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Effect of sodium butyrate on performance, immune status, microarchitecture of small intestinal mucosa and lymphoid organs in broiler chickens.

INTRODUCTION

After imposition of ban on using some dietary antibiotics as growth enhancers by the European Union [1], the focused area in the poultry and animal sector is to find/create safe feed items which are free of antibiotics [2]. Subsequently, the research focused on development of optimal alternatives with the aim to maintain functions of gut and immune system. Organic acids (commonly known as acidifiers) and their salts are generally considered as harmless and have been approved by most technologically advanced countries to be used as a feed additive for animals. The acidifiers, including sodium butyrate (SB) is known for decreasing the gut mucosal pH, thus creating an acidic environment for the growth of normal commensals [3]. They overcome the development and proliferation of some Salmonella spp. [4]. There were noteworthy improvements in weight gain, carcass characteristics and increase in size of intestinal villi in the butyric acid-supplemented birds [5]. Chamba et al [6] used SB in broilers and observed its growth promoter effect. Enriched IgG concentration in serum and total IgA+ in jejunum was observed in SB treated piglets [7], and the immunostimultory property of SB has also been highlighted in chicken [8] by inducing host defense peptides. Hence the SB was registered as an immune modulator [9]. It has been observed that the microencapsulated (coated with fatty acid matrix) type of organic acid was more effective than an antibiotic growth promoter (Enramycin) in rising growth performance in broilers [10]. The microencapsulated butyrate delivered portion of the butyrate to be free further distal in the intestinal tract because of slow release during digestion [4] and causes mucosal modulation in the gut [6]. Its use led to a tendency towards better growth performance, lower colonization and fecal shedding of Salmonella compared to the non-protected feed supplements [4]. To- date, limited reports have been published to evaluate the effects of SB on humoral and cellular immune status [9] and microarchitecture of visceral organs, including the segments of small intestine, bursa of Fabricius, thymus and spleen in broiler chickens. A comprehensive study was, therefore, needed to assess such effects in commercial chickens.

The aim of the current study was to evaluate whether the gut development and immune system of broiler chickens is influenced by feeding microencapsulated SB from day-1.

MATERIALS AND METHODS

Experimental chicks, husbandry and ration

A total of 120 1-d-old M77 Hubbard broiler chicks (as hatched) were reared for a total period of 35 days in environmentally controlled shed, where the temperature and relative humidity (RH) were maintained at 32[degrees]C [+ or -] 1[degrees]C and 70% [+ or -] 5%, respectively at the beginning. The temperature was reduced by 3[degrees]C per week till it reached 24[degrees]C [+ or -] 2[degrees]C, with RH closed to 65% [+ or -] 5%; thereafter, the temperature was maintained till d-35. Broiler chicks at the age of d-1 were weighed and divided into the following four groups: corn-soy based basal diet (BD) with no supplement (control), BD supplemented with 10% zinc bacitracin (ZnB, Hubei Yuancheng Tech. Develop. Co. Ltd., Wuhan, China added 0.01% of feed) or 1.0 g/kg SB (SB-1) or 0.5 g/kg SB (SB-0.5), respectively. Each group had 30 birds with three replicates (n = 10) each. The birds in each group were offered a BD prepared locally in feed mill without antibiotics (Table 1) in starter (d-1 to d-21) and grower (d-22 to d-35) phases, respectively. Feed was formulated accordingly to meet the energy and nutrient requirements of broilers and was supplemented with SB (CM3000, Hangzhou King Techina Feed Company Ltd., Hangzhou 311112, China) as described elsewhere [11] with the following minor modification. The microencapsulated SB was weighed and was thoroughly homogenized with 1 kg mesh form of BD. To ensure its proper mixing, the resulting mixture was then mixed in vertical mixer with the control mesh form of BD. Availability of diet and water was ad libitum throughout the study. Six broiler chickens/group were killed on d-21 and d-35, and samples including blood and organs were collected.

Growth performance

The average body weight (ABW), weekly weight gain (WWG), average feed intake (AFI) and FCR were studied to record weekly growth performance [12].

Immune response evaluation

Three in-vivo tests in which cutaneous response to phytohemagglutinin-P (PHA-P) and serum antibody level produced in response to Newcastle disease vaccine and sheep red blood cells (SRBCs) were measured to evaluate the immune competence in broiler chickens treated with SB.

Cell-mediated immunity

Cell-mediated immunity was assessed through injection of PHA-P (Sigma-Aldrich, St. Louis, MO, USA) as reported by Corrier et al [13] with the following slight modification. Briefly, two birds per replicate (n = 6 per group) were randomly selected on d-17 and the PHA-P solution (prepared in sterile phosphate-buffered saline [PBS]) was injected intradermally (100 [micro]g/100 [micro]L/chicken) between the 3rd and 4th digits of the right foot. The left foot served as control and was injected with 100 [micro]L of PBS. The net increase in thickness of the injected sites was evaluated on 24, 48, and 72 hours post-injection using pressure sensitive micrometer. The immune response (foot web index) to PHA-P was measured by subtracting the left foot thickness from that of the right foot.

Humoral immunity

The antibody titer or antibody production level is attributed to humoral immune response in animals. Humoral immune responses against Newcastle disease virus (NDV) and sheep RBCs antigens were evaluated through serological titration as defined previously [14]. Briefly, all the chicks were vaccinated with ND vaccine (Nobilis ND LaSota, Intervet International B.V. Boxmeer, Holland) on d-1 and d-9 via the ocular route and boosted on d-16 and d-23 via drinking water. And in case of SRBCs, 2 birds per treatment replicate (n = 6 per group) were randomly selected, wing banded and injected bilaterally with 5% SRBCs antigen (sheep blood collected in Alsevier's solution, washed thrice and suspended in phosphate buffer saline) in two parts (0.5 mL each, intramuscularly in both sides of the Musculus pectoralis) on d-14. Booster dose was given on d-21. Blood samples were collected on seven days post-primary injection and on d-35. Sera separated (2,000 x g for 10 minutes) and were stored at -20[degrees]C till analysis. The antibody responses to NDV and SRBCs were measured using micro-titer hemagglutination inhibition and hemagglutination (HA) assays, respectively as mentioned by King [14].

Relative weight of lymphoid organs

Two chickens per treatment replicate were randomly selected, weighed and killed on d-21 and 35. Small intestine, liver, spleen, thymus and bursa of Fabricius were isolated from the carcass and their relative weights were evaluated. Representative samples of the organs were then subsequently collected in 10% neutral buffered formalin for histological processing.

Microarchitecture of immune organs

Thymus: Histomorphometry of the thymus was carried out as described Madej et al [15] with slight modifications; briefly, the thymic lobules were split into 4 sections by two lines crossing each other at right angle in the center of the medulla. All the lines expressing overall widths of the cortex were evaluated and the average was represented as cortical thickness. Area of the medulla was evaluated by uniting the measurements of two lines expressing the length and width of the medulla. Afterward, the ratios of cortex to medulla were evaluated in three well-oriented lobules per section, and the mean values obtained from three sections per bird were calculated.

Spleen: Well-oriented germinal center areas in the spleen were combined together and were noted as a percentage of the total field of view at 10x [15]. Later the average of three sections values was determined.

Bursa: Total numbers of lymphoid follicles in one microscopic field were recorded. Length, width and area of 5 well-oriented bursal follicles per section (3 sections per sample at 4x) was measured [16]. Later, average value of the three sections was noted.

