Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

  • Print ISSN 0367-6722

  • Online ISSN 0976-0555

  • NAAS Rating 6.50

  • SJR 0.263

  • Impact Factor 0.4 (2024)

Frequency :
Monthly (January, February, March, April, May, June, July, August, September, October, November and December)
Indexing Services :
Science Citation Index Expanded, BIOSIS Preview, ISI Citation Index, Biological Abstracts, Scopus, AGRICOLA, Google Scholar, CrossRef, CAB Abstracting Journals, Chemical Abstracts, Indian Science Abstracts, EBSCO Indexing Services, Index Copernicus
Submit

Research Article

Indian Journal of Animal Research, volume 55 issue 10 (october 2021) : 1192-1199

Impact of Probiotic and Zinc on Brush-border Enzyme and Histoenzymatic Profile in the Small Intestine of Pre and Post-weaned Piglets

Arup Kalita1,*, M. Talukdar1, K. Sarma1, P.C. Kalita1, J.M. Gali2, S. Tamuli2, O.P. Choudhary1, P.J. Doley1, S. Debroy1, Keneisenuo1
1Department of Veterinary Anatomy and Histology, College of Veterinary Sciences and Animal Husbandry, Central Agricultural University (I), Selesih, Aizawl-796 015, Mizoram, India.
2Department of Veterinary Anatomy and Histology, College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati-781 022, Assam, India.
Cite article:- Kalita Arup, Talukdar M., Sarma K., Kalita P.C., Gali J.M., Tamuli S., Choudhary O.P., Doley P.J., Debroy S., Keneisenuo (2021). Impact of Probiotic and Zinc on Brush-border Enzyme and Histoenzymatic Profile in the Small Intestine of Pre and Post-weaned Piglets . Indian Journal of Animal Research. 55(10): 1192-1199. doi: 10.18805/IJAR.B-4211.

ABSTRACT

Background: Probiotics and zinc are commonly used and beneficial in pig production. This work aimed to assess the effect of probiotic and zinc on brush-border enzyme activity and histoenzymatic study of the small intestine in pre and post-weaned piglets.  

Methods: Eighteen LWY piglets were divided equally into control and treatment groups. The piglets were maintained in standard management conditions and were weaned at 28 days of age. The treatment group of piglets fed a mixture of probiotics orally @ 1.25 x 109 CFU/day and zinc @ 2000 ppm/day from birth to 10 days of age. At three different age-groups viz. day 20 (pre-weaning), day 30 (weaning) and day 60 (post-weaning), the animals were sacrificed. For disaccharidase enzyme estimation, the mucosal brush border of the small intestine was scrapped off and the experiment was conducted. For histoenzymatic assay, the small intestine samples were preserved in liquid nitrogen at -196ºC immediately after sacrifice. They were sectioned at 10µm thickness maintained at -20ºC and stained for different histochemical staining. The statistical analysis of the data using the appropriate statistical tests was also conducted. 

Result: The activity of different brush-border enzymes such as maltase, sucrase and lactase was more in the treatment group of piglets. The activity of different histochemical enzymes such as alkaline phosphatase, acid phosphatase, adenosine tri-phosphatase and non-specific esterase was increased in the treated group of piglets.

INTRODUCTION

Neonatal and post-weaning piglet mortality is a major concern in pig husbandry, which causes huge economic losses to pig farmers. Weaning is a major critical period because of increased susceptibility to gut disorders, infections and diarrhoea due to dietary changes. The major effect of weaning is a reduction of feed (energy) intake leading to undernutrition and retarded growth. At this time, there is an alteration of the intestine that includes changes in villus/crypt morphology and in brush-border enzyme activities that further aggravate the conditions. Previous researches illustrated that probiotics enhance the growth performance of swine (Wang et al., 2012), modulate the immune system (Gonzalez-Ortiz et al., 2013) and promote intestinal health (Tojo et al., 2014). Probiotics are the micro-organisms that have potential benefits on the host health (Djurasevic et al., 2017). Probiotics are also capable of improving the microecology of the intestine to provide a beneficial effect (Kohler et al., 2003). Feeding of zinc is well recognized to have growth-promoting effects in weaned pigs (Poulsen, 1995, Case and Carlson, 2002) and reduce the problems with post-weaning diarrhoea (Poulsen, 1989). Supplementing the zinc in pig diet may promote the processes of tissue repair in the small intestine and stimulate the synthesis of brush-border and other digestive enzymes, resulting in better digestion and absorption of nutrients and potentially improving growth performance.  
       
