ETHYL ALCOHOL REDUCTION OF SALMONELLA TYPHIMURIUM IN POULTRY FEED

This study was designed to evaluate the effect of ethyl alcohol as a potential treatment for reduction of Salmonella populations in poultry feed. Growth rate of S. typhimurium in tryptic soy broth was significantly reduced by addition of greater than 0.3% volume/volume of ethyl alcohol and growth was completely inhibited by addition of 5% ethyl alcohol. Ethyl alcohol concentrations of 20% volume/weight and greater significantly reduced initial S.typhimurium populations in poultry feed (for 20% treated, 2.31 ± 0.31 vs 3.39 ± 0.29 for untreated; P < 0.05). When feed treatment was administered either before or after inoculation of S. typhimurium with 60% ethyl alcohol or 0.04% buffered propionic acid, populations in feeds treated after inoculation were decreased to a nondetection level (< 1.0 log₁₀ CFU/g) by ethyl alcohol treatment but not by other treatments. Ethyl alcohol treatment may have the potential for reducing Salmonella spp. contamination in poultry feed.     

Survivability of indigenous microflora and a Salmonella typhimurium marker strain in poultry mash treated with buffered propionic acid

Buffered propionic acid (BPA) was evaluated as a potential treatment for the elimination of Salmonella spp. in poultry mash. A primary poultry isolate marker strain of Salmonella typhimurium was added as either a broth or in a dry chalk carrier form to poultry mash containing soybean meal as a protein supplement. The mash was supplemented with buffered propionic acid at 2, 4, 6, 8, 10, 20, 30, 50 and 100 g kg⁻¹ diet and samples were enumerated for indigenous aerobic bacteria, fungi and the S. typhimurium marker strain. Total indigenous aerobic bacteria and fungal populations were generally decreased by addition of more than 20 g BPA kg⁻¹, but an addition of 100 g BPA kg⁻¹ mash was usually required to achieve reductions of approximately 90% of indigenous aerobic bacteria and 99% of indigenous fungi. After 7 days of storage, 8 g BPA kg⁻¹ mash also reduced S. typhimurium populations by more than 90% in mash inoculated via chalk, while at least 50 g BPA kg⁻¹ mash was required to provide the same level of reduction in mash inoculated with a liquid culture of S. typhimurium. Although BPA does not appear to be an overly effective antimicrobial agent with respect to indigenous aerobic bacterial populations in animal feed, higher concentrations may have the potential for reducing fungal and Salmonella spp. contamination in poultry mash.     

SURVIVAL OF AN UNIRRADIATED SALMONELLA TYPHIMURIUM MARKER STRAIN INOCULATED IN POULTRY FEEDS AFTER IRRADIATION

The present study was designed to compare unirradiated Salmonella typhimurium survival during storage after inoculation in either irradiated or unirradiated poultry feed. The effects of irradiation (5 kGy) on the indigenous feed microflora and on the survival of marker strain of S. typhimurium contaminated after irradiation treatment were determined during 56 days of storage of either soybean meal (SBM) or meat and bone meal (MBM) based feeds. The initial aerobic bacterial populations were reduced more than 90% in both SBM (4.96 to 4.08 ± 0.03 log₁₀ CPU/g feed) and MBM (5.12 to 3.90 ± 0.03) by irradiation. Irradiation treatment reduced the average fungal counts during 56 days of storage in both SBM (4.24 to 2.74 ± 0.03) and MBM (4.38 to 2.15 ± 0.03) containing feeds. However, unirradiated S. typhimurium populations inoculated after irradiation of the feed were not different in either irradiated or nonirradiated SBM and MBM based feeds. Therefore, the differences in fungal versus bacterial sensitivity among the feed types and storage times suggests that gamma irradiation can alter the makeup of indigenous microbial populations in feed but this does not appear to have a discernible influence on subsequent survival of unirradiated S. typhimurium added as a dry inoculum after irradiation.     

