Effect of Quantum phytase on nutrient digestibility and bone ash in White Leghorn laying hens fed corn-soybean meal-based diets.

The efficacy of an Escherichia coli 6-phytase supplementation (Quantum) on nutrient digestibility-retention and bone ash in laying hens fed corn-soybean meal (CSM) diets was investigated. White Leghorn hens (Shaver and Bovan strains) were fed CSM diets containing 0.35% (positive control, PC), 0.25% (negative control 1, NC1), or 0.15% (negative control 2, NC2) nonphytate P from 21 to 61 wk of age. Six more diets were manufactured by supplementing the negative control diets with 200, 400, and 600 units per kilogram of exogenous phytase resulting in a total of 9 treatments. Each dietary treatment x strain subclass was replicated twice with 6 hens per replication. Fecal and ileal digesta samples were collected at 42 wk of age to determine apparent nutrient digestibility or retention. Left tibiae were collected at 42 and 61 wk of age to determine bone ash. The coefficients for ileal digestibility and fecal retention for protein were higher (P < 0.05) for the unsupplemented negative control treatments compared with the PC. A linear reduction in phytate digestibility and ileal protein digestibility was reported with increasing levels of phytase to the NC1 diet. Phytase addition to the NC1 treatment resulted in a linear decrease in the digestibility of amino acids except for methionine and proline. Significantly higher phytate digestibility was demonstrated with the NC2 treatment containing 400 units per kilogram of phytase compared with the PC. Tibial bone ash percentage was higher (P < 0.05) in 61-wk-old hens fed 200 or 400 units per kilogram of phytase-supplemented NC2 diets. Significantly higher diet AME and fecal protein retention were demonstrated for Shaver hens in comparison to the Bovan hens. Overall, the Quantum phytase was not efficacious at improving nutrient digestibility-retention in laying hens fed CSM diets deficient in nonphytate P.

Whole blood chemiluminescence response in broiler chickens on different experimental diets and challenged with Clostridium perfringens.

UNLABELLED: 1. OBJECTIVES: To validate the whole blood chemiluminescence (WBCL) assay in chickens, a simple and rapid method of measuring production of reactive oxygen species by circulating polymorphonuclear (PMN) leukocytes. To determine the physiological response and innate immune response associated with oral challenge with Clostridium perfringens in broiler chickens under different nutritional conditions. 2. In Experiment 1, birds were orally challenged with C. perfringens 1. type A or sham-challenged saline on days 14-21 post-hatch and fed protein-balanced diets containing 160 or 180 g crude protein/kg and 0.98 or 1.75% glycine in a 2 x 2 x 2 factorial arrangement of treatments. 2. Challenged birds had higher WBCL responses and more severe intestinal lesions than unchallenged birds. Birds fed diets containing 1.75% glycine had more intestinal lesions than those fed 0.98% glycine. 3. In Experiment 2 birds were fed protein-balanced diets containing 0.76, 2.10, 3.43 or 4.77% glycine. The birds fed 0.76% glycine diet had lower WBCL responses compared to birds fed the other three diets. Intestinal lesions were worse in the birds fed the highest, 4.77% glycine diet than in those fed the 0.76 or 2.10% glycine diets. 4. We conclude that the WBCL assay is a practical and sensitive means of assessing innate immune function in birds. The results suggest that both bacterial challenge and glycine content of chickens' diet influence their lesion scores and innate immune function.

The efficacy of quantum phytase in a forty-week production trial using white leghorn laying hens fed corn-soybean meal-based diets.