Keeping these points in view, the present study was undertaken to investigate the combined effect of probiotic and zinc on brush-border enzyme and histoenzymatic profiles in the small intestine of pre and post-weaned piglets.

MATERIALS AND METHODS

Animals
 
Eighteen healthy large white yorkshire (LWY) piglets, irrespective of sex obtained from three sows were utilizedfor the study. Care and management of the animals were provided in Instructional Pig Farm, College of Veterinary Sciences and Animal Husbandry, Central Agricultural University (I), Selesih, Aizawl, Mizoram, India. The Institutional Animal Ethics Committee (IAEC) ethically approved the animals used for the experiment vide Approval No. 770/ac/CPCSEA/FVSc/AAU/IAEC/17-18/490 dated 09.08.2017.
 
Selection, dose and period of treatment
 
A mixture of probiotic consisted with Lactobacillus acidophilus (650 million), Lactobacillus rhamnosus (400 million) and Bifidobacterium longum (200 million) was orally administered to the treatment group of piglets @ 1.25 × 109 CFU (1 gm powder dissolved in 3 ml of sterilized saline solution) once a day from birth to 10 days of age (Liu et al., 2014). The ZnO was given orally to the treatment group of piglets @ 2000 ppm once a day from birth to 10 days of age (Case and Carlson 2002). The piglets of the control group were given the same volume of sterilized saline solution.
 
Experimental design
 
Each of 6 (six) numbers of piglets was selected from 3 (three) sows at different stages of development as age-group of 20, 30 and 60 days. Out of the 6 (six) piglets, 3 (three) piglets from each litter were used as the control group (C) with basal diet and the other 3 (three) piglets were fed with combined probiotic and zinc supplement and used as treatment group (T). The combined probiotic and zinc were supplemented orally to the treatment group piglets along with the basal diet. The basal diet used in this experiment was in pellet form and was formulated to provide the nutrient requirements recommended by the (NRC) National Research Council (1998). The piglets were weaned at 28 days of age.
 
Sample preparation
 
The experimental animals were first anesthetized using diazepam @ 2mg/kg body weight followed by ketamine @ 10 mg/kg body weight intravenously and then exsanguinated the animals. The animals were sacrificed at day 20, 30 and 60 from both the groups. Subsequent to sacrifice, the abdominal cavity of the animal was exposed by reflecting the skin, fascia, abdominal muscles and peritoneum and parts of the small intestine were observed. The abdominal cavity was after that opened and the parts of the small intestine were then dissected out as per the method of Habel (1964). Tissue samples were taken immediately after sacrifice from the duodenum (5 cm caudal to the pylorus), jejunum (In the middle of the jejunum) and ileum (5 cm cranial to the ileocaecal valve).
 
Preparation for brush-border (disaccharidase) enzyme activity examination
 
The activity of disaccharidase enzymes in the small intestine was estimated by the method described by Dahlqvist (1964, 1968 and 1984) with slight modifications. The mucosal brush border of the small intestine was scrapped off using glass slide and stored at -80°C till further use. Mucosal tissue (100 mg) was placed in 5 ml centrifuge tube, homogenized with the help of micro pestle and added 900 µl of chilled normal saline solution (NSS). Tubes were vortexed and the volume of the suspension was increased to three times with chilled NSS. To 0.1 ml of mucosal tissue suspension, 0.1 ml of appropriate substrate solution (maltose/sucrose/lactose) was added. To this mixture, 2 µl of toluene was added and incubated the tubes at 37°C for 1 hr. A sample blank was prepared with the same composition and immersed the tubes in boiling water immediately after mixing of the enzyme and substrate. Estimation of glucose was carried out using glucose (GO) assay kit (Sigma, MI, USA) as per the manufacturer’s instruction. Glucose concentration in unknown samples was estimated using the standard curve prepared with glucose standards provided with the kit. The disaccharidase activity of the test samples was obtained by the following formula:
 
