Research Note: Influence of monocalcium phosphate and phytase in the diet on phytate degradation in cecectomized laying hens.

This study investigated the effects of phytase and monocalcium phosphate supplementation on the dephosphorylation of phytic acid [myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate); InsP6] in cecectomized laying hens using total excreta collection. Four corn-soybean meal-rapeseed meal-based diets were mixed with or without 6 g of monocalcium phosphate/kg, with or without supplementation of 1,500 FTU phytase/kg, and had the same calcium concentration at 39 g/kg of feed. Each diet was tested in 5 replicates using a row-column design with 10 cecectomized laying hens in 2 periods. The hens received 120 g/d of feed while being housed individually in metabolism units, and total excreta were collected for a period of 4 d. The monocalcium phosphate × phytase interaction was not significant for InsP6 degradation (P = 0.054). Phytase increased InsP6 disappearance from 13% to 83% (P < 0.001), whereas monocalcium phosphate had no effect. Concentrations of most of the lower inositol phosphate isomers in excreta were higher when monocalcium phosphate was added to the diets. The concentration of Ins(1,2,5,6)P4 in excreta was the highest among the studied partially dephosphorylated inositol phosphates with phytase supplementation and was higher than in diets without phytase supplementation (P < 0.001). Supplementation with phytase increased myo-inositol concentration in excreta (P = 0.002), whereas monocalcium phosphate had no effect. Phosphorus utilization ranged from 4% to 18% and was not significantly affected by the treatments. These results suggest that phytase supplementation markedly increased InsP6 degradation in laying hens. The cecectomized laying hen assay may be suitable for studying the effects of phytase supplementation on phytate dephosphorylation under dietary conditions when performance and phosphorus excretion are unlikely to be affected.

Phytate degradation and phosphorus utilisation by broiler chickens fed diets containing wheat with increased phytase activity.

1. The objective of this study was to investigate wheat genotypes bred for increased intrinsic phytase activity for InsP6 disappearance and the formation of lower inositol phosphates in such wheat-fed broiler chickens. The influence of monocalcium phosphate (MCP) supplementation on these characteristics and the utilisation of P and Ca were also determined. A three-step in vitro assay and a broiler trial were performed.2. In the 63 wheat genotypes tested in vitro, phytase activity varied from 1900 FTU/kg to 5200 FTU/kg, and InsP6 disappearance increased with higher phytase activity of wheat in a linear manner. The addition of MCP significantly reduced in vitro InsP6 disappearance by one-third, independent of the inclusion level of wheat in the feed. When exogenous phytase was added to wheat, in vitro InsP6 disappearance increased independently of the phytase activity of the wheat used.3. In the broiler trial, four wheat genotypes with phytase activities between 2400 and 3700 FTU/kg were included at 400 g/kg in diets with and without MCP. The diets were not pelleted. Separately, wheat 1, without MCP, was tested with the addition of exogenous phytase. Unsexed Ross 308 broiler chickens were allocated to 72 metabolic units of 10 birds each and assigned one of the nine diets. Mineral utilisation was measured based on excreta collection from 20 to 23 d of age. Digesta from the crop and terminal ileum were collected on d 24.4. In the crop and ileum, InsP6 disappearance was not affected by the wheat genotypes, but the addition of MCP significantly decreased InsP6 disappearance. Precaecal P disappearance was significantly reduced by the addition of MCP, with wheat genotypes also exerting an effect. Wheat genotypes and the addition of exogenous phytase significantly affected P utilisation. Exogenous phytase had no effect on InsP6 disappearance in the crop but did up to the terminal ileum, the precaecal InsP6 and P disappearance increased with the addition of exogenous phytase.5. Although the intrinsic wheat phytase activity exerted distinct effects on in vitro InsP6 disappearance, no such effect was found in the broiler trial. The addition of MCP significantly inhibited InsP6 degradation in vitro and in vivo.

Phytate degradation and phosphorus digestibility in broilers and turkeys fed different corn sources with or without added phytase.