Microbial phytase is a prominent feed enzyme used in animal feeds, but there is relatively little information on its use in laying hen diets. In this experiment, an Escherichia coli 6-phytase (Quantum) was evaluated for its efficacy in a 40-wk laying hen production trial. A total of 1,080 White Leghorn hens (540 each of Shaver and Bovan strains) were fed mash corn-soybean meal-based diets containing 0.35% (positive control, PC), 0.25% (negative control, NC1), or 0.15% (NC2) nonphytate phosphorus (NPP). Six more diets were manufactured by supplementing the negative control diets with 200, 400, and 600 U/kg of exogenous phytase, resulting in a total of 9 treatments. Each dietary treatment x strain subclass was replicated 4 times with 5 adjoining cages per replicate (3 hens per cage) in a randomized complete block design. Production performance was measured from 21 to 61 wk of age. Only minor differences in production characteristics were found between the PC and NC1 treatments regardless of phytase addition, indicating that 0.25% NPP resulted in P intake that was at or above the hen's requirement. In contrast, the hens fed 0.15% NPP diet without phytase supplementation had significantly (P < 0.05) reduced total hen housed egg production and body weight at 61 wk of age in comparison to the PC treatment, whereas the incidence of soft-shelled, cracked, and broken eggs was increased significantly (P < 0.05) in hens fed the NC2 diet. Addition of phytase to the NC2 diet improved these production characteristics to levels equal or better than the PC diet. The results indicated that Quantum phytase was efficacious in corn-soybean meal-based diets fed to White Leghorn laying hens and can be used to reduce diet supplementation with inorganic phosphorus.

The effect of dietary sinapic acid (4-hydroxy-3, 5-dimethoxy-cinnamic acid) on gastrointestinal tract microbial fermentation, nutrient utilization, and egg quality in laying hens.

Plant-based simple phenolics are natural antimicrobial and antioxidant compounds that may possess biological activity when included in animal diets. Previous work in our lab has shown that sinapic acid (SA) found in canola meal improves nutrient retention and alters gut microbiota when fed at low levels (0 to 0.10%) to broiler chickens. It was of interest to investigate the effect of dietary SA in laying hens that possess a more mature intestinal microbiota. Therefore, 2 experiments were conducted to determine the effects of dietary SA on performance, nutrient digestibility, and egg quality in laying hens. Sinapic acid (Sigma-Aldrich, 98% purity) was included in diets at 0, 0.025, 0.050, and 0.075% in experiment 1, and 0 and 0.5% in experiment 2. Both experiments were conducted for 21 d. Dietary treatment did not affect production traits and gastrointestinal tract volatile fatty acid production in either experiment. Apparent metabolizable energy and protein digestibility were also unaffected by treatment in experiment 1, but in experiment 2, feeding 0.5% SA increased both AME(n) and protein digestibility in comparison to the unsupplemented control. Egg weight, albumen weight, yolk weight, egg specific gravity, shell thickness, and Haugh units were not significantly affected by dietary treatments in either of the experiments. Sinapic acid was not detected in eggs from experiment 1 but in experiment 2, low levels of SA were detected in both the yolk and albumen of eggs from hens fed 0.5% SA. Further research is needed to more completely define the relationship between dietary SA level and nutrient utilization as well as the accumulation of SA in egg components.

Nutritional and physiological effects of dietary sinapic acid (4-hydroxy-3,5-dimethoxy-cinnamic acid) in broiler chickens and its metabolism in the digestive tract.

