Aspergillus Oryzae Fermentation Extract Research Articles

Interactions between maternal parity and feed additives drive the composition of pig gut microbiomes in the post-weaning period

BackgroundNursery pigs undergo stressors in the post-weaning period that result in production and welfare challenges. These challenges disproportionately impact the offspring of primiparous sows compared to those of multiparous counterparts. Little is known regarding potential interactions between parity and feed additives in the post-weaning period and their effects on nursery pig microbiomes. Therefore, the objective of this study was to investigate the effects of maternal parity on sow and offspring microbiomes and the influence of sow parity on pig fecal microbiome and performance in response to a prebiotic post-weaning. At weaning, piglets were allotted into three treatment groups: a standard nursery diet including pharmacological doses of Zn and Cu (Con), a group fed a commercial prebiotic only (Preb) based on an Aspergillus oryzae fermentation extract, and a group fed the same prebiotic plus Zn and Cu (Preb + ZnCu).ResultsAlthough there were no differences in vaginal microbiome composition between primiparous and multiparous sows, fecal microbiome composition was different (R2 = 0.02, P = 0.03). The fecal microbiomes of primiparous offspring displayed significantly higher bacterial diversity compared to multiparous offspring at d 0 and d 21 postweaning (P < 0.01), with differences in community composition observed at d 21 (R2 = 0.03, P = 0.04). When analyzing the effects of maternal parity within each treatment, only the Preb diet triggered significant microbiome distinctions between primiparous and multiparous offspring (d 21: R2 = 0.13, P = 0.01; d 42: R2 = 0.19, P = 0.001). Compositional differences in pig fecal microbiomes between treatments were observed only at d 21 (R2 = 0.12, P = 0.001). Pigs in the Con group gained significantly more weight throughout the nursery period when compared to those in the Preb + ZnCu group.ConclusionsNursery pig gut microbiome composition was influenced by supplementation with an Aspergillus oryzae fermentation extract, with varying effects on performance when combined with pharmacological levels of Zn and Cu or for offspring of different maternal parity groups. These results indicate that the development of nursery pig gut microbiomes is shaped by maternal parity and potential interactions with the effects of dietary feed additives.

In vitro gas production kinetics are influenced by grain processing, flake density, starch retrogradation, and Aspergillus oryzae fermentation extract containing α-amylase activity.

Grain processing such as particle size, flake density, or starch retrogradation can influence ruminal degradability characteristics; however, it is unclear how exogenous α-amylase supplementation interacts with different processed grains. Four experiments were conducted to compare the effects of Aspergillus oryzae fermentation extract (Amaize; Alltech Biotechnology Inc., Nicholasville, KY) supplementation on in vitro gas production kinetics of grain substrates with different processing methods that are common in the feedlot industry. In experiment 1, corn processing (dry-rolled, high-moisture, steam-flaked) and Amaize supplementation (0 or 15 U α-amylase activity/100 mL) were evaluated in a 3 × 2 factorial arrangement of treatments. The rate of gas production for dry-rolled corn was higher (P < 0.001) with Amaize supplementation. In experiment 2, flake density (296, 322, 348, 373, and 399 g/L) and starch retrogradation (storage in heat-sealed foil bags for 3 d at 23 or 55°C) were evaluated in a 5 × 2 factorial arrangement of treatments. There was a flake density × starch retrogradation interaction (P < 0.01) for the rate of gas production because the decrease in the rate of gas production with starch retrogradation was greater at lighter flake densities compared with heavier flake densities. In experiment 3, Amaize supplementation was evaluated across flake densities of nonretrograded steam-flaked corn (stored at 23°C) used in experiment 2. There was a flake density × Amaize interaction (P < 0.01) for the rate of gas production where Amaize supplementation resulted in a lower rate of gas production at lighter flake densities (296, 322, and 348 g/L) but a higher rate of gas production at heavier flake densities (373 and 399 g/L). In experiment 4, Amaize supplementation was evaluated across flake densities of retrograded steam-flaked corn (stored at 55°C) used in experiment 2. Gas production was lower after 24 h with Amaize supplementation for retrograded flakes produced to a density of 322 and 399 g/L while Amaize supplementation did not influence gas production at 24 h at other flake densities. There was a flake density × Amaize interaction for the rate of gas production because Amaize supplementation resulted in a faster (P < 0.01) rate of gas production for all flake densities except retrograded flakes produced to a density of 296 g/L. Enzymatic starch availability was positively correlated with the rate of gas production. These data demonstrate that supplementation of 15 U/100 mL of Amaize resulted in greater rates of gas production for dry-rolled corn, corn steam-flaked to heavier densities, and retrograded steam-flaked corn.

