Fate of Escherichia coli during Ensiling of Wheat and Corn

A recombinant Escherichia coli strain carrying a plasmid with an antibiotic resistance marker and expressing the green fluorescent protein was inoculated at a concentration of 3.8 × 10⁸ CFU/g into direct-cut wheat (348 g of dry matter kg⁻¹), wilted wheat (450 g of dry matter kg⁻¹), and corn (375 g of dry matter kg⁻¹). The forages were ensiled in mini-silos. The treatments included control (no E. coli added), application of tagged E. coli, and delayed sealing of the inoculated wheat. Three silos per treatment were sampled on predetermined dates, and the numbers of E. coli were determined on Chromocult TBX medium with or without kanamycin. Colonies presumptively identified as E. coli were also tested for fluorescence activity. Addition of E. coli at the time of ensiling resulted in a more rapid decrease in the pH but had almost no effect on the chemical composition of the final silages or their aerobic stability. E. coli disappeared from the silages when the pH decreased below 5.0. It persisted longer in silages of wilted wheat, in which the pH declined more slowly. Control silages of all crops also contained bacteria, presumptively identified as E. coli, that were resistant to the antibiotic, which suggests that some epiphytic strains are naturally resistant to antibiotics.     

The effect of applying lactic bacteria at ensilage on the chemical and microbiological composition of vetch, wheat and alfalfa silages

The effect of applying a commercial lactic acid bacterial inoculant, at 5.6 × 10⁴ cfu/g fresh material, to vetch, wheat, direct-cut and wilted alfalfa silages has been studied under laboratory conditions, and on wheat also under farm conditions. Dry matter losses in the inoculated vetch and alfalfa silages were smaller than in the control silages, due to improved fermentation in the former as indicated by a faster and larger pH decrease and by a faster and larger lactic acid build-up. Volatile fatty acid analysis also indicated more efficient fermentation patterns in the inoculated vetch and alfalfa silages with less ethanol, acetic and butyric acids compared with the respective control silages. The inoculant suppressed enterobacteria and clostridia in the inoculated direct-cut alfalfa silage. The inoculant did not have a great effect on the wheat silages.     

Survival of <Emphasis Type="Italic">Listeria monocytogenes</Emphasis> in Wilted and Additive-Treated Grass Silage

Grass was field-dried to 3 different dry matter (DM) levels (200, 430 and 540 g/kg) and inoculated with 10⁶–10⁷ cfu/g of a Listeria monocytogenes strain sharing a phagovar occasionally involved in food-borne outbreaks of listeriosis. Formic acid (3 ml/kg) or lactic acid bacteria (8·10⁵/g) with cellulolytic enzymes were applied only to forages with low and intermediate DM levels. Forages were ensiled in laboratory silos (1700 ml) and were stored at 25°C for 30 or 90 days. After 90 days of storage, L. monocytogenes could not be detected in any silo, except one with the high dry matter grass without additive. After 30 days of storage, between 10² and 10⁶ cfu L. monocytogenes/g silage were isolated from the untreated silages. Increasing the DM content from 200 to 540 g/kg did not reduce listeria counts possibly because of the lower production of fermentation acids (higher pH). In silages treated with additives, counts of L. monocytogenes were always lower than in silages without additive. In wet silages (DM 200 g/kg) both additives were effective, but in the wilted silages (DM 430 g/kg) only the bacterial additive reduced listeria counts below detection level. Listeria counts were highly correlated to silage pH (r = 0.92), the concentration of lactic acid (r = -0.80) and the pooled amount of undissociated acids (r = -0.83).     

