Intestinal IgA+ Cell Numbers as well as IgA, IgG, and IgM Contents Correlate with Mucosal Humoral Immunity of Broilers During Supplementation with High Fluorine in the Diets

Fluoride (F), a well-recognized harmful substance, is easily absorbed by the intestinal mucosa. The intestinal mucosal immune system is equipped with unique innate and adaptive defense mechanisms that provide a first line of protection against infectious agents. Meanwhile, immunoglobulins are the major secretory products of the adaptive immune system and their levels can be a strong indicator of a disease or condition. In this study, therefore, we investigated the effects of high dietary fluorine on the numbers of immunoglobulin A-positive (IgA⁺) cells in the lamina propria of intestines (duodenum, jejunum and ileum) by immunohistochemistry as well as on the contents of immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) in the mucosa of intestines (duodenum, jejunum, and ileum) by enzyme-linked immunosorbent assay (ELISA). A total of 280 1-day-old healthy avian broilers were randomly divided into four groups and fed on a corn–soybean basal diet as control diet (fluorine 22.6 mg/kg) or the same basal diet supplemented with 400, 800, and 1,200 mg/kg fluorine (high fluorine groups I, II, and III) in the form of sodium fluoride (NaF) for 42 days. The experimental data showed that the numbers of IgA⁺ cells as well as the IgA, IgG, and IgM contents were significantly decreased (P?<?0.01 or P?<?0.05) in the high fluorine groups II and III when compared with those of the control group. It was concluded that dietary fluorine in the range of 800–1,200 mg/kg significantly reduced the numbers of the IgA⁺ cells and the contents of aforementioned immunoglobulins in the intestines (duodenum, jejunum, and ileum) of broilers, which could finally impact the mucosal humoral immune function in the intestines by a way that reduces the lymphocyte population and/or lymphocyte activation.     

Effects of High Dietary Fluorine on Erythrocytes and Erythrocyte Immune Adherence Function in Broiler Chickens

Fluoride can exert toxic effects on soft tissues, giving rise to a broad array of symptoms and pathological changes. The aim of this study was to investigate on erythrocytes and erythrocyte immune adherence function in broiler chickens fed with high fluorine (F) diets by measuring the total erythrocyte count (TEC), the contents of hemoglobin (Hb), packed cell volumn (PCV), erythrocyte osmotic fragility (EOF), erythrocyte C_3b receptor rosette rate (E-C_3bRR), and erythrocyte immune complex rosette rate (E-ICRR). A total of 280 1-day-old healthy avian broiler chickens were randomly allotted into four equal groups of 70 birds each and fed with a corn–soybean basal diet containing 22.6 mg F/kg (control group) or same basal diets supplemented with 400, 800, and 1,200 mg F/kg (high F groups I, II, and III) in the form of sodium fluoride for 42 days. Blood samples were collected for the abovementioned parameters analysis at 14, 28, and 42 days of age during the experiment. The experimental results indicated that TEC, Hb, and PCV were significantly lower (p?<?0.05 or p?<?0.01), and EOF was higher (p?<?0.05 or p?<?0.01) in the high F groups II and III than that in the control group from 14 to 42 days of age. The E-C_3bRR was significantly decreased (p?<?0.01) in the three high F groups, whereas the E-ICRR was markedly increased (p?<?0.01) in the high F groups II and III from 14 to 42 days of age. It was concluded that dietary F in the range of 800 to 1, 200 mg/kg could significantly cause anemia and impair the integrity of erythrocyte membrane, the transport capacity of oxygen and carbon dioxide, and erythrocyte immune adherence function in broiler chickens.     