The effect of Saccharomyces cerevisiae live cells and Aspergillus oryzae fermentation extract on the lactational performance of dairy cows

Eighty multiparous lactating Holstein cows (635 ± 33 kg) were used to study the effect of feeding Saccharomyces cerevisiae and/or Aspergillus oryzae on lactational performance for 14 weeks. Cows were assigned in a completely randomized experimental design, with repeated measures into four treatments, and were fed a basal diet of concentrates and forage at a ratio of 592:408, respectively. The treatments were: (1) the basal diet with no additive (Control treatment); (2) the basal diet supplemented with 3.5 g of live S. cerevisiae/cow daily (SC treatment); (3) the basal diet supplemented with 3.5 g A. oryzae fermentation extract/cow daily (AO treatment); and (4) the basal diet supplemented with 3.5 g of live S. cerevisiae + 3.5 g A. oryzae fermentation extract/cow daily (AOSC treatment). The AO and AOSC treatments increased (p < .05) feed intake and daily milk production, with a low milk fat content for the AO treatment. Feeding SC treatment decreased (p = .002) serum glucose concentration, while the AOSC treatment increased serum glutamate-oxaloacetate transaminase concentration. It is concluded that S. cerevisiae supplementation did not enhance milk production; however, A. oryzae fermentation extract improved feed intake and milk production.

Growth, ruminal measurements, and health characteristics of Holstein bull calves fed an Aspergillus oryzae fermentation extract

A fermentation extract of the fungus Aspergillus oryzae can be used as a prebiotic. The objective was to determine if dietary inclusion of a fermentation extract of A. oryzae as well as calf age would alter growth, health, performance parameters, and the growth and development of the rumen in Holstein calves from birth thru 1 wk postweaning; it was hypothesized that it would. Purchased bull calves (n=52) that originated from 1 of 13 farms were used in this experiment. All calves had serum IgG greater than 10 mg/mL. Calves were randomly assigned to a slaughter age, 4 (n=16) or 8 wk (n=36), and treatment, control (n=27) or fermentation extract of A. oryzae (AMF; n=25). Calves were housed and fed individually; no bedding was used and no forage was fed. Calves assigned to AMF were fed 2 g of AMF daily. Liquid AMF was delivered in milk replacer for the first 4 wk of the study; solid AMF was top-dressed on texturized starter thereafter. Calves were fed nonmedicated milk replacer twice daily (22.0% crude protein, 20.0% fat, dry matter basis; 680 g/d) and were weaned upon consumption of 0.91 kg of starter (20% crude protein, 2.0% fat; medicated with decoquinate) for 3 consecutive days or on d 45 of the study, whichever came first. Calves had ad libitum access to starter and water throughout the study. Feed intake as well as fecal and respiratory scores were recorded daily; body weight, withers height, and hip height were recorded weekly. Gross rumen measurements and rumen samples for future gross and histological analyses were taken at 4 and 8 wk. All calves grew similarly; weaning age averaged 40.39±0.77 d. Lifetime average daily gain was 0.60±0.05 kg/d and lifetime gain-to-feed ratio was 0.56±0.05. Milk replacer, starter, total dry matter intake, gross and histological rumen measurements, rumen pH, fecal and respiratory scores, and total medical costs were not affected by treatment. Despite total medical costs not differing by treatment, a lower percentage of AMF calves were treated for respiratory ailments and respiratory treatment cost was lower for AMF compared with control. Several factors may have contributed to an overall lack of observed treatment effect in this study, including include the use of a low-forage (compared with concentrate) diet, immunocompetent calves, and the selected product dose and routes of delivery. Dietary inclusion (2 g/d) of an extract of A. oryzae did not affect calf growth, intake, and ruminal or health measurements.

