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PubMed1980

Evaluation by electron microscopy and anaerobic culture of types of rumen bacteria associated with digestion of forage cell walls.

Different morphological types of rumen bacteria which degraded cell walls of forage grasses with various in vitro digestibilities were evaluated with electron microscopy. The majority of these bacteria (i.e., about 70% or more) consisted of two distinct types: (i) encapsulated cocci and (ii) irregularly shaped bacteria, resembling major fiber digesters found in the rumen. Each type was capable of degrading structurally intact cell walls. Differences (P less than or equal to 0.02) in the percent ratio of encapsulated cocci to irregularly shaped bacteria were observed between Bermuda grass and fescue; the ratio of encapsulated cocci to irregularly shaped bacteria between Bermuda grass and orchard grass was similar and variations were high. The proportion of irregularly shaped bacteria usually increased with increased time of digestion. Differences (P greater than 0.1) were not found in the percentage ratio of encapsulated cocci to irregularly shaped bacteria attached to specific tissue types in either Bermuda grass or fescue. However, encapsulated cocci tended to be more prevalent on sclerenchyma than other tissues in Bermuda grass, but less prevalent on sclerenchyma than other tissues in fescue. Transmission electron microscopy of tissue digestion of rapidly degraded orchard grass blades revealed that mesophyll, parenchyma bundle sheath, and parts of the epidermal cell wall apparently were degraded without direct attachment of bacteria although bacteria were near the cell walls undergoing digestion. Anaerobic growth studies showed that the total culturable bacteria developing on medium 10 and media containing carbohydrates similar to those in forage cell walls (i.e., pectin, xylan, and cellobiose) were 80% higher from rumen bacterial populations adapted in vitro to cell walls of orchard grass compared to those from Bermuda grass; the number of colonies from the orchard grass-adapted population was significantly (P less than or equal to 0.05) greater on the medium containing xylan. Filter paper tests showed that the cellulolytic activity of populations adapted to fescue was greater than that of orchard grass or Bermuda grass.

Anaerobiosis↗
PubMed1983

Response of early weaned rabbits to source and level of dietary fiber.

Growth, cecal volatile fatty acids, mineral profile of plasma, liver weight and gastrointestinal weights were measured in rabbits fed isonitrogenous, equicaloric diets containing 3, 6, 9 or 12% dietary crude fiber from either alfalfa or Coastal Bermuda grass. Rabbits were fed from 4 to 8 wk of age. Alfalfa diets improved (P less than .05) average daily gain (20.5 vs 11.9 g/d) and livability (67.5 vs 42.5%). Feed intake, average daily gain and livability increased linearly with increasing fiber content within the alfalfa diets only. Source of fiber had no effect on total or individual cecal volatile fatty acids, except for an increased proportion of isobutyrate in rabbits fed Coastal Bermuda grass. Increased fiber level did not affect total volatile fatty acids, but did increase the proportion of butyrate and the butyrate:propionate ratio for both sources. Decreased propionate and valerate proportions were observed in cecal contents of rabbits fed Coastal Bermuda grass. Increased fiber levels decreased cecal pH in Coastal Bermuda grass-fed, but not alfalfa-fed rabbits. Relative cecal weights decreased as fiber levels increased for both sources. Coastal Bermuda grass diets resulted in larger stomach weights. Neither dietary fiber source nor level caused a consistent alteration in plasma mineral profile. The butyrate:propionate ratio reflected to a greater extent the value of feeds in fattening rabbits than did the acetate:propionate ratio normally calculated for ruminants and horses. It is concluded that the growing rabbit requires greater than 9% dietary crude fiber and that Coastal Bermuda grass is not a suitable source of fiber.

Animals↗
PubMed2001

Allergenicity and cross-reactivity of buffalo grass (Stenotaphrum secundatum).