Dietary Nickel Chloride Restrains the Development of Small Intestine in Broilers

The aim of this study was to determine the effects of dietary NiCl₂ on the development of the small intestine in broilers by the methods of light microscopy, histochemistry and enzyme-linked immunosorbent assay. A total of 240 one-day-old avian broilers were divided into four groups and fed on a corn–soybean basal diet or the same basal diet supplemented with 300, 600 and 900 mg/kg of nickel chloride (NiCl₂) for 42 days. Results showed that the small intestinal villus height, crypt depth and villus/crypt ratio were significantly decreased, and also the small intestinal goblet cells numbers and insulin-like growth factor-1 (IGF-1) and epidermal growth factor (EGF) contents were significantly decreased in the 300-, 600- and 900-mg/kg groups when compared with those of the control group. In conclusion, dietary NiCl₂ in excess of 300 mg/kg reduced the villus height, crypt depth, the goblet cells population and the IGF-1 and EGF contents in the small intestine, indicating that the normal development and function of the small intestine were finally impaired in broilers. This study firstly provided the new experimental information for future studies on the effects of NiCl₂ on the intestinal function in humans and other animals.     

Efficacy of dietary supplemental insoluble fibrous materials in ameliorating adverse effects of coccidial challenge in broiler chickens

ABSTRACT

The current experiment was designed to examine effects of dietary supplemental sunflower hulls (SH) and rice hulls (RH) on growth performance, carcass traits, intestinal morphology, lesion score and oocyst shedding in broiler chickens exposed to coccidial challenge. A total of 540 broiler chickens (Ross 308) were assigned to six dietary treatments based on a factorial arrangement (2 × 3) across 1–14, 14–28 and 28-42-d periods. Experimental treatments consisted of broiler chickens without or with coccidial challenge each offered with three different diets: a basal diet or basal diet supplemented with either RH or SH at 40 g/kg diet, respectively. Infection with Eimeria impaired daily weight gain (DWG) and feed conversion ratio (FCR) of broiler chickens during growing period (p < 0.05) while supplementation of SH or RH reduced the adverse effect of coccidiosis so that birds had similar DWG to those fed the basal diet without infection. However, only dietary SH improved the FCR of broilers challenged with coccidiosis. Regardless of coccidial challenge, dietary access to insoluble fibre improved performance of broilers across the growing period (p < 0.05); however, this effect was not observed during the entire rearing period. Relative weights of liver and pancreas were increased in birds subjected to coccidial challenge on d 21 of age (p < 0.05). Moreover, relative weights of the intestinal segments were enhanced (p < 0.05). Furthermore, gizzard weights were higher in birds receiving diets added with fibre (p < 0.05). Infection with coccidiosis decreased villus height and villus height to crypt depth ratio in duodenum of broilers which received the basal diet compared with those fed the same feed without coccidial challenge (p < 0.05). However, supplemental SH could decrease the negative effect of infection on the noted intestinal morphometric attributes. Similarly, a marked reduction was observed for lesion score and faecal oocyst excretion of challenged broilers fed on dietary supplemental fibre (p < 0.05). In conclusion, supplementation of insoluble fibre could ameliorate negative effects of coccidial challenge on DWG of broiler chickens and inclusion of SH in diet of birds exposed to Eimeria infection could be recommended.     

The Association Between Cytokines and Intestinal Mucosal Immunity Among Broilers Fed on Diets Supplemented with Fluorine

Fluorine (F) bioaccumulation has been reported in the organs and tissues of organisms, including intestine. The intestinal mucosa is very important to the immune development. Meanwhile, cytokines are present in the normal intestinal mucosal and play an important role in the immune function. Thus, changes of the cytokine contents are related to the state of intestinal mucosal immunity. In this study, we investigated the changes in contents of cytokines such as interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-6 (IL-6), interferon gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) induced by dietary high F in the mucosa of different parts of intestines (duodenum, jejunum, and ileum) by enzyme-linked immunosorbent assay. A total of 280 one-day-old healthy avian broilers were randomly divided into four groups and fed on a corn–soybean basal diet as control diet (F 22.6 mg/kg) or the same basal diet supplemented with 400, 800, and 1,200 mg?F/kg (high F groups I, II, and III) in the form of sodium fluoride for 42 days. The experimental data showed that the contents of IL-2, IL-4, IL-6, IFN-γ, and TNF-α in the intestinal mucosa were significantly decreased in the high F groups II and III when compared with those of the control group from 14 to 42 days of age. It was concluded that dietary F in the range of 800–1,200 mg/kg significantly reduced the contents of aforementioned cytokines in the intestinal mucosa of broilers, which could impact the function of intestinal mucosal immunity through the pathways that decreased the lymphocyte population and/or lymphocyte activation.     