Salmonella typhimurium poultry isolate growth response to propionic acid and sodium propionate under aerobic and anaerobic conditions

Propionic acid and its sodium salt have long been used as additives in poultry feed to reduce microbial populations, including Salmonella spp. Propionic acids in poultry feed may have a potential role in inhibiting growth of Salmonella in the chicken intestine. In this study, we determined growth response of a Salmonella typhimurium poultry isolate to propionic acid and sodium propionate under aerobic and anaerobic conditions. Growth rate consistently decreased with the addition of greater concentrations of either propionic acid or sodium propionate. The extent of growth inhibition was much greater with propionic acid than the sodium form. Media pH decreased only with addition of propionic acid. Growth inhibition was more effective under anaerobic growth conditions with either propionic acid or sodium propionate. When determined at the same pH level, growth rate was significantly lowered by addition of 25 mM of either propionate or sodium propionate alone, and also by the decrease in pH levels (P<0.05). These results showed that growth inhibition of S. typhimurium by propionic acid or sodium propionate is greatly enhanced by pH decrease, and to lesser extent by anaerobiosis. We also found that sodium propionate was more inhibitory for growth of S. typhimurium than propionic acid when compared at the same pH levels.     

Viability of VA-mycorrhizal fungi following soil solarization and fumigation

Two field experiments were conducted to examine the effect of soil solarization on the survival of arbuscular mycorrhizal (AM) fungi and root colonization of three crops. The experiments were carried out in a loamy sand soil (Rehovot) and a silty soil (Bet She'an Valley). For both experiments, assessment of indigenous AM fungal populations by the most probable number (MPN) method indicated that populations were reduced to zero after 2 or 4 weeks of solarization treatment. However, Glomus intraradices inoculum applied to the soil prior to solarization remained viable even after 8 weeks of solarization. After soil fumigation with methyl bromide both indigenous and applied AM fungi were nondetectable. Percentage root colonization by the indigenous AM fungal populations, together with plant-growth parameters, were assessed for three crops: onion and wheat (Rehovot), and carrot (Bet She'an). When sown on solarized field plots, onion and carrot seedlings showed a plant growth retardation, whereas wheat showed an increased growth response. Root colonization by indigenous AM fungi was not evident until 6 weeks after seedling emergence. Fumigation with methyl bromide reduced root colonization by indigenous AM populations, and reduced onion and wheat plant development at early growth stages. In a laboratory experiment, a temperature of 45° C for up to 24 h did not affect AM spore viability, indicating that temperatures reached during the solarization treatment cannot solely account for the reduced AM fungi viability in the field. Apparently, soil solarization temporarily delays root colonization by indigenous AM fungi until 6-8 weeks after plant emergence.     

LOW NUTRIENT R2A CULTURE MEDIUM FOR BACTERIAL ENUMERATION FROM POULTRY FEEDS

Poultry feed matrices can be particularly stressful to bacterial populations and limit recovery and accurate enumeration of viable organisms. The objectives in this study were to enumerate bacterial populations with either tryptic soy plate medium or an R2A minimal nutrient medium from commercial poultry and turkey dry feeds and compare enumerations from the two media with feed composition. Total population estimates from tryptic soy plates varied between 3.7 and log₁₀ 5.4 CFU per g feed and depended upon poultry feed source (P < 0.05). R2A plate counts varied between 2.7 and log₁₀ 5.4 CFU per g feed, but were not significantly different among poultry feed sources. Feed R2A plate count populations were significantly correlated (P < 0.01) with tryptic soy plate count populations enumerated from the feed sources, but not protein and fat content of the feed source. It is apparent that bacterial counts recovered by minimal R2A medium are similar to enumerations using tryptic soy plate medium and that R2A medium could be substituted for tryptic soy plate medium for bacterial enumeration in poultry feeds.     

Effect of 2,4-Diacetylphloroglucinol Producing,Overproducing, and Nonproducing Pseudomonas fluorescens F113 in the Rhizosphere of Pea