The aim of the present study was to test whether different dietary corn sources and phytase supplementation affect the prececal phosphorus digestibility (pcdP) and appearance of inositol phosphates in the lower ileum of growing broiler chickens and turkeys. Two experiments were conducted, one with broiler chickens and one with turkeys. Four corn diets were provided; these were formulated to contain low P and calcium (Ca) contents and incorporated 43% of one of the four different corn sources. Diets were either unsupplemented or supplemented with 500 FTU of an Escherichia coli-derived phytase/kg feed. Experimental diets were fed ad libitum from day 20 post-hatch. At 28 d of age, digesta were sampled from the lower ileum of animals to determine pcdP and pc myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) degradation and to analyze the concentrations of lower inositol phosphate isomers. The pcdP of non-supplemented diets ranged from 51 to 60% and from 22 to 28% in broilers and turkeys, respectively. A negative correlation was observed between the InsP6 content of the corn source and the pcdP of diets in broilers only. Without phytase supplementation, pc InsP6 degradation ranged from 64 to 76% in broilers and from 6 to 15% in turkeys. Phytase increased the pcdP by around 15% in broilers (P < 0.001) and 9 to 17% in turkeys (P < 0.001). In turkeys, phytase efficacy was greatest when the diets contained corn with higher contents of ether extract and InsP6. An effect of corn source on the appearance of lower InsPs in the ileal digesta was found in broilers only. These results suggest that broilers possess a greater capacity for InsP6 degradation and hydrolysis of lower InsPs compared with turkeys. Furthermore, the results are influenced by the corn source used. Further research is needed to identify the factors responsible for the low level of phytate degradation in turkeys in order to improve the availability of InsP6-P and the efficacy of phytase.

Effect of supplemental phytase and xylanase in wheat-based diets on prececal phosphorus digestibility and phytate degradation in young turkeys.

This study aimed to investigate the effect of phytase and a combination of phytase and xylanase on the prececal phosphorus digestibility (pcdP) of wheat-based diets in turkeys. A low-P basal diet (BD) based on cornstarch and soybean meal, and 2 diets containing 43% of different wheat genotypes (genotype diets GD6 or GD7) were fed to turkeys from 20 to 27 d of age. Diets were fed either without enzyme supplementation or supplemented with phytase (500 FTU/kg) or a combination of phytase and xylanase (16,000 BXU/kg). At 27 d of age, digesta were sampled from the lower ileum of animals to determine pcdP and pc myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) disappearance, and to analyze the concentrations of lower inositol phosphate isomers. Similar pcdP was observed in non-supplemented BD and GD (∼36%). Phytase alone increased the pcdP in all diets by 8 to 12%, but a beneficial effect of xylanase was found only for BD. Similar results were found for pc InsP6 disappearance, although xylanase addition compared to phytase alone decreased pc InsP6 disappearance in GD7 compared to phytase alone. Animals fed GD7 performed better than those fed GD6; however, these differences could not be linked to the pcdP. The pattern of lower inositol phosphates in digesta also changed with enzyme supplementation, resulting in lower proportions of InsP5 and higher proportions of InsP4. Phytase alone decreased Ins(1,2,3,4,6)P5 but increased D-Ins(1,2,3,4,5)P5 and D-Ins(1,2,5,6)P4 concentrations. An additional increase in D-Ins(1,2,3,4,5)P5 and D-Ins(1,2,5,6)P4 concentrations was achieved with xylanase, although for the former isomer, this was observed only with GD. These results indicate that enzyme supplementation alters the pc degradation of InsP6, and that combining both enzymes had a minor additional effect on the pcdP from wheat-based diets when compared to phytase alone.

Influence of phytase or myo-inositol supplements on performance and phytate degradation products in the crop, ileum, and blood of broiler chickens.

The objective of this study was to investigate the effects of supplementation with free myo-inositol (MI) or graded levels of phytase on inositol phosphate (InsP) degradation, concentrations of MI in the digestive tract and blood, bone mineralization, and prececal digestibility of amino acids (AA). Ross 308 broiler hatchlings were allocated to 40 pens with 11 birds each and assigned to one of 5 treatments. The birds were fed a starter diet until d 11 and a grower diet from d 11 to d 22. All diets were based on wheat, soybean meal, and corn. Birds were fed a control diet, calculated to contain adequate levels of all nutrients without (C) or with MI supplementation (C+MI), or one of 3 experimental diets that differed in phytase level (modified E. coli-derived 6-phytase; Phy500, Phy1500, or Phy3000 FTU/kg), with P and Ca levels adapted to the recommendations of the phytase supplier for a phytase level of 500 FTU/kg. The gain:feed ratio (G:F) was increased by MI or phytase in the starter+grower phase by 0.02 g/g. Prececal P and Ca digestibility, P and Ca concentration in blood serum, and tibia ash weight did not differ among treatments (P > 0.05). MI supplementation led to the highest MI concentration in the crop, ileum, and blood plasma across treatments. Phytase supplementation increased MI concentrations in the crop and ileum digesta in a dose-dependent manner and in plasma without any dose effect (P > 0.05). Prececal digestibility of some AA was increased by phytase. These outcomes indicate that MI might have been a relevant cause for the increase in G:F. Therefore, it is likely that the release of MI after complete dephosphorylation of phytate is one of the beneficial effects of phytase, along with the release of P and improvement in digestibility of other nutrients. Simultaneously, MI seems to have no diminishing effects on InsP degradation.