Effect of Dietary High Molybdenum on the Cell Cycle and Apoptosis of Kidney in Broilers

The experiment was conducted with the objective of examining the effects of high molybdenum on the cell cycle and apoptosis of kidney in broilers by the methods of flow cytometry. Three hundred 1-day-old Avian broilers were randomly divided into four groups, and fed on diets as follows: control diet (Mo 13 mg/kg) and high molybdenum diets (Mo 500 mg/kg, high molybdenum group I; Mo 1,000 mg/kg, high molybdenum group II; Mo 1,500 mg/kg, high molybdenum group III) for 6 weeks. The results showed that the relative weight of kidney were higher (P?<?0.05 or P?<?0.01), and the cellular percentages of G₀/G₁ phase were lower, and cellular percentages of S phase and the proliferating index were higher in high molybdenum groups II and III than in control group (P?<?0.01). The percentage of renal cell apoptosis was increased in high molybdenum groups II and III when compared with that of control group (P?<?0.01). Immunohistochemical test showed that there were increased frequencies of positive cells containing Bax protein and decreased frequencies of positive cells containing Bcl-2 protein in high molybdenum groups II and III. It was concluded that dietary high molybdenum (1,000 mg/kg and 1,500 mg/kg) impaired the progression of renal cells from S phase to G₂M phase obviously and induced renal cell apoptosis.     

Changes of the Serum Cytokine Contents in Broilers Fed on Diets Supplemented with Nickel Chloride

Cytokines are immunoregulatory proteins which play an important role in the immune system. The purpose of this study was to examine the serum cytokine contents including interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-10 (IL-10), interferon gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) induced by dietary nickel chloride in broilers by enzyme-linked immunospecific assay. A total of 240 one-day-old avian broilers were divided into four groups and fed on a corn–soybean basal diet as control diet or the same basal diet supplemented with 300, 600, and 900 mg/kg of nickel chloride. During the experimental period of 42 days, the results showed that the serum IL-2, IL-4, IL-6, IL-10, IFN-γ and TNF-α contents were lower (p?<?0.05 or p?<?0.01) in the 300, 600, 900 mg/kg groups than those in the control group. It was concluded that dietary nickel chloride in the range of 300 to 900 mg/kg could reduce the serum cytokine contents, which could finally impact the immune function in broilers.     

Effects of dietary l-lysine intake on the intestinal mucosa and expression of CAT genes in weaned piglets

The objective of this study was to evaluate effects of dietary l-lysine on the intestinal mucosa and expression of cationic amino acid transporters (CAT) in weaned piglets. Twenty-eight piglets weaned at 21 days of age (Duroc × Landrace × Yorkshire; 6.51 ± 0.65 kg body weight) were assigned randomly into one of the four groups: Zein + LYS (zein-based diet + 1.35 % supplemental lysine), Zein ? LYS (zein-based diet), NF (nitrogen-free diet), and CON (basal diet). The experiment lasted for 3 weeks, during which food intake and body weight were recorded. At the end of the trial, blood was collected from the jugular vein of all pigs, followed by their euthanasia. Dietary supplementation with lysine enhanced villus height and crypt depth in the jejunum (P < 0.05). Jejunal mRNA levels for the b^0,+-AT, y⁺LAT1 and CAT1 genes were greater (P < 0.05) in the Zein + LYS group than in the control, and the opposite was observed for CAT1. Dietary content of lysine differentially affected intestinal CAT expression to modulate absorption of lysine and other basic amino acids. Thus, transport of these nutrients is a key regulatory step in utilization of dietary protein by growing pigs and lysine in the diet influences the expression of amino acid transporters in the small intestine.     