Influence of dietary ingredients on in vitro inflammatory response of intestinal porcine epithelial cells challenged by an enterotoxigenic Escherichia coli (K88)

Enterotoxigenic Escherichia coli (ETEC) K88 is the main bacterial cause of diarrhea in piglets around weaning and the adhesion of ETEC to the intestinal mucosa is a prerequisite step for its colonization. In this study, the adhesion of a fimbriated ETEC and a non-fimbriated E. coli (NFEC) to the intestinal cells and the activation of the innate immune system were evaluated using a porcine intestinal epithelial cell line (IPEC-J2). The impact of several feedstuffs (wheat bran (WB); casein glycomacropeptide (CGMP); mannan-oligosaccharides (MOS); locust bean extract (LB) and Aspergillus oryzae fermentation extract (AO)) on ETEC attachment and the inflammatory response were also studied. The gene expression of TLR-4; TLR-5; IL-1β; IL-8; IL-10 and TNF-α were quantified using Cyclophilin-A, as a reference gene, and related to a non-challenged treatment. The fimbriated strain was markedly better than the non-fimbriated strain at adherence to intestinal cells and inducing an inflammatory response. All the feedstuffs studied were able to reduce the adhesion of ETEC, with the greatest decrease with CGMP or MOS at highest concentration. Regarding the inflammatory response, the highest dose of WB promoted the lowest relative expression of cytokines and chemokines. All tested feedstuffs were able to reduce the adhesion of ETEC to IPEC-J2 and interfere on the innate inflammatory response; however WB should be further studied according to the beneficial results on the intestinal inflammatory process evidenced in this study.

Effects of Aspergillus oryzae extract and a Saccharomyces cerevisiae fermentation product on intake, body weight gain and digestibility in buffalo calves

An Aspergillus oryzae fermentation extract and a Saccharomyces cerevisiae culture product were added to a pelleted calf starter, respectively, at 6.0 and 26 g/kg of dry matter (DM) to determine effects on faecal score, DM intake, body weight (BW) gain and in vivo digestibility. Treatments were control groups with the control starter alone (CSt) or with free access to ryegrass hay (CStH) and combinations with the fungal supplemented starter (ExpSt and ExpStH). Forty buffalo calves (12 male, 28 female; 10 calves per treatment) were started on the experiment at 10 days of age and for 12 weeks. Faecal scoring was conducted twice weekly, and DM intake was measured weekly. Every 2 weeks, BW gain was recorded and grab faecal samples were collected to examine for the presence of intestinal parasites and Salmonella. At the end of the experimental period, on a subset of 20 calves ( i.e., five per treatment), nutrient in vivo digestibility was measured by using acid-insoluble ash as internal indigestibility marker. Starter DM intake was unaffected by fungal supplementation, but inclusion of fungal additives in the starter resulted in increased apparent total tract digestibility, regardless of the presence of hay in the diet. Calves fed ExpStH diet versus the CStH and ExpSt diets had improved faecal consistency, higher average daily gain and higher total tract digestibility of fibre. Supplementation of starter with A. oryzae fermentation extract and S. cerevisiae culture may improve calf digestive efficiency if forage is in the diet.

Characterization of Aspergillus oryzae fermentation extract effects on the rumen fungus Neocallimastix frontalis, EB 188. Part 1. Zoospore development and physiology.

Experiments were performed to determine the effect of Aspergillus oryzae (AO) fermentation extract on zoospore development in the rumen fungus Neocallimastix frontalis EB 188. Powdered product, or liquid extract prepared from such powder, was added at the recommended value for supplementation in dairy cattle. Stationary and stirred cultures were periodically sampled and assayed for extracellular and intracellular protein and enzymes, gas production, zoospore production and maturation, and carbon source utilization. Soluble extract increased fungal physiology when grown in stirred vessels or stationary cultures. Treated cultures produced higher levels of enzymes (nearly double). Mobile zoospores matured into germination entities more rapidly in treated cultures, and when powdered product was used, nearly 3 times more motile zoospores were produced at 56 h of fungal growth. Levels of the intracellular enzyme malate dehydrogenase increased by 6-fold in the presence of powdered product. Product wheat bran carrier used as soluble extract or powder had very little effect on fungal cultures. Medium cellulose was completely hydrolyzed in all cultures but this occurred earlier in those containing AO treatment.