Microarchitecture of small intestinal mucosa

Representative samples of approximately 2 cm (length) segments were excised from duodenum (10 cm distal to the junction between duodenum and gizzard), jejunum (5 cm proximal to the Meckel's diverticulum) and ileum (5 cm proximal to the ileo-cecal junction). Intestinal samples were processed using paraffin embedding technique, sectioned at 5 gm using a microtome (AMOS Scientific AEM-450, St. Veit/Glan, Austria), and stained with H&E [17] similar to the immue organs, except for histochemical differentiation of goblet cells for which the slides were stained using combined alcian blue periodic acid-Schiff technique [18].

Gut mucosal histomorphometry. Three sections were collected (one section after every 10 sections) from each intestinal sample. From each section, five well-orientated villi having intact lamina propria were selected randomly for examination. Consequently an average of 15 values was analyzed for each sample. Finally, the mean values from six chickens were noted as mean values for one treatment. Slides were examined under a light microscope (Olympus CX31, Olympus, Hicksville, New York, USA) at 4x magnification, supported with digitalized live image analysis program (Olympus DP20, Olympus, USA). The variables calculated for histomorphological modulations were villus height, VH; villus width, VW; villus surface area, VSA; crypt depth, CD; villus height to crypts depth ratio, VH:C D [19, 12].

Intraepithelial lymphocytes count: Sections already used for morphometry were also used for intraepithelial lymphocytes (IELs) count as described previously by Ashraf et al [19]. The IELs are described as small (7 to 10 gm) and large lymphocytes, (10 to 20 gm) having intensely stained round to oval nucleus surrounded by tiny cytoplasm and is positioned along the columnar epithelium [20].

Goblet cell histochemistry: Slides prepared and processed earlier were subjected to alcian blue periodic acid Shiff (AB-PAS) staining. Three sections were obtained from each intestinal segment and goblet cells were counted in 5 villi/section. Thus an average of 15 values was calculated for each sample. The histochemical differentiation on the basis of acidic and mixed (acidic and neutral) mucins was noted according to the methods described elsewhere by Ashraf et al [19]. Goblet cells containing acidic mucin were stained blue by the AB, whereas mixed mucin were stained purple by PAS staining.

Statistical analysis

The normal distribution of the data was confirmed using Kolmogorov-Smirnov test and data were presented as means [+ or -] standard error of the mean. The data were analyzed using one-way analysis of variance (SPSS for windows version 20, Chicago, IL, USA). Statistical differences among treatment means were determined through Duncan's multiple range tests. In all statistical analyses, p<0.05 was considered significant.

Ethical note

This research was approved by the Ethical Review Committee for the use of laboratory animals of the University (reference no.: DR/257 dated 13-04-15).

RESULTS

Growth performance

During the first week, the feed intake decreased (p<0.05) in SB-1 chicks compared to control and FCR was noted to be lower (p< 0.05) in ZnB compared to SB-0.5 and control groups. The ABW and WWG in the 4th week, and ABW in 5th week were noted to be greater (p<0.05) in the treated groups compared to control (Table 2). During 5th week the WWG increased (p<0.05) in SB-1 compared to control, and AFI was higher (p<0.05) in both SB-offered groups compared to ZnB and control. The collective average of WWG per week increased (p<0.05), and that of FCR decreased (p<0.05) in all the supplemented groups compared to control. The AFI per week was higher (p<0.05) in both SB-offered groups compared to ZnB and control.

Immunological responses

No significant difference was noted between mean values of different groups at 24 and 72 h. However, higher (p<0.05) skin thickness was observed in SB-1 at 48 h post PHA-P injection compared to ZnB and control groups (Table 3).

Immune responses on day-21 were non-significant among the groups; however, the SB-1 group registered a tendency towards better response compared to other groups. On day-35, antibody titer against ND and SRBCs registered higher (p<0.05) in SB-1 compared with ZnB and control (Table 4).

Relative organ's weight

Bursa weighed more (p<0.05) in both SB-offered groups, while thymus weighed more (p<0.05) in SB-1 group compared to control on d-21 (Table 5). On d-35, spleen registered more (p<0.05) weight in SB-1, while thymus weighed more (p<0.05) in all the supplemented groups compared to control.

Histomorphometry of immune organs

On d-21 thickness of thymus medulla increased (p<0.05) in SB-1 compared to other groups. Germinal center area of spleen in SB-1 increased (p<0.05) compared to ZnB and control (Table 6). On d-35 germinal center area in spleen was greater (p<0.05) in both SB-offered groups compared to ZnB and control (Table 7).

Mucosal histomorphometry of small intestine

Starter phase (D-21): On day-21, VH and VSA in duodenum and jejunum of both SB-offered groups and VH in ileum of SB-1 group chickens was found enhanced (p<0.05) compared to ZnB and control. The VH:CD was increased (p<0.05) in duodenum and ileum of SB-1 group, and in jejunum of both SB groups com pared to ZnB and control (Table 8).

Grower phase (D-35): SB-1 group showed increased (p<0.05) VH and VSA in duodenum and jejunum compared to ZnB and control, and increased (p<0.05) VH and VSA in ileum compared to control. Villus width and VH:CD was higher (p<0.05) in duodenum of SB-1 compared to control (Table 9).

Intraepithelial lymphocytes count: The IEL count did not vary statistically among groups (Table 10).

Goblet cell histochemistry. On d-21, number of goblet cells containing acidic mucins was greater (p<0.05) in all the segments of small intestine in SB-1 group compered to control (Table 11). On d-35, the acidic-natured goblet cells increased (p<0.05) in ileum of SB-1, compared to all other treatment groups (Table 12).

DISCUSSION

The growth performance of SB-1 group was numerically better compared to control group (Table 2), linked well with the observations of Chamba et al [6] and Dehghani-Tafti and Jahanian [21]. The variability in FCR between control and the treatment groups is due to unidentified factors, however, it is assumed that better performance may be due to the creation of the acidic environment in the gut after SB consumption [3], which in turns minimizes the load of pathogens [10]. Average weekly feed intake was noted higher (p<0.05) in both SB supplemented groups compared to ZnB and control. Zinc bacitracin used in the current study because this antibiotic is famous for having growth promoting properties and are being practiced in local poultry sector. The infeed SB may improve the intraluminal digestibility of mineral and proteins which may result in improved weight gain in SB offered groups as mentioned by Zhang et al [22].