Effects of Zinc Glycine Chelate on Oxidative Stress,Contents of Trace Elements,and Intestinal Morphology in Broilers

Three hundred and sixty healthy Ross × Ross 1-day-old broilers were used to study the effects of zinc glycine chelate (Zn-Gly) on oxidative stress, contents of trace elements, and intestinal morphology. All broilers were randomly assigned to six treatment groups, which replicates three times. Diets were as follows: (1) control (containing 29.3 mg zinc (Zn)/kg basic diet (0–21 days) and 27.8 mg Zn/kg (22–42 days)); (2) basic diet plus 30 mg Zn/kg from Zn-Gly; (3) basic diet plus 60 mg Zn/kg from Zn-Gly; (4) basic diet plus 90 mg Zn/kg from Zn-Gly; (5) basic diet plus 120 mg Zn/kg from Zn-Gly; and (6) positive control, basic diet plus 120 mg Zn/kg from zinc sulfate (ZnSO₄). The results showed that the addition of 90 or 120 mg/kg Zn-Gly led to an improvement of activity of Cu/Zn superoxide dismutase and glutathione peroxidase and a reduction of malondialdehyde content in livers at 21 and 42 days. With 90 mg/kg Zn-Gly, the content of sera zinc increased by 17.55% (P < 0.05) in 21-day broilers and 10.77% (P > 0.05) in 42-day broilers compared with that of the control. Adding 120 mg/kg Zn-Gly or ZnSO₄ to broilers' diets greatly enhanced the content of zinc in feces at 21 days (P < 0.05) and at 42 days (P < 0.05). For 42-day chickens, increased villus height and decreased crypt depth of the jejunum could be observed in the second growth stage of broilers fed with 90 mg/kg Zn-Gly. Also, intestinal wall thickness decreased (P < 0.05). In addition, adding 90 mg/kg Zn-Gly to the diet markedly elevated villus length of duodenum and decreased crypt depth of ileum (P < 0.05) in 42-day broilers.     

Dietary High Fluorine Induces Apoptosis and Alters Bcl-2, Bax, and Caspase-3 Protein Expression in the Cecal Tonsil Lymphocytes of Broilers

Long-term excessive fluoride intake is known to be toxic and can lead to fluorosis and bone pathologies. However, the cellular mechanisms underlying sodium fluoride-induced cytotoxicity in the cecal tonsil lymphocytes are not well understood. The aims of this study were to investigate the effects of high dietary fluorine on apoptosis and the expression of the Bcl-2, Bax, and caspase-3 in the cecal tonsil lymphocytes of broilers. The broilers were fed on high-fluorine diets containing 0, 400, 800, and 1,200 mg/kg fluorine. As measured by flow cytometry, the percentage of apoptotic lymphocytes was significantly increased in the high-fluorine groups II and III when compared with those in the control group. Meanwhile, immunohistochemical tests showed that the Bcl-2 protein expression decreased, and the Bax and caspase-3 protein expression increased in the high-fluorine groups II and III. In conclusion, dietary fluorine in the range of 800–1,200 mg/kg increased lymphocyte apoptosis in the cecal tonsil of broilers, suggesting that the lymphocyte apoptosis in the cecal tonsil was mediated by direct effects of fluoride on the expression of Bcl-2, Bax, and caspase-3.     