Disaccharidase activity (units/ml) = (a - b) × d/n × 540
 
Where,
a = Amount of glucose (µg) found in an aliquot of the incubated sample.
b = Amount of glucose (µg) found in an aliquot of the corresponding blank.
d = Dilution factor of the enzyme solution used for mixing with the substrate. 
n= Number of glucose molecules that were liberated per substrate (disaccharide) molecule hydrolyzed.
The maltose was composed of two glucose molecules (n=2).
 
The sucrose and lactose, which contain one molecule of glucose and one of fructose or galactose, respectively (n=1).
Units of activity = µmoles disaccharide hydrolyzed/ minute Preparation for the histochemical examination.
       
For histoenzymatic assay, the small intestine samples were preserved in liquid nitrogen at -196°C immediately after sacrifice. They were sectioned at10µm thickness in cryostat microtome (Shandon Finesse) maintained at -20°C. They were temporarily stored at -22°C and then treated for histochemical staining with the following methods:
 
1.     Gomori’s alkaline phosphatase cobalt method (Singh and Sulochana, 1978).
2.     Gomori’s method for acid phosphatase (Singh and Sulochana, 1978).
3.     Lead method for ATPase (Bancroft, 2008).
4.     Gomori’s method for non-specific esterase (Bancroft, 2008).
 
Statistical analysis
 
The data obtained were analyzed using statistical package SPSS version 20. General linear model of two way ANOVA based on fisher’s least significant difference method was used to determine the significant difference among days (20, 30 and 60 days) for control and treatment groups. The significant values in the ANOVA were further tested through the Duncan multiple range test. Results are presented as mean ± SEM and differences were considered significantwhen P<0.05. An independent sample t-test has been applied between groups (Control and treatment) on different days to see the significant changes.

RESULTS AND DISCUSSION

Brush-border (disaccharidase) enzyme profile
 
The activity of brush border enzymes such as maltase, sucrase and lactase in control (basal diet) and treated (basal diet + probiotic + zinc) piglets were estimated per gm of tissue based on the capacity of these enzymes to convert their respective substrates into glucose and the results are shown in Table 1 and Fig 1 to Fig 3.
 

Table 1: Brush-border enzyme activity in small intestine of piglets fed with probiotic and zinc.


 

Fig 1: Activity of maltase enzyme.


 

Fig 2: Activity of lactase enzyme.


 

Fig 3: Activity of sucrase enzyme.


 
Maltase
 
The activity of maltase increased towards the advancement of age in both the groups, being significant (P<0.05) in the control group and highly significant (P<0.01) in the treatment group (Fig 1a). The increased activity of this disaccharidase was also reported by Hornbuckle et al., (2008). In the present study, the maltase activity was lower as compared to the activity of lactase and sucrase. This was due to the procedure of estimation in a one-step method caused by interference between maltose and glucose oxidase, i.e., with the assay system and not with the disaccharidase activity. The lower estimation value of maltase activity was alsorecorded by Dahlqvist (1968). In the current study, the maltase activity for the conversion of maltose into two molecules of glucose as end products of starch digestion was higher in the treatment group of piglets than control in all the age-groups (Fig 1b). Moreover, it was significantly increased at day 30 (P<0.01) and day 60 (P<0.05) in treated piglets. Hedemann et al., (2006) and Hu et al., (2018) reported significantly higher maltase activity in piglets after feeding 100 ppm Zn (P<0.01) and probiotic (P<0.05) which were in close agreement with the present findings.
 
