rod-shaped bacteria were identified in approximately 60% of the enterocytes that contained developmental stages of E. scabra. No such bacteria were observed in cells where coccidia were absent. Gamonts of cryptosporidia were also observed within the microvillous zone of the enterocytes in the distal jejunum of weaned pigs on DPI 9 with E. scabra. Cryptosporidia were present only in enterocytes harbouring stages of E. scabra. Chlamydial particles were also found in the cytoplasm of enterocytes 7 DPI with E. scabra. The presence of other enteropathogens exclusively in the enterocytes containing developmental stages of coccidia suggests that the coccidium E. scabra facilitates the invasion and development of bacteria, cryptosporidia and chlamydia in the enterocytes.     

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.     

Intestinal lesions caused by a strain of Chlamydia suis in weanling pigs infected at 21 days of age.

The objective of this study was to determine whether a strain of Chlamydia suis shown previously to be an intestinal pathogen in gnotobiotic piglets could cause diarrhea and intestinal lesions in young weanling pigs. Pigs from 2 sows were randomly assigned to 2 groups. Group 1 included 13 pigs that were weaned at 24 hours of age and then housed in isolator units and fed milk replacer and unmedicated starter ration. Group 2 included 8 pigs that nursed their respective sows, consumed unmedicated starter ration, and were weaned at 21 days of age. Ten pigs in group 1 and 6 pigs in group 2 were inoculated orally with 4 x 108 inclusion-forming units of C. suis strain R27 at 21 days of age. Control pigs were inoculated with sham inoculum. The pigs were necropsied 5-14 days postinoculation (DPI). None of the Chlamydia-infected pigs developed diarrhea. Villus atrophy was seen histologically in sections of ileum from Chlamydia-infected pigs in both groups 5 and 7 days DPI. Lymphangitis and multiple lymphohistiocytic and neutrophilic aggregates were seen in the submucosa, tunica muscularis, and serosa of the distal jejunum, ileum, and colon from Chlamydia-infected pigs in both groups 5-14 DPI. Immunostaining of sections of distal jejunum, ileum, and colon from infected pigs revealed chlamydial antigen in intestinal epithelium and in foci of lymphangitis/inflammation. The results indicated that C. suis strain R27 can cause intestinal lesions in young weanling pigs, and the lesions are similar to those seen in gnotobiotic piglets. The results also indicated that strain R27 causes asymptomatic intestinal infections in young weanling pigs, at least under the conditions of this study.     

Experimental caprine coccidiosis caused by Eimeria arloingi: Morphopathologic and electron microscopic studies

The progressive morphohistopathologic changes, distribution pattern of lesions and ultrastructural characteristics in Eimeria arloingi infection were investigated in experimentally infected kids. The 18 newborn animals allocated to 3 equal groups. Two of groups, A, B were inoculated with a single dose of 1 × 10³ and1 × 10⁵ sporulated oocysts of E. arloingi, respectively. At 7, 14, 21, 28, 35, and 42 days postinoculation (DPI), 1 kid from each group was necropsied for pathologic and ultrastructural studies. Progressive lesions were present at 21, 28, 35 and 42 DPI in the jejunum, ileum, cecum with fewer in the duodenum and proximal colon. The oocysts shedding begin between 16 to 18 DPI. Grossly, minimal changes were observed at 21 DPI as few whitish plaques or nodules and advanced lesions at 42 DPI as pseudoadenomatous pattern in the mucosa and a cerebriform pattern on the serosal surface of jejunum and ileum. Early histopathologic lesions due to schizogony phase were including presence of intracytoplasmic developmental stages of the parasite such as trophozoites, immature to mature schizonts and mild infiltration of inflammatory cells. In late lesions due to various stages of gametogony, the histological pattern was mainly remarkable hyperplasia of the villi and crypts epithelial cells, eventually developed into papillary projections of reactive epithelium. The mesenteric lymph nodes showed a few numbers of large schizonts in the cortical lacteals. This study showed E. arloingi as a highly pathogenic species for kids, the incubation period was 16–18 days and the main target organ was jejunum with characteristic morphohistopathologic lesions.     

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).     

Activity of selected dehydrogenases and monoamine oxidases in the small intestine of gnotobiotic piglets infected with the coccidium Isospora suis

In the small intestine of 16 gnotobiotic piglets infected a day post partum (DPP) by Isospora suis coccidia the activities were studied of selected dehydrogenases and monoaminoxidase (O2 oxidoreductase, MAOx, EC 1.4.3.4.). The following dehydrogenases were investigated: succinate dehydrogenase (SDH, EC 1.3.99.1.), glycerol-3-phosphate dehydrogenase (glycerol-3-phosphate:menadion oxidoreductase, GPOX, EC 1.1.99.5.) and tetrazolium oxidoreductase (NADH, ES 1.6.99.3.). The activities of NADH and GPOX were found to decrease, a decrease being somewhat milder in MAOx, at a high infection dose of I. suis oocysts (750,000 oocysts), in comparison with the control, already on the first day after infection (DAI). The SDH levels did not change. In piglets infected by a low infection dose of I. suis oocysts a double marked decrease (negative to slightly positive finding) was recorded in the period from the third to the eighth day after infection (DAI). A similar pattern with a longer time interval between the decreases was observed in GPOX (4th to 11th day after infection). The findings of SDH and MAOx activities were different. The SDH activity is maintained at the same level (++) for the whole period of investigation and there occurs a decrease (+) only on the 9th day after infection, persisting until 11 DAI. The MAOx activity and its change correspond to the SDH activity; the difference being that in the second group the starting level is high ( ) and on the eleventh day after infection it is low or medium (0-++), in comparison with the standard. This variability is discernible from 8th DAI.     

断奶仔猪小肠黏膜脂肪酸结合蛋白和二肽转运载体1mRNA表达发育性变化及谷氨酰胺对其的影响

本试验旨在研究断奶仔猪小肠黏膜脂肪酸结合蛋白( Ⅰ-FABP)和二肽转运载体1( PEPT1) mRNA表达的发育性变化及谷氨酰胺对其的影响.以69头(21±3)日龄断奶杜×长×大仔猪为试验动物,断奶当天选取3头猪进行屠宰,剩余66头随机分为2组,每组3个重复,每个重复11头.对照组饲喂基础饲粮,试验组饲喂基础饲粮+1％谷氨酰胺.断奶后第3、5、7、14天试验组和对照组分别选取3头猪进行屠宰(共计27头),取十二指肠、空肠和回肠黏膜组织样品,通过实时定量PCR法测定Ⅰ-FABP和PEPT1 mRNA的表达量.结果表明:1)Ⅰ-FABP和PEPT1 mRNA的表达量各肠段间无显著差异(P＞0.05);2)Ⅰ-FABP和PEPT1 mRNA在十二指肠、空肠和回肠的表达量均在断奶后急剧下降,断奶第3天的表达量最低,显著低于断奶当天(P＜0.05),而后逐渐升高,第14天达到峰值;3)试验组Ⅰ-FABP和PEPT1 mRNA表达量与对照组无显著差异(P＞0.05),但试验组表现出促使十二指肠、空肠、回肠黏膜的Ⅰ-FABP和十二指肠PEPT1 mRNA表达提前恢复至断奶前水平的趋势.结果提示,断奶仔猪Ⅰ-FABP和PEPT1 mRNA表达量随时间而变化,谷氨酰胺对断奶后Ⅰ-FABP和PEPT1 mRNA表达量的恢复有一定的促进作用.