Effects of aromatic amino acids, phenylacetate and phenylpropionate on fermentation of xylan by the rumen anaerobic fungi, Neocallimastix frontalis and Piromyces communis.

AIMS: Anaerobic fungi are important members of the fibrolytic community of the rumen. The aim of this study was to study their requirement for aromatic amino acids (AA) and related phenyl acids (phenylpropionic and phenylacetic acids) for optimal xylan fermentation. METHODS AND RESULTS: Neocallimastix frontalis RE1 and Piromyces communis P were grown in a defined medium containing oat spelts xylan as the sole energy source, plus one of the following N sources: ammonia; ammonia plus a complete mixture of 20 AA commonly found in protein; ammonia plus complete AA mixture minus aromatic AA; ammonia plus phenyl acids; ammonia plus complete AA mixture without aromatic AA plus phenyl acids. Both species grew in all the media, indicating no absolute requirement for AA. The complete AA mixture increased (P<0.05) acetate concentration by 18% and 15%, sugar utilization by 33% and 22% and microbial yield by about 22% and 15% in N. frontalis and P. communis, respectively, in comparison with the treatments that had ammonia as the only N source. Neither the supply of aromatic AA or phenol acids, nor their deletion from the complete AA mixture, affected the fermentation rate, products or yield of either species. CONCLUSIONS: AA were not essential for N. frontalis and P. communis, but their growth on xylan was stimulated. The effects could not be explained in terms of aromatic AA alone. SIGNIFICANCE AND IMPACT OF THE STUDY: Ruminant diets should contain sufficient protein to sustain optimal fibre digestion by ruminal fungi. Aromatic AA or phenyl acids alone cannot replace the complete AA mixture.

Use of stable isotopes to measure de novo synthesis and turnover of amino acid-C and -N in mixed micro-organisms from the sheep rumen in vitro.

Protein synthesis and turnover in ruminal micro-organisms were assessed by stable-isotope methods in order to follow independently the fate of amino acid (AA)-C and -N in different AA. Rumen fluid taken from sheep receiving a grass hay-concentrate diet were strained and incubated in vitro with starch-cellobiose-xylose in the presence of NH3 and 5 g algal protein hydrolysate (APH)/l, in incubations where the labels were (15)NH3, [(15)N]APH or [(13)C]APH. Total (15)N incorporation was calculated from separate incubations with (15)NH3 and [(15)N]APH, and net N synthesis from the increase in AA in protein-bound material. The large difference between total and net AA synthesis indicated that substantial turnover of microbial protein occurred, averaging 3.5 %/h. Soluble AA-N was incorporated on average more extensively than soluble AA-C (70 v. 50 % respectively, P=0.001); however, incorporation of individual AA varied. Ninety percent of phenylalanine-C was derived from the C-skeleton of soluble AA, whereas the incorporation of phenylalanine-N was 72 %. In contrast, only 15 % aspartate-C+asparagine-C was incorporated, while 45 % aspartate-N+asparagine-N was incorporated. Deconvolution analysis of mass spectra indicated substantial exchange of carboxyl groups in several AA before incorporation and a condensation of unidentified C2 and C4 intermediates during isoleucine metabolism. The present results demonstrate that differential labelling with stable isotopes is a way in which fluxes of AA synthesis and degradation, their biosynthetic routes, and separate fates of AA-C and -N can be determined in a mixed microbial population.

Detection of subclinical mastitis in dromedary camels (Camelus dromedarius) using somatic cell counts and the N-acetyl-beta-D-glucosaminidase test.

Somatic cell counts, N-acetyl-beta-D-glucosaminidase (NAGase) activity and the infection status of the udder were determined in quarter milk samples (n = 86) from 22 multiparous, clinically healthy camels, traditionally managed by Bedouin nomads in the Negev desert, Israel. Seventy (81.4%) of the 86 samples examined contained bacteria, of which 35 (40.7%) gave mixed isolations of two or more bacteria, suggesting the existence of subclinical mastitis in the camel herds studied. Sixteen samples (18.6%) yielded no growth of bacteria. Staphylococcus aureus, Micrococcus spp., Bacillus spp., Streptococcus dysgalactiae and Escherichia coli were the main organisms isolated. The somatic cell count (SCC) ranged from 1.01 x 105 to 11.78 x 106 cells/ml. NAGase values were between 41.4 and 372 NAGase units. Quarter milk samples that contained bacteria had significantly (p < 0.01) higher mean values for SCC but the mean NAGase levels were not significantly different for the bacteriologically negative and positive samples. There was a low correlation coefficient (r2 = 0.097) between the SCC and NAGase in the quarter milk samples from which bacteria were not isolated (n = 16) and a low negative correlation (r2 = -0.038) with the samples that contained bacteria (n = 70). The type of bacteria had a significant effect (p < 0.01) on the SCC but not on the NAGase activity. Quarter samples from which Staphylococcus aureus (coagulase positive) was isolated showed the highest mean SCC and this organism is therefore suspected to be the underlying cause of the subclinical mastitis. The SCC gave a better indication of the presence of pathogenic microorganisms in milk samples than did NAGase.