Lactase
 
In the current investigation, the lactase activity for the conversion of lactose (galactose-glucose) was decreased as per the advancement of age, being highest at day 20 and lowest at day 60 in both the groups (Fig 2a). The present finding was closely similar to the findings of Hornbuckle et al., (2008). The relatively high lactase activity in the younger animal might be an advantage in utilizing large quantities of lactose present in their diets. In this present study, the lactase activity was moderately higher (P>0.05) in the treatment group of piglets in comparison to the control animal (Fig 2b), which might indicate better conversion of lactose into galactose and glucose. The increased lactase activity was also recorded in piglets after feeding 100 ppm of Zn (Hedemann et al., 2006) and probiotics (Hu et al., 2018) in the diets. The above findings were in accordance with the present study.
 
Sucrase
 
In the present study, the sucrase activity for the conversion of sucrose (fructose-glucose) was increased as the age advanced in both the groups (Fig 3a). This finding was similar to the findings of Hornbuckle et al., (2008). In this current study, the activity of sucrase was higher in the treated piglets than that of control animals in all age-groups without any significant difference (Fig 3b). This finding might suggest the effective conversion of sucrose into fructose and glucose in the treatment group of piglets. The enhanced sucrase and lactase activity in intestinal mucosa after dietary inclusion of probiotic were also reported by Southcott et al., (2008) and Goyal et al., (2013) in rats and Hu et al., (2018) in piglets. 
       
Carbohydrates were one of the major components of the diet. In the gastrointestinal tract, carbohydrates were mainly digested by salivary and pancreatic amylases, further broken down into monosaccharides by disaccharidase, such as maltase, lactase and sucrase, which secreted by enterocytes of the brush border and then were absorbed (Zhen et al., 2018) in chicken. Thus, higher conversion of disaccharides to monosaccharides in treatment group piglets might be indicative of more absorption of glucose from the available carbohydrate present in the intestine and resulted in better growth and development in this group of piglets.
 
Histoenzymatic profile
 
The cryosections from all segments of the small intestine of control and treated piglets were subjected to histochemical staining by respective protocols and the results are illustrated in Table 2 to Table 5 and in Fig 4 to Fig 7. The different histochemical activities were observed in absorptive epithelium, glandular and, follicular and interfollicular areas. The gradation for intensity of histochemical reaction is: NA, Not Available; -, Negative; +, Weak; ++, Moderate; +++, Strong; ++++, Intense.   
 

Table 2: Alkaline phosphatase activity in histocompartments of the small intestine in piglets fed with probiotic and zinc.


 

Table 3: Acid phosphatase activity in histocompartments of the small intestine in piglets fed with probiotic and zinc.


 

Table 4: Adenosine tri-phosphatase activity in histocompartments of the small intestine in piglets fed with probiotic and zinc.


 

Table 5: Non-specific esterase activity in histocompartments of the small intestine in piglets fed with probiotic and zinc.


 

Fig 4: Photomicrographs showing histochemical staining of alkaline phosphatase activity (arrow) in cryosections of 30 days old treated piglet (Gomori’s, x100)


 

Fig 5: Photomicrographs showing histochemical staining of acid phosphatase activity (arrow) in cryosections.


 

Fig 6: Photomicrographs showing histochemical staining of adenosine-tri-phosphatase activity (arrow) in cryosections of 30 days old treated piglet.


 

Fig 7: Photomicrographs showing histochemical staining of non-specific esterase activity (arrow) in cryosections.


 
Alkaline phosphatase
 
In the present study, the alkaline phosphatase activity in the absorptive epithelium was intense in jejunum and ileum and strong in the duodenum in the treatment group of piglets (Fig 4a). However, this activity was moderate in duodenum and strong in jejunum and ileum of control animals (Table 2). Sprague et al., (1963) reported increasing alkaline phosphatase activity from cranial and middle sections of jejunum and ileum in new-born pigs, which was in agreement with the present finding. In the absorptive epithelium of the present study, this enzyme activity was slightly lower at day 30 in the control group of piglets, which might be due to the weaning effect as also revealed by Melo et al., (2016). However, in the treatment group piglets, the decreased activity of this enzyme at day 30 was not observed in the current study. The increased alkaline phosphatase activity in the absorptive epithelium recorded in the treated animals might be correlated with more ionic movements across the epithelium and increased number of matured enterocytes in the villi as reported by Kapoor and Singh (2017) in buffalo.
       