Interactive effects of phosphorus, calcium, and phytase supplements on products of phytate degradation in the digestive tract of broiler chickens.

This study aimed to distinguish between the single and interactive effects of phosphorus (P), calcium (Ca), and phytase on products of phytate degradation, including the disappearance of myo-inositol (MI), P, Ca, and amino acids (AA) in different segments of the digestive tract in broiler chickens. Additionally, all dephosphorylation steps from myo-inositol 1,2,3,4,5,6-hexakis (dihydrogen phosphate) (InsP6) to MI were investigated in the digesta of the terminal ileum. Unsexed Ross 308 broiler chickens were allocated to 56 pens with 19 birds per pen, and assigned to one of 8 dietary treatments. The dietary treatments included diets without (P-, 4.1 g/kg DM) or with (P+, 6.9 g/kg DM) monosodium phosphate supplementation, without (Ca-, 6.2 g/kg DM) or with (Ca+, 10.3 g/kg DM) additional fine limestone supplementation, and without or with 1,500 FTU phytase/kg feed in a factorial design. Adding Ca or P had no effect on InsP6 disappearance in the crop when phytase was added. InsP6 disappearance up to the terminal ileum (P-Ca- 56%) was decreased in P+Ca- (40%), and even more so in P+Ca+ (21%), when no phytase was added. Adding phytase removed all effects of P and Ca (77 to 87%); however, P+Ca+ increased the concentrations of lower InsP esters and reduced free MI in the ileum, even in the presence of phytase. These results indicate that mineral supplements, especially P and Ca combined, reduce the efficacy of endogenous microbial or epithelial phosphatases. Supplementation with phytase increased, while supplementation with Ca decreased the concentration of MI in all segments of the digestive tract and in blood plasma, demonstrating the ability of broilers to fully degrade phytate and absorb released MI. While AA disappearance was not affected by P or Ca, or an interaction among P, Ca, and phytase, it increased with the addition of phytase by 2 to 6%. This demonstrates the potential of the phytase used to increase AA digestibility, likely independent of P and Ca supply.

Ruminal phytate degradation of maize grain and rapeseed meal in vitro and as affected by phytate content in donor animal diets and inorganic phosphorus in the buffer.

The ruminal disappearance of phytate phosphorus (InsP6 -P) from maize grain and rapeseed meal (RSM) was determined in two in vitro studies. In experiment 1, two diets differing in phosphorus (P) and InsP6 -P concentration were fed to the donor animals of rumen fluid (diet HP: 0.49% P in dry matter, diet LP: 0.29% P). Maize grain and RSM were incubated in a rumen fluid/saliva mixture for 3, 6, 12 and 24 h. In experiment 2, a diet similar to diet HP was fed, and the rumen fluid was mixed with artificial saliva containing 120 mg inorganic P/l (Pi) or no inorganic P (P0). Maize grain and RSM were incubated with either buffer for 3, 6, 12 and 24 h. Total P (tP) and InsP6 concentration were analysed in the fermenter fluids and feed residues. The disappearance of InsP6 -P from maize was completed after 12 h of incubation in both experiments. From RSM, 93% (diet LP) and 99% (diet HP) of the InsP6 -P in experiment 1 and 80% (Pi) and 89% (P0) in experiment 2 had disappeared after 24 h of incubation. InsP6 -P disappearance was higher when diet HP was fed (maize: 3 and 6 h; RSM: 6 and 24 h of incubation) and when rumen fluid was mixed with buffer P0 (maize: 6 h; RSM: 12 and 24 h of incubation). InsP6 -P concentration in the fermenter fluids was higher for maize, but no accumulation of InsP6 -P occurred, indicating a prompt degradation of soluble InsP6 . These results confirmed the capability of rumen micro-organisms to efficiently degrade InsP6 . However, differences between the feedstuffs and diet composition as well as the presence of inorganic P in the in vitro system influenced the degradation process. Further studies are required to understand how these factors affect InsP6 degradation and their respective relevance in vivo.