Pseudomonas fluorescens F113lacZY and modified strains carrying different function modifications were assessed for their impact in the rhizosphere of pea. Strain F113lacZY naturally produces the anti-fungal metabolite 2,4-diacetylphloroglucinol (Phl) useful in plant disease control. The first modified strain of F113 was repressed in production of Phl, creating the Phl negative strain F113G22. The second was a plasmid based overproducer of Phl (F113Rif (pCUGP)). Both the F113lacZY and the F113Rif (pCUGP) strains increased the rhizoplane fungal populations, whereas the same strains reduced the rhizosphere soil fungal populations with respect to the control. Similar results were found with the rhizoplane and rhizosphere soil bacterial populations. The F113G22 treatment resulted in a significantly greater indigenous fluorescent Pseudomonas population than the F113lacZY and F113Rif (pCUGP) treatments and a greater total Pseudomonas population than the control, F113lacZY, and F113Rif (pCUGP) treatments. Overproduction of Phl did not affect the establishment of the introduced Pseudomonas population. None of the inocula displaced the indigenous populations, but the F113G22 inocula had an additive effect on the total Pseudomonas population. P (phosphatase), S (sulphatase), and N (urease) cycle enzyme activities were increased while C (glucosidase, NAGase) cycle activities were decreased by the F113lacZY and F113Rif (pCUGP) treatments, suggesting C leakage from the roots. Overall, most effects of inoculation compared to the wild type were found with the non-Phl-producing strain. Overproduction of Phl had little environmental effect in relation to wild-type inocula.     

Influence of antimicrobial agents on the spoilage of a meat-based entomophage diet

The microbial decomposition of a meat-based entomophage diet presented in Parafilm packets was investigated. Considerable bacteria but not fungi were associated with components used to prepare the diet (i.e., hens' eggs, liver, and ground beef). At the initial sampling time, there were no differences among diet treatments in the size of bacterial or fungal populations. Bacterial populations in diets not containing antibacterial agents rapidly increased and reached an asymptote by 24 h (approximately 10(10) colony-forming units per gram). Bacterial populations also increased in diets containing antibacterial agents, but they were significantly smaller than in diets not containing antibacterial agents. The most prevalent bacteria isolated were Carnobacterium piscicola, Carnobacterium divergens, Lactobacillus curvatus, Lactobacillus sakei, Leuconostoc mesenteroides, and Enterococcus spp., regardless of the antibacterial treatment used. The proliferation of fungi was delayed relative to bacteria, but significant differences were observed among the diet treatments. Fungi were most inhibited by sorbic acid and propionic acid in the absence of antibacterial agents. The most common fungi isolated were the yeasts Candida zeylanoides, Torulaspora globosa, and Yarrowia lipolytica. The pH of diets not containing antibacterial agents decreased rapidly and was highly correlated with increases in bacteria but not fungi. The results of this study demonstrate that antimicrobial agents significantly inhibit spoilage microorganisms in a meat-based diet and that alternative management strategies to delay the decomposition of such diets presented in Parafilm packets should target lactic acid spoilage bacteria, particularly Carnobacterium and Lactobacillus species.     

CYCLOHEXIMIDE AS A MEDIA AMENDMENT FOR ENUMERATING BACTERIAL POPULATIONS IN ANIMAL FEEDS

The present study was designed to evaluate cycloheximide as a potential media amendment for differential bacterial and fungal enumeration of animal feeds. The objectives of this study were to determine the effect of cycloheximide on bacterial growth rates and to evaluate its efficacy for the reduction of indigenous spreading fungi when enumerating bacterial populations in three types of feeds and after short or long-term storage. Escherichia coli, Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes and Pseudomonas fluorescens were grown in tryptic soy broth containing cycloheximide to determine its effect on bacterial specific growth rates. Growth rates of B. cereus and S. aureus were significantly decreased by the addition of 600 and 1000 mg/L cycloheximide respectively, but other pure cultures were not significantly influenced by cycloheximide addition. Intrinsic bacterial populations from feed were not significantly affected by cycloheximide additions at concentrations from 10 to 300 mg/L, but the indigenous spreading molds from feeds were significantly decreased by these cycloheximide concentrations and were decreased below detection levels by 300 mg/L of cycloheximide. The addition of 300 mg/L of cycloheximide effectively eliminates fungal growth for accurate enumeration of bacterial populations in feeds.     

