Capacity for digestive hydrolysis and amino acid absorption in Atlantic salmon (Salmo salar) fed diets with soybean meal or inulin with or without addition of antibiotics

This experiment was done to study the effects of dietary soybean meal (SBM) and inulin (a prebiotic) on the capacity for digestive hydrolysis and amino acid absorption by Atlantic salmon, and how a dietary supplement of the broad-spectrum antibiotic oxytetracycline (OTC) modulated these responses. A control diet (FM) was made from fish meal, fish oil and extruded wheat. Two similar diets were made with 250 g soybean meal (SBM) or 75 g inulin kg^− 1. Each diet was made with or without a supplement of 3 g OTC kg^− 1. All six diets contained yttrium oxide for estimation of apparent nutrient absorption. Each diet was fed to two groups of 172 g salmon kept in 1 m² tanks with 9 °C saltwater for 3 weeks. Intestinal organs were then sampled and weighed. Gastrointestinal tracts (GIT) were sectioned for analyses of brush border alkaline phosphatase (ALP) and leucine aminopeptidase (LAP) activities. Tissue from the distal intestine (DI) was also fixed for histological examination. Digesta from the different sections were freeze dried for estimation of trypsin and amylase activities, and of apparent absorption of amino acids (AA), nitrogen (N), and sulphur (S). About 85% of the trypsin activity, 70% of the amylase activity, 85% of the ALP activity, and 82% of the LAP activity were found in the proximal (PI) and mid (MI) intestine of fish with functional DI, and the absorption of AA, N, and S was quantitatively completed in the MI. Dietary OTC resulted in lower relative liver weight, but apart from increased ALP and LAP activities in DI when feeding OTC in combination with inulin, OTC did not modify the responses to dietary SBM or inulin. Dietary SBM resulted in lower relative liver weight, and induced pathomorphological changes in the DI mucosa, thus lower the ALP and LAP activities in the DI. SBM also stimulated absorption of AA, N, and S in the PI, but at the same time increased the activities of trypsin and amylase in the DI, indicating reduced re-absorption and increased faecal losses of these endogenous enzymes. Dietary inulin did not damage the DI, and stimulated intestinal growth and higher relative mass of the GIT. Inulin without OTC did not affect the hydrolytic and absorptive capacity of the salmon GIT.     

Characterization of Shiga toxin – producing Escherichia coli infections in beef feeder calves and the effectiveness of a prebiotic in alleviating Shiga toxin - producing Escherichia coli infections

Background

In the less-sensitive mouse model, Shiga toxin-producing Escherichia coli (STEC) challenges result in shedding that reflect the amount of infection and the expression of virulence factors such as Shiga toxins (Stx). The purpose of this study was to characterize the contribution of STEC diversity and Stx expression to shedding in beef feeder calves and to evaluate the effectiveness of a prebiotic, Celmanax®, to alleviate STEC shedding. Fecal samples were collected from calves at entry and after 35 days in the feedlot in spring and summer. STECs were evaluated using selective media, biochemical profile, serotyping and Stx detection. Statistical analysis was performed using repeated measures ANOVA and logistic regression.

Results

At entry, non-O157 STEC were dominant in shedding calves. In spring, 21%, 14% and 14% of calves acquired O157, non-O157 and mixed STEC infections, respectively. In contrast, 45%, 48% and 46% of calves in summer acquired O157, non-O157 and mixed STEC infections, respectively. Treatment with a prebiotic, Celmanax®, in spring significantly reduced 50% of the O157 STEC infections, 50% of the non-O157 STEC infections and 36% of the STEC co-infections (P = 0.037). In summer, there was no significant effect of the prebiotic on STEC infections. The amount of shedding at entry was significantly related to the number and type of STECs present and Stx expression (r² = 0.82). The same relationship was found for shedding at day 35 (r² = 0.85), but it was also related to the number and type of STECs present at entry. Stx - producing STEC infections resulted in 100 to 1000 × higher shedding in calves compared with Stx-negative STECs.