Effect of polyphenols extracted from tamarind (Tamarindus indica L.) seed coat on pathophysiological changes and red blood cell glutathione peroxidase activity in heat-stressed broilers

The purpose of this study was to determine the effect of polyphenols extracted from the tamarind seed coat (PETSC) on glutathione peroxidase (GPx) activity, red blood cell parameters and bilirubin in heat-stressed broilers. One hundred forty-seven broilers, 18-days old were divided into two groups. In group 1, broilers were maintained at an environmental temperature of 26?±?2 °C throughout the experimental period. In group 2, the broilers were maintained at 38?±?2 °C (cyclic temperature: 26?±?2 °C; ?38?±?2 °C; and ?26?±?2 °C, and broilers were maintained at 38?±?2 °C for 6 h/ day) and received PETSC at a concentration of 0, 100, 200, 300, 400 or 500 mg/kg in their diet ad libitum. Parameters were investigated on days 1, 7, 14 and 21 of the experimental period. Results showed that GPx activity of heat-stressed broilers that received 100 mg/kg of PETSC in their diet was lower (P?<?0.05) than that in broilers fed the other concentrations. The mean total red blood cell count and hemoglobin concentration of heat-stressed broilers that received 100 mg/kg PETSC was higher (P?<?0.05) than those in broilers in group 1 and those fed the other concentrations. The mean bilirubin level in the excreta of heat-stressed broilers that received 100 mg/kg of PETSC was lower (P?<?0.05) than that in broilers that received 0, 300, 400 and 500 mg/kg of PETSC. This showed that PETSC could reduce GPx activity and bilirubin in feces, and increase red blood cell parameters in heat-stressed broilers.     

Influences of Copper/Zinc-Loaded Montmorillonite on Growth Performance,Mineral Retention,Intestinal Morphology,Mucosa Antioxidant Capacity,and Cytokine Contents in Weaned Piglets

The effects of copper/zinc-loaded montmorillonite (Cu/Zn-Mt) on growth performance, mineral retention, intestinal morphology, mucosa antioxidant capacity, and cytokine contents in weaned piglets were investigated in the present study. One hundred eight piglets weaned at 21?±?1 days of age (Duroc × Landrace× Yorkshire; average initial weight of 6.36 kg) were allotted to three treatments for 2 weeks. The three treatments were as follows: (1) control group: basal diet; (2) Cu/Zn-Mt group: basal diet?+?39 mg/kg Cu and 75 mg/kg Zn as Cu/Zn-Mt; (3) Cu?+?Zn?+?Mt group: basal diet?+?mixture of CuSO₄, ZnSO₄, and Mt (equal amount of Cu, Zn, and Mt to the Cu/Zn-Mt group). Each treatment had six pens of six piglets. The results showed that as compared with the control group and the Cu?+?Zn?+?Mt group, Cu/Zn-Mt supplementation increased (P?<?0.05) the average daily gain and the gain/feed ratio; Cu/Zn-Mt supplementation increased (P?<?0.05) the Cu and Zn concentrations in serum, jejunum, and ileum mucosa, villus height, the ratio of villus height to crypt depth, and the activities of SOD, GSH-Px, and IL-10 levels, and decreased the malondialdehyde concentrations in the jejunum and ileum, and intestinal IL-1β, IL-6, and TNF-α levels. Moreover, supplementation with the mixture of CuSO₄, ZnSO₄, and Mt had no effect on the growth performance, but increased the mucosa Cu and Zn concentrations, intestinal morphology, antioxidant capacity, and immune function in the duodenum, while it had no effect on the above indexes in the jejunum and ileum. The results indicated that Mt could be used as a controlled carrier for Cu and Zn, which made Cu/Zn-Mt have better biological activities in the intestine than the mixture of Cu, Zn, and Mt.     

Effect of High Fluorine on the Cell Cycle and Apoptosis of Renal Cells in Chickens

The experiment was conducted with the objective of evaluating the effect of dietary high fluorine (F) on cell cycle and apoptosis of kidney in chickens by the methods of flow cytometry. Three hundred 1-day-old Avian broilers were divided into four groups and fed on control diet (F 23 mg/kg) and high F diets (400 mg/kg, high F group I; 800 mg/kg, high F group II; 1,200 mg/kg, high F group III) for 6 weeks. As tested by flow cytometry, the percentage of renal cell apoptosis was increased with increasing of dietary F, and it obviously rose in three high F groups when compared with that of control group. Renal cells in G₀/G₁ phase were much higher, and renal cells in S phase, G₂+M phase, and proliferation index value were much lower in high F groups I, II, and III than in control group. The results showed that excess dietary F in the range of 400–1,200 mg/kg caused G₀/G₁ arrest and increased cellular apoptosis in the kidney, which might finally interfere with the excretion and retention of fluoride in chickens.     