In the interfollicular area of jejunum and ileum, the alkaline phosphatase activity was moderate in control animals, while a strong reaction was observed in the treated piglets (Fig 4b) in all age-groups. The current observationmight be an indicator of the presence of more energy-dependent active transport mechanisms at these sites. Gautam (2015) recorded strong alkaline phosphatase reactions in these areas in growing piglets, which was similar to the present study. A weak to negative activity was observed in the glandular epithelium (Fig 4c) throughout all segments of the small intestine in both the groups, as also reported by Gautam (2015) in growing piglets. 
 
Acid phosphatase
 
The acid phosphatase activity in the current study was found to be moderate in the absorptive epithelium irrespective of segments of intestine and age in both the groups (Table 3). In the glandular epithelium, it was observed intense in the treated piglets (Fig 5a) and strong in the control animal (Fig 5b). The intense reaction of this enzyme in the glandular epithelium of treatment group piglets might indicate the presence of more lysosomal activity in the respective sites as depicted by Gautam (2015) in growing piglets.  
       
In the present study, the activity of acid phosphatase was strong and moderate in the follicular and interfollicularareas of jejunum and ileum of treated (Fig 5c) and control group (Fig 5d) of piglets, respectively in all age-groups. A reticular pattern acid phosphatase activity was seen in the interfollicular regions in both groups. This finding was also tuned to the finding of Halleraker et al., (1990), who reported reticular pattern acid phosphatase activity in the interfollicular regions of PP in calf, kid and lamb.
 
Adenosine-tri-phosphatase
 
In the present investigation, the activity of the adenosine- tri-phosphatase (ATPase) enzyme is presented in Table 4. The follicular and interfollicular areas of PP showed strong reaction in treated piglets (Fig 6a) and moderate reaction in control piglets for adenosine tri-phosphatase activity. This observation was in accordance to the findings of Halleraker et al., (1990), who opined that follicular dendritic cells and B-cell area were positive for magnesium-dependent ATPase activity. The increased activity of ATPase in the follicular area of treated piglets might suggest induced B-cell area for better immunity in this group of piglets. This finding was also confirmed by immunofluorescence observation in this study. 
       
The ATPase enzyme activity in the present observation was strong in the absorptive epithelium of treated piglets (Fig 6b) and moderate in the control group animals. However, this activity was moderate in the glandular region (Fig 6c) in both the groups irrespective of segments of intestine and age. The present findings were almost similar to the findings of Gautam (2015) in growing piglets. 
 
Non-specific esterase
 
In the present study, the non-specific esterase enzyme activity has been presented in Table 5. The dietary inclusion of probiotic and zinc revealed strong non-specific esterase activity in the absorptive epithelium (Fig 7a) and, follicular and interfollicular areas of PP (Fig7b). However, this activity was found to be moderate in these areas of control animals (Fig 7c). The follicles of PP showed positively stained macrophages (Fig 7d), which were predominant in the treatment group of piglets. The strong non-specific esterase reaction in PP area was also recorded by Halleraker et al., (1990), who described a positive reticular reaction in the T-cell area of the follicles and detected macrophages stained positive with non-specific esterase in ruminants. The sites of non-specific esterase activity were found to be a T-cell rich area, which was demonstrated by the APAP technique by Mishra (1998) and Rajkhowa (2003) in the pig.
       
The non-specific esterase activity in the glandular area of present observation was found to be moderate in all segments of the small intestine irrespective of treatment and age-groups.