Conclusions

STEC infections in beef feeder calves reflect the number and type of STECs involved in the infection and STEC expression of Stx. Application of Celmanax® reduced O157 and non-O157 STEC shedding by calves but further research is required to determine appropriate dosages to manage STEC infections.     

Effect of prebiotic xylooligosaccharides on growth performances and digestive enzyme activities of allogynogenetic crucian carp (<Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>)

The effect of prebiotic xylooligosaccharides (XOS) on the growth performance and digestive enzyme activities of the allogynogenetic crucian carp, Carassius auratus gibelio, was investigated. XOS was added to fish basal semi-purified diets at three concentrations by dry feed weight: diet 1, 50 mg kg⁻¹; diet 2, 100 mg kg⁻¹; diet 3, 200 mg kg⁻¹, respectively. Twelve aquaria (n = 20) with three replicates for each treatment group (diets 1–3) and control treated without XOS were used. Weights of all collected carp from each aquarium were determined at the initial phase and at the end of the experiment, and the carp survival was also determined by counting the individuals in each aquarium. After 45 days, there were significant differences (P < 0.05) in the relative gain rate (RGR), and daily weight gain (DWG) of diets 1–3 were compared with the control. However, the survival rate was not affected (P > 0.05) by the dietary treatments. For enzymatic analysis, dissection produced a crude mixture of intestine and hepatopancreas of each segment to measure. The protease activity in the intestine and hepatopancreas content of fish in diet 2 (487.37 ± 20.58 U g⁻¹ and 20.52 ± 1.93 U g⁻¹) were significantly different (P < 0.05) from that in the control (428.13 ± 23.26 U g⁻¹ and 12.81 ± 1.52 U g⁻¹) and diet 3 (428.00 ± 23.78 U g⁻¹ and 14.04 ± 1.59 U g⁻¹). Amylase activity in the intestine was significantly higher for diet 2 compared to diet 1 and the control. As for amylase in the hepatopancreas, assays showed higher activity in diet 2 (P < 0.05) compared to the rest.     

Comparison of conventional and molecular techniques to investigate the intestinal microflora of rainbow trout (Oncorhynchus mykiss)

The concept of the use of probiotic organisms or prebiotic compounds to modify the fish gut microflora is becoming a popular topic for investigation. A major flaw in many such studies is a failure to consider fully the nature of the established microflora, which is to be modified pre-, or probiotically. Since it is widely accepted that a large proportion of bacteria are non-culturable, the use of conventional bacteriological (culture) techniques alone to investigate fish intestinal microflora may be expected to bias results. We report a study designed to investigate the normal intestinal microflora of rainbow trout (Oncorhynchus mykiss) using both conventional bacteriological and molecular methods.Over an 18 month period, the intestinal microflora of a single population of laboratory-raised rainbow trout was investigated. Bacteria isolated using bacteriological techniques were identified using the BiOLOG system and 16S rRNA gene sequencing. Dominant bacteria consistently were Aeromonas sp. and Carnobacterium piscicola, demonstrating that the microflora is stable in fish kept in defined conditions. Restriction Fragment Length Polymorphism (RFLP) and 16S rRNA gene sequence analysis was used to investigate anaerobic and non-culturable bacteria. An obligate anaerobe, Clostridium gasigenes, was shown to be among the dominating intestinal microflora.     