An l-Arginine supplement improves broiler hypertensive response and gut function in broiler chickens reared at high altitude

An experiment was carried out to examine the effects of supplemental dietary arginine (ARG) on growth, hypertensive response, and gut function in broilers reared at high altitude (2,100 m). A total of 120 day-old male broilers (Cobb 500) were divided equally into two treatment groups. Treatments included a control basal diet composed of corn and soybean meal and an experimental diet to which an l-ARG supplement was added at 10 g/kg. The trial lasted for 42 days. There were no treatment differences with regard to feed intake, body weight gain, or feed conversion ratio. However ARG supplementation did increase the plasma concentration of nitric oxide, a potent vasodilator (P?<?0.05), and attenuated indices of pulmonary hypertension as reflected by reductions in the hematocrit and the right to total ventricular weight ratio (P?<?0.05). Significantly enhanced intestinal mucosal development was observed in broilers receiving ARG supplement when compared with controls (P?<?0.05), suggesting that ARG supplementation increased the absorptive surface area of the jejunum and ileum. In conclusion, broiler diets supplemented with ARG beneficially improved pulmonary hemodynamics and appeared to enhance gut function.     

Effects of Dietary Supplementation of Recombinant Plectasin on Growth Performance,Intestinal Health and Innate Immunity Response in Broilers

The present study was conducted to evaluate the effects of dietary supplementation of recombinant plectasin (Ple) on the growth performance, intestinal health, and serum immune parameters in broilers. A total of 288 1-day-old male broilers (Arbor Acres) were randomly allotted to four dietary treatments including the basal diet (NC) and basal diet supplemented with 10 mg enramycin/kg (PC), 100 mg Ple/kg (LPle), and 200 mg Ple/kg (HPle) diets. The results indicated Ple increased (P < 0.01) average daily gain and decreased (P ≤ 0.02) feed to gain ratio of broilers. In addition, the supplementation of Ple in the diets increased (P ≤ 0.01) duodenal lipase (day 21) and trypsin (day 42) activities compared with the NC group. Similar as the supplementation of enramycin, Ple also increased villus height and decreased crypt depth in jejunum (day 21), and thus the villus height to crypt depth ratio (P < 0.01) was increased compared to the NC group on day 42. The serum immunoglobulin M (days 21 and 42), immunoglobulin G (day 42), complement 3 (day 21), and complement 4 (days 21 and 42) were significantly increased (P ≤ 0.02) due to the supplementation of Ple and enramycin, while the concentration of malondialdehyde in jejunum was decreased (P < 0.01) in PC, LPle, and HPle groups on day 21 compared with those in the NC group. Furthermore, Ple reduced (P < 0.01) Escherichia coli and total aerobic bacteria population in ileum and cecum of birds on days 21 and 42. These results indicate that the recombinant plectasin has beneficial effects on growth performance, intestinal health, and innate immunity in broilers.

     

Effects of Aflatoxin B1 Exposure and Sodium Selenite Supplementation on the Histology,Cell Proliferation,and Cell Cycle of Jejunum in Broilers