Effect of dietary mineral phosphorus and phytate on in situ ruminal phytate disappearance from different concentrates in dairy cows.

The first objective of this study was to determine the influence of dietary composition on the in situ disappearance of phytate (InsP6) from wheat, corn, soybean meal, and rapeseed meal [solvent-extracted, without (RSM) or with (hRSM) heat treatment] in the rumen of dairy cows. The second objective was to assess the primary degradation products of InsP6 in the rumen. Three diets differing in phosphorus and InsP6 concentration (basal diet = 0.38% P in dry matter; high-P diet = 0.56% P; high-InsP6 diet = 0.39% P) were fed to 3 ruminally fistulated lactating Jersey cows in a 3 × 3 Latin square. Ground concentrates (sieve size = 2 mm) were incubated in polyester bags in the rumen for 2, 4, 8, 16, and 24 h. The bag residues were analyzed for P, InsP6, isomers of lower inositol phosphates (InsP5, InsP4, InsP3), and crude protein. The InsP6 disappeared more rapidly from cereal grains than from oilseed meals; however, after 24 h of incubation ≥95% InsP6 had disappeared from all concentrates except hRSM (57%; diet average). Feeding the high-InsP6 diet increased InsP6 disappearance for oilseed meals, but not for corn and wheat. The predominant InsP5 isomer in all bag residues was Ins(1,2,4,5,6)P5 followed by Ins(1,2,3,4,5)P5 and Ins(1,2,3,4,6)P5. A further InsP5 isomer [Ins(1,3,4,5,6)P5] was detected in both rapeseed meal bag residues. Feeding the high-InsP6 diet led to lower concentrations of Ins(1,2,4,5,6)P5 and Ins(1,2,3,4,5)P5, whereas an interaction between diet, concentrate, and time occurred for Ins(1,2,3,4,6)P5 and Ins(1,3,4,5,6)P5. The results confirm the high potential of rumen microorganisms to hydrolyze InsP6; however, increasing the amount of InsP6 in the diet can further enhance InsP6 hydrolysis, which may be relevant when concentrates with slowly degradable InsP6, such as RSM or heat-treated concentrates, are fed to dairy cows. Based on the concentrations of InsP5 isomers, 3 and 6 phytases appear to play a major role in the rumen. Conversely, intrinsic plant phytase activity appears to be less relevant as the percentage of its primary hydrolysis product, Ins(1,2,3,4,5)P5, changed only slightly upon using wheat known for high intrinsic phytase activity instead of the other concentrates. Additional information regarding the factors influencing the extent of ruminal InsP6 disappearance will require further studies to determine the phytase activity of rumen microorganisms and the characteristics of their respective phytases.

Effect of glycine supplementation in low protein diets with amino acids from soy protein isolate or free amino acids on broiler growth and nitrogen utilisation.

Here, it was investigated whether substitution of amino acids (AA) from soy protein isolate with free AA in low crude protein diets influences the growth performance and N utilisation in broilers, and whether interactions with dietary glycine equivalent (Glyequi) concentration exist. Birds were distributed in two 2 × 2 factorial arrangements of 48 floor pens containing 10 birds each, plus 48 metabolism cages containing two birds each. Experimental feed was provided for ad libitum consumption from d 7 to 22. Diets contained either a soy protein isolate at 79 g/kg or a mix of free AA, which supplied the same amount of 18 proteinogenic AA. A mix of free glycine and l-serine was used to obtain low and high (12.0 and 20.5 g/kg dry matter) levels of dietary Glyequi. Substitution of soy protein isolate with free AA reduced the average daily gain and feed efficiency, mainly due to reduced feed intake. Efficiency of N accretion was not influenced by the AA source or Glyequi concentration on d 21, possibly due to the lower AA digestibility of soy protein isolate and higher urinary excretion of nitrogenous substances in the treatments with the AA mix. The average daily weight gain of the treatments with high Glyequi concentration was higher for both AA sources. This increase was due to higher average daily feed intake by broilers in the treatments with soy protein isolate and due to the increased feed efficiency in the treatments with the AA mix. Broilers exhibited different growth responses to dietary Glyequi between the AA sources; however, these responses could not be attributed to the different utilisation of Glyequi for uric acid synthesis.