Scientists take interest in using in-vivo T cell dependent immune function test to PHA-P [23], and antibody response to heterologous erythrocytes [24]. For immune-competence of commercial chicken fed diet containing SB, we used these tests. T-cell mitogenic PHA-P was injected intradermally to evaluate the cell-mediated immunity. We found the net increase in swelling of the PHA-P injected area in SB-1 at 48 h compared to ZnB and control groups (Table 3), which indicates better immune response in that group [25]. The cell mediated immune response might be due to delayed type of hypersensitivity. Sheep RBCs (SRBCs) act as Thymus-dependent immunogens and are used for antibody response evaluation in chickens [24]. We observed higher (p<0.05) titer results against ND and SRBCs in SB-1 group on day-35 (Table 4). This indicates that SB may modulate the function of B and T cells in later stages of the antigenic exposure and can regulate the host immunity [9,23]. Park et al [26] noted that short chain fatty acids including butyrates, are commonly synthesized in the gut which support the regulation and growth of Th1 and Th17 effector cells as well as interleukin-10 (IL-10) regulatory T-cells. These cells maintain the immune system framework. The effect of butyrate on macrophage cell line was reported by Zhou et al [27]. They noted that butyrate inhibited nitric oxide production, and diminished the cytokines expression, including IL-6, IL-10, interferon gamma, and IL-1p, which controlled inflammation and maintained immune homeostasis. Similar to our findings, Eshak et al [28] reported high antibody titer against ND in SB treated chickens. The butyrate regulates the macrophage activities in intestine, and the macrophage effectuates the function of T cells and dendritic cells in the gut [29]. The latter cells also have a role in host immunity. The supplemented SB increased number of IgA+ cells which later on produced secretory immunoglobulin-A in piglets jejunum [7]. The sIgA contributes in improvement of one of the first line of mucosal defense. It was published that butyrate activated the immunomodulatory property through the production of host defense peptides [8] and has no effect on provoking inflammation [3].

In healthy animals the increase in weight of immune organs is correlated with improved immune responses of the body. Bursa, thymus and spleen are the key players of the immune system and their weights in the current study increased (p<0.05) in SB-offered chickens (Table 5). Alike our finding, Eshak et al [28] reported that bursa weighed more (p<0.05) in chickens treated with SB. The increased weights may be due to increased thickness of the parenchymal areas.

The greater sized thymus medulla and germinal center area in spleen in SB treated chickens (Table 6), and an increasing trend of bursal parameters observed in SB-1 group indicated that SB may effectuate the systemic immune systems in broilers also. To the best of our knowledge, no such study is available in literature highlighting the effect of SB on compartmental changes in immune organs to which we may compare our results.

Small intestine is the site for absorption in which the available nutrients are taken up through epithelial cells and drained into the general circulation. Architectural modulation of the small intestine is assumed to have a relationship with production performance of animals. We noted that small intestinal parameters including VH, VSA, and VH:CD in SB-offered groups improved significantly (Tables 8, 10). Butyrate acts as a rich source of energy for the enterocytes [9], and it may possibly increase the cell mitosis in the crypts. The SB may protect the mucosal epithelium from injury and alleviate the enteropathic stress [19] by increasing thyroid hormone in the circulation (data not reported). We found improved histomorphometrics in SB offered groups in duodenum and jejunum compared to ZnB and control. These findings proposed that the incoming ingesta containing SB at ileum had earlier been presented to utmost absorption in the former gut lumen and displayed better effect there.

The increased villus length and surface area could predict the gain in weight [19]. As the ingredient status of the diets in all groups was almost similar, the apparent enhancement in growth performance of the SB groups compared to control was assumed to be the result of the mucosal architectural modulations in those groups. Similar to our observation, various scientists reported that organic acid supplementation markedly increased the intestinal absorption area by promoting villus growth in height [5,11].

IELs are a mucosal portion of gut-associated lymphoid tissue and are expected to play important role in early contacts with antigens. Increased number of IEL is reported to have a role in immune modulations in infected animals offered Enterococcus faecium NCIMB 10415 [20]. In the current study the IELs population among groups was not dissimilar statistically (Table 10).

The mucus in the goblet cells acts as lubricant, source of nutrition for the normal commensals and protection of the gut from pathogens [30]. We found increased (p<0.05) acidic natured goblet cells in all the segments of small intestines in SB-1 group (Table 11). It is assumed that SB may uphold the goblet cells activities by regulating its mucin gene and may positively contribute to the protective mechanisms in the gut. On d-35 the underlying mechanism involved in increased acidic natured goblet cell population in the ileum is not clear; however, it may be in response to the high microbial population in that segment compared to the upper segments. In conclusion, diets supplemented with 1.0 g/kg SB stimulated a positive influence on the animal health by modulation of mucosal morphology of small intestine in birds. These microarchitecture modulations have a relationship with improved immunity, which suggests that SB at 1 g/kg feed may be a proper replacement for the antibiotics.

CONFLICT OF INTEREST

We certify that there is no conflict of interest with any financial organization regarding the material discussed in the manuscript.

https://doi.org/10.5713/ajas.16.0824

ACKNOWLEDGMENTS

Funds for this study were sponsored by Higher Education Commission, Pakistan under PhD indigenous scholarship HEC 2a-v1-147. The authors would like to thank Prof. Dr. Ashiq Hussain Cheema for his guidance in the study.

REFERENCES

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Arbab Sikandar (1), *, Hafsa Zaneb (2), *, Muhammad Younus (1), Saima Masood (2), Asim Aslam (2), Farina Khattak (3), Saima Ashraf (2), Muhammad Shahbaz Yousaf (2), and Habib Rehman (2)

* Corresponding Authors: Arbab Sikandar

Tel: +92-334-4038376, Fax: +92-47-7671262; E-mail: arbab.sikandar@uvas.edu.pk

Hafsa Zaneb

Tel: +92-42-9921-1449 (238), E-mail: hafsa.zaneb@uvas.edu.pk

(1) Sub-campus, Jhang, University of Veterinary and Animal Sciences, Lahore 35200, Pakistan

(2) University of Veterinary and Animal Sciences, Lahore 54000, Pakistan

(3) Monogastric Science Research Centre, Scotland's Rural College, Ayr KA6 5HW, UK

Submitted Oct 25, 2016; Revised Nov 22, 2016; Accepted Jan 3, 2017
Table 1. Control diet ingredients and calculated analysis

Items                       Starter phase   Grower phase

Ingredient (%)
  Corn                          40.15          57.57
  Rice broken                   15.0             --
  Soy meal                      11.54           9.60
  Sunflower meal                12.00          13.00
  Canola meal                   9.00            5.00
  Rapeseed meal                 5.00            7.60
  Rice polish                    --             4.00
  Guar meal                     1.00             --
  Wheat bran                    1.34             --
  Molasses                      2.00             --
  Sodium bicarbonate            0.03           0.065
  Sodium chloride               0.21            0.21
  Di-calcium Phosphate          1.33            1.49
  L-lysine                      0.30            0.35
  DL-Methionine                 0.10            0.12
  Vit-mineral premix (1)        1.00            1.00
Nutrient composition
  Calculated ME (kcal/kg)       2,750          2,850
  CP (%)                        19.6            18.5
  DM (%)                        87.0             88
  Crude fiber (%)               6.05            6.35
  Crude fat (%)                 2.16            2.35
  Total ash (%)                 5.77            5.40

(1) Vitamin mineral premix (each kg contained): ascorbic acid,
26,000 IU; retinol, 200,000 IU; cholecalciferol, 80,000 IU;
tocopherol, 1,072 IU; thiamine, 11,666 IU; pyridoxine, 33,333 IU;
menadione, 1 1,333 IU; riboflavin, 54,000 IU ; niacin, 5,36,000 IU;
folic acid, 13,600 IU; methylcobalamin, 223 IU; biotin, 1,340 IU;
Ca, 195 g; K, 70 g; Na, 18 g; Mg, 6 g; Fe, 2,000 mg; Zn, 1,200 mg;
Mn, 1,200 mg; Cu, 400 mg; I, 40 mg; Co, 20 mg; and Se, 8 mg.