Characterization of Aspergillus oryzae fermentation extract effects on the rumen fungus Neocallimastix frontalis , EB 188. Part 2. Carbon source utilization and effects on zoospore production

The effect of a commercial Aspergillus oryzae fermentation extract on the utilization of carbon source and zoospore production by the rumen fungus Neocallimastix frontalis EB 188 was determined. In addition, the composition of a soluble extract prepared from the commercial product was analyzed. This extract was added to N. frontalis EB 188 cultures grown on a variety of substrates and periodically assayed for protein, enzymes, zoospore production, and carbon source utilization. The powdered product contained 93% dry matter, more than 3,000 A. oryzaespores per gram, and did not contain strong buffers or high concentrations of salt. Measurable concentrations of DNA, protein, carbohydrate and several enzymes including cellulase and amylase were also found. Soluble extract increased fungal physiology and treated cultures produced significantly higher levels of supernatant protein and enzymes including amylase, cellulase and beta-glucosidase. The fungal response depended on culture carbon source. However, culture zoospore production was increased regardless of substrate provided. Culture utilization of glucose was more rapid in treated cultures, yet high levels of the extract greatly inhibited glucose utilization.

Effects of Aspergillus oryzae Fermentation Extract on Performance of Lactating Cows in the Summer and Winter in Taiwan

The aims of this study is to evaluate the effect of Aspergillus oryzae Fermentation Extract (AFE) on the performance of lactating cows in summer (May to July) and winter (December to February). The experiment was a completely randomized design (CRD) and dietary treatments were 1) basal diet without AFE, 2) basal plus 3 g/d AFE into the basal total mixed ration (TMR), 3) basal plus 45.4 mg AFE/kg the ensiling corn silage and 4) AFE inclusion in silage and TMR. Twenty-eight cows from each trial were selected and randomly allocated into the four treatment groups, confined in individual pens, and fed ad libitum for 8 weeks in both seasons of feeding trials. Results showed that AFE inclusion in corn silage significantly improved DM intake by 4.4% and milk yield by 3.1% (p<0.05) during summer. In the winter season, AFE inclusion in the diet significantly improved milk yield by 10%. Direct addition of AFE to the TMR even further significantly improved milk yield over the addition through corn silage by 7.4% in winter (p<0.05). An additive effect of AFE inclusion into TMR and through corn silage was also demonstrated in the winter-feeding. AFE inclusion however, did not improve DM intake during the winter trial. In the summer trial, inclusion of AFE showed an adverse effect on the percentage of milk fat, but did not impact on the milk fat yield. Adding AFE through corn silage showed a trend towards alleviating the negative effects of milk fat from direct AFE inclusion in TMR. The similar trend occurred in the winter trial. The inclusion of AFE through corn silage significantly lowered the milk protein content over direct AFE addition, but did not significantly impacted the milk protein yield in summer. AFE supplementation during the winter season significantly increased milk protein content. Adding AFE to the corn silage significantly increased milk protein content over direct AFE addition in winter although inclusion of AFE significantly decreased total milk solid content in the summer (p<0.005). During the winter season, inclusion of AFE required less DM to produce a unit of milk. Inclusion of AFE into corn silage required less DM, energy and protein to produce a unit of milk. But inclusion of AFE did not alleviate heat stress on the lactating cows. (Asian-Aust. J. Anim. Sci. 2002. Vol 15, No. 3 : 382-389)

Intake, digestion, and digestive characteristics of Neotyphodium coenophialum-infected and uninfected fescue by heifers offered hay diets supplemented with Aspergillus oryzae fermentation extract or laidlomycin propionate.