Water vapor, aqueous ethyl alcohol, and heat activation of Bacillus megaterium spore germination

Dormant spores of Bacillus megaterium were activated for germination on glucose by heating them in aqueous suspension (but not if heated dry), by treating them with aqueous ethyl alcohol at 30 C, or by exposing them to water vapor at room temperature. The degree of water vapor activation depended upon the relative humidity, the time, and the temperature of exposure. Activation increased the extent and rate of glucose-induced germination and decreased the average microlag. Extended water vapor treatment also activated spores for germination induced by KI and by l-alanine. Spores activated by any of the three treatments were deactivated by treatment at 66 C, either for 18 hr in 100% ethyl alcohol or for 40 hr over P(2)O(5). Deactivated spores were reactivated by heat, by 5 m ethyl alcohol, or by water vapor. It is postulated that heating and ethyl alcohol may change the structure of liquid water, so that it is more like water vapor and can more readily penetrate to and hydrate a critical (enzymatic?) spore site, leading to activation.     

Gas-phase enzyme catalysis using immobilized lipase for ester production

Bench-scale reactors were operated in continuous recycle and single-pass modes using immobilized porcine lipase to catalyze gas-phase esterification of ethyl alcohol with two carboxylic acids (acetic acid and propionic acid). Approximately one order of magnitude increases (over uncatalyzed reactions) in conversion were achieved; produc-tion concentrations ranged from 0.1 to 0.5 mM in air, and were affected strongly by substrate concentration and acid-induced enzyme inactivation.     

Survey of <Emphasis Type="Italic">Aspergillus</Emphasis> and <Emphasis Type="Italic">Fusarium</Emphasis> species and their mycotoxins in raw materials and poultry feeds from Córdoba,Argentina

The aims of the present work were: (1) to determine both mycobiota in raw materials and finisher poultry feed, as well as the ability to produce aflatoxin B₁ by A. flavus strains, and (2) to evaluate the natural co-occurrence of aflatoxins (AFs), fumonisins (FBs), gliotoxin, diacetoxyscirpenol (DAS), HT-2 toxin, and T-2 toxin in poultry feed by LC-MS/MS. Nineteen percent of raw materials and 79% of finisher poultry feed samples exceeded the maximum allowed total fungal count (1?×?10⁴ CFU g^?1) to ensure hygienic quality. Aspergillus flavus was the only species belonging to section Flavi which was isolated while Fusarium verticilliodes was the prevalent species. Forty-seven percent of A. flavus strains were aflatoxin B₁ producers and the highest frequency of aflatoxigenic strains was isolated from finisher poultry feeds. Principal component analysis showed that corn grains are closely related with total fungal and Fusarium counts. This positive relationship suggests that total fungal and Fusarium spp. counts in poultry feed might come mainly from corn grains. Regarding poultry feeds, in ground finisher type, Aspergillus spp. counts increased as water activity (a_w) diminished. A positive relationship among a_w, total fungal and Fusarium spp. counts was observed in both ground finisher and ground starter feed. Several mycotoxins were monitored in feeds by applying the LC MS/MS technique. One hundred percent of poultry samples were contaminated with FB₁, and the highest levels were detected in pelleted finisher poultry. AFB₁, gliotoxin, DAS, HT-2 toxin, and T-2 toxin were not detected in any poultry feed. The scarcity of available mycotoxicological studies from Argentinean poultry feed using a multitoxin analysis technique enhances the contribution of the findings of this report.     

Survival of a Salmonella typhimurium poultry isolate in the presence of propionic acid under aerobic and anaerobic conditions