Table 2. Effect of Sodium butyrate and zinc bacitracin on
performance in broilers

                                         Treatments
Growth   Parameters
phase                          SB-1                    SB-0.5

1st      Avg. body     151.13 [+ or -] 3.45     148.67 [+ or -] 1.43
week     weight (g)
         Weekly        110.13 [+ or -] 3.45     107.00 [+ or -] 1.43
         weight
         gain (g)
         Avg. feed     130.87 [+ or -] 2.30     129.57 [+ or -] 1.34
         intake (g)             (b)                      (ab)
         FCR             1.18 [+ or -] 0.02       1.21 [+ or -] 0.02
                                 (ab)                     (a)
2nd      Avg. body     420.50 [+ or -] 4.59     422.76 [+ or -] 7.73
week     weight (g)
         Weekly        269.37 [+ or -] 5.64     274.77 [+ or -] 8.29
         weight
         gain (g)
         Avg. feed     340.73 [+ or -] 8.79     341.30 [+ or -] 1.14
         intake (g)
         FCR             1.27 [+ or -] 0.06       1.24 [+ or -] 0.04
3rd      Avg. body     755.43 [+ or -] 4.14     736.66 [+ or -] 11.21
week     weight (g)
         Weekly        334.93 [+ or -] 8.50     313.90 [+ or -] 12.66
         weight
         gain (g)
         Avg. feed     521.73 [+ or -] 10.76    512.83 [+ or -] 8.87
         intake (g)
         FCR             1.56 [+ or -] 0.03       1.64 [+ or -] 0.08
4th      Avg. body   1,191.25 [+ or -] 4.65   1,156.04 [+ or -] 8.40
week     weight (g)            (a)                      (a)
         Weekly        435.82 [+ or -] 8.21     419.37 [+ or -] 6.10
         weight                 (a)                      (a)
         gain (g)
         Avg. feed     749.17 [+ or -] 11.51    744.49 [+ or -] 4.46
         intake (g)
         FCR             1.72 [+ or -] 0.00       1.78 [+ or -] 0.07
                                 (b)                      (b)
5th      Avg. body   1,741.83 [+ or -] 16.69  1,689.42 [+ or -] 34.54
week     weight (g)            (a)                      (a)
         Weekly        550.58 [+ or -] 16.78    533.38 [+ or -] 43.52
         weight                 (a)                      (ab)
         gain (g)
         Avg. feed   1,153.17 [+ or -] 5.88   1,165.54 [+ or -] 2.58
         intake (g)            (a)                      (a)
         FCR             2.09 [+ or -] 0.06       2.21 [+ or -] 0.18

                                         Treatments
Growth   Parameters
phase                          ZnB                    Control

1st      Avg. body     151.40 [+ or -] 1.31     152.03 [+ or -] 1.13
week     weight (g)
         Weekly        110.40 [+ or -] 1.31     111.03 [+ or -] 1.13
         weight
         gain (g)
         Avg. feed     127.27 [+ or -] 1.19     134.10 [+ or -] 5.06
         intake (g)             (ab)                     (a)
         FCR             1.15 [+ or -] 0.03       1.20 [+ or -] 0.02
                                 (b)                      (a)
2nd      Avg. body     412.37 [+ or -] 2.02     410.43 [+ or -] 3.81
week     weight (g)
         Weekly        260.96 [+ or -] 0.84     258.40 [+ or -] 2.91
         weight
         gain (g)
         Avg. feed     348.70 [+ or -] 1.01     350.23 [+ or -] 0.98
         intake (g)
         FCR             1.34 [+ or -] 0.01       1.36 [+ or -] 0.02
3rd      Avg. body     735.50 [+ or -] 3.62     729.00 [+ or -] 4.04
week     weight (g)
         Weekly        323.13 [+ or -] 5.21     318.56 [+ or -] 7.71
         weight
         gain (g)
         Avg. feed     513.93 [+ or -] 1.19     523.57 [+ or -] 5.06
         intake (g)
         FCR             1.58 [+ or -] 0.03       1.64 [+ or -] 0.02
4th      Avg. body   1,138.20 [+ or -] 6.55   1,039.21 [+ or -] 10.43
week     weight (g)            (a)                      (b)
         Weekly        402.71 [+ or -] 8.11     310.21 [+ or -] 10.78
         weight                 (a)                      (b)
         gain (g)
         Avg. feed     741.17 [+ or -] 6.12     733.62 [+ or -] 3.95
         intake (g)
         FCR             1.84 [+ or -] 0.02       2.37 [+ or -] 0.09
                                 (b)                      (a)
5th      Avg. body   1,672.21 [+ or -] 14.36  1,485.88 [+ or -] 19.83
week     weight (g)            (a)                      (b)
         Weekly        534.00 [+ or -] 11.38    446.67 [+ or -] 28.81
         weight                 (ab)                     (b)
         gain (g)
         Avg. feed   1,062.38 [+ or -] 2.67   1,039.46 [+ or -] 4.78
         intake (g)            (b)                      (c)
         FCR             1.99 [+ or -] 0.04       2.35 [+ or -] 0.17

Growth   Parameters  p-value
phase

1st      Avg. body    0.545
week     weight (g)
         Weekly       0.545
         weight
         gain (g)
         Avg. feed    0.042
         intake (g)
         FCR          0.092

2nd      Avg. body    0.296
week     weight (g)
         Weekly       0.181
         weight
         gain (g)
         Avg. feed    0.369
         intake (g)
         FCR          0.172
3rd      Avg. body    0.095
week     weight (g)
         Weekly       0.433
         weight
         gain (g)
         Avg. feed    0.674
         intake (g)
         FCR          0.532
4th      Avg. body    0.003
week     weight (g)
         Weekly       0.001
         weight
         gain (g)
         Avg. feed    0.513
         intake (g)
         FCR          0.001

5th      Avg. body    0.000
week     weight (g)
         Weekly       0.107
         weight
         gain (g)
         Avg. feed    0.000
         intake (g)
         FCR          0.301

SB-1, SB-0.5, sodium butyrate 1.0 g/kg, 0.5 g/kg); ZnB, zinc
bacitracin 0.01% of feed; FCR, feed conversion ratio.

Means within a row marked with different superscripts were
significantly different (p<0.05).

Values represent mean [+ or -] standard error of the mean.

Table 3. Effect of sodium butyrate and zinc bacitracin on cell
mediated immune response against PHA-P in broilers

Time                 Skin thickness (mm) after PHA-P
interval               injection in various groups
post
injection             SB-1                     SB-0.5
(h)

24           0.71 [+ or -] 0.04        0.66 [+ or -] 0.04
48           0.65 [+ or -] 0.02 (a)    0.55 [+ or -] 0.03 (ab)
72           0.48 [+ or -] 0.05        0.45 [+ or -] 0.05

Time                 Skin thickness (mm) after PHA-P
interval               injection in various groups
post                                                           p-value
injection             ZnB                    Control
(h)

24           0.65 [+ or -] 0.04       0.57 [+ or -] 0.05        0.209
48           0.51 [+ or -] 0.03 (b)   0.49 [+ or -] 0.06 (b)    0.029
72           0.39 [+ or -] 0.02       0.38 [+ or -] 0.04        0.314

PHA-P, phytohemagglutinin-P; SB-1, SB-0.5, sodium butyrate 1.0 g/kg,
0.5 g/kg; ZnB, zinc bacitracin 0.01% of feed.