Hulless barley and β-glucanase affect ileal digesta soluble beta-glucan molecular weight and digestive tract characteristics of coccidiosis-vaccinated broilers

Exogenous β-glucanase (BGase) in barley-based feed has been shown to reduce digesta viscosity in chickens, and thereby improve performance. Less well studied is the potential for BGase to convert barley β-glucan into low molecular weight carbohydrates, which might influence digestive tract function and enteric disease. Coccidiosis-vaccinated broiler chickens were fed graded levels of hulless barley (HB) and BGase to determine their effects on β-glucan depolymerization and digestive tract characteristics. Broilers were fed high β-glucan HB (0%, 30% and 60% replacing wheat) and BGase (0%, 0.01% and 0.1%) in a 3 × 3 factorial arrangement. A total of 5,346 broilers were raised in litter floor pens and vaccinated for coccidiosis on d 5. Each treatment was assigned to 1 pen in each of 9 rooms. The significance level was set at P ≤ 0.05. At both 11 and 33 d of broiler ages, peak molecular weight of β-glucan in ileal digesta decreased with increasing BGase for 30% and 60% HB. The maximum molecular weight for the smallest 10% β-glucan molecules (MW-10%) decreased with BGase at both ages for 30% and 60% HB; for birds fed 0% HB, only 0.1% BGase decreased MW-10%. The 0.1% BGase increased caecal short chain fatty acids (SCFA) compared to the 0.01% BGase at d 11 only for the 60% HB. Ileal pH increased with increasing HB and BGase at d 11 and 33. Caecal pH was lower for 0.1% BGase than 0% BGase for 60% HB at d 11. Relative mRNA expression of interleukin 6 (IL-6) and IL-8 in the ileum increased with 0.1% BGase at d 11 and 33, respectively, whereas expression of ileal mucin 2 (MUC2) decreased with 0.1% BGase at d 33. In the caeca, interactions between HB and BGase were significant for monocarboxylate transporter 1 (MCT1) and mucin 5AC (MUC5AC) on d 11, but no treatment effects were found at d 33. In conclusion, BGase depolymerized high molecular weight β-glucan in HB in a dose-dependent manner. Hulless barley and BGase did not increase SCFA concentrations (except for 60% HB with 0.1% BGase at d 11) and caused minor effects on digestive tract histomorphological measurements and relative mRNA gene expression.     

Impact of Dietary Cellobiose on the Fecal Microbiota of Horses

Cellobiose is a disaccharide with potential prebiotic effects, as demonstrated in different animal species, but not yet in horses. It was, therefore, the aim of the present study to evaluate the impact of dietary cellobiose on the fecal microbiota of horses. Eight healthy adult horses and two ponies were included in this study. The animals received a diet without or with 10 g and 20 g cellobiose per day for 14 days each. At the end of the feeding periods, fresh fecal samples were collected to measure bacterial metabolites and the microbial composition. For the microbiota analysis, 16S rRNA gene sequencing was used. Cellobiose was well accepted and tolerated by the animals. The lowest fecal concentrations of D-lactate, propionic acid, i-valeric acid, and total short-chain fatty acids were measured at the dose 10 g cellobiose per horse per day (quadratic effect: P < .05). A dose-dependent increase of the relative abundance of Firmicutes (P = .049), Coriobacteriales (P < .001), and Clostridium (P = .031) could be detected. In addition, a dose-dependent decrease of the relative abundance of Bacteroidetes (P = .035) was observed. In conclusion, the increase of Coriobacteriales and Clostridium indicates a bacterial fermentation of cellobiose in the equine intestine, as members of both groups exert saccharolytic activity. As clostridia have previously been assumed to be a key component of the intestinal microbiota in horses, the observed increase of Clostridium in the feces might indicate beneficial and potentially prebiotic effects of cellobiose in horses. However, this finding requires further investigation, particularly with regard to the Clostridium species that have been promoted by dietary cellobiose.     