Tarentaise heifers fitted with a rumen cannula (539 +/- 7.5 and 487 +/- 15.7 kg avg initial BW in Exp. 1 and 2, respectively) were used in two Latin square metabolism experiments having 2 x 2 factorial treatment arrangements to determine the effects of supplementation with Aspergillus oryzae fermentation extract (AO) or laidlomycin propionate (LP) on intake, digestion, and digestive characteristics of Neotyphodium coenophialum-infected (IF) or uninfected (FF) tall fescue (Festuca arundinacea) hay diets consumed ad libitum. Heifers were housed in individual stanchions in a metabolism facility with ambient temperatures controlled to range between 26.7 and 32.2 degrees C daily. Total feces and urine were collected for 5 d following a 21-d dietary adaptation period. In situ DM and NDF disappearance and ruminal fermentation characteristics were also determined. In Exp. 1, DMI was 24% greater (P < 0.01) by heifers offered FF than by those offered IF (6.7 vs 5.4 kg/d). Heifers fed 2 g/d AO tended (P = 0.09) to consume 4% more DM than those fed a diet without AO. Degradable DM and NDF fractions of IF were greater (P < 0.01) than those of FF, but AO supplementation did not affect situ disappearance (P > or = 0.42). In Exp. 2, DMI was 18.9% greater (P < 0.01) by heifers offered FF than by those offered IF (6.6 vs 5.5 kg/d). Heifers fed LP (50 mg/d) consumed 10.6% less (P < 0.05) DM than those not fed LP (5.7 vs 6/5 kg/d). Digestibility of NDF tended to be greater (P = 0.08) and digestibility of ADF was greater (P < 0.05) from FF than from IF. Conversely, apparent N absorption (%) was greater (P < 0.05) from IF than from FF. Heifers fed LP had lower (P < 0.05) ADF digestibility than those not fed LP. In situ degradable DM and NDF fractions were greater (P < 0.01) from IF than from FF. Diets supplemented with LP had higher (P < 0.01) indigestible DM and NDF fractions than those without LP. Propionic acid and total VFA concentrations were greater (P < 0.05) from heifers offered FF than from those offered IF and from heifers fed LP than from those not fed LP. Therefore, it appears the major effect of N. coenophialum was a reduction in forage intake and total-tract fiber digestibility in certain situations. Response to the feed additives was similar whether heifers were offered IF or FF and no evidence was apparent that either additive would improve performance substantially by animals consuming low-quality fescue hay diets.

Feeding of Aspergillus oryzae fermentation culture (AOFC) to growing sheep: 2. Growth rate and feed efficiency

Utilization of yeast and or filamentous fungi as feed additive to ruminants has been of interest since the late 1980’s. Two fungi species have been commercially produced in the United States, (1) Yea-Sacc containing living cells of Saccharomyces cerevisiae and (2) Amaferm bearing Aspergillus oryzae fermentation extract. It has been demonstrated and proven that the cultures can enhance rumen development and function in young ruminants. This paper concerns the use of Aspergillus oryzae fermentation culture (AOFC) as feed additive for young-growing male ‘Garut’ sheep. The A. oryzae was cultured in a media made of mineral-enriched ‘onggok’ flour, a material of tapioca processing waste. The AOFC was prepared gradually by incubating the fungus at room temperature (26 – 300C) for 5 days, dried at 400C and ground. The AOFC was added to a commercial concentrate (GT-03) at 0, 5 and 10% (w/w) levels, as treatment C0 (control), C1, and C2, respectively. Fifteen growing ‘Garut’ sheep were used and the concentrate feed treatments were randomly allotted based on a randomized block design. Drinking water was available at all time. The amount of feed offered (chopped King grass and concentrates) and their refusals were weighed daily and live-weight of sheep was measured once a week in the morning. Daily feces was collected and weighed in the last 10 days of the 14-week experimental period. All feed and fecal samples were analyzed for dry matter, crude protein, total fiber (NDF), and ash. AOFC supplementation resulted in higher weight gains (P<0.05), which were 94.81; 122.08; and 140.52 g/d for C0, C1, and C2 treatments, respectively. Dry and organic matter, as well as protein intake was also significantly increased by inclusion of AOFC into concentrate diet (P<0.05). The increment in nutrient intake was from increased consumption of concentrates, and not from King grass, however, there was no effect of AOFC supplementation on feed efficiency. Key words: A. oryzae , sheep, gain, efficiency

Fermentation extract effects on the morphology and metabolism of the rumen fungus Neocallimastix frontalis EB188.