Effect of wilting and the additives,straw, molasses and urea on the fermentation pattern of maize silage

The process of ensiling was studied in fresh maize (15% dry matter (DM)), wilted maize (18 and 24% DM) and maize mixed with 5–20% of wheat straw (18, 25 and 29% DM). Silages with 24% DM were preserved better than those with lower dry matter content. There was a significant change, with time, in pH, titrable acidity, volatile fatty acids, lactic acid, number of lactic acid bacteria, volatile nitrogen and soluble sugars in all the treatments. There was a significant decline in volatile fatty acids (P<0.05) and ammonia (P<0.01) production, and a significant increase in soluble sugar (P<0.01) in silages made after wilting. A significant decline in titrable acidity (P<0.01), volatile fatty acid production (p<0.05) and ammonia nitrogen (P<0.01), and a significant increase in pH (P<0.01) were found in silages of maize mixed with wheat straw. The overall rate of fermentation decreased during the first few days of fermentation in wilted and wheat straw silages, but the final products had characteristics of a good silage. In the second experiment the effect of urea and molasses was studied on wheat straw plus maize (15:85) silage with an initial DM content of 31–34%. Three levels of molasses (0, 3 and 6% of fresh weight) and two levels of urea (0 and 0.5% of fresh weight) were studied. Urea treatment with 3% molasses was found to be the best on the basis of silage characteristics.     

Fermentative profile and lactic acid bacterial dynamics in non-wilted and wilted alfalfa silage in tropical conditions

This study was conducted to evaluate the fermentative profile and microbial populations of wilted and non-wilted alfalfa silages ensiled with or without inoculant and the population dynamics of lactic acid bacteria (LAB) of wilted alfalfa plant and theirs silage. A 2?×?2?×?6 factorial arrangement was used, with the absence or presence of wilting (W), with and without bacterial inoculant (I) and six fermentation periods (P) (1, 3, 7, 14, 28 and 56 days), in a completely randomized design, with three replicates. The alfalfa was slightly wilted for 6 h and increased the dry matter content from 133.9 to 233.4 g/kg. It was performed the cultivation, followed by the isolation of LAB from samples of alfalfa forage before ensiling and its silage only in non-inoculated silages, after different fermentation periods. DNA was extracted from the isolated strains of LAB; the 16S rRNA gene sequences were amplified by PCR and the sequences were compared to those available from the GenBank database. Wilting provided silages with lower pH, ammonia nitrogen and acetic acid concentrations. The wilting process did not alter the amount of LAB; however, it affected the LAB diversity of the silages. The Lactobacillus plantarum was the predominant species in non-wilted and wilted silages.

     

Fate of Escherichia coli O157 and detection of stx phage during fermentation of maize, an animal feedstuff

AIMS: The fate of inoculated Escherichia coli O157, stx phages and the physico-chemical properties of maize were studied during laboratory-scale fermentation by naturally occurring lactic acid bacteria. METHODS AND RESULTS: Before fermentation, chopped maize was inoculated with 6.2 log(10) CFU g(-1) of a five-isolate mixture of E. coli O157. After fermentation, the silage contained 70.6 g kg(-1) dry matter (DM) lactic acid and 12.8 g kg(-1) DM acetic acid and was pH 4.0. Levels of E. coli O157 fell rapidly, and none of the five isolates could be recovered from the fermenting maize after 8 days. Using a resuscitation step did not consistently enhance recovery of E. coli O157. Stx phages were not isolated from the fermenting maize at any time. Pulsed-field gel electrophoresis (PFGE) genotyping showed that E. coli O157 2975 and 64a/01 survived better than the other three isolates studied. Escherichia coli O157 isolate 1474/00 was particularly sensitive to the laboratory procedures used to harvest the inocula and contaminate the maize. Some colonies recovered during the fermentation had one to three band alterations compared with the initial PFGE genotypes. SIGNIFICANCE AND IMPACT OF THE STUDY: None of the five different E. coli O157 genotypes survived maize fermentation. Fermentation of maize produces an animal feedstuff that is unlikely to contain E. coli O157 or stx phages.     

Effects of maturity stage, wilting and acid treatment on crude protein fractions and chemical composition of whole crop pea silages (Pisum sativum L.)