The aim of this study was to investigate the effects of aflatoxin B₁ (AFB₁) exposure and sodium selenite supplementation on the histology, cell proliferation and cell cycle of jejunum in broilers. A total of 240 1-day-old male AA broilers were divided into four groups of 60 each, fed with basal diet (control group), 0.3 mg/kg AFB₁ (AFB₁ group), 0.4 mg/kg supplement Se (Se group), and 0.3 mg/kg AFB₁?+?0.4 mg/kg supplement Se (AFB₁ + Se group) for 21 days, respectively. Compared with the control group, decreased jejunal villus height, villus height/crypt ratio, and proliferation cell nuclear antigen (PCNA)-positive cells, and G₂/M phase arrest and shedded epithelial cells on the tip of jejunal villus were observed in AFB₁ groups at 7 and 14 days of age. However, the villus/crypt ratio, PCNA-positive cells and cell percentage of G₀/G₁, S, and G₂/M phases had no significant differences between AFB₁ group and control group at 21 days. Simultaneous supplementation with sodium selenite restored these parameters to be close to those in control group. In conclusion, 0.3 mg/kg AFB₁ in the diet inhibits the development of broiler’s jejunum by reducing cellular proliferation and inducing G₂/M arrest during only the first 2 weeks after hatching. Supplementation of dietary sodium selenite at the concentration of 0.4 mg/kg Se had protective action against these toxic effects of AFB₁.     

Effects of dose and formulation of carvacrol and thymol on bacteria and some functional traits of the gut in piglets after weaning

Two trials were conducted to study the effects of dose and formulation of carvacrol and thymol on bacterial counts, metabolites and functional traits of the gut in weaned piglets. In the first experiment (Exp. I), 25 piglets (28 d, 6.59 ± 0.48 kg BW) were allocated to five dietary treatments: a control diet, or the same diet supplemented with either carvacrol or thymol at doses of 500 and 2000 mg kg^?1. In the second experiment (Exp. II), 35 piglets (28 d, 7.99 ± 0.73 kg BW) were assigned to seven dietary treatments: the same control diet as in Exp. I, or this diet supplemented with thymol in one of three formulations (on celite, on alphacel or microencapsulated) at doses of 500 and 2000 mg kg^?1. At 11/12 days post-weaning piglets were euthanised, and digesta from stomach, proximal and distal small intestine were sampled for bacteriological and biochemical analysis. Small intestinal tissue was sampled for histo-morphological determinations. In none of the experiments or sections of the gut was the number of bacteria lowered by the carvacrol or thymol supplementation. In Exp. I, the villus/crypt ratio at the distal small intestine for the experimental diets (1.30–1.32) was higher than for the control diet (1.24) (p < 0.05). Thymol fed animals in Exp. II had a lower number of intra-epithelial lymphocytes at the proximal (p < 0.05) and at the distal (p < 0.1) small intestine as compared to control animals. Mean concentration of the active ingredient in the stomach and proximal small intestine for the 2000 mg kg^?1 carvacrol diet was 521 and 5 mg kg^?1 fresh digesta, respectively, and for the 2000 mg kg^?1 thymol diets it ranged between 475 and 647 and between 13 and 24 mg kg^?1 fresh digesta, respectively. Cumulative absorption in the proximal small intestine was higher than 90% for all treatments and was not affected by formulation type. These data suggest that carvacrol and thymol can improve gut health, but evidence for clear antimicrobial effects towards the major culturable bacteria of the pig foregut is limited.     

Growth performance,intestinal histomorphology,gut microflora and ghrelin gene expression analysis of broiler by supplementing natural growth promoters: A nutrigenomics approach

In an epoch of escalating number of antibiotic-resistance bacteria, there is a dire need to develop efficient and novel feeding strategies for animal nutrition as alternatives to antibiotics. Here, implicating nutrigenomic approach, phytobiotics and organic acids were used to evaluate ghrelin gene expression levels, gut microflora composition, performance parameters and intestinal histomorphological changes in broiler chickens. One-day-old chicks (n = 315) were reared for 42 days and distributed randomly into five experimental groups; each with three replicates (21 birds per replicate). Experimental groups were control: basal diet only, antimicrobial growth promoter: 40 g/metric ton of basal diet (virginiamycin), organic acids: 4 kg/metric ton of basal diet, phytobiotics: 3 kg/metric ton of basal diet, combination: 7 kg/metric ton of basal diet (organic acids 4 kg and phytobiotics 3 kg metric ton of feed). Growth performance, histological and ghrelin gene expression analysis were executed on 21 and 42 days while, quantitative bacterial analysis of cecum and ileum was performed on day 42. Increased feed intake and body weight (p < 0.05) were noticed in phytobiotics group. Addition of phytobiotics significantly improved (p < 0.05) villus height and ratio of villus height/crypt depth in ileum, jejunum, and duodenum and down-regulated ghrelin gene expression levels. Total coliform and Escherichia coli in cecal and ileal digesta were decreased significantly (p < 0.05) in organic acids group. Correlation analysis revealed Lactobacillus spp. were positively correlated to villus height/crypt depth ration in duodenum. The findings indicated the importance of gene-nutrient-microbiota interactions based on nutrigenomics approach. Hence, phytobiotics and organic acids might be suitable alternatives to antibiotics for improved performance and immunity, along with healthier meat production in poultry.     