Propionic acid is commonly found as a fermentation product in the gastrointestinal tracts of food animals and has also been used to limit the microbial contaminants in animal feeds. Because propionic acid is known to have antibacterial activity, the propionic acid encountered by foodborne pathogens during their life cycles may play an important role in inhibiting the survival of the pathogens. The survival patterns of Salmonella typhimurium poultry isolate were determined both in aerobic and anaerobic tryptic soy broth (TSB; pH 5.0 or 7.0) containing various concentrations of propionic acid (0-200 mM). The levels of recovered cells were consistently greater at pH 7.0 compared to those at pH 5.0. For the first 4 days, the levels were significantly decreased by incubation under anaerobic conditions as compared to aerobic condition at pH 7.0 (P<0.05). However, there were fluctuations of cell populations with different patterns depending on both concentrations and growth conditions. To characterize the nature of the capability which allowed the cell multiplication following decreases in cell population during incubation at pH 7.0, the cells isolated from the outgrowth cultures were tested for survival in aerobic or anaerobic TSB (pH 5.0 or pH 7.0) containing propionic acid (50 mM). The outgrowth isolates did not show significant differences in the level of recovered cells in the presence of propionic acid when compared to the wild type strain (P>0.05), suggesting that the cells in the outgrowth cultures did not harbour mutation(s) conferring increased resistance to propionic acid. In addition, the level of recovered cells of isogenic rpoS mutant strain of S. typhimurium was not significantly different from that of the wild type strain in the same assay conditions (P<0.05). The results of this study show that the bactericidal activity of propionic acid on S. typhimurium can be affected by environmental conditions such as acidic pH levels and anaerobiosis in food materials and gastrointestinal tracts. However, S. typhimurium is also able to multiply in the presence of sublethal concentrations of propionic acid at neutral pH during prolonged incubation under both aerobic and anaerobic conditions.     

A potent feed preservative candidate produced by Calcarisporium sp., an endophyte residing in stargrass (Cynodon dactylon)

AIMS: The cultures of an endophytic fungus Calcarisporium sp. were screened for inhibitors on the growth of feed-associated moulds and on the aflatoxin biosynthesis to find a safe and effective feed preservative. METHODS AND RESULTS: Eight test fungi were isolated from the spoiled poultry feed. The endophytic fungus Calcarisporium sp. was separated from the Chinese coastal grass Cynodon dactylon. The antifungal action concerning the endophytic culture extract (ECE) was performed with propionic acid (PPA) as the corresponding reference. The ECE had a similar antifungal efficacy to PPA in a concentration-dependent manner. The susceptibility order of the ECE to the test fungi was found to be Fusarium sp. > Aspergillus spp. > Penicillium spp. Furthermore, the application of the ECE in pelleted-layer duck feed as a preservative was carried out at a humidity of 10, 15 and 20%. It has been discerned that mould growth and aflatoxin biosynthesis could be co-inhibited almost completely by ECE at concentrations higher than 1.0% (w/w). The LD50 of the ECE on mice was shown to be higher than 28 g kg-1. CONCLUSIONS: The ECE can be selected as an inhibitor to preserve poultry feed on inhibiting the growth of mould and aflatoxin biosynthesis during feed storage. SIGNIFICANCE AND IMPACT OF THE STUDY: The ECE may be an effective and biosafe antifungal ingredient for poultry feed and holds a potential market prospect in feed industry.     

Regulation of Fusarium oxysporum population introduced into soil: the amoebal predation hypothesis

Abstract Inoculation of fungi into soil has been suggested for biological control of plant diseases. The aim of our work was to test the ability of protozoa to reduce the density of introduced fungal populations. The survival of Fusarium oxysporum in non-sterile soil was studied after introduction at densities of: 1 × 10⁴, 1 × 10⁶ and 5 × 10⁷ cfu/g soil. The dynamics of protozoa were also followed. The fungal populations remained close to the initial inoculation densities and did not induce the growth of indigenous protozoa. A bacterial population ( Enterobacter aerogenes ) was used to promote and stimulate the predatory activity of amoebae. Then, after simultaneous inoculation with bacteria and fungi, the density of protozoa increased but this had no effect on the fungal population, although some amoebae are able to feed on small fungal propagules such as conidia. The physiological state of Fusarium in soil and intraspecific competition seem to be more important in regulating introduced fungal populations than amoebal predation. We conclude that the regulation of bacterial and fungal populations in soil depend on different mechanisms.     

