J. Böhm

Effects of dietary inclusion of probiotic and synbiotic on growth performance, organ weights, and intestinal histomorphology of broiler chickens

A feeding trial was conducted to investigate the effects of dietary supplementations of synbiotic and probiotic on broiler performance, carcass yield, organs weights, and histomorphological measurements of small intestine. Six hundred 1-d-old broiler chicks were randomly assigned to 1 of 3 dietary treatments for 5 wk. The dietary treatments were 1) control, 2) basal diets supplemented with synbiotic (1 kg of Biomin IMBO/ ton of the starter diets and 0.5 kg/ton of the grower diets), 3) basal diets supplemented with probiotic (1 kg of a homofermentative and a heterofermentative Lacto-bacillus sp./ton of feed). The BW, average daily weight gain, carcass yield percentage, and feed conversion rate were significantly (P < 0.05) increased by the dietary inclusion of the synbiotic compared with the control and probiotic-fed broilers. Moreover, a slight improvement in performance traits was observed in broilers fed the probiotic compared with control birds. The absolute and relative weight of spleen and thymus tended to be greater (P < 0.1) for the probiotic-supplemented group compared with the synbiotic-supplemented group. The relative liver weight was greater (P < 0.05) for probiotic-fed birds compared with synbiotic-fed birds. Additionally, the weight of small intestine was greater for either probiotic- (3.17) or synbiotic-fed birds (3.11) than the controls (2.89). Furthermore, dietary treatments influenced the histomorphological measurements of small intestinal villi. The addition of either probiotic or synbiotic increased (P < 0.05) the villus height:crypt depth ratio and villus height in both duodenum and ileum. The duodenal crypt depth remained unaffected (P > 0.05). However, the ileal crypt depth was decreased by dietary supplementations compared with control. In conclusion, synbiotic or probiotic displayed a greater efficacy as growth promoters for broilers. Furthermore, the dietary supplementations resulted in an increase in the villus height and crypt depth of intestinal mucosa of broilers. The increase in the villus height and villus height:crypt depth ratio was associated with improvement of growth performance for both synbiotic and probiotic. This indicates that the synbiotic and probiotic can be used as a growth promoter in broiler diets and can improve the gut health. These products show promising effects as alternatives for antibiotics as pressure to eliminate growth-promotant antibiotic use increases.

Aflatoxin B1 in Affecting Broiler’s Performance, Immunity, and Gastrointestinal Tract: A Review of History and Contemporary Issues

Aflatoxin B1 is a common contaminant of poultry feeds in tropical and subtropical climates. Research during the last five decades has well established the negative effects of the mycotoxin on health of poultry. However, the last ten years of relevant data have accentuated the potential of low levels of aflatoxin B1 to deteriorate broiler performance. In this regard, any attempt to establish a dose-effect relationship between aflatoxin B1 level and broiler performance is also complicated due to differences in types of broilers and length of exposure to the mycotoxin in different studies. Contrary to the prevalent notion regarding literature saturation with respect to aflatoxicosis of chicken, many areas of aflatoxicosis still need to be explored. Literature regarding effects of the mycotoxin on the gastrointestinal tract in this regard is particular scanty and non-conclusive. In addition to these issues, the metabolism of aflatoxin B1 and recently proposed hypotheses regarding biphasic effects of the mycotoxin in broilers are briefly discussed.

Intestinal Structure and Function of Broiler Chickens on Diets Supplemented with a Synbiotic Containing Enterococcus faecium and Oligosaccharides

A feeding trial was conducted on broiler chickens to study the effects of the synbiotic BIOMIN IMBO [a combination of Enterococcus faecium, a prebiotic (derived from chicory) and immune modulating substances (derived from sea algae)], with a dose of 1 kg/ton of the starter diets and 0.5 kg/ton of the grower diets on the intestinal morphometry and nutrient absorption. The general performance was improved (P < 0.05) by the dietary inclusion of synbiotic compared with the controls. Furthermore, the addition of synbiotic increased (P < 0.001) the villus height/crypt depth ratio and villus height in ileum. However, the ileal crypt depth was decreased by dietary supplementation of synbiotic compared with control. The addition of glucose in Ussing chamber produced a significant increase (P ≤ 0.001) in short-circuit current (Isc) in jejunum and colon relative to the basal values in both synbiotic and control groups. However, in jejunum the percentage of Isc increase after glucose addition was higher for synbiotic group (333 %) than control group (45 %). In conclusion, dietary inclusion of synbiotic BIOMIN IMBO increased the growth performance and improved intestinal morphology and nutrient absorption.