Probiotic bacteria maintain normal growth mechanisms of heat stressed broiler chickens

Probiotics have growth promoting effects even under periods of heat stress challenge. More information is needed to understand the mechanisms by which probiotics maintain the growth performance. The aim of this study was to evaluate the effect of a probiotic based on Bacillus subtilis bacteria on growth related mechanisms of broilers under heat stress conditions. Specifically, growth performance, skeletal bone characteristics, skeletal muscles size, intestinal villus-crypt structure, intestinal bacteria, growth hormone (GH), insulin-like growth factor-1 (IGF-1), cholesterol, and glucose. A total of 1200 one day old Ross 308 male broilers were randomly distributed into 4 treatments, with 12 replicates per treatment and 25 birds per replicate. A 2 × 2 factorial arrangement was used; the main factors were environmental temperature (thermoneutral or heat stress) and diet (control or control + B. subtilis; 3 × 10⁷ cfu/kg of feed). From d 22 to 35 of age, birds were either exposed to thermoneutral conditions (21 °C) or chronic heat stress (30 °C). During the same period, each group was divided into 2 subgroups and fed either the control diet or the B. subtilis supplemented diet. The results demonstrated that B. subtilis had positive effects (P < 0.05) on the body weight gain, feed conversion ratio, villus height, crypt depth, villus surface area, absorptive epithelial cell area and viable counts of intestinal beneficial bacteria. B. subtilis increased (P < 0.05) serum GH, IGF-1 and maintain normal levels of cholesterol and glucose under heat stress conditions. In addition, broilers fed B. subtilis under heat stress conditions exhibited higher (P < 0.05) skeletal muscles size and improved (P < 0.05) tibia traits and lower (P < 0.05) abdominal fat pads deposition compared with the controls. B. subtilis had no effect on rectal temperature under thermoneutral or heat stress conditions. It is concluded that B. subtilis can be used as growth promoters in broilers, particularly during the periods of heat stress conditions.     

Downregulation of TLR4 and 7 mRNA Expression Levels in Broiler’s Spleen Caused by Diets Supplemented with Nickel Chloride

Toll-like receptors (TLRs) are important immune receptors in discriminating self from nonself and in initiating the innate and adaptive immune response. TLR4 and TLR7 have been proven to be highly expressed in chicken’s spleen. Thus, this study was to evaluate the TLR4 and TLR7 messenger RNA (mRNA) expression levels in the spleen of broilers fed diets supplemented with nickel chloride (NiCl₂) using the methods of quantitative real-time PCR (qRT-PCR). Two hundred forty-one-day-old avian broilers were equally divided into 4 groups and fed on a corn-soybean basal diet as control diet or the same basal diet supplemented with 300, 600, and 900 mg/kg of NiCl₂ for 42 days. Results showed that TLR4 and TLR7 mRNA expression levels in the spleen were lower (P?<?0.05 or P?<?0.01) in the 300, 600, and 900 mg/kg groups than those in the control group. It was concluded that dietary NiCl₂ in excess of 300 mg/kg could lower TLR4 and TLR7 mRNA expression levels in the spleen of broilers, implying that NiCl₂ could impair the innate and adaptive immunity in spleen by injuring immunocytes and/or decreasing the content of cytokines through TLRs.