BACKGROUND: In the subtropical climate of South Africa, grasses of the subfamily Panicoideae are predominant. Bermuda grass has previously been shown to be an important local allergen, and immunoglobulin E (IgE) epitopes of Bermuda grass extracts are known to be distinct from those of the Pooid pollen extracts. Following our demonstration of sensitivity in 43% of patients grass-allergic to the Panicoid, Kikuyu grass, we have studied the closely related buffalo grass, Stenotaphrum secundatum, indigenous to the Western Cape region, the east coast of Africa and the oceanic islands such as Mauritius; and Eragrostis, another common indigenous grass with a wide distribution. OBJECTIVE: To partially characterise the allergens of buffalo pollen, and examine its immunological relationships with local common grasses such as Eragrostis and Kikuyu. METHODS: Grass-allergic patients were evaluated clinically, and skin prick tests (SPTs) and radio-allergosorbent tests (RASTs) to Bermuda and grass mix were performed. Sera of timothy grass-sensitive patients from Belgium were also included in this study. Pollen extract from buffalo grass was characterised by specific IgE binding by means of immunoblotting and enzyme-linked immunosorbent assay (ELISA). Cross-reactivity between the grasses was studied by means of inhibition of IgE binding. RESULTS: More than 90% of grass-sensitive patients were found to have IgE antibodies to Buffalo and Eragrostis pollen. Inhibition of ELISA and immunoblots revealed that extracts of these grass pollens could significant inhibit IgE binding to the local grass pollens, Kikuyu, buffalo, Eragrostis and Bermuda on solid phase, but 100% inhibition was never achieved, indicating that cross-reactive but also unique epitopes are present. We also identified a subset of patients with negative RASTs to Bermuda, and minimal inhibition by Bermuda pollen extract. CONCLUSION: Buffalo and Eragrostis are important aeroallergens in the Cape, dispersed during the long dry, windy summer. Our data suggest that the local grasses are major sensitisers, and that South African diagnostic panels should include extracts of buffalo and Eragrostis grasses.

Cross Reactions↗
PubMed1984

Rumen fungi and forage fiber degradation.

The role of anaerobic rumen fungi in in vitro forage fiber degradation was determined in a two forage x two inoculum source x five treatment factorial design. Forages used as substrates for rumen microorganisms were Coastal bermuda grass and alfalfa; inoculum sources were rumen fluid samples from a steer fed Coastal bermuda grass hay or alfalfa hay; treatments were whole rumen fluid (WRF), WRF plus streptomycin (0.2 mg/ml of rumen fluid) and penicillin (1.25 mg/ml of fluid), WRF plus cycloheximide (0.5 mg/ml of fluid), WRF plus streptomycin, penicillin, and cycloheximide, and McDougall buffer. Populations of fungi as shown by sporangial development were greater on bermuda grass leaves than on alfalfa leaflets regardless of inoculum source. However, endogenous fungal populations were greater from the alfalfa hay inoculum. Cycloheximide inhibited the fungi, whereas streptomycin and penicillin, which inhibit bacterial populations, resulted in an increase in numbers of sporangia in the alfalfa inoculum, suggesting an interaction between bacteria and fungi. Bacteria (i.e., WRF plus cycloheximide) were equal to the total population in degrading dry matter, neutral-detergent fiber (NDF), acid-detergent fiber (ADF), and cellulose for both inocula and both forages. Degradation of dry matter, NDF, ADF, and cellulose by anaerobic fungi (i.e., WRF plus streptomycin and penicillin) was less than that due to the total population or bacteria alone. However, NDF, ADF, and cellulose digestion was 1.3, 2.4, and 7.9 percentage units higher, respectively, for bermuda grass substrate with the alfalfa versus bermuda grass inoculum, suggesting a slight benefit by rumen fungi. No substantial loss of lignin (72% H(2)SO(4) method) occurred due to fungal degradation. The most active fiber-digesting population in the rumen was the bacteria, even when streptomycin and penicillin treatment resulted in an increase in rumen fungi over untreated WRF. The development of large numbers of sporangia on fiber may not indicate a substantial role as digesters of forage.