Simultaneous quantification of A-trichothecene mycotoxins in grains using liquid chromatography–atmospheric pressure chemical ionisation mass spectrometry

An approach for simultaneous determination of the main type A-trichothecenes by liquid chromatography and atmospheric pressure chemical ionization mass spectrometry is described. Parameters for coupling of LC-MS such as cone voltage, nebulizing temperature and the LC flow-rate, were optimized to provide detection of mycotoxins with maximum sensitivity. Furthermore, the effects of cone voltage and temperature on the fragmentation pattern of the tested toxins were studied. Main type A-trichothecenes such as T-2 Toxin, HT-2 Toxin, acetyl T-2 Toxin, diacetoxyscirpenol, monoacetoxyscirpenol (15-acetoxyscirpenol) and neosolaniol were separated on a reversed-phase narrow bore C18 column, using a linear gradient and a flow-rate of 0.3 ml/min. Mass spectra were obtained in positive ion mode for confirmation and quantitation. The method involves extraction and purification of toxins by using multifunctional Mycosep columns. Deuterated T-2 Toxin was used as an internal standard. A linear working range between 80 and 500 microg/kg in matrix with an acceptable correlation coefficient was observed. The developed method was validated by using a blank oats sample. The detection limit in the matrix was found to be between 50 and 85 microg/kg in selected ion mode for all tested A-trichothecenes. Recovery data were found to be between 77 and 101%. Within run and day-to-day precision were determined as having comparable levels to those found using GC methods. Furthermore, the matrix effect was investigated by comparing the internal standard versus the external standard method in quantification studies. In addition, the developed method was applied for the analysis of naturally contaminated oats, maize, barley and wheat samples.

Decontamination and detoxification strategies for the Fusarium mycotoxin deoxynivalenol in animal feed and the effectiveness of microbial biodegradation

Trichothecenes are a group of mycotoxins mainly produced by fungi of the Fusarium genus. Deoxynivalenol (DON) is one of the most abundant and important trichothecenes in food and feed, and is a significant contaminants due to its frequent occurrence in toxicologically relevant concentrations worldwide. Since toxin production depends strongly on environmental conditions, such as temperature and humidity, Fusarium toxin contamination can not be avoided completely. Therefore, exposure to this toxin is a permanent health risk for both humans and farm animals. As cereal crops are commonly contaminated with DON and animal diets consist mainly of cereals, it can be assumed that animals are frequently exposed to DON-contaminated feeds. Many strategies can be undertaken to reduce the toxic effect of DON. In addition to the general necessity for minimizing all risk factors that might influence the contamination of cereals with DON, such as the so-called field toxins before harvest, several post-harvest strategies can be applied to counteract possible deleterious effects of this mycotoxin in farm animals. Another approach for decontamination in feedstuffs is the use of adsorbent materials. Adsorbent materials may bind mycotoxins in the gastrointestinal tract and reduce absorption and systemic toxicity. It has been shown that some adsorbents are suitable to alleviate the toxic effects of specific mycotoxins, but its efficacy against trichothecenes is practically zero. Therefore, alternative strategies to reduce animal and human health risk are needed. The use of microbial additives is a method which uses microorganisms having the capability to detoxify mycotoxins by metabolism or degradation prior to their resorption in the gastrointestinal tract. DON has been reported to be completely transformed to de-epoxy-DON by ruminal and intestinal microflora. Eubacterium BBSH 797 was capable of DON degradation and counteracted the toxic effects of DON in animals. This review focuses on the efficacy of microbial feed additives in ameliorating the toxic effects of DON. According to the results of experiments to date, it appears that microorganisms are the main living organisms suitable for this mycotoxin biodegradation. However, the use of this approach depends on its effectiveness from both a practical and economic perspective.