CONCLUSION

In conclusion, the activity of brush border enzymes such as maltase, sucrase and lactase was more in treated piglets that indicate increased capacity of these enzymes to convert their respective substrates to glucose compared to control animals. Thus, higher conversion of disaccharides to monosaccharides in the treatment group of piglets might be indicative of more absorption of glucose from theavailable carbohydrate present in the intestine and resulted in better growth and development in this group of piglets. Similarly, the activity of alkaline phosphatase, acid phosphatase, adenosine tri-phosphatase and non-specific esterase was increased in treated group of piglets that might be correlated with increased ionic movements across the epithelium, higher lysosomal enzyme activity, localization of more number of B cells for better immunity and more concentration of T cells, respectively.

REFERENCES


  1. Bancroft, J.D. (2008). Theory and Practice of Histological Techniques. 6th Edition, Churchill Livingstone, Elsevier Health Sciences.

  2. Case, C.L. and Carlson, M.S. (2002). Effect of feeding organic and inorganic sources of additional zinc on growth performance and zinc balance in nursery pigs. J. Anim. Sci. 80: 1917-1924.

  3. Dahlqvist, A. (1961). Intestinal carbohydrases of a new-born pig. Nature. 190: 31-32.

  4. Dahlqvist, A. (1964). Method for assay of intestinal disaccharidases. Analytical Biochemistry. 7: 18-25.

  5. Dahlqvist, A. (1968). Assay of intestinal disaccharidases. Analytical Biochemistry, 22: 99-107.

  6. Dahlqvist, A. (1984). Assay of intestinal disaccharidases. Scand. J. Clin. Lab. Invest. 44: 169-172.

  7. Djurasevic, S., Jama, A., Jasnic, N., Vujovic, P., Jovanovic, M., Mitic-Culafic, D., Knezevic-Vukcevic, J., Cakic- Milosevic, M., Ilijevic, K. and Djordjevic, J. (2017). The Protective Effects of Probiotic Bacteria on Cadmium Toxicity in Rats. J. Med. Food., 20(2): 189-196.

  8. Gautam, C.K. (2015). Antigen uptake and comparative histomorphology and ultrastructure of the histocompartments of Peyer’s patches and solitary lymphoid nodules of intestine of growing piglet. Ph. D. Thesis, Assam Agricultural University, Khanapara, Guwahati, Assam, India.

  9. Gonzalez, L.M., Williamson, I., Piedrahita, J.A., Blikslager, A.T. and Magness, S.T. (2013). Cell lineage identification and stem cell culture in a porcine model for the study of intestinal epithelial regeneration. PLOS One. 8(6): 1-18.

  10. Goyal, N., Rishi, P. and Shukla, G. (2013). Lactobacillus rhamnosus GG antagonizes Giardia intestinalis induced oxidative stress and intestinal disaccharidases: an experimental study. World J. Microbiol. Biotechnol. 29: 1049-1057.

  11. Habel, R.E. (1964). Guide to the dissection of domestic ruminants. Edwards Brother Inc., Ann. Arbor, Michigan.

  12. Halleraker, M., Landsverk, T. and Nicander, L. (1990). Organization of ruminant Peyer’s patches as seen with enzyme histochemical markers of stromal and accessory cells. Vet. Immunol. Immunopathol. 26: 93-104.

  13. Hedemann, M.S., Jensen, B.B. and Poulsen, H.D. (2006). Influence of dietary zinc on digestive enzyme activity and intestinal morphology in weaned pigs. J. Anim. Sci. 84: 3310-3320.

  14. Hornbuckle, W.E., Simpson, K.W. and Tennant, B.C. (2008). ‘Gastrointestinal function’. In: [Clinical Biochemistry of Domestic Animals. Chapter 14, 6th Edn.] Elsevier. pp. 413-457.

  15. Hu, S., Cao, X., Wu, Y., Mei, X., Xu, H., Wang, Y., Zhang, X., Gong, L. and Li, W. (2018). Effects of probiotic Bacillus as an alternative of antibiotics on digestive enzymes activity and intestinal integrity of piglets. Frontiers in Microbiology 9: 1-9.