Means within a row marked with different superscripts were
significantly different (p<0.05).

Values represent mean [+ or -] standard error of the mean.

Table 4. Effect of sodium butyrate and zinc bacitracin on humoral
immune response in broilers

              Antibody titre ([log.sub.2]) on various days

Treatments                       NDV

                   Day-21                 Day-35

SB-1         2.19 [+ or -] 0.12   2.31 [+ or -] 0.08 (a)
SB-0.5       2.16 [+ or -] 0.07   2.21 [+ or -] 0.07 (ab)
ZnB          2.09 [+ or -] 0.12   1.98 [+ or -] 0.09 (b)
Control      1.98 [+ or -] 0.11   1.98 [+ or -] 0.09 (b)
p-value            0.290                   0.026

              Antibody titre ([log.sub.2]) on various days

Treatments                         SRBC

                   Day-21                 Day-35

SB-1         6.00 [+ or -] 0.37   8.67 [+ or -] 0.42 (a)
SB-0.5       5.50 [+ or -] 0.22   7.17 [+ or -] 0.37 (ab)
ZnB          5.66 [+ or -] 0.33   7.16 [+ or -] 0.30 (bc)
Control      5.16 [+ or -] 0.30   6.00 [+ or -] 0.36 (c)
p-value            0.322                   0.000

NDV, Newcastle disease virus; SRBC, sheep red blood cells; SB-1,
SB-0.5, sodium butyrate 1.0 g/kg, 0.5 g/kg; ZnB, zinc bacitracin
0.01% of feed.

Means within a column marked with different superscripts were
significantly different (p < 0.05).

Values represent mean [+ or -] standard error of the mean.

Table 5. Effect of sodium butyrate and zinc bacitracin on the
relative organs weight in broilers

                       Relative organs weight (1)

Treatments                      At day-21

                     Liver                 Spleen

SB-1         2.2272 [+ or -] 0.19   0.1027 [+ or -] 0.01
SB-0.5       2.1582 [+ or -] 0.15   0.0974 [+ or -] 0.02
ZnB          2.1210 [+ or -] 0.14   0.0953 [+ or -] 0.03
Control      1.8735 [+ or -] 0.09    0.078 [+ or -] 0.01
p-value              0.395                  0.199

                       Relative organs weight (1)

Treatments                      At day-21

                       Thymus                      Bursa

SB-1         0.6206 [+ or -] 0.03 (a)    0.3125 [+ or -] 0.03 (a)
SB-0.5       0.5895 [+ or -] 0.03 (ab)   0.2807 [+ or -] 0.03 (a)
ZnB          0.5221 [+ or -] 0.02 (ab)   0.2700 [+ or -] 0.04 (ab)
Control      0.5479 [+ or -] 0.04 (b)    0.1804 [+ or -] 0.02 (b)
p-value                0.146                       0.044

                       Relative organs weight (1)

Treatments                       At day-35

                     Liver                    Spleen

SB-1         1.7085 [+ or -] 0.02   0.1646 [+ or -] 0.02 (a)
SB-0.5       1.6320 [+ or -] 0.05   0.1418 [+ or -] 0.00 (ab)
ZnB          1.5971 [+ or -] 0.05   0.1435 [+ or -] 0.02 (ab)
Control      1.5753 [+ or -] 0.05   0.0960 [+ or -] 0.01 (b)
p-value              0.172                    0.065

                       Relative organs weight (1)

Treatments                       At day-35

                      Thymus                    Bursa

SB-1         0.3861 [+ or -] 0.00 (a)   0.1667 [+ or -] 0.02
SB-0.5       0.3721 [+ or -] 0.00 (a)   0.1550 [+ or -] 0.02
ZnB          0.3857 [+ or -] 0.00 (a)   0.1617 [+ or -] 0.02
Control      0.3184 [+ or -] 0.01 (b)   0.1167 [+ or -] 0.02
p-value                0.000            0.295

SB/1, SB/0.5, sodium butyrate 1.0 g/kg, 0.5 g/kg; ZnB, zinc
bacitracin 0.01% of feed.

(1) Relative organs weight = organ weight/body weight x 100.

Means within a column marked with different superscripts were
significantly different (p < 0.05).

Mean [+ or -] standard error of the mean.

Table 6. Effect of sodium butyrate and zinc bacitracin on immune
organ morphology of broilers during starter phase

                                        Treatments
Organs  Parameters
                              SB-1                    SB-0.5

Thymus  Thymic        308.33 [+ or -] 24.71     292.33 [+ or -] 18.65
        cortex
        ([micro]m)
        Thymic        604.50 [+ or -] 21.13     522.00 [+ or -] 17.02
        medulla                (a)                       (b)
        ([micro]
        [m.sup.2])
        Cortex:       0.5050 [+ or -] .03       0.5633 [+ or -] .04
        medulla

Spleen  Germinal      0.7636 [+ or -] 0.02      0.7361 [+ or -] 0.01
        center/                (a)                       (ab)
        field area
        (%)

Bursa   Bursal        508.33 [+ or -] 61.15     500.67 [+ or -] 21.78
        follicular
        length
        ([micro]m)
        Bursal        161.17 [+ or -] 7.53      151.17 [+ or -] 9.55
        follicular
        width
        ([micro]m)
        Bursal      81,648.6 [+ or -] 3562.2  75,901.3 [+ or -] 6365.3
        follicular
        area
        ([micro]
        [m.sup.2])
        Bursal          9.50 [+ or -] 0.43        8.33 [+ or -] 0.42
        follicular
        number

                                        Treatments
Organs  Parameters
                              ZnB                   Control

Thymus  Thymic        295.83 [+ or -] 12.24   288.17 [+ or -] 11.49
        cortex
        ([micro]m)
        Thymic        523.67 [+ or -] 26.29   494.00 [+ or -] 25.26
        medulla                (b)                     (b)
        ([micro]
        [m.sup.2])
        Cortex:       0.5741 [+ or -] .04     0.5912 [+ or -] .04
        medulla

Spleen  Germinal      0.6465 [+ or -] 0.04    0.6449 [+ or -] 0.03
        center/                (b)                     (b)
        field area
        (%)

Bursa   Bursal        485.50 [+ or -] 29.20   468.67 [+ or -] 44.10
        follicular
        length
        ([micro]m)
        Bursal        155.50 [+ or -] 8.29    142.67 [+ or -] 8.75
        follicular
        width
        ([micro]m)
        Bursal      75,004.3 [+ or -] 5042.7  66,691 [+ or -] 7330.1
        follicular
        area
        ([micro]
        [m.sup.2])
        Bursal          8.83 [+ or -] 0.48      8.67 [+ or -] 0.71
        follicular
        number

Organs  Parameters   p-value

Thymus  Thymic       0.865
        cortex
        ([micro]m)
        Thymic       0.016
        medulla
        ([micro]
        [m.sup.2])
        Cortex:      0.464
        medulla

Spleen  Germinal     0.032
        center/
        field area
        (%)

Bursa   Bursal       0.791
        follicular
        length
        ([micro]m)
        Bursal       0.493
        follicular
        width
        ([micro]m)
        Bursal       0.354
        follicular
        area
        ([micro]
        [m.sup.2])
        Bursal       0.470
        follicular
        number

SB-1, SB-0.5, sodium butyrate 1.0 g/kg, 0.5 g/kg; ZnB, zinc
bacitracin 0.01% of feed.