The effects of Aspergillus oryzae fermentation extract, Amaferm, on the rumen fungus Neocallimastix frontalis EB188 were studied. The secretion of cellulase was increased by 67% and rhyzoid development was increased 3.8-fold in the presence of extract. Strength of fungal response increased in a dose-dependent manner and demonstrated a positive correlation between cell surface area and enzyme secretion. Above certain concentrations of extract, however, the development of the fungus and enzyme secretions remained at control values or slightly diminished. Supernatant fluid appearance of the intracellular enzyme, malate dehydrogenase, paralleled the secretion of cellulase both in the presence and absence of extract. Ether solubilization of extract demonstrated that the active component(s) possessed a moderately polar value between 2.7 and 2.8. Thin layer chromatography separated extract into inert, inhibitory and intensely stimulating fractions. These results support the idea that by accelerating fungal growth and metabolism, Amaferm increases the rate (or extent) of fibre degradation caused by rumen fungi and that this, in turn, may contribute to enhanced animal performance.

Effect of Aspergillus oryzae Extract Alone or in Combination with Antimicrobial Compounds on Ruminal Bacteria

The effect of an Aspergillus oryzae fermentation extract on the growth rates of pure cultures of ruminal bacteria was determined. Bacteria were grown in an anaerobic ruminal fluid and carbohydrate medium. A sterile filtrate made with 10% A. oryzae was added to the medium at 2 or 5% (vol/vol) to provide a final A. oryzae concentration of 2 or 5mg/ml, respectively. The filtrate had no effect on the growth rates of 10 of the 19 ruminal bacteria tested; however, the filtrate increased the growth rates of the bacteria that digest fiber, Ruminococcus albus and Fibrobacter succinogenes, and the bacteria that utilize lactate, Megasphaera elsdenii, Selenomonas lactilytica, and Selenomonas ruminantium. No differences in growth rate were detected between the two concentrations of A. oryzae filtrate.We also investigated the interactions between A. oryzae and antimicrobial compounds on the growth rates of six species of ruminal bacteria that had shown positive responses or no response to the filtrate. The addition of A. oryzae filtrate to medium containing chlortetracycline or neomycin tended to diminish the negative effects of those compounds on the growth rates of some ruminal bacteria, although the bacteria had no positive growth response to the filtrate alone. In contrast, the combination of A. oryzae filtrate and tylosin decreased the growth rate of Sel. ruminantium D. These results indicated that A. oryzae stimulates growth of some bacteria that digest fiber and ferment lactate in the rumen and interacts positively or negatively with certain antimicrobial feed additives.

Growth Performance and Ruminal Fermentation Characteristics of Steers Fed High Forage Diets Supplemented with Aspergillus oryzae Fermentation Extract (Amaferm)

In a completely randomized design, 100 steers (269 kg) were allotted to 20 pens and fed a corn stalk/wheat middlings diet for 104 d. Treatments were no supplementation or supplementation with Aspergillus oryzae fermentation extract (AO) at 2 g/head/d. Feed intake, daily gain, and feed efficiency were unaffected by treatment (P>0.22). Ruminal fluid was collected with a vacuum pump via the esophagus near the end of the feeding period. Ruminal VFA and ammonia concentrations were similar across treatments (P>0.21) with the exception that the concentration of isobutyrate was greater (P=0.05) in steers supplemented with AO. Enumeration of ruminal bacteria showed greater counts of cellulolytic bacteria (P=0.07) in steers supplemented with AO. However, supplementation with AO did not affect ruminal fermentation end products or steer growth performance.

Influence of Tallow and Aspergillus oryzae Fermentation Extract in Dairy Cattle Rations

Objectives were to determine the effects of adding 3 g/d of Aspergillus oryzae fermentation extract to diets with or without 5.6% added tallow. Twenty-eight Holstein cows (X¯ = 98 d of lactation) were assigned to a randomized block experiment in a 2 × 2 factorial arrangement of treatments. Treatments were the basal diet 1) without tallow or extract, 2) with extract but no tallow, 3) with tallow but no extract, and 4) with tallow and extract. Milk production, dry matter intake, 3.5% fat corrected milk, digestibility of neutral detergent fiber in the total tract were depressed for cows fed tallow. Addition of fermentation extract did not stimulate fiber digestion or milk production of cows fed diets with or without fat. Addition of extract did not overcome depression of fiber digestibility by cows fed tallow.