Sinapic acid (SA) is a major free phenolic acid in rapeseed meal, with the majority found in the esterified form of sinapine. Two experiments were conducted to delineate the effect of dietary SA on broiler chickens, in terms of performance, toxicity, and nutrient digestibility. In the first experiment, 80 male broiler chicks were randomly assigned to 5 diets adequate in all nutrients and containing 0, 0.05, 0.10, 0.15, or 0.20% dietary SA. Performance from 0 to 18 d of age and the relative size of all the internal organs and intestines were not affected (all P>0.05) by dietary treatment. In addition, dietary SA had no effect (P>0.05) on the serum activity of creatine kinase and lactate dehydrogenase, which indicated that there was no damage to skeletal muscle, heart muscle, liver, kidneys, or brain. The objectives of the second experiment were to investigate the effect of SA on nutrient retention and to study the retention of SA in the digestive tract. Male broiler chicks (96) were randomly assigned into 4 treatments containing dietary SA at 0, 0.025, 0.05, and 0.10%. Dietary SA at the 0.025% level increased feed intake compared with control (P<0.05). Regression analysis indicated a quadratic response for both weight gain and feed intake (P<0.05) as SA increased in the diet. Again, dietary SA did not affect the relative weights of bursa of Fabricius, liver, kidney, or digestive tract. Nitrogen-corrected AME and protein digestibility were not affected by SA level. A negative linear relationship was found between dietary SA level and apparent ileal digestibility of Met, Thr, Ser, Pro, Gly, Ala, and Phe. Sinapic acid disappearance in the ileum was above 97% for all SA levels, whereas apparent SA retention in excreta ranged from 64 to 79%. Sinapic acid at dietary levels investigated in this research is apparently not toxic to broiler chickens but may have a negative effect on amino acid digestibility at higher levels.

Effect of different dietary methionine sources on intestinal microbial populations in broiler chickens.

Two experiments were conducted to study the effect of various levels of DL-Met or 2-hydroxy-4-methylthiobutanoic acid (MHA-FA) on Clostridium perfringens and other intestinal bacteria in broiler chickens. In each experiment, 2 cages of 6 birds (14 d posthatch) were assigned to 1 of 7 different diets in a 2 x 4 factorial arrangement. The main effects were Met source (either DL-Met or MHA-FA) and Met level (0, 0.2, 0.4, or 0.8% dl-Met or 0, 0.227, 0.454, and 0.908% MHA-FA, thus providing 4 corresponding equimolar levels of each Met source). All birds were orally gavaged with a C. perfringens type A broth culture on d 1 and on d 14 to 20 and killed on d 28. Intestinal populations of C. perfringens, lactobacilli, Streptococcus group D, and coliforms were enumerated in the ileum and cecum, and necrotic enteritis intestinal lesions were scored. There was a significant reduction (P < 0.05) in C. perfringens populations in birds fed either Met source in the cecum (experiment 1) or the ileum and cecum (experiment 2). In experiment 2, the lactobacillus populations were significantly higher (P < 0.05) in the ceca of birds receiving 0.8% Met than in the birds given diets with the other levels of Met tested. Significantly lower populations (P < 0.05) of coliforms and Streptococcus group D were enumerated in the ileum of birds fed the 0.8% Met-supplemented diet than in the other dietary treatments. The effect of Met source on intestinal bacteria was not significant, suggesting that both DL-Met and MHA-FA have similar antibacterial properties. Last, there were no significant differences in intestinal lesion scores or the performance of birds fed different Met sources and concentrations. The results suggest that both DL-Met and MHA-FA may reduce intestinal populations of C. perfringens in broiler chickens when used in relatively high concentrations, and may reduce the risk of necrotic enteritis. Thus, feeding low-protein diets supplemented with crystalline amino acids might be beneficial in terms of the growth of various enteric pathogens.

The impact of nutrient density, feed form, and photoperiod on the walking ability and skeletal quality of broiler chickens.