Effects of different forms and origins of oilseeds on dynamics of ruminal biohydrogenation of long-chain fatty acids in vitro.

Dietary unsaturated fatty acids (FA) are intensively hydrogenated in the rumen, resulting in reduced amount of poly-unsaturated fatty acids (PUFA) and accumulation of several biohydrogenation (BH) products. In this study, BH of PUFA originating from different oilseeds (linseed, soya beans, sunflower seed and rapeseed) present in crushed oilseeds or their free oils were assessed in vitro. The assay substrates were incubated in buffered rumen fluid for 0, 6, 12 and 24 h. After incubation, the FA pattern of the incubated samples was analysed using gas chromatography. Biohydrogenation is defined as disappearance of double bonds (DB) calculated from the contents of unsaturated FA. After 24-h incubation, the DB contents of all oilseeds were reduced (p < 0.001) by 40-60%. The reduction was higher (p < 0.001) for the crushed form compared with the oil form. In addition, linseed and sunflower seed known as oilseeds with high contents of linolenic acid C18:3 c9,12,15 (LNA) and linoleic acid C18:2 c9,12 (LA), respectively, showed a higher (p < 0.001) accumulation of the BH intermediates conjugated linoleic acid (CLA, isomer C18:2 c9t11) and vaccenic acid (C18:1 t11) for the crushed form, when compared with the oil. These results suggest an inherent effect of the physical form of the assay oilseeds on in vitro BH. Changes in FA pattern during BH in vitro can be attributed to both source and physical form of the assay oilseeds. However, further investigations are warranted to ensure whether the observed in vitro effects on ruminal BH can be confirmed in vivo.

Effects of the composition of the basal diet on the evaluation of mineral phosphorus sources and interactions with phytate hydrolysis in broilers.

The objectives of this study were to determine the availability of P from mineral phosphate sources by using different basal diets and measurement of P retention and prececal (pc) P digestibility as well as pc myo-inositol phosphate (InsP) degradation in broilers. Semi-synthetic and corn-soybean meal-based basal diets were used in experiment 1, and corn-based and wheat-based basal diets were used in experiment 2. Anhydrous monosodium phosphate (MSPa) or monocalcium phosphate monohydrate (MCPh) was supplemented to increment the P concentration by 0.05, 0.10, and 0.15% or by 0.075 and 0.150% in experiments 1 and 2, respectively. Titanium dioxide was used as an indigestible marker. Diets were pelleted through a 3-mm screen. In experiment 1, retention was measured based on total excreta collection from 20 to 24 d of age using 7 replicated birds per diet. In experiment 2, digesta from the terminal ileum was collected from 22-d-old broilers penned in groups of 19 with 5 replicated pens per diet. The P retention response to supplemented MSPa did not differ between the 2 basal diets in experiment 1. The response in pc P digestibility to MCPh supplements also did not differ between the 2 basal diets in experiment 2, as calculated by linear regression analysis. Hydrolysis of InsP6 measured on both the excreta and pc levels was high in the basal diets without a mineral P supplement. Mineral P supplementation significantly decreased (P < 0.05) InsP6 hydrolysis from the InsP-containing diets in both experiments. Thus, the choice of the basal diet did not affect the evaluation of the supplemented mineral P source. However, calculated values for mineral P sources need to be adjusted for the decline in hydrolysis of InsP contained in the basal diet that results from the P supplement.

Effects of long-term supplementation of chestnut and valonea extracts on methane release, digestibility and nitrogen excretion in sheep.