Influence of biotic and abiotic factors on the persistence of a Beauveria bassiana biopesticide in laboratory and high-rise poultry house settings

Recent studies suggest the potential for use of oil formulations of the entomopathogenic fungus, Beauveria bassiana, as residual sprays for control of house flies in poultry production facilities. The current study investigated the influence of biotic and abiotic factors on biopesticide persistence. We found that flies physically removed and deactivated conidia, with higher fly densities and greater cumulative exposure hastening the decline. Nonetheless, very low densities of viable conidia were still able to cause rapid mortality, suggesting the potential for relatively long re-treatment intervals as fly populations are controlled. Considering abiotic factors, we found that fungal spray treatments remained viable for up to 13 weeks under laboratory conditions. Periodic exposure of flies to the spray residue showed high levels of mortality, with very little decline in mortality rate over time. Equivalent treatments placed in a commercial poultry house showed much more rapid decline. One trial at the end of summer showed conidia to remain viable up to seven weeks. However, repeats during the winter months revealed decay in 1–2 weeks, with fly mortality rates influenced accordingly. The exact reasons for the more rapid decay remain unclear but could be linked to high concentrations of ammonia in the basement areas, especially during winter when ventilation is minimal. The combined data suggest the potential for adaptive treatment regimes with weekly spray intervals in conditions with very high fly populations and/or high ammonia levels, and potentially monthly spray intervals when fly populations and ammonia levels are reduced.     

Effects of Pseudomonas fluorescens F113 on Ecological Functions in the Pea Rhizosphere Are Dependent on pH

Abstract The aim of this microcosm study was to determine influence of the antibiotic 2,4-diacetylphloroglucinol (DAPG) on the effect of wild-type and functionally modified Pseudomonas fluorescens F113 strains in a sandy loam soil of pH 5.4 planted with pea (Pisum sativum var Montana). The functional modification of strain F113 was a repressed production of DAPG, useful in plant disease control, creating the DAPG negative strain F113 G22; both were marked with a lacZY gene cassette. Lowering the soil pH to 4.4 significantly reduced the plant shoot and root weights and the root length, whereas the bacterial inocula had no significant effect. Both inocula significantly reduced the shoot/root ratio at pH 5.4, but this effect was not evident at the lowered or elevated (6.4) pH levels. The decrease in pH significantly increased the fungal and yeast colony-forming units from the rhizosphere (root extract), but did not affect the total bacterial c.f.u.'s. Inoculatioin with strain F113 in the pH 4.4 soil resulted in a significantly greater total bacterial population. The fungal and yeast c.f.u.'s were not significantly affected by the inocula at any pH studied. Increasing the pH significantly increased the indigenous Pseudomonas population in comparison to the reduced pH treatment and significantly increased both the introduced and total Pseudomonas populations. The antibiotic producing strain significantly reduced the total bacterial population and the NAGase activity (related to fungal activity) at pH 6.4 where the inocula population was the greatest. Alkaline phosphatase, phosphodiesterase, aryl sulfatase, β-glucosidase, alkaline β-galactosidase, and NAGase activities significantly increased with increasing in pH. The F113 inocula reduced the acid phosphatase activity at pH 5.4 and increased the acid β-galactosidase activity over all the pH treatments. The results presented illustrate the variation in impact with soil pH, with implications for variability in efficacy of Pseudomonas fluorescens biocontrol agents with soil pH. Received: 26 June 1998; Accepted: 1 February 1999     

The effects of grain treatment, grain feed level and grass silage feed value on the performance of and meat quality from, finishing beef cattle

A completely randomised design study involving 132 continental crossbred beef steers was undertaken to evaluate the effects of method of grain treatment and feed level, and grass silage feed value on animal performance, carcass characteristics and meat quality of beef cattle. Winter wheat was harvested and the grain was stored either ensiled crimped and treated with 4.5 l/t of a proprietary acid-based additive (crimped), ensiled whole and treated with 20 kg feed-grade urea per t (urea) or stored conventionally in an open bin treated with 3 l propionic acid per t. Two grass silages, of contrasting feed value (L and H) were ensiled. For the conventional, crimped and urea treatments, grain dry matter (DM) concentrations were 802, 658 and 640 g/kg, respectively. For the L- and H-feed value silages, DM concentrations were 192 and 240 g/kg and D values were 671 and 730 g/kg DM, respectively. The silages were offered as the sole forage supplemented with either conventional, crimped or urea-treated grain-based concentrate at either 3.5 or 6.0 kg DM per steer per day. The grain supplement consisted of 850 and 150 g/kg DM of grain and citrus pulp, respectively. For the conventional, urea and crimped treatments, DM intakes were 8.85, 9.43 and 9.04 kg/day (standard error (s.e.) = 0.129); estimated carcass gains were 0.60, 0.55 and 0.61 kg/day (s.e. = 0.020), respectively. For the low- and high- feed value grass silages, estimated carcass gains were 0.56 and 0.61 kg/day (s.e. = 0.014), respectively. For the low and high grain feed levels, estimated carcass gains were 0.56 and 0.61 kg/day, respectively. Grain treatment, grain feed level or silage feed value did not alter (P > 0.05) meat quality, lean colour or fat colour. There were significant silage feed value × grain feed level interactions (P < 0.05) for final live weight (LW) and daily live-weight gain (DLWG). Increasing grain feed level increased final LW and DLWG when offered with the low-feed value silage, however, grain feed level had no effect on final LW or DLWG when offered with the high-feed value silage. It is concluded that urea treatment of grain increased silage intake and feed conversion ratio (kg DM intake per kg carcass) and tended to decrease carcass gain. Crimping provides a biologically equally effective method to store grain as conventional methods. Improving grass silage feed value had a greater impact on animal performance than increasing grain feed level by 2.4 kg DM per day.     