Journal Article↗
PubMed1985

Cross allergenicity among grasses determined by tissue threshold changes.

Twenty-seven volunteers with grass-pollen allergy who had not received prior immunotherapy had titrated prick tests with fivefold dilutions of 10 grasses (timothy, Bermuda, smooth brome, grama, salt, quack, western wheat, Johnson, June, and Bahia) to a final dilution of 1:3,906,250 w/v. Ten subjects served as untreated control subjects. Seventeen subjects received immunotherapy with an extract containing equal parts of timothy and Bermuda. Twelve months later subjects had repeat titrated skin testing with reconstituted aliquots of the same lyophilized extract, at the same time of day, and at the same location on the back as in the initial testing period. Nine treated subjects and eight control subjects completed the study protocol. The skin test areas with all 10 grass extracts were significantly decreased in the treated group compared to the control group (p less than 0.01), and there was no difference in the extent of the reduction among the 10 grasses by analysis of variance. We conclude that immunotherapy to timothy and Bermuda alone is capable of reducing to a similar degree the clinical sensitivity to all 10 grasses studied. The results of this study suggest that treatment with timothy and Bermuda alone may be sufficient in most grass-sensitive individuals.

Allergens↗
PubMed1996

Studies of allergen extract stability: the effects of dilution and mixing.

BACKGROUND: However potent the allergy extracts provided by manufacturers, they are subject to deterioration with storage, especially after dilution or mixture with other extracts. OBJECTIVE: This study was performed to assess separately the deterioration during storage in allergen extract potency caused by dilution or by mixture with allergen extracts that have been reported to contain proteases. METHODS: To assess the effect of dilution, three serial 10-fold dilutions of cat, short ragweed, Bermuda grass, and Dermatophagoides farinae extracts were prepared alone or combined with other extracts. They were stored at 4 degrees C for 3 and 12 months. To assess the effect of mixing with other extracts that have been reported to contain proteases, extracts of timothy grass, Bermuda grass, short ragweed, Russian thistle, white oak, box elder, D. farinae, and cat were stored alone or combined with one or more extracts of American cockroach, Alternaria spp., Cladosporium spp., Penicillium spp., and a house dust mite mix for 3 months at 4 degrees C. RESULTS: Bermuda grass, cat, and house dust mite extracts incurred significant loss of potency at all dilutions with storage. Short ragweed was stable at all dilutions. Potency of extracts of timothy grass, Bermuda grass, Russian thistle, white oak, box elder, and cat were all reduced by combination with one or more extracts potentially containing proteases. Only short ragweed and D. farinae, which was in a final concentration of 25% glycerin, were resistant. Alternaria extract was most frequently responsible for loss of potency, followed by cockroach and Cladosporium extracts. Combination with extracts of Penicillium and a house dust mite mix did not reduce the potency of any extract. CONCLUSIONS: Both dilution alone and mixture with extracts reported to contain proteases caused loss of potency of most extracts tested. Ragweed was uniquely resistant under both conditions of storage.

Allergens↗
PubMed2000

Late summer and fall (March-May) pollen allergy and respiratory disease in Northern New South Wales, Australia.

BACKGROUND: Many people in the subtropical Northern Rivers area of New South Wales, Australia, blame the pollen of Tibouchina tree, which flowers at the same time as ragweed, Bahia grass and Bermuda grass, for hayfever and asthma exacerbations during fall between March and May. OBJECTIVES: To determine whether Tibouchina pollen is allergenic. To determine whether airborne ragweed pollen is present in this region for sufficient length of time and concentration to cause fall respiratory symptoms, and to determine if Bahia grass and Bermuda grass are associated with fall respiratory symptoms. METHODS: Pollen and Alternaria spores were monitored using a Burkard 7-day spore trap. Two hundred and six volunteers in the Northern Rivers area filled in questionnaires before skin prick tests (SPT) were performed with a panel of skin testing extracts. RESULTS: One hundred fifty-three (74.3%) subjects were atopic and reacted to one or more aeroallergens. Seventy were SPT positive to ragweed, OR 3.36 (CI 1.03 to 12.15) and 11 to Tibouchina (OR incalculable). Fifty of the 70 ragweed-positive subjects had fall hayfever or exacerbations of hayfever and/or asthma, OR 23.4 (CI 8.90 to 64.00). Eleven subjects were SPT positive to Tibouchina extract. There was a statistical association between Bermuda grass and hayfever, but not asthma OR 13.44 (CI 1.85 to 27.04). CONCLUSIONS: Ragweed pollen was present for a sufficient length of time and concentration to sensitize and provoke fall hayfever and asthma exacerbations. Tibouchina pollen is an aeroallergen causing mild-to-moderate allergic symptoms in a few people. There is an association between Bahia grass and asthma in children, and between Bermuda grass and allergic rhinitis in adults.