The long-term effects of adding chestnut (CHE; Castanea sativa) and valonea (VAL; Quercus valonea) tannin-rich extracts to sheep feed were investigated. In Experiment 1, sheep (65 kg BW) were fed 842 g/day of a ryegrass-based hay. The control-treated animals (CON) received 464 g/day of concentrate, and tannin-treated animals received the same amount of concentrate additionally containing 20 g of the respective tannin-rich extract. Hay and concentrates were offered together in one meal. After the onset of treatment, methane release was measured in respiration chambers for 23.5-h intervals (nine times) in a 190-days period. Faeces and urine were collected three times (including once before the onset of the tannin treatment) to assess digestibility and urinary excretion of purine derivatives. Based on the results obtained from Experiment 1, a second experiment (Experiment 2) was initiated, in which the daily tannin dosage was almost doubled (from 0.9 (Experiment 1) to 1.7 g/kg BW0.75). With the exception of the dosage and duration of the treatment (85 days), Experiment 2 followed the same design as Experiment 1, with the same measurements. In an attempt to compare in vitro and in vivo effects of tannin supplementation, the same substrates and tannin treatments were examined in the Hohenheim gas test. In vitro methane production was not significantly different between treatments. None of the tannin-rich extract doses induced a reduction in methane in the sheep experiments. On the 1st day of tannin feeding in both experiments, tannin inclusion tended to decrease methane release, but this trend disappeared by day 14 in both experiments. In balance period 3 of Experiment 1, lower dry matter and organic matter digestibility was noted for tannin treatments. The digestibility of CP, but not NDF or ADF, was reduced in both experiments. A significant shift in N excretion from urine to faeces was observed for both tannin-rich extracts in both experiments, particularly in Experiment 2. In balance period 2 of Experiment 2, an increased intake of metabolisable energy for VAL was observed. The urinary excretion of purine derivatives was not significantly different between treatments, indicating that microbial protein synthesis was equal for all treatments. Thus, we concluded that both tannin-rich extracts temporary affect processes in the rumen but did not alter methane release over a longer period.

Effects of different tannin-rich extracts and rapeseed tannin monomers on methane formation and microbial protein synthesis in vitro.

Tannins, polyphenolic compounds found in plants, are known to complex with proteins of feed and rumen bacteria. This group of substances has the potential to reduce methane production either with or without negative effects on digestibility and microbial yield. In the first step of this study, 10 tannin-rich extracts from chestnut, mimosa, myrabolan, quebracho, sumach, tara, valonea, oak, cocoa and grape seed, and four rapeseed tannin monomers (pelargonidin, catechin, cyanidin and sinapinic acid) were used in a series of in vitro trials using the Hohenheim gas test, with grass silage as substrate. The objective was to screen the potential of various tannin-rich extracts to reduce methane production without a significant effect on total gas production (GP). Supplementation with pelargonidin and cyanidin did not reduce methane production; however, catechin and sinapinic acid reduced methane production without altering GP. All tannin-rich extracts, except for tara extract, significantly reduced methane production by 8% to 28% without altering GP. On the basis of these results, five tannin-rich extracts were selected and further investigated in a second step using a Rusitec system. Each tannin-rich extract (1.5 g) was supplemented to grass silage (15 g). In this experiment, nutrient degradation, microbial protein synthesis and volatile fatty acid production were used as additional response criteria. Chestnut extract caused the greatest reduction in methane production followed by valonea, grape seed and sumach, whereas myrabolan extract did not reduce methane production. Whereas chestnut extract reduced acetate production by 19%, supplementation with grape seed or myrabolan extract increased acetate production. However, degradation of fibre fractions was reduced in all tannin treatments. Degradation of dry matter and organic matter was also reduced by tannin supplementation, and no differences were found between the tannin-rich extracts. CP degradation and ammonia-N accumulation in the Rusitec were reduced by tannin treatment. The amount and efficiency of microbial protein synthesis were not significantly affected by tannin supplementation. The results of this study indicated that some tannin-rich extracts are able to reduce methane production without altering microbial protein synthesis. We hypothesized that chestnut and valonea extract have the greatest potential to reduce methane production without negative side effects.

Genetic background of host-pathogen interaction between Cucumis sativus L. and Pseudomonas syringae pv. lachrymans.

The interplay of plant resistance mechanisms and bacterial pathogenicity is very complex. This applies also to the interaction that takes place between the pathogen Pseudomonas syringae pv. lachrymans (Smith et Bryan) and the cucumber (Cucumis sativus L.) as its host plant. Research on P. syringae pv. lachrymans has led to the discovery of specific factors produced during pathogenesis, i.e. toxins or enzymes. Similarly, studies on cucumber have identified the specific types of plant resistance expressed, namely Systemic Acquired Resistance (SAR) or Induced Systemic Resistance (ISR). This paper presents a summary of the current state of knowledge about this particular host-pathogen interaction, with reference to general information about interactions of P. syringae pathovars with host plants.

Effects of Fusarium toxin-contaminated wheat and feed intake level on the biotransformation and carry-over of deoxynivalenol in dairy cows.