The aim of this study was to examine the main and interaction effects of 3 dietary nutrient densities (low-, medium-, high-density), 2 feed forms (mash, pellet), and 2 lighting programs (20L:4D, 12L:12D) on the walking ability and skeletal quality of broiler chickens raised to a young age (35 d). Forty-eight birds per treatment (nutrient density x feed form x lighting program subclass) were randomly selected (0 d) for assessment of their gait score (GS) and BW at 11, 18, 25, and 32 d. Samples of birds from various GS classifications were selected at 32 d for radiographs and histology of the femur and tibiotarsus. Bone ash (BA) content was evaluated at 35 d from the right tibiotarsus to assess bone quality. Overall, the mean GS values were low (GS <1). An interaction existed between lighting and sex. Males provided with 20L:4D had a higher GS (0.74) than females (0.45), and this score was greater than for broilers provided with 12L:12D (0.34 and 0.26 for males and females, respectively). Feeding mash reduced the GS (0.29) compared with pellet rations (0.62). The GS increased with age, and by 32 d 2.43% of birds had a GS >/=3. Broilers fed mash had a higher BA content (50.6%) than birds fed pellet diets (49.8%), and the 12L:12D value (50.5%) was greater than for 20L:4D (48.9%). Last, males had a lower BA content (49.8%) than females (50.6%). A positive correlation existed between BW and GS based on sex, where BW at 11, 18, and 25 d affected bird mobility at 32 d (r(2) = 0.39, 0.49, and 0.50 for males; r(2) = 0.34, 0.37, and 0.36 for females, respectively). Radiography and histology were unaffected by GS. This study confirmed that a reduced growth rate improved GS but also demonstrated that overall bird mobility was good and the incidence of skeletal disease was low.

Influence of dietary nutrient density, feed form, and lighting on growth and meat yield of broiler chickens.

The objective of this study was to examine main and interactive effects of nutrient density (ND), feed form (FF; mash, pellet), and lighting program (12L:12D, 20L:4D) on production characteristics and meat yield of broilers raised to 35 d of age. Diets (starter, grower, and finisher) were formulated so that amino acid levels were in proportion to the dietary energy level. Lighting programs were initiated at 4 d of age. Body weight was not affected by ND when diets were fed in a pellet form but decreased in a linear manner with lower ND when fed as a mash. Final BW of birds fed mash were less than those of birds fed pellet diets. Feed to gain ratio decreased with increasing ND but was not affected by FF. Feed intake decreased with increasing ND and was lower for birds fed mash. The effect of ND on feed intake was less when birds were fed mash in contrast to pellet diets (P(ND x F) < 0.0001). Dietary ND had no effect on mortality, but feeding mash decreased mortality (3.8%) compared with feeding pelleted feed (5.6%). Lighting programs affected production characteristics independently of ND and FF. Use of 12L:12D reduced BW, feed to gain ratio, feed intake, and mortality compared with 20L:4D. Similarly, carcass components were not affected by ND when fed in pellet form but decreased with lower ND when fed as a mash. Overall, carcass yields were reduced when broilers were fed mash or provided with 12L:12D. Female birds had higher carcass yields and increased proportional breast meat deposition compared with males.

Dietary encapsulated glycine influences Clostridium perfringens and Lactobacilli growth in the gastrointestinal tract of broiler chickens.

Three experiments were conducted to determine whether there is a causative relation between dietary glycine concentration and intestinal Clostridium perfringens growth in broiler chickens. Expt. 1 showed that glycine concentrations were higher (P < 0.05) in jejunum and ileum of birds fed fat-encapsulated glycine compared with crystalline glycine. In Expt. 2, 2 cages of 6 birds were assigned to 1 of 6 experimental diets formulated to contain 7.6 and 10.6, 17.8 and 40.6, 27.8 and 30.6, 37.8 and 20.6, 47.7 and 10.6, and 7.8 and 50.6 g/kg total glycine and proline, respectively, provided primarily by supplementation with encapsulated glycine or proline as required. In Expt. 3, 12 groups of 6 birds were fed 4 different diets supplemented with encapsulated glycine to achieve 7.6, 21.0, 34.3, or 47.7 g/kg total glycine. The birds were orally challenged with C. perfringens type A on d 1 and d 14-21 and killed on d 28. In Expt. 2, C. perfringens populations were higher (P < 0.05) in ileum and cecum of birds, which received either 37.8 or 47.7 g/kg total glycine compared with those fed 7.6 g/kg glycine. In Expt. 3, C. perfringens numbers were higher (P < 0.05) in ileum of birds fed either 34.3 or 47.7 g/kg dietary glycine than those given either 7.6 or 21.0 g/kg glycine. Conversely, lactobacilli counts in ileum and cecum were significantly lower in birds fed the higher levels of glycine in both experiments. High C. perfringens colonization and high intestinal lesion scores were associated with reduced performance (P < 0.05). We conclude that glycine is an important determinant of C. perfringens growth in the intestinal tract of broiler chickens.