The effect of Aspergillus oryzae fermentation extract on the anaerobic fungus Neocallimastix frontalis EB 188: effects on physiology

Aspergillus oryzae fermentation extract (Amaferm) was used to stimulate the in vitro growth of the cellulolytic fungus Neocallimastix frontalis EB 188. Soluble and filter-sterilized extract was added either at the start or throughout culture growth. Culture mass, protein secretion and cellulase secretion were measured in stationary test-tubes or round-bottom flasks and a stirred (desktop) fermenter. The soluble extract increased culture mass and protein and cellulase secretion in a dose-dependent manner. Maximum stimulation caused the supernatant cellulase to nearly double (i.e., 87% over controls; P<0.05), cell mass increased by 27% (P<0.05) over controls and secreted protein increased 37% (P<0.05) over controls. The timing of extract addition did not alter the culture response and suggested a recycling of components. The robustness of fungal zoospores used as inoculum, however, greatly influenced the effectiveness of the extract to stimulate secretions. Extracts did not directly influence the pH of the culture medium or the endogenous levels of enzymes. The rate of carbon source utilization and morphology of the fungus were unchanged by soluble-extract additions at any level tested. The extract was inhibitory when added to concentrations exceeding an amount equivalent to 20 g animal-1 day-1.

Performance by Stocker Cattle Offered Feedstuffs Containing Acremonium coenophialum and Supplemented with Aspergillus oryzae Fermentation Extract

An experiment was conducted to evaluate the effects of supplemental Aspergillus oryzae fermentation extract (AO) on intake, gain, temperature, and serum prolactin levels of steers fed seed screenings with and without Acremonium coenophialum. Sixty-four Angus and Angus × Hereford crossbred steers were allotted randomly by weight into one of 16 groups and fed an A. coenophialum-free hay basal diet offered for ad libitum consumption. Groups were assigned randomly to one of four treatments in a 2 × 2 factorial arrangement Treatments consisted of 2.0 kg/d seed screenings from either bromegrass (B) or A. coenophialum-infected fescue (F; 677 μg/kg ergovaline). Within each screenings type, 0.9 kg/d of a soybean hull-mineral supplement was offered either with 2g per head per d AO or no AO (C) for 90 d beginning on June 30. Steers fed B gained 15.6kg more (P<0.01) weight, consumed 0.7 kg/d more (P<0.01) DM, and converted DM to gain more efficiently (P<0.01) than those fed F. A screening type × sampling date interaction was detected (P<0.05) for rectal temperatures and serum prolactin levels. Steers fed B had lower (P<0.01) rectal temperatures on July 27 and tended (P<0.10) to have lower temperatures on September 7 and 21 than those fed F. Serum prolactin levels were higher (P<0.05) on September 21 and 28, and tended (P<0.10) to be higher on July 27 from cattle receiving B than from those receiving F. Supplementation with AO did not affect gain, intake, or feed conversion. Rectal temperatures from steers fed AO tended (P<0.10) to be higher than temperatures of those fed C on July 13 and August 11. Therefore, consumption of A. coenophialum-infected tall fescue screenings resulted in typical fescue toxicosis symptoms, but A. oryzae fermentation extract did not overcome these symptoms.

Response to Various Amounts of Aspergillus oryzae Fermentation Extract on Ruminal Metabolism in Cattle

The objective of this study was to determine whether Aspergillus oryzae fermentation extract stimulated or inhibited ruminal fermentation when fed at higher than recommended doses (3 g/d). Four dietary treatments of A. oryzae fermentation extract were fed daily to six cows fitted with ruminal cannulas. For each of four periods, bromegrass hay (6% CP) with and without extract was fed for 28 d. Dacron® bags containing bromegrass cell walls were ruminally incubated to determine ruminal fiber degradation. The A. oryzue fermentation extract did not affect degradation of cell walls, cellulose, or hemicellulose. Total ruminal anaerobic or cellulolytic bacteria were not different among treatments; neither were the proportions of cellulolytic species, Butyrivibrio sp., Ruminococcus albus, or Ruminococcus flavefaciens. Ruminal ammonia was not different; however, total VFA were higher, and pH tended to be lower, when 27 g/d of A. oryzae fermentation extract was fed. The proportion of VFA was not different among treatments. The A. oryzae fermentation extract fed at nine times the recommended dosage did not produce any stimulatory effects, except for total VFA, and was not inhibitory or toxic to ruminal metabolism and forage fiber degradation.