An enzyme-linked immunosorbent assay for monitoring of Aspergillus ochraceus growth in coffee powder, chilli powder and poultry feed

AIMS: The work was carried out to develop an immunoassay for estimation of Aspergillus ochraceus biomass on solid substrate. METHODS AND RESULTS: An indirect noncompetitive enzyme-linked immunosorbent assay (ELISA) was developed for determination of fungal biomass in food commodities using antibody raised against A. ochraceus mycelial antigen. The sensitivity of the assay was linear in the range of 10-160 microg fungal biomass per millilitre extract of coffee (R(2)=0.989), poultry feed (R(2)=0.987) and chilli (R(2)=0.989). The growth of A. ochraceus in the food commodities like chilli, coffee beans and poultry feed, under the influence of two levels of moisture (20% and 30%) were monitored by the ELISA. The maximum fungal colonization was observed in poultry feed (9.8 and 11.8 mg g(-1)) followed by coffee beans (6.8 and 11.3 mg g(-1)) and chilli (5.1 and 6.3 mg g(-1)) at 20% and 30% moisture after 20 days of incubation. Similarly the fungus produced maximum ochratoxin A in poultry feed (25 and 120 microg g(-1)) followed by coffee beans (8 and 24 microg g(-1)) and chilli (0.2 and 0.45 microg g(-1)) at 20% and 30% moisture after 20 days of incubation. CONCLUSIONS: The method can be used for quantitative estimation of fungal biomass and comparison of fungal colonization in food substrates varying in composition. SIGNIFICANCE AND IMPACT OF THE STUDY: The method can be adapted for studying the fungal colonization in different solid substrates under different culture condition. The method is sensitive to mould colonization of >or=0.02% (w/w) and can be used for early detection of specific fungal infestation in food commodities.     

Fed-batch fermentation with and without on-line extraction for propionic and acetic acid production byPropionibacterium acidipropionici

Fed-batch propionic and acetic acid fermentations were performed in semi-defined laboratory medium and in corn steep liquor withPropionibacterium acidipropionici strain P9. On average, over four experiments, 34.5 g/l propionic acid and 12.8 g/l acetic acid were obtained in about 146 h in laboratory medium with 79 g/l glucose added over five feeding periods. The highest concentration of propionic acid, 45 g/l, was obtained when the glucose concentration was not allowed to drop to zero. In corn steep liquor 35 g/l propionic acid and 11 g/l acetic acid were produced in 108 h from 59.4 g/l total lactic acid provided as seven feedings of corn steep liquor. Extractive fed-batch fermentations were conducted in semi-defined medium using either flat-sheet-supported liquid membranes or hollow-fiber membrane extraction to remove organic acids from the culture medium. As operated during the course of the fermentation, these systems extracted 25% and 22% of the acetic acid and 36.5% and 44.5% of the propionic acid, respectively, produced in the fermentation. Total amounts of acids produced were about the same as in comparable nonextractive fermentations: 30–37 g/l propionic acid and 13 g/l acetic acid were produced in 150 h. Limitations on acid production can be attributed to limited substrate feed, not to failure of the extraction system.Journal paper J-16303 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Project 3122.