Means within a row marked with different superscripts were
significantly different (p<0.05).

Values represent mean [+ or -] standard error of the mean.

Table 7. Effect of sodium butyrate and zinc bacitracin on immune
organ morphology in broilers during grower phase

                                        Treatments
Organs  Parameters
                              SB-1                     SB-0.5

Thymus  Thymic        271.83 [+ or -] 12.84    242.67 [+ or -] 13.29
        cortex
        ([micro]m)
        Thymic        432.50 [+ or -] 26.39    398.83 [+ or -] 18.66
        medulla
        ([micro]
        [m.sup.2])
        Cortex:       0.6406 [+ or -] 0.05     0.6151 [+ or -] 0.04
        medulla

Spleen  Germinal      0.8876 [+ or -] 0.08     0.8619 [+ or -] 0.08 (a)
        center/       (a)
        field area
        (%)

Bursa   Bursal        398.50 [+ or -] 23.37    389.50 [+ or -] 12.48
        follicular
        length
        ([micro]m)
        Bursal        142.83 [+ or -] 13.79    124.50 [+ or -] 15.89
        follicular
        width
        ([micro]m)
        Bursal      56,326.3 [+ or -]        47,595.5 [+ or -]
        follicular          5,067.40                  4,839.20
        area
        ([micro]
        [m.sup.2])
        Bursal          8.17 [+ or -] 0.98       7.17 [+ or -] 1.83
        follicular
        number

                                        Treatments
Organs  Parameters
                              ZnB                    Control

Thymus  Thymic        257.67 [+ or -] 12.96    234.83 [+ or -] 17.91
        cortex
        ([micro]m)
        Thymic        404.67 [+ or -] 27.33    364.50 [+ or -] 14.18
        medulla
        ([micro]
        [m.sup.2])
        Cortex:       0.6473 [+ or -] 0.04     0.6449 [+ or -] 0.04
        medulla

Spleen  Germinal      0.7550 [+ or -] 0.05     0.5329 [+ or -] 0.09
        center/       (ab)                     (b)
        field area
        (%)

Bursa   Bursal        373.67 [+ or -] 21.53    373.17 [+ or -] 17.56
        follicular
        length
        ([micro]m)
        Bursal        134.50 [+ or -] 18.24    104.67 [+ or -] 9.97
        follicular
        width
        ([micro]m)
        Bursal      50,285.0 [+ or -]        38,022.5 [+ or -]
        follicular          7,721.80                 3,932.60
        area
        ([micro]
        [m.sup.2])
        Bursal          7.50 [+ or -] 0.67       7.00 [+ or -] 0.58
        follicular
        number

                     p-value
Organs  Parameters

Thymus  Thymic       0.303
        cortex
        ([micro]m)
        Thymic       0.234
        medulla
        ([micro]
        [m.sup.2])
        Cortex:      0.950
        medulla

Spleen  Germinal     0.018
        center/
        field area
        (%)

Bursa   Bursal       0.730
        follicular
        length
        ([micro]m)
        Bursal       0.691
        follicular
        width
        ([micro]m)
        Bursal       0.509
        follicular
        area
        ([micro]
        [m.sup.2])
        Bursal       0.559
        follicular
        number

SB/1, SB/0.5, sodium butyrate 1.0 g/kg, 0.5 g/kg; ZnB, zinc
bacitracin 0.01% of feed.

Means within a row marked with different superscripts were
significantly different (p<0.05).

Values represent mean [+ or -] standard error of the mean.

Table 8. Effect of sodium butyrate and zinc bacitracin on small
intestine morphology of broilers on d-21

                                            Treatments
Intestina  Parameters
segments                        SB-1                    SB-0.5

Duodenum   Villus      1,321.12 [+ or -] 29.17  1,316.29 [+ or -] 28.89
           height                (a)                      (a)
           ([micro]m)
           Villus        138.76 [+ or -] 7.73     133.20 [+ or -] 9.52
           width
           ([micro]m)
           Villus         0.578 [+ or -] 0.04      0.552 [+ or -] 0.05
           surface               (a)                      (a)
           area
           ([mm.sup.
           2])
           Crypt         136.50 [+ or -] 6.07     143.67 [+ or -] 3.13
           depth                (ab)                      (a)
           ([micro]m)
           VH:CD           9.73 [+ or -] 0.24       9.17 [+ or -] 0.18
                                 (a)                     (ab)

Jejunum    Villus        928.17 [+ or -] 16.73    899.00 [+ or -] 18.73
           height                (a)                      (a)
           ([micro]m)
           Villus        105.37 [+ or -] 3.63     105.61 [+ or -] 4.17
           width
           ([micro]m)
           Villus         0.307 [+ or -] 0.09      0.297 [+ or -] 0.08
           surface               (a)                      (a)
           area
           ([mm.sup.
           2])
           Crypt         106.13 [+ or -] 2.42     106.55 [+ or -] 3.79
           depth
           ([micro]m)
           VH:CD           8.77 [+ or -] 0.30       8.47 [+ or -] 0.22
                                 (a)                      (a)

Ileum      Villus        494.10 [+ or -] 19.55    474.08 [+ or -] 18.07
           height                (a)                     (ab)
           ([micro]m)
           Villus        104.93 [+ or -] 2.50     104.87 [+ or -] 2.75
           width
           ([micro]m)
           Villus         0.163 [+ or -] 0.09      0.156 [+ or -] 0.082
           surface               (a)                     (ab)
           area
           ([mm.sup.
           2])
           Crypt         112.30 [+ or -] 2.56     111.05 [+ or -] 3.11
           depth
           ([micro]m)
           VH:CD           4.40 [+ or -] 0.16       4.27 [+ or -] 0.14
                                   (a)                     (ab)

                                            Treatments
Intestina  Parameters
segments                         ZnB                    Control

Duodenum   Villus      l,019.86 [+ or -] 17.35  1,033.25 [+ or -] 14.83
           height                (b)                      (b)
           ([micro]m)
           Villus        138.76 [+ or -] 17.27    119.62 [+ or -] 6.49
           width
           ([micro]m)
           Villus         0.453 [+ or -] 0.05      0.388 [+ or -] 0.02
           surface              (ab)                      (b)
           area
           ([mm.sup.
           2])
           Crypt         118.30 [+ or -] 2.52     126.58 [+ or -] 4.19
           depth                 (c)                     (bc)
           ([micro]m)
           VH:CD           8.63 [+ or -] 0.18       8.21 [+ or -] 0.33
                                (bc)                      (c)

Jejunum    Villus        686.00 [+ or -] 10.77    658.00 [+ or -] 17.64
           height                (b)                      (b)
           ([micro]m)
           Villus        107.17 [+ or -] 4.29     107.53 [+ or -] 1.94
           width
           ([micro]m)
           Villus         0.232 [+ or -] 0.07      0.222 [+ or -] 0.04
           surface               (b)                      (b)
           area
           ([mm.sup.
           2])
           Crypt         104.07 [+ or -] 6.35     106.68 [+ or -] 2.75
           depth
           ([micro]m)
           VH:CD           6.76 [+ or -] 0.56       6.19 [+ or -] 0.22
                                 (b)                      (b)