An experiment was carried out to examine the effects of feeding Fusarium toxin-contaminated wheat (8.21 mg deoxynivalenol (DON) and 0.09 mg zearalenone (ZON) per kg dry matter) at different feed intake levels on the biotransformation and carry-over of DON in dairy cows. For this purpose, 14 ruminal and duodenal fistulated dairy cows were fed a diet containing 60% concentrate with a wheat portion of 55% (Fusarium toxin-contaminated wheat (mycotoxin period) or control wheat (control period)) and the ration was completed with maize- and grass silage (50 : 50) on a dry matter basis. Daily DON intakes ranged from 16.6 to 75.6 mg in the mycotoxin period at dry matter intakes of 5.6-20.5 kg. DON was almost completely biotransformed to de-epoxy DON (94-99%) independent of the DON/feed intake, and the flow of DON and de-epoxy DON at the duodenum related to DON intake ranged from 12 to 77% when the Fusarium toxin-contaminated wheat was fed. In the serum samples, de-epoxy DON was detected in the range of 4-28 ng ml-1 in the mycotoxin period, while concentrations of DON were all below the detection limit. The daily excretion of DON and de-epoxy DON in the milk of cows fed the contaminated wheat varied between 1 and 10 microg and between 14 and 104 microg, respectively. The total carry-over rates as the ratio between the daily excretion of DON and de-epoxy DON into milk and DON intake were in the ranges of 0.0001-0.0002 and 0.0004-0.0024, respectively. Total carry-over rates of DON as DON and de-epoxy DON into the milk increased significantly with increasing milk yield. In the urine samples, de-epoxy DON was the predominant substance as compared with DON with a portion of the total DON plus de-epoxy DON concentration to 96% when the Fusarium toxin-contaminated wheat was fed, whereas the total residues of DON plus de-epoxy DON in faeces ranged between 2 and 18% of DON intake in the mycotoxin period. The degree of glucuronidation of de-epoxy DON was found to be approximately 100% in serum. From 33 to 80% of DON and from 73 to 92% of de-epoxy DON, and from 21 to 92% of DON and from 86 to 100% of de-epoxy DON were glucuronidated in the milk and urine, respectively. It is concluded that DON is very rapidly biotransformed to de-epoxy DON in the rumen and only negligible amounts of DON and de-epoxy DON were transmitted into the milk within the range of 5.6-20.5 kg day-1 dry matter intake and milk yields (fat corrected milk) between 10 and 42 kg day-1.

Subacute effects of moderate feed loads of isolated Fusarium toxin deoxynivalenol on selected parameters of metabolism in weaned growing piglets.

In 36 female weaned piglets, the effect of different dosages (0, 300, 600 and 1200 microg/kg feed) of isolated, pure Fusarium toxin deoxynivalenol (DON) was examined during a period of 8 weeks. Standardised trial conditions were provided. Pigs were fed restrictively to allow a complete feed intake of all animals. Parameters of liver integrity, haematological data and blood concentrations of some selected metabolic components of energy and protein metabolism were examined weekly. Enzyme aspartate aminotransferase was affected subclinically by age and significantly by dosage, which was proved by Wald F-test. Some additional enzymes, for instance alanine aminotransferase, gamma glutamyl transferase, glutamate dehydrogenase and sorbit dehydrogenase, showed no clear systematic effect. Urea and glucose in the blood were inter-related. Depending on DON load with increasing glucose concentrations, the urea level declined. Albumin and total protein in serum showed no significant DON-related effect. Haemoglobin in blood was found to be significantly affected by DON, which was proved by the Wald F-test, where the effect was more pronounced with 600 microg DON/kg diet compared to 1 200 microg DON/kg. An obvious DON-related affection of liver, N-metabolism and stimulation of haematopoiesis depending on dosage and time is discussed.

Relationship between thiamine concentration and fermentation patterns in the rumen fluid of dairy cows fed with graded concentrate levels.

The objective of this study was to investigate the relationship between the thiamine concentration and the fermentation patterns [pH, concentration of short-chain fatty acids (SCFA)] in the free liquid (FRL) and particle-associated liquid (PARL) of the rumen in dairy cows fed with graded concentrate levels in the diet. Four ruminally cannulated Holstein cows in mid lactation were fed [semi-ad libitum, 18 kg dry matter (DM)/day] diets consisting of hay (H) and slowly degradable concentrate (C), offered in five different H:C ratios (% DM basis) in the following sequence: period 1, 30:70; period 2, 40:60; period 3, 50:50; period 4, 60:40 and period 5, 75:25. A negative quadratic relationship was observed between thiamine concentration in FRL and intake of dietary thiamine (R(2) = 0.36), of concentrate (R(2) = 0.38) and of digestible non-fibre carbohydrates (R(2) = 0.37). The thiamine concentration in PARL was higher (p < or = 0.05) compared to FRL. The ruminal thiamine concentration correlated negatively to pH and positively to concentrations of SCFA and propionate in the rumen. R(2) of these relationships varied markedly (0.002 and 0.77), depending on time after feeding and fluid digesta compartment. The higher R(2) were observed at 8 and 11 h after the morning feeding for FRL (R(2) 0.46-0.71) and PARL (R(2) 0.41-0.77), respectively.