  16. Kapoor, K. and Singh, O. (2017). Histoenzymic distribution of phosphatases, dehydrogenases and esterases in ileal Peyer’s patches of neonatal buffalo calves. Indian J. Anim. Sci. 51(4): 670-675.

  17. Kohler, H., McCormick, B.A. and Walker, W.A. (2003). Bacterial-enterocyte crosstalk: cellular mechanisms in health and disease. J Pediatr. 36: 175-185.

  18. Liu, H., Zhang, J., Zhang, S., Yang, F., Thacker, P.A., Zhang, G., Qiao, S. and Ma, X. (2014). Oral administration of Lactobacillus fermentum I5007 favors intestinal development and alters the intestinal microbiota in formula-fed piglets. J. Agricul. Food Chem. 62: 860-866.

  19. Melo, A.D.B., Silveira, H., Luciano, F.B. andrade, C., Costa, L.B. and Rostagno, M.H. (2016). Intestinal alkaline phosphatase: Potential roles in promoting gut health in weanling piglets and its modulation by feed additives - a review. Asian Australas. J. Anim. Sci. 29(1): 16-22.

  20. Mishra, S. (1998). Immunohistological study of gut associated lymphoid tissues in pigs. M.V.Sc. Thesis, Assam Agricultural University, Assam, India.

  21. N.R.C. (1998). Nutrient Requirements of Swine. In: Computer Model Program for Predicting Nutrient Requirements. 10th Edn., National Academy of Sciences, Washington, DC, USA.

  22. Poulsen, H.D. (1989). Zinc oxide for pigs during the weaning period. Publication no. 746. Danish Institute of Animal Science, Tjele, Denmark.

  23. Poulsen, H.D. (1995). Zinc oxide for weanling piglets. Acta Agric. Scand. Sect. A. Animal Sci. 45: 159-167.

  24. Rajkhowa, J. (2003). Development of small intestine of piglets during preweaning period. M.V.Sc. Thesis, Assam Agricultural University, Assam, India.

  25. Singh, U.B. and Sulochana, S. (1978). A laboratory manual of histological and histochemical technique. 1st Edn., Kothari Medical Publishing House, Bombay. pp. 50-54.

  26. Southcott, E., Tooley, K.L., Howarth, G.S., Davidson, G.P. and Butler, R.N. (2008). Yoghurts containing probiotics reduce disruption of the small intestinal barrier in methotrexate-treated rats. Dig. Dis. Dci. 53: 1837-1841.

  27. Sprague, J.I., Ullrey, D.E., Waddill, D.G., Miller, E.R., Zutaut, C.L. and Hoefer, J.A. (1963). Intestinal lactase, alkaline and acid phosphatase in the swine fetus and new-born pig. J. Anim. Sci. 22(1): 121-124.

  28. Tojo, R., Suarez, A., Clemente, M.G., Reyes-Gavilan, C.G., Margolles, A., Gueimonde, M. and Ruas-Madiedo, P. (2014). Intestinal microbiota in health and disease: Role of bifidobacteria in gut homeostasis. World J. Gastroenterol. 20(41): 15163-15176.

  29. Wang, X., Yang, F., Liu, C., Zhou, H., Wu, G., Qiao, S., Li, D. and Wang, J. (2012). Dietary supplementation with the probiotic Lactobacillus fermentum I5007 and the antibiotic aureomycin differentially affects the small intestinal proteomes of weanling piglets. J Nutr. 142: 7-13.

  30. Zhen, W., Shao, Y., Gong, X., Wu, Y., Geng, Y., Wang, Z. and Guo, Y. (2018). Effect of dietary Bacillus coagulans supplementation on growth performance and immune responses of broiler chickens challenged by Salmonella enteritidis. Poul. Sci. 97: 2654-2666.
Reviewed By

Download Article
In this Article
    Contact us
    Follow us

    Editorial Board

    View all (0)