In order to determine if maturity stage, and wilting or acid treatment, change the crude protein (CP) fraction distribution (determined according to the Cornell Net Carbohydrate and Protein System) of whole crop pea silages, a pea with variegated flowers (Pisum sativum ssp. arvense L., cv Timo) was compared to a white-flowered, semi-leafless pea (P. sativum ssp. hortense L, cv Capella). Herbage was harvested at three maturity stages being: pod set, pod swell and full pod, and either acid-treated or wilted. Timo was acid-treated using 4 (acid4), 6 (acid6) or 8 (acid8) L/tonne fresh matter (FM) with a 2:1 mixture of formic and propionic acid, or wilted to a dry matter (DM) content of about 400 g/kg. Capella was treated with acid6 or wilted. Herbage was ensiled for 103 days in 10 kg laboratory silos. Despite differences in wilting conditions, all wilted herbages had similar protein fraction distributions. In the Capella silages the soluble CP content was lower in the later maturity stages, but this was not the case in the Timo silages. The amount of acid added only affected the B1 CP fraction content, which decreased with increasing acid. At pod set and pod swell for Timo, and at pod set for Capella, the direct-harvested herbages were difficult to ensile because of the high buffering capacity and low level of water soluble carbohydrates. Wilting improved ensilability. Acid treatment reduced proteolysis, but crops with DM contents below 150 g/kg must be acid treated with at least 6 L/tonne FM to ensure stable fermentation. Timo silages were more prone to malfermentation, probably caused by lodging, which made Capella the preferred cultivar for producing pea silages harvested at the pod swell stage or later. Proteolysis and the amount of soluble CP in the silage were lower in later maturity stages in the Capella, but not the Timo, cultivar.     

The effect of a propionic acid bacterial inoculant applied at ensiling on the aerobic stability of wheat and sorghum silages

The effect of a new strain ofPropionibacterium shermanii (PAB), applied at ensiling, on the aerobic stability of wheat and sorghum silages was studied in several experiments under laboratory conditions. In the one experiment with wheat and in those with sorghum a lactic acid bacteria (LAB) inoculant (Lactobacillus plantarum andPediococcus cerevisiae) was also included. After treatment, the chopped forages were ensiled in 1.5-L anaerobic jars which were sampled in triplicate on predetermined dates to follow fermentation dynamics. At the end of the experiments, the silages were subjected to an aerobic stability test. The PAB inoculant improved the aerobic stability only in one experiment with wheat, in which the decrease in pH was very slow; the final pH remained relatively high (4.5). The PAB-treated silages contained 19.5±2.0 g of propionic acid per kg of dry matter. In the experiments with sorghum, the control and PAB-inoculated silages were stable, whereas LAB-inoculated silages deteriorated. The results suggest that PAB can survive in and improve the aerobic stability of only slow-fermenting silages which are prone to aerobic deterioration.     

Fatty Acid and Isoprenoid Quinone Composition in the classification of Bacteroides melaninogenicus and Related Taxa

Aerobic deterioration of lucerne, maize and wheat silages was characterized by rapid increases in yeast and mould flora which oxidized lactic and volatile acids resulting in increased temperature and pH. While populations of yeasts and moulds were similar, temperature increases were slightly greater for silages inoculated with Lactobacillus acidophilus and Candida spp. After 48 h the pH of the inoculated silages was higher in general and concentrations of acids were lower than controls. Bacterial growth was slight although continued lactic acid production was probable. In contrast to lucerne and maize silages, the pH of wheat silage remained stable during this period because of high butyric levels, but temperature and yeast populations increased. After 48 h the pH rose above 5 in maize and lucerne, and bacterial growth and metabolic activity resumed resulting in volatile and non-volatile acid production from carbohydrate fermentation and deamination of amino acids. During this phase of aerobic deterioration yeast growth slowed or stopped, but temperatures remained high and pH continued to climb probably because of production of ammonia. The changes in gross composition of the silages did not follow any particular pattern. Losses in dry matter were small (2.5–4.0%) and changes in individual components probably reflect this loss rather than substantial changes. Protein availability in the lucerne silages undoubtedly decreased, as protein losses were high. It is concluded that the aerobic deterioration of silage is enhanced by the addition of L. acidophilus and Candida spp. at ensiling.     