Immunogenicity of major cell surface protein(s) of Brucella melitensis Rev 1.

Brucella melitensis Rev 1 organisms were salt-extracted and the cell surface proteins (BCSPs) were found to be mainly 39-42 kDa (group 2 porin proteins) in addition to 31.6, 32.5, 58.5 and 14.7 kDa proteins. DEAE-Sephadex anion-exchange column chromatography of BCSPs yielded fraction 1, which contained one major protein (39.8-42.0 kDa) and a minor protein (31.6 kDa). All these proteins were found to be immunogenic by Western blotting. Fraction 1 along with monophosphoryl lipid A and trehalose dicorynomycolate adjuvants as well as BCSPs alone induced significant (p < or = 0.05) protection in BALB/c mice. Both these immunizing agents produced almost equivalent protection to live B. melitensis Rev 1 vaccine at 15 and 30 days post challenge. Lymphocyte stimulation test as well as delayed-type hypersensitivity reaction revealed that both these preparations induced cell-mediated immune response. These preparations also induced humoral immune response as indicated by indirect ELISA. Neither of the immune responses was significantly less (p < or = 0.05) than that with live B. melitensis Rev 1 vaccine, except that their duration was short.

Dietary glycine concentration affects intestinal Clostridium perfringens and lactobacilli populations in broiler chickens.

Previous studies have reported that intestinal populations of Clostridium perfringens, the causative agent of necrotic enteritis (NE), are correlated with diets high in glycine. To establish a direct causative link, 3 trials were conducted to examine the effect of dietary glycine levels on gut populations of C. perfringens, alpha-toxin production, and NE lesion scores in broiler chickens. In trials 1 and 2, 12 groups of 4 birds were fed 4 different ideal protein-balanced diets formulated to contain 0.75, 1.58, 3.04, or 4.21% glycine from d 14 to 28 of age. In trial 3, 24 groups of 4 birds were given 6 different ideal protein-balanced diets formulated to contain 0.50, 0.75, 1.00, 1.50, 2.00, or 4.00% glycine. All birds were orally challenged with a broth culture of C. perfringens type A on d 1 and between d 14 and 21 of age and killed on d 28. The majority of birds showed clinical signs of NE with 4.16 to 8.33% mortality in the 3 trials. The highest mortality and intestinal lesion scores were observed in chickens receiving 3.04% glycine in trials 1 and 2, and 4.00% glycine in trial 3. Clostridium perfringens populations in the cecum varied quadratically with increasing dietary glycine, with the maximal response seen at 3.30,3.89, and 3.51% dietary glycine in trials 1, 2, and 3, respectively. Numbers of lactobacilli in cecum declined significantly (P < 0.05) with increasing levels of glycine. The results suggest that dietary glycine level has a significant effect on C. perfringens and lactobacilli populations and may be a predisposing factor for NE in broiler chickens.

Biotransformation of cholesterol using Lactobacillus bulgaricus in a glucose-controlled bioreactor.

A novel single-step microbial transformation process for the production of testosterone from cholestrol by Lactobacillus bulgaricus in an aerated fermenter was investigated. The metabolism of glucose possibly supplying reducing power as NADH was necessary for the reduction of androst-4-en-3,17-dione (AD) to testosterone (TS). The growth period for the accumulation of testosterone in maximal amount and the residual glucose increased in parallel with the amount of glucose supplement in growth cultures. After the glucose in the fermentation culture was completely exhausted, most of the testosterone was oxidized to AD. Addition of a larger amount of glucose could prevent oxidation of testosterone to AD. The biotransformation of cholestrol was significantly increased in the presence of cyclodextrin (CD) in the fermenting medium. The addition of 0.1% CD to the growth medium facilitated the transport of the steroid substrate through the microbial cell wall.