In Vitro Stimulation of Forage Fiber Degradation by Ruminal Microorganisms with Aspergillus oryzae Fermentation Extract

Aspergillus oryzae fermentation extract (Amaferm) was evaluated for its ability to influence degradation of brome grass and switchgrass fiber fractions by mixed ruminal microorganisms in vitro. Addition of Amaferm at a concentration of 0.067 mg/ml, which is approximately the concentration found in the rumen ecosystem (0.06 mg/ml), increased the degradation of brome grass neutral detergent fiber (NDF) by 28% after fermentation for 12 h (P < 0.01), but had no effect after fermentation for 24 or 48 h. The levels of degradation of both the cellulose and hemicellulose fractions were increased after fermentation for 12 h (P < 0.01). Additions of 0.08 and 8% (vol/vol) Amaferm filtrate (12.5 g/100 ml) stimulated degradation of switchgrass NDF by 12 and 24% (P < 0.01), respectively, after fermentation for 12 h; when 80% filtrate was added, degradation was decreased by 38%. The concentrations of total anaerobes in culture tubes containing 80% filtrate were 5 times greater than the concentrations in the controls; however, the concentrations of cellulolytic organisms were 3.5 times lower than the concentrations in the controls (P < 0.05). These results suggested that the filtrate contained high concentrations of soluble substrate which did not allow the cellulolytic organisms to compete well with other populations. The remaining concentrations of esterified p-coumaric and ferulic acids were lower at 12 h in NDF residues obtained from fermentation mixtures supplemented with Amaferm. Because the total anaerobes were not inhibited in fermentation mixtures containing Amaferm, antibiotics are unlikely to be involved as a mode of action for increasing NDF degradation. The possibility that Amaferm contains enzymes (possibly esterases) that may play a role in stimulating the rate of fiber degradation by mixed ruminal microorganisms by removal of plant cell wall phenolic acid esters is discussed.

Effects of an Aspergillus oryzae fermentation extract and other factors on lactate utilization by the ruminal bacterium Megasphaera elsdenii.

The objective of this study was to determine the effects of an Aspergillus oryzae fermentation extract (Amaferm) as well as other factors on lactate utilization by the ruminal bacterium Megasphaera elsdenii B159. Addition of Amaferm or a filter-sterilized Amaferm filtrate stimulated L-lactate uptake by both M. elsdenii and the ruminal selenomonad strain H18. Growth of M. elsdenii in medium that contained DL-lactate (2 g/L), Trypticase, and yeast extract was only slightly stimulated by the addition of 5% (vol/vol) Amaferm filtrate after 24 h. However, growth of M. elsdenii in a similar medium lacking Trypticase and yeast extract was increased over twofold by the addition of either 2 or 5% (vol/vol) Amaferm filtrate. These results suggest that Amaferm provides growth factors (i.e., amino acids, B vitamins) to support growth of M. elsdenii on lactate. There was no inhibition of L-lactate uptake when lactate-grown cells of M. elsdenii were incubated with excess (10 mM) glucose, sucrose, or maltose. In addition, when cells were grown on glucose, sucrose, or maltose rather than lactate there was little difference in L-lactate uptake, suggesting that L-lactate transport in M. elsdenii is not subject to catabolite repression by these soluble sugars. Both K+ and Na+ had little effect on L-lactate uptake. Uptake was unaffected at extracellular pH values between 6.0 and 8.0, whereas pH values of 5.0 and 4.0 increased uptake. In addition, L-lactate uptake was inhibited between 34 and 61% by protonophores. These results suggest that protons may be involved in the uptake of L-lactate by M. elsdenii B159.