Ileum      Villus        422.03 [+ or -] 16.43    417.20 [+ or -] 17.87
           height               (bc)                      (c)
           ([micro]m)
           Villus        102.48 [+ or -] 2.19      99.82 [+ or -] 3.33
           width
           ([micro]m)
           Villus         0.136 [+ or -] 0.06      0.131 [+ or -] 0.09
           surface               (b)                      (b)
           area
           ([mm.sup.
           2])
           Crypt         109.88 [+ or -] 3.75     110.93 [+ or -] 2.99
           depth
           ([micro]m)
           VH:CD           3.85 [+ or -] 0.17       3.79 [+ or -] 0.26
                                  (ab)                      (b)

Intestina  Parameters   p-value
segments

Duodenum   Villus       0.000
           height
           ([micro]m)
           Villus       0.521
           width
           ([micro]m)
           Villus       0.016
           surface
           area
           ([mm.sup.
           2])
           Crypt        0.002
           depth
           ([micro]m)
           VH:CD        0.002

Jejunum    Villus       0.000
           height
           ([micro]m)
           Villus       0.935
           width
           ([micro]m)
           Villus       0.000
           surface
           area
           ([mm.sup.
           2])
           Crypt        0.960
           depth
           ([micro]m)
           VH:CD        0.000

Ileum      Villus       0.015
           height
           ([micro]m)
           Villus       0.513
           width
           ([micro]m)
           Villus       0.031
           surface
           area
           ([mm.sup.
           2])
           Crypt        0.959
           depth
           ([micro]m)
           VH:CD        0.083

SB-1, SB-0.5, sodium butyrate 1.0 g/kg, 0.5 g/kg; ZnB, zinc
bacitracin 0.01% of feed; VH, villus height; CD, crypt depth.

Means within a row marked with different superscripts were
significantly different (p<0.05).

Values represent mean + standard error of the mean.

Table 9. Effect of sodium butyrate and zinc bacitracin on small
intestine morphology in broilers on d-35

                                         Treatments
Intestinal  Parameters
segments                       SB-1                  SB-0.5

Duodenum    Villus      1,470.62 [+ or -]      1,290.63 [+ or -]
            height           33.99 (a)              17.22 (b)
            ([micro]m)
            Villus      180.44 [+ or -] 6.39   159.87 [+ or -] 5.39
            width                (a)                   (ab)
            ([micro]m)
            Villus       0.832 [+ or -] 0.03    0.607 [+ or -] 0.02
            surface              (a)                    (b)
            area
            ([mm.sup.
            2])
            Crypt       137.92 [+ or -] 9.49   140.49 [+ or -] 9.20
            depth
            ([micro]m)
            VH:CD        10.87 [+ or -] 0.64     9.18 [+ or -] 0.59
                                 (a)                     (b)

Jejunum     Villus      835.06 [+ or -] 43.73  772.19 [+ or -] 2495
            height               (a)                   (ab)
            ([micro]m)
            Villus      127.12 [+ or -] 6.41   103.22 [+ or -] 3.91
            width                (a)                    (b)
            ([micro]m)
            Villus      0.3362 [+ or -] 0.03   0.2493 [+ or -] 0.01
            surface              (a)                    (b)
            area
            ([mm.sup.
            2])
            Crypt       130.55 [+ or -] 3.94   128.41 [+ or -] 3.04
            depth
            ([micro]m)
            VH:CD         6.39 [+ or -] 0.27     6.03 [+ or -] 0.27

Ileum       Villus      460.67 [+ or -] 14.63  451.86 [+ or -] 6.96
            height               (a)                   (ab)
            ([micro]m)
            Villus      114.34 [+ or -] 3.01   107.37 [+ or -] 6.01
            width
            ([micro]m)
            Villus      0.1652 [+ or -] 0.01   0.1520 [+ or -] 0.01
            surface              (a)                   (ab)
            area
            ([mm.sup.
            2])
            Crypt       105.22 [+ or -] 2.26   111.05 [+ or -] 2.69
            depth
            ([micro]m)
            VH:CD         4.38 [+ or -] 0.09     4.08 [+ or -] 0.08

                                           Treatments
Intestinal  Parameters
segments                        ZnB                   Control

Duodenum    Villus       1,116.70 [+ or -]       1,102.41 [+ or -]
            height           27.02 (bc)              44.49 (c)
            ([micro]m)
            Villus      151.60 [+ or -] 19.34   129.62 [+ or -] 6.48
            width                (ab)                    (b)
            ([micro]m)
            Villus       0.532 [+ or -] 0.07     0.449 [+ or -] 0.03
            surface              (bc)                    (c)
            area
            ([mm.sup.
            2])
            Crypt       113.30 [+ or -] 2.20    119.85 [+ or -] 12.93
            depth
            ([micro]m)
            VH:CD         9.87 [+ or -] 0.25      9.34 [+ or -] 0.59
                                  (ab)                    (b)

Jejunum     Villus      643.30 [+ or -] 30.77  685.1 5 [+ or -] 14.61
            height               (c)                    (bc)
            ([micro]m)
            Villus      112.85 [+ or -] 4.64    110.70 [+ or -] 5.69
            width                (ab)                    (b)
            ([micro]m)
            Villus      0.2280 [+ or -] 0.01    0.2387 [+ or -] 0.01
            surface              (b)                     (b)
            area
            ([mm.sup.
            2])
            Crypt       121.79 [+ or -] 5.02    124.76 [+ or -] 9.62
            depth
            ([micro]m)
            VH:CD         5.37 [+ or -] 0.46      5.75 [+ or -] 0.52

Ileum       Villus      436.17 [+ or -] 8.09    429.44 [+ or -] 5.66
            height               (ab)                    (b)
            ([micro]m)
            Villus      104.85 [+ or -] 7.14    102.39 [+ or -] 5.18
            width
            ([micro]m)
            Villus      0.1432 [+ or -] 0.01    0.1383 [+ or -] 0.01
            surface              (ab)                    (b)
            area
            ([mm.sup.
            2])
            Crypt       105.36 [+ or -] 3.18    104.78 [+ or -] 1.94
            depth
            ([micro]m)
            VH:CD         4.16 [+ or -] 0.14      4.10 [+ or -] 0.09

Intestinal  Parameters   p-value
segments

Duodenum    Villus       0.000
            height
            ([micro]m)
            Villus       0.030
            width
            ([micro]m)
            Villus       0.000
            surface
            area
            ([mm.sup.
            2])
            Crypt        0.174
            depth
            ([micro]m)
            VH:CD        0.015

Jejunum     Villus       0.001
            height
            ([micro]m)
            Villus       0.031
            width
            ([micro]m)
            Villus       0.003
            surface
            area
            ([mm.sup.
            2])
            Crypt        0.689
            depth
            ([micro]m)
            VH:CD        0.330

Ileum       Villus       0.112
            height
            ([micro]m)
            Villus       0.477
            width
            ([micro]m)
            Villus       0.111
            surface
            area
            ([mm.sup.
            2])
            Crypt        0.264
            depth
            ([micro]m)
            VH:CD        0.203

SB-1, SB-0.5, sodium butyrate 1.0 g/kg, 0.5 g/kg; ZnB, zinc
bacitracin 0.01% of feed; VH, villus height; CD, crypt depth.

Means within a row marked with different superscripts were
significantly different (p< 0.05).

Values represent mean [+ or -] standard error of the mean.