Soluble arabinoxylans extracted from soft and hard wheat show a differential prebiotic effect in vitro and in vivo

Arabinoxylans (AX) are part of dietary fiber. They are currently under study due to their potential prebiotic effect. Wheat whole grain flours contain all the grain layers and, therefore, present a higher arabinoxylan content than white flour. It is known that the chemical structure of these compounds varies with the type of wheat cultivar and the tissue from which they are extracted. In this work, water soluble extractable arabinoxylans (WE-AX) from two types of wheat whole flours (hard and soft) were extracted. We characterized the molecular size distribution and the potential prebiotic effect of those extracts. The prebiotic effect was evaluated in vitro and confirmed in vivo. Bacterial group abundance (Lactobacillus, Bifidobacterium, Clostridium, Enterococcus, Bacteriodes and total bacteria) was determined by quantitative RT-PCR. The molecular size of AX from hard wheats was significantly higher than AX from soft wheats. Both extracts showed potential prebiotic activity by increasing the growth of beneficial bacteria in vitro and in vivo, decreasing the pathogens in the profile of intestinal microorganisms and increasing the amount of short chain fatty acids in the intestine. WE-AX from hard wheat showed a higher prebiotic activity. Prebiotic effect assessed in vitro and in vivo assays showed a significant correlation between both types of analysis. This finding suggests that the in vitro indices performed allow predicting the potential prebiotic effect in vivo.     

Growth performance,survival, body composition,hematological parameters,intestinal histomorphology,and digestive enzymes’ activity in juvenile rainbow trout (Oncorhynchus mykiss) fed dietary Immunogen®

The aim of present study was to evaluate the effects of dietary Immunogen® supplementation on juvenile rainbow trout, where 120 fish (81.7 ± 2.23 g) were fed diets supplemented with either 0.0 (control) or 2 g Immunogen® kg^?1 for 7 weeks. The fish fed the Immunogen®-supplemented diet showed an improvement in growth performance, feed utilization, and protein efficiency ratio and a decrease in feed conversion ratio. The fish fed the Immunogen®-supplemented diet showed higher body protein content. There were no significant differences in red blood cell count, eosinophil, or lymphocyte between the treatments. However, white blood cells, neutrophil, and monocyte were higher in the fish fed fish with the Immunogen®-supplemented diet. Supplementation of Immunogen® has increased villi height and tunica muscularis thickness as well as gut protease and lipase activities. The present study revealed that supplementation of Immunogen® can improve intestinal morphology and increase digestive enzyme activity and consequently increase trout growth and feed efficiency.     

Effects of a prebiotic supplement on health of neonatal dairy calves

Calf morbidity and mortality represent major financial losses for many dairy producers. The primary cause of calf mortality in the U.S. is diarrhea caused by intestinal bacterial infections. Traditionally, formulated antibiotics in milk replacer have been used as preventive treatments. However, as consumer concern about antibiotic resistance has increased, interest in alternative supplements has grown. Prebiotics, which are food ingredients not digestible by animal enzymes, and probiotics, which are beneficial bacteria, are two proposed alternatives for antibiotics to reduce diarrhea and improve immunity in calves. In this trial 75 Holstein heifer calves were randomly assigned to one of 3 treatments: control (C; no additive), antibiotic treatment (A; oxytetracycline 0.220 g/kg and neomycin base 0.441 g/kg), and prebiotic supplement (PB; 20 g/day Prebio Support™, Meiji Feed Co., Ltd. Tokyo, Japan). Calf scours, respiratory, and general appearance scores were not affected by treatment. No statistical differences were detected in any of the health data collected; however, treatment C had nearly twice as many calves that scoured and refused feed than treatments A and PB. Populations of beneficial and pathogenic bacteria in feces were generally not affected by treatments. However, PB treatment calves had an overall trend for more Lactobacilli in their feces than calves fed A (P = 0.08), and Enterobacter population was greater in C calves than in calves on treatment A in week 4 (P < 0.05). There was a tendency for the PB treatment to have elevated populations of beneficial species of bacteria during week 2. Populations of lymphocytes carrying CD4 and CD8 markers were not affected by treatment. Fecal IgA concentration decreased as calf age increased, with the greatest concentration in week 2 (P < 0.01) for all calves. Fecal IgA levels were numerically greater for the PB group in weeks 2 and 4. Both PB and A treatments affected fecal parameters with PB having positive effects in several areas. Since calves in this study were generally healthy, observable benefits from the treatments were likely minimized.