Nutritive value of ensiled broiler litter for cattle

Broiler litter, consisting of bedding material (chopped wheat straw or rice hulls), excreta, wasted feed and feathers was ensiled at 40 or 50% moisture for 42 days. Cheddar cheese whey was added to adjust the moisture level in some of the silage. The dry matter digestibility in vitro (IVDMD), after 21 days of ensiling was greater for silage containing the wheat straw base than for that with the rice hull base. Whey improved the IVDMD of the wheat straw base silage but not that of the rice hull base silage. Moisture levels did not influence the IVDMD. In a second trial, broiler litter consisting of chopped wheat straw bedding, excreta, wasted feed and feathers was ensiled for 28 days at approximately 45% dry matter. The litter was ensiled as: (1) litter alone; (2) litter plus Irish potato cannery waste (IPW); (3) litter plus ground maize, and (4) litter plus ground grain sorghum. The materials added supplied 33% of the dry matter of the silages. The pH of the silages 1 and 4 weeks after ensiling was (1) 6.00, 5.83; (2) 5.00, 4.56; (3) 4.96, 4.80; and (4) 4.92, 4.78. Total faecal and urine collection trials were conducted using 12 Holstein steers having an average body weight of 200 kg. Dry matter digestibility was greater (P < 0.01) for silages 2, 3 and 4. Digestible energy and protein were 61.2, 70.6; 65.3, 74.6; 65.2, 71.2; and 68.2, 76.4% for silages 1 through 4.     

Changes during aerobic exposure of wheat silages

Aerobic stability is an important characteristic of silages because they are exposed to air during storage and feedout. The objective of the current study was to investigate changes that occur in wheat silages during aerobic exposure. Silages of whole crop wheat harvested at the flowering, milk and dough stages of maturity were prepared in 1.5 L anaerobic glass jars. Three months after ensiling, silages were subjected to a 7-day aerobic stability test. The silages of wheat harvested at the flowering stage were the most stable upon aerobic exposure, but had the largest fermentation losses. Silages of milk-stage wheat were unstable upon aerobic exposure, and had large amounts of CO₂ and heating, large yeast populations, decreased amounts of fermentation products and decreased dry matter (DM) and neutral detergent fiber (NDF) digestibility (from 667 to 572 g/kg DM and from 597 to 558 g/kg DM, following 7-day aerobic exposure, respectively). Silages of dough-stage wheat had relatively low fermentation losses and were quite stable for at least 4 days of aerobic exposure. Silage samples from the center of commercial bunker silos, and from areas adjacent to the walls of the silos, were judged to be of good quality. Samples from the shoulders of bunker silos were spoiled with higher pH (P<0.05), higher ash content and lower DM and NDF digestibility compared with samples from the center of the silos and areas near the walls (6.8 vs. 4.0, 19.0 vs. 7.8, 477 g/kg DM vs. 634 g/kg DM and 230 g/kg DM vs. 487 g/kg DM, respectively).     

Growth of Escherichia coli O157 in poorly fermented laboratory silage: a possible environmental dimension in the epidemiology of E. coli O157

Laboratory silages, inoculated with either c. 1000 cfu g-1, an atoxigenic Escherichia coli O157 or a toxigenic E. coli O157 isolate, were made in plastic bags which permitted limited aerobic spoilage. Replicate bags of each treatment were opened at weekly intervals after incubation at 20 degrees C. In all silages the fermentation was slow and aerobic spoilage with visible moulding ocurred at the tie ends after 7 d. In all the aerobically spoiled silages Enterobacteriaceae reached over 107 cfu g-1 within 1 week. The E. coli in control silages increased from barely detectable levels to 104 cfu g-1 within 13 d; over the same period both strains of E. coli O157 increased from 103 to 107 cfu g-1. The increases in the poorly fermented interior of the silage bags were initially similar but declined slightly as the pH fell. It is suggested that faecal contamination of grass followed by poor silage management may be a factor in the persistence of E. coli O157 carriage in ruminants.     