Decolourisation of molasses wastewater by cells of Pseudomonas fluorescens immobilised on porous cellulose carrier.

Pseudomonas fluorescens isolated from soil samples contaminated with molasses, decolourised molasses wastewater (MWW) samples up to 76% under non-sterile conditions in four days at 30 degrees C. Immobilised cells could be reused for decolourisation activity. However, in subsequent cycles, this was found to decrease from 76% to 50% and from 50% to 24%. Decolourisation activity was regenerated from 30% to 45% by recultivating the immobilised cells in a fresh growth medium. Cellulose carrier coated with collagen was found to be most effective carrier, which produced the highest decolourisation activity of 94% in a 4-day process. This carrier could be reused with 50% of the decolourisation activity retained until the seventh day.

Decolourisation of synthetic and spentwash melanoidins using the white-rot fungus Phanerochaete chrysosporium JAG-40.

Phanerochaete chrysosporium JAG-40 was isolated from the soil samples saturated with spilled molasses collected from a sugar mill. This isolate decolourised synthetic and natural melanoidins present in spentwash in liquid fermentation; up to 80% in 6 days at 30 degrees C under aerobic conditions. A large inoculum size stimulated fungal biomass production, but this gave less decolourisation of pigment; 5% w/v (dry weight) mycelial suspension was found optimum for maximum decolourisation in melanoidin medium supplemented with glucose and peptone. Gel-filtration chromatography showed that larger molecular weight fractions of melanoidin were decolourised more rapidly than small molecular weight fractions.

Simultaneous raw starch hydrolysis and ethanol fermentation by glucoamylase from Rhizoctonia solani and Saccharomyces cerevisiae.

Crude glucoamylase preparation from Rhizoctonia solani was used to saccharify raw and cooked starch. Various concentrations of potato starch and wheat flour from 10-40%, w/v were used for mashing but 30% was found to be the optimal and economical. The saccharified mash yielded 5.89%, v/v ethanol in a simultaneous saccharification and fermentation process using a yeast strain Saccharomyces cerevisiae (SC-39) at 35 degrees C for 4 days. Removal of inhibitory substances from the fermenting broth through dialysis caused considerable increase in ethanol production.

Transformation of Progesterone by Traemetes hispida.

TRAEMETES HISPIDA was found to convert progesterone into a mixture of various products. Five products were isolated in sufficient amounts by HPLC analysis for structural identification and characterization. Three products were found to have two hydroxy moieties on axial and equatorial positions at 7beta,15beta, 6beta, 11alpha, and 11alpha, 15beta sites. The other two products had an extra 17alpha-hydroxy on the above dihydroxy derivatives to give trihydroxyprogesterone. The hydroxylating steps caused by this strain were greatly influenced by the 17beta side chain, especially by the C-20 moiety in the steroidal skeleton.

Sucrose: a constitutive elicitor of phytoalexin synthesis.

Extracts of seeds and leaves of the tropical legume Cajanus cajan (L.) Millsp. (the pigeon pea) elicited the accumulation of three phytoalexins when applied as droplets to superficially wounded leaves of the plant. The active component was purified and identified as sucrose. Phytoalexin accumulation was proportional to the logarithm of the concentration of sucrose applied, with maxima ranging from 338 to 455 micrograms per gram (fresh weight) of leaf tissue. The sucrose concentrations required to elicit half these amounts ranged from 20 to 35 micrograms per milliliter, but other sugars had little effect even at 1000 micrograms per milliliter. The elicitor activity of sucrose was abolished by actinomycin D, puromycin, and cycloheximide at a concentration of 10 micrograms per milliliter or greater, suggesting that gene derepression is required for expression of the phytoalexin response.