Table 10. Effect of sodium butyrate and zinc bacitracin on
intraepithelial lymphocytes count

                                 Day

Treatments                       21

                   Duodenum              Jejunum

SB-1         41.17 [+ or -] 2.26   44.33 [+ or -] 1.67
SB-0.5       43.83 [+ or -] 1.64   40.83 [+ or -] 2.21
ZnB          43.66 [+ or -] 2.84   47.33 [+ or -] 1.52
Control      43.00 [+ or -] 4.52   39.50 [+ or -] 5.56
p-value             0.919                 0.327

                                 Day

Treatments            21                    35

                    Ileum                Duodenum

SB-1         45.83 [+ or -] 2.17   68.83 [+ or -] 3.07
SB-0.5       45.67 [+ or -] 1.41   66.50 [+ or -] 1.67
ZnB          44.33 [+ or -] 2.15   65.83 [+ or -] 2.65
Control      42.00 [+ or -] 1.93   63.66 [+ or -] 6.99
p-value             0.491                 0.850

                                 Day

Treatments                        35

                   Jejunum                Ileum

SB-1         56.17 [+ or -] 2.25   56.17 [+ or -] 2.06
SB-0.5       46.83 [+ or -] 4.06   51.50 [+ or -] 2.87
ZnB          57.16 [+ or -] 3.62   60.50 [+ or -] 2.53
Control      49.33 [+ or -] 4.49   48.66 [+ or -] 4.30
p-value             0.187                 0.060

SB-1, SB-0.5, sodium butyrate 1.0 g/kg, 0.5 g/kg; ZnB, zinc
bacitracin 0.01% of feed.

Means within a column marked with different superscripts were
significantly different (p < 0.05).

Values represent mean [+ or -] standard error of the mean.

Table 11. Effect of sodium butyrate and zinc bacitracin on
differentiated goblet cells count in small intestine of broilers on
d-21

Intestinal   Goblet                    Treatments
segments      cell
             mucin            SB-1                 SB-0.5
              type

Duodenum     Acidic   47.50 [+ or -] 2.94   44.00 [+ or -] 3.88
                              (a)                   (ab)
             Mixed    38.00 [+ or -] 3.88   38.67 [+ or -] 4.51
             Total    85.50 [+ or -] 4.02   82.67 [+ or -] 6.46
Jejunum      Acidic   50.67 [+ or -] 3.60   44.00 [+ or -] 3.88
                              (a)                   (ab)
             Mixed    38.83 [+ or -] 4.66   38.67 [+ or -] 4.50
             Total    89.50 [+ or -] 4.48   82.67 [+ or -] 6.46
Ileum        Acidic   45.00 [+ or -] 2.73   43.67 [+ or -] 1.56
                              (a)                   (ab)
             Mixed    39.17 [+ or -] 2.36   37.83 [+ or -] 1.64
             Total    82.83 [+ or -] 2.44   82.83 [+ or -] 3.96

Intestinal   Goblet                    Treatments
segments      cell
             mucin              ZnB                  Control
              type

Duodenum     Acidic   40.50 [+ or -] 4.22      34.67 [+ or -] 1.16
                               (ab)                    (b)
             Mixed    41.00 [+ or -] 3.62      42.17 [+ or -] 3.37
             Total    81.50 [+ or -] 4.77      76.83 [+ or -] 3.98
Jejunum      Acidic   40.50 [+ or -] .4.22     36.33 [+ or -] .80
                               (ab)                    (b)
             Mixed    41.00 [+ or -] 3.62      42.16 [+ or -] 3.47
             Total    81.50 [+ or -] 4.77      78.50 [+ or -] 3.89
Ileum        Acidic   44.17 [+ or -] 2.24      38.17 [+ or -] 174
                               (ab)                    (b)
             Mixed    38.83 [+ or -] 2.73.62   41.83 [+ or -] 2.55
             Total    83.00 [+ or -] 2.52      80.00 [+ or -] 4.06

             Goblet
Intestinal    cell
segments     mucin    p-value
              type

Duodenum     Acidic    0.066
             Mixed     0.857
             Total     0.661
Jejunum      Acidic    0.047
             Mixed     0.912
             Total     0.474
Ileum        Acidic    0.126
             Mixed     0.669
             Total     0.903

Means within a row marked with different superscripts were
significantly different (p<0.05).

Values represent mean [+ or -] standard error of the mean.

SB/1, SB/0.5, sodium butyrate 1.0 g/kg, 0.5 g/kg; ZnB, zinc
bacitracin 0.01% of feed.

Table 12. Effect of sodium butyrate and zinc bacitracin on
differentiated goblet cells count in small intestine of broilers on
d-35

             Goblet
Intestinal    cell                     Treatments
segments     mucin
              type            SB-1                 SB-0.5

Duodenum     Acidic   53.67 [+ or -] 1.81   53.83 [+ or -] 9.17
             Mixed    37.33 [+ or -] 2.17   38.83 [+ or -] 3.34
             Total    91.83 [+ or -] 3.43   92.67 [+ or -] 7.65
Jejunum      Acidic   51.33 [+ or -] 4.26   50.83 [+ or -] 5.54
             Mixed    42.33 [+ or -] 1.28   45.67 [+ or -] 3.68
             Total    93.67 [+ or -] 3.40   96.50 [+ or -] 4.19
Ileum        Acidic   53.17 [+ or -] 3.02   45.67 [+ or -] 2.12
                              (a)                   (b)
             Mixed    32.83 [+ or -] 2.14   39.33 [+ or -] 2.89
             Total    86.00 [+ or -] 4.82   85.00 [+ or -] 4.02

             Goblet
Intestinal    cell                     Treatments
segments     mucin
              type            ZnB                 Control

Duodenum     Acidic   51.16 [+ or -] 3.55   47.83 [+ or -] 232
             Mixed    42.00 [+ or -] 4.13   45.16 [+ or -] 3.96
             Total    93.16 [+ or -] 4.41   93.00 [+ or -] 5.48
Jejunum      Acidic   44.00 [+ or -] 3.94   39.50 [+ or -] 1.94
             Mixed    42.67 [+ or -] 2.66   47.66 [+ or -] 2.61
             Total    86.67 [+ or -] 4.63   87.17 [+ or -] 4.35
Ileum        Acidic   43.50 [+ or -] 1.89   39.17 [+ or -] 2.04
                              (b)                   (b)
             Mixed    35.33 [+ or -] 2.79   36.17 [+ or -] 2.12
             Total    78.83 [+ or -] 3.38   75.33 [+ or -] 1.15

             Goblet
Intestinal    cell
segments     mucin
              type    p-value

Duodenum     Acidic    0.826
             Mixed     0.414
             Total     0.992
Jejunum      Acidic    0.161
             Mixed     0.465
             Total     0.284
Ileum        Acidic    0.003
             Mixed     0.356
             Total     0.149

SB-1, SB-0.5, sodium butyrate 1.0 g/kg, 0.5 g/kg; ZnB, zinc
bacitracin 0.01% of feed.

Means within a row marked with different superscripts were
significantly different (p<0.05).

Values represent mean [+ or -] standard error of the mean.
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Author:Sikandar, Arbab; Zaneb, Hafsa; Younus, Muhammad; Masood, Saima; Aslam, Asim; Khattak, Farina; Ashraf
Publication:Asian - Australasian Journal of Animal Sciences
Article Type:Report
Date:May 1, 2017
Words:9155
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