Host effects of glandular-haired alfalfa on alfalfa weevil (Coleoptera: Curculionidae) and potato leafhopper (Homoptera: Cicadellidae) populations in Virginia

Cultivars of glandular-haired alfalfa, Medicago sativa L., such as '54H69', are currently available and marketed as being resistant to potato leafhopper, Empoasca fabae (Harris). Between 2000 and 2002, studies were conducted to compare the effects of '54H69' and a standard, nonglandular-haired alfalfa cultivar, 'Choice', on alfalfa weevil, Hypera postica (Gyllenhal), and potato leafhopper populations at Campbell and Montgomery counties, Virginia. '54H69' had no effect on alfalfa weevil populations. At each location, densities of alfalfa weevil