Adolescent↗
PubMed2003

Mass spectrometric analysis of electrophoretically separated allergens and proteases in grass pollen diffusates.

BACKGROUND: Pollens are important triggers for allergic asthma and seasonal rhinitis, and proteases released by major allergenic pollens can injure airway epithelial cells in vitro. Disruption of mucosal epithelial integrity by proteases released by inhaled pollens could promote allergic sensitisation. METHODS: Pollen diffusates from Kentucky blue grass (Poa pratensis), rye grass (Lolium perenne) and Bermuda grass (Cynodon dactylon) were assessed for peptidase activity using a fluorogenic substrate, as well as by gelatin zymography. Following one- or two-dimensional gel electrophoresis, Coomassie-stained individual bands/spots were excised, subjected to tryptic digestion and analysed by mass spectrometry, either MALDI reflectron TOF or microcapillary liquid chromatography MS-MS. Database searches were used to identify allergens and other plant proteins in pollen diffusates. RESULTS: All pollen diffusates tested exhibited peptidase activity. Gelatin zymography revealed high Mr proteolytic activity at approximately 95,000 in all diffusates and additional proteolytic bands in rye and Bermuda grass diffusates, which appeared to be serine proteases on the basis of inhibition studies. A proteolytic band at Mr approximately 35,000 in Bermuda grass diffusate, which corresponded to an intense band detected by Western blotting using a monoclonal antibody to the timothy grass (Phleum pratense) group 1 allergen Phl p 1, was identified by mass spectrometric analysis as the group 1 allergen Cyn d 1. Two-dimensional analysis similarly demonstrated proteolytic activity corresponding to protein spots identified as Cyn d 1. CONCLUSION: One- and two-dimensional electrophoretic separation, combined with analysis by mass spectrometry, is useful for rapid determination of the identities of pollen proteins. A component of the proteolytic activity in Bermuda grass diffusate is likely to be related to the allergen Cyn d 1.

Antigens, Plant↗
PubMed2002

Factors associated with development of ileal impaction in horses with surgical colic: 78 cases (1986-2000).

Deal impaction is prevalent in the south-eastern USA, where feeding of Coastal Bermuda hay has been implicated as a risk factor. Alternatively, infection with the tapeworm Anoplocephala perfoliata has been identified as a risk factor for ileal impaction in the UK. We hypothesised that feeding Coastal Bermuda hay and failure to administer routinely an anthelmintic with efficacy against tapeworms would place horses at risk of developing ileal impaction in the USA. Seventy-eight horses, with surgically confirmed ileal impaction and 100 horses admitted for colic that did not have an ileal impaction, were selected retrospectively for logistic regression analysis. Using odds ratios (OR) as an index of risk, feeding Coastal Bermuda hay (OR = 2.9) and failure to administer a pyrantel salt within 3 months of admission (OR = 3.1) placed horses at risk of development of ileal impaction. This study confirms the belief that feeding Coastal Bermuda hay places horses at risk of ileal impaction, although the quality of the hay may also play a role. Periodic administration of anthelmintics with efficacy against tapeworms should be considered to reduce risk of ileal impaction.

Animal Feed↗