Accumulation of 1,2-propanediol and enhancement of aerobic stability in whole crop maize silage inoculated with Lactobacillus buchneri

Aims: To assess the effects of inoculation of Lactobacillus buchneri on the ensiling properties and aerobic stability of maize silage. Methods and Results: Chopped whole crop maize was ensiled in 0.5 litre airtight polyethylene bottles (0.4 kg per bottle) and in double-layered, thin polyethylene bags (15 kg per bag), with or without inoculation of Lact. buchneri. The silos were stored for two to four months and the chemical composition, microbial numbers and aerobic stability were determined. Inoculation lowered lactic acid and yeasts, and increased acetic acid and pH value, resulting in improved aerobic stability of the silages. Inoculated silages produced 1,2-propanediol, the content of which increased as ensiling was prolonged, and nearly 50 g kg-1 dry matter had accumulated after four months of storage. The effects of inoculation, however, were much less pronounced in silages prepared in bags. Mannitol was found in all silages; the production was lowered by Lact. buchneri treatment and appeared to be unrelated to the accumulation of 1,2-propanediol. Conclusions: Inoculation of Lact. buchneri occasionally causes accumulation of 1,2-propanediol in silages without further degradation into propionic acid and 1-propanol. Significance and Impact of the Study: Substantial amounts of 1,2-propanediol could be consumed by ruminants when fed on silages inoculated with Lact. buchneri. In addition to increasing acetic acid, attention needs to be paid to 1,2-propanediol because the two fermentation products might affect the intake and utilization of silage-based diets.     

The effect of Lactobacillus buchneri on the fermentation, aerobic stability and ruminal degradability of maize silage

AIMS: To evaluate the effect of Lactobacillus buchneri, heterofermentative lactic acid bacteria (LAB), on the fermentation, aerobic stability and ruminal degradability of whole-crop maize silages under laboratory conditions. Two homofermentative LAB were tested for the purpose of comparison. METHODS AND RESULTS: Maize was harvested at early dent [290 g kg(-1) dry matter (DM)] and one-half milk line (355 g kg(-1) DM) stages. Both homofermentative LAB were applied at 1 x 10(5) CFU g(-1) of fresh forage. Lactobacillus buchneri was applied at 1 x 10(5), 5 x 10(5) and 1 x 10(6) CFU g(-1) of fresh forage. Silages with no additives served as control. After treatment, the chopped forages were ensiled in 1.5-l anaerobic jars. Three jars per treatment were sampled on day 60. After 60 days of storage, silages were subjected to an aerobic stability test lasting for 5 days, in which CO(2) production, as well as chemical and microbiological parameters, was measured to determine the extent of aerobic deterioration. Both homofermentative LAB increased the concentration of lactic acid and the numbers of yeasts, and decreased the concentration of acetic acid and impaired the aerobic stability of silages. In contrast, applying L. buchneri decreased the concentration of lactic acid and increased the concentration of acetic acid of the silages. Under aerobic conditions, silages treated with 5 x 10(5) and 1 x 10(6) CFU g(-1) of L. buchneri, had lower pH, CO(2) production and the numbers of yeasts than the silages treated with 1 x 10(5) CFU g(-1) of L. buchneri (P < 0.05). However, all doses of L. buchneri and both homofermentative LAB did not affect in situ rumen DM, organic matter and neutral detergent fibre degradability of the silages. CONCLUSIONS: Lactobacillus buchneri was very effective in protecting maize silages exposed to air under laboratory conditions. All doses of L. buchneri, especially 5 x 10(5) CFU g(-1) or more, markedly decreased the numbers of yeasts and improved the aerobic stability of silages. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of L. buchneri, as a silage inoculant, can improve the aerobic stability of maize silages by inhibition of yeast activity.     

The effects of lactic acid bacteria inoculants and formic acid on the formation of biogenic amines in grass silages

Silages were prepared in six laboratory experiments from four direct-cut grassland swards and pure swards of perennial ryegrass and false oat with dry matter contents ranging between 180 and 325 g/kg. Grass was fermented at 22 degrees C and silages were stored at the same temperature for 4 months. Untreated silages (negative control) and silages preserved with 3 g/kg of formic acid (positive control) were compared with silages inoculated with commercial strains of Lactobacillus plantarum, Lactobacillus buchneri and a mixed preparation Microsil. The inoculants were applied at a dose of 5.10(6) CFU/g of grass. Seven biogenic amines were extracted from silages with perchloric acid and determined as N-benzamides by micellar electrokinetic capillary chromatography. Common chemical quality parameters of silages were also determined. Tyramine, cadaverine and putrescine were the amines occurring at the highest concentration. As compared to untreated silages, formic acid was most effective to suppress formation of the main amines. Also the inoculants often decreased amine contents significantly (P < 0.05). The inoculants decreased levels of polyamine spermidine more efficiently than formic acid. Contents of histamine, tryptamine and polyamine spermine were very low, commonly below the detection limits.     