Determination of nivalenol and deoxynivalenol in wheat using liquid chromatography–mass spectrometry with negative ion atmospheric pressure chemical ionisation

A new, rapid and sensitive method has been developed for the determination of nivalenol (NIV) and deoxynivalenol (DON) by using HPLC in combination with an atmospheric pressure chemical ionization (APCI)-interface and a single quadrupole mass spectrometer. Different LC and MS parameters have been optimized prior to this in order to obtain better results and sensitivity. The effect of nebulizing temperature on the sensitivity and fragmentation of NIV and DON in an APCI interface was investigated. Also, the influence of the cone voltage on the fragmentation pattern was studied, which was shown to have a tremendous effect. Furthermore, the effect of modifiers such as ammonium acetate, acetic acid and ammonia on the ionisation yield of the above substances have been investigated. The extraction was carried out using acetonitrile-water. A two step purification was then applied on two different Mycosep clean up columns. We have used a modified, rapid and isocratic HPLC method combined with a negative ion APCI-MS for the separation and quantitative determination of NIV and DON in wheat extract. An RP C18 column was used for the separation of selected compounds in wheat extract with water-acetonitrile-methanol (82:9:9, v/v/v) at a flow-rate of 1 ml/min without a split. Calibration curves show good linearity and reproducibility. The detection limit and precision were determined for NIV and DON. Both compounds could be detected down to microg/kg level in wheat using selected ion monitoring of the [M-H]- ions and the main fragments.

Contamination of aflatoxins in herbal medicinal products in Thailand.

Twenty-eight herbal medicinal products from Thailand were investigated for aflatoxin (AF) contaminations by employing a specific HPLC assay for the determination of AFB1, B2, G1 and G2. The samples were extracted with 80% (v/v) methanol in water before further cleaned up with an immunoaffinity column and followed by the detection of AFs by using an electrochemically post-column derivatization with iodine and fluorescence detector. The extraction procedure was optimized in order to obtain the best recovery. The method was successfully carried out with all the herbal products diversified as to compositions and dosage forms. The results revealed that five (18%) of herbal samples were contaminated with detectable amount of the total AFs ranging from 1.7 to 14.3 ng/g. The association between particular herbal/plant and the AF contaminated could not be determined due to the low frequency of positive samples. The contaminated products were those in tablet (4) and capsule (1) dosage forms. It was possible that the original fungal infection of these products may have been derived from either the crude herbal or other ingredients making these preparations, such as starch. In conclusion, none of the AF contaminated level found was above the current legislative level permissible in Thailand (20 ng/g). A word of caution, however, exporting some high AF-contaminated herbal products to countries where more stringent permissable level of aflatoxins exist could result in trade Barriers.

Simultaneous determination of major B-trichothecenes and the de-epoxy-metabolite of deoxynivalenol in pig urine and maize using high-performance liquid chromatography-mass spectrometry.

A selective analytical method based on high-performance liquid chromatography (HPLC), combined with atmospheric pressure chemical ionisation (APCI-) mass spectrometry (MS), has been developed for simultaneous determination of B-trichothecenes and the major metabolites of deoxynivalenol. The method allows simultaneous analysis of nivalenol (NIV), deoxynivalenol (DON), 15-acetyldeoxynivalenol (15-AcDON), 3-acetyldeoxynivalenol (3-AcDON), fusarenon X (Fus-X) and de-epoxydeoxynivalenol (DOM-1). The method is based on one-step sample clean-up using a multifunctional MycoSep column. A linear gradient mobile phase system, consisting of water:acetonitrile:methanol (H2O:ACN:MeOH) at a flow-rate of 1 ml/min, and a Polar-RP C18 column, were utilised to obtain the best resolution of all tested compounds along with column and equilibrating within 30 min. Dexamethasone (Dex) was used as internal standard. The developed method shows good repeatability for inter- and intra-day precisions as well as good linearity of calibration curves (r2 ranged from 0.9936 to 0.9998). Average recoveries for tested compounds in both matrices have been determined ranging from 63.7 to 102.3% and limit of quantification (LOQ) ranged from 25 to 150 ng/g. The utility and practical impact of the method is demonstrated using contaminated pig urine and maize samples.