Mycotoxins in food systems in Sub Saharan Africa: A review.

Mycotoxins, toxic secondary metabolites of fungi are now recognised as major cause of food intoxications in Sub Saharan Africa (SSA). Aflatoxins, the most important of the group have been implicated in acute aflatoxicoses, carcinogenicity, growth retardation, neonatal jaundice and immunological suppression in SSA. The hot and humid tropical climate provides ideal condition for growth of toxigenicAspergillus spp, making food contamination to be widespread in SSA, with maize and groundnuts being the most contaminated. The available data suggests that cassava products (the most important African food) are not prone to aflatoxin contamination. Recent data on ochratoxin A produced by species ofAspergillus on grains have indicated the necessity for it to be monitored in SSA. Fumonisins represent the most importantFusarium mycotoxins in SSA, and surveillance data indicate very high contamination rates of almost 100% in maize samples from West Africa. Limited information exists on the occurrence of trichothecenes, while the data currently available suggest that zearalenone contamination seems not to be a problem in SSA. The strategies under investigation to mitigate the mycotoxin problem in SSA include education of the people on the danger of consuming mouldy foods, pre and post harvest management strategies with emphasis on biological control, use of plant products to arrest fungal growth during storage, enterosorbent clay technology, and the search for traditional techniques that could reduce/detoxify mycotoxins during food processing.

On the effects of a chronic deoxynivalenol intoxication on performance, haematological and serum parameters of pigs when diets are offered either for ad libitum consumption or fed restrictively.

Wheat infected naturally with Fusarium, contaminated mainly with deoxynivalenol (DON) (16.6 mg DON/kg), was added to a total constant wheat content of 400 g/kg diet. To distinguish between differences in feed intake and specific effects of the DON contaminated diet, control and DON contaminated feed was administered for 11 weeks under ad libitum and restrictive feeding conditions to 48 pigs of both sexes, which were randomly divided into four groups (n = 12 per group). Feed intake was 2.90 kg/day, live weight gain 987 g/day and feed to gain ratio 2.77 kg/kg for the ad libitum fed control group. The group fed DON contaminated wheat ad libitum significantly consumed 15% less feed and gained 13% less live weight, while the feed to gain ratio was unaffected. Moreover, it was concluded that the lower growth performance by DON contaminated feed resulted mainly from the lower voluntary feed intake, because there were no differences in live weight gain between the groups with the restrictive feeding regimen. On the contrary, metabolizable energy, nitrogen retention digestibility of organic matter, crude protein, crude fat and crude fibre were significantly increased by 3, 10, 3, 6, 9 and 20% in the DON group respectively. Animals fed DON contaminated diets needed more time to consume the restrictive ration than the control group. For example in the first hour after feeding 85% of the control pigs had consumed all feed, but only 39% of the DON group had. There were only few differences in haematological and serum parameters, which were characterized by a high variation between individuals. DON and IgA concentrations in serum were significantly influenced by DON exposure.

Fusarium toxin contents of maize and maize products purchased in the years 2000 and 2001 in Germany.

Samples (n=106) of maize and maize products were analysed for 13 trichothecene toxins and zearalenone (ZON). All 14 toxins examined were detected, although with varying frequency. Cooccurrence of two or more toxins was observed in 96% of samples. The toxins of the scirpenol group scirpentriol, 15-monoacetoxyscirpenol and diacetoxyscirpenol were detected in 14, 27 and 3% of the samples analysed, the toxins of the T-2 group T-2 toxin, HT-2 toxin, T-2 triol und T-2 tetraol were found in 33, 66, 2 and 7%. Toxin content was higher in feeds than in foods (semolina and flour). In food samples, the German regulatory level for DON (500 µg/kg) was not exceeded, three samples of maize flour contained ZON above the regulatory level (50 µg/kg).