Survival of Escherichia coli O157:H7 in feces from corn- and barley-fed steers

The survival of Escherichia coli O157:H7 in feces from steers fed corn (CO) or barley (BA) was evaluated at -10, +4 and +22 degrees C. Fecal pats were inoculated with a four-strain mixture of nalidixic-acid resistant E. coli O157:H7 at two levels: 10(3) CFU g(-1) (low, L) and 105 CFU g(-1) (high, H). At -10 degrees C, duration of survival of E. coli O157:H7 was reduced (p < 0.05) in CO-L (35 days) compared to BA-L (49 days), likely due to the effects of fecal volatile fatty acids in combination with a fecal pH of <6.5. At 4 degrees C, E. coli O157:H7 was detected in BA-H, CO-H, CO-L and BA-L for 77, 77, 56 and 63 days, respectively, with no difference (p > 0.05) observed in the duration of survival or rate of decline of E. coli O157:H7 among treatments. Survival of E. coli O157:H7 was twice as likely (p < 0.05) at 22 degrees C than at 4 degrees C and -10 degrees C. While pH and dry matter content increased, and volatile fatty acid concentrations decreased over 84 days at all three temperatures, these changes were most pronounced at 22 degrees C. Survival of E. coli O157:H7 for extended periods of time in feces from both corn- and barley-fed animals was demonstrated, thus fecal material may serve as a vector for the transmission of the organism. The greater survival of E. coli O157:H7 at 22 degrees C suggests that temperature may play a role in the seasonality of transmission and prevalence of this bacterium in feedlot cattle. The reported greater prevalence of E. coli O157:H7 in cattle fed barley as compared to those fed corn does not appear to be related to elevated risk of transmission arising from differential survival of the bacterium in feces.     

Energy and nitrogen balance,intake and growth rate of lambs fed on unwilted grass silages treated with a formaldehyde—Acid mixture or untreated

Wether lambs were fed on precision-chopped first-cut ryegrass silage ad libitum in an intake trial, and a maintenance and 1.5 times maintenance in balance trials.The untreated and treated silages had pH values of 4.72 and 4.40, mean dry matter (DM) contents of 176 and 184 g kg^?1 and mean gross energy (GE) contents of 18.8 and 19.0 MJ/kg DM, respectively.Mean digestibility coefficients of DM (0.787 and 0.783), organic matter (OM) (0.827 and 0.820) and GE (0.794 and 0.793), for the treated and untreated silages respectively, were high. The metabolisable energy (ME) contents of the untreated and treated silages were 12.52 and 12.76 MJ/kg DM at maintenance and 11.94 and 12.56 MJ/kg DM at 1.5 times maintenance, respectively. The mean efficiency of utilisation of ME of the untreated and treated silages was 0.65 and 0.66 for maintenance (k_m) and 0.34 ± 0.134 and 0.40 ± 0.069 for growth (k_g), respectively; the k_g values were lower than expected.Dry matter intakes of these silages when given ad libitum were 27.9 and 28.8 g/kg W per day and produced live weight gains of 129 and 140 g day^?1 for the untreated and treated silages, respectively. These gains were similar to predicted values for live weight gain only when the experimentally determined k_g and k_m values were substituted in the equation of the Agricultural Research Council (1980) used for calculating the daily metabolisable energy requirements for live weight gain.     