Evaluating the efficacy of an avian-specific probiotic to reduce the colonization of Campylobacter jejuni in broiler chickens

Campylobacteriosis is the most frequent zoonotic disease in humans worldwide, and the contaminated poultry meat by Campylobacter jejuni can be considered one of the important sources of enteric infections in humans. The use of probiotics, which can help to improve the natural defense of animals against pathogenic bacteria, is an alternative and effective approach to antibiotic administration for livestock to reduce bacterial contamination. In vitro experiments showed that Enterococcus faecium, Pediococcus acidilactici, Lactobacillus salivarius, and Lactobacillus reuteri isolated from healthy chicken gut inhibited the growth of C. jejuni. To demonstrate this effect in vivo, 1-d-old broiler chicks received 2 mg/bird per day of a multispecies probiotic product via the drinking water. Controls received no probiotic treatment, and all chicks were infected with C. jejuni orally. Results showed that the cecal colonization by C. jejuni was significantly reduced by probiotic treatment at both 8 and 15 d postchallenge. To confirm this effect, in a second in vivo experiment, 1-d-old broiler chicks received the same dose of the same probiotic via the drinking water and controls received no probiotic, and all chicks were infected with C. jejuni orally. Similarly, probiotic treatment reduced (P=0.001) cecal colonization by C. jejuni at both 8 and 15 d postchallenge. The results of our in vivo experiments conclude that probiotic administration reduced the colonization of C. jejuni in broiler chickens.

Effect of addition of a probiotic micro-organism to broiler diet on intestinal mucosal architecture and electrophysiological parameters.

Probiotics might be one of the solutions to reduce the effects of the recent ban on antimicrobial growth promoters in feed. However, the mode of action of probiotics still not fully understood. Therefore, evaluating probiotics (microbial feed additives) is essential. Thus the objective of this work was to investigate the efficacy of a new microbial feed additive (Lactobacillus salivarius and Lactobacillus reuteri) in broiler nutrition. The body weight (BW), average daily weight gain was relatively increased by the dietary inclusion of Lactobacillus sp. in broiler diets. Furthermore, the Lactobacillus feed additive influenced the histomorphological measurements of small intestinal villi. The addition of Lactobacillus sp. increased (p < 0.05) the villus height (VH)/crypt depth ratio and the VH was numerically increased in duodenum. The duodenal crypt depth remained unaffected (p > 0.05), while the ileal crypt depth was decreased by dietary supplementation of Lactobacillus sp. compared with the control. At the end of the feeding period, the basal and glucose stimulated short-circuit current (Isc) and electrical tissue conductivity were measured in the isolated gut mucosa to characterize the electrical properties of the gut. The addition of glucose on the mucosal side in Ussing chamber produced a significant increase (p = 0.001) in Isc in both jejunum and colon relative to the basal values in Lactobacillus probiotic group. This increase in Isc for probiotic group in jejunum is equivalent to an increase of about two times that for the basal values, while in the control group is about half fold that for the basal value. In addition, the DeltaIsc after glucose addition to the large intestine was greater than the DeltaIsc in the small intestine in both control and probiotic group. Moreover in both jejunum and colon, the increase in Isc for birds fed Lactobacillus was higher than their control counterparts (p < or = 0.1). This result suggests that the addition of Lactobacillus sp. to broiler diets increased the glucose transport. Additionally, the results indicated that the conductivity of jejunal and colonic tissues remained unaffected by the dietary inclusion of Lactobacillus and support the concept that this additive enhances the maintenance and function of the epithelial barrier. In conclusion, dietary inclusion of a microbial feed additive (L. salivarius and L. reuteri) slightly increased the growth performance and improved intestinal nutrient absorption which was in association with the intestinal architecture improvement.