Characterization of protein fractions and amino acids in ensiled alfalfa treated with different chemical additives

Formic acid, formaldehyde, tannic acid or mixtures of two were studied on their effects on ensiled alfalfa (Medicago sativa L.) amino acids and N fractions by the Cornell Net Carbohydrate and Protein System (CNCPS). The alfalfa forage was a second cut and was wilted to a mean over-dry dry matter (DM) content of 330 g/kg. All silages were prepared as mini-silos using 100 ml polypropylene centrifuge tubes (50 g) on a small laboratory-scale, with the additives added in 20 ml aliquots/kg herbage fresh weight (FW). After 35 d of ensiling, most of forage true protein was converted to fraction A and all of the added additives reduced fraction A content in the ensiled forages (P<0.05). The content of fraction B₁ in all of the additive-treated silages was higher (P<0.05) than that in control silage. Large proportions of true protein in the tannic acid/formaldehyde- and formic acid/formaldehyde-treated silages were fractions B₂ and B₃, respectively. No difference was observed on fraction C content between the control silage and silages treated with additives except for the formaldehyde or tannic acid-treated silages. Amino acids were well preserved in additive-treated silages compared with the control silage. Concentration of total amino acid was higher in formic acid-treated silages than that in the control and the other additive-treated silages (P<0.05). The pattern of changes in individual amino acid in all of the silages indicated that branched chain amino acids and methionine were relatively well preserved during fermentation but the basic and acidic amino acids were not.     

Fate of inoculated Escherichia coli in hay

AIMS: To monitor the fate of inoculated Escherichia coli in dry and moist hay of various types, under laboratory conditions. METHODS AND RESULTS: Wheat, vetch and clover hay were used as received or wetted to 250-300 g kg(-1) moisture. The hay was inoculated at about 10(6) CFU g(-1) with a kanamycin-resistant E. coli strain that expresses the green fluorescence protein, and was stored in small open glass jars that were covered with aluminium foil. Three jars per treatment were sampled on days 1 and 3, or 4 and 7, or 8, 20 and 50, respectively, after the initiation of the experiments, and the numbers of E. coli in the hay were determined. The results indicated that E. coli disappeared from both dry and moist hay by 7-8 days after inoculation. However, in a few cases colonies that were presumed to be E. coli developed after incubation in Luria broth medium. CONCLUSIONS: The tagged E. coli strain usually disappeared rapidly from both the dry and the moist hay, in spite of the high level of inocula used. However, in some cases a few, possibly injured E. coli might have persisted, and could be detected after incubation in a rich growth medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is part of a risk assessment associated with sewage irrigation of forage crops in Israel. The results indicate that E. coli added to the hay is not likely to pose a health risk to cattle or to humans. Nevertheless, more research with natural strains of E. coli and other enteric pathogens that might be more adapted to forage conditions is warranted in order to ensure the safety of sewage-irrigated crops.     

Fermentation characteristics and feeding value of ensiled poultry litter containing wheat straw,bagasse or sawdust

Poultry excreta substantially increased the crude protein (CP) content and the calcium and phosporus content of the base bedding materials. The crude fibre (CF) content of sawdust (52.3%) was higher than that of wheat straw (38.9%) and bagasse (30.2%). Dry matter digestibility in vitro (IVDMD) of wheat straw, bagasse and sawdust poultry litters (PL) was 65.4, 64.5 and 48.1%, respectively. Green sorghum fodder when ensiled alone or with 20% wheat straw PL, sawdust PL or bagasse PL on fresh basis contained 4.67, 7.80, 10.00 and 7.55% CP, respectively. Nitrogen-free extract (NFE) content of PL silages was lower than that of the control. Apart from wheat straw PL, all silages accumulated considerable amounts of lactic acid. The total volatile fatty acids (TVFA) concentrations were similar for all silages. The addition of PL caused an increase in the proportion of ammonia nitrogen. A feeding trial with crossbred adult male cattle revealed no significant difference in dry matter (DM), CP and ether extract (EE) digestibility of wheat straw and bagasse PL silages. The CF digestibility was similar for all the silages. Sawdust PL silage, however, was significantly lower (P < 0.05) in digestibility of DM, EE and NFE compared to other PL based silages. The DCP and TDN values for the control, wheat straw, bagasse and sawdust PL silages were 2.0, 60.1; 4.3, 45.3; 6.1, 50.3 and 2.9, 41.9 kg/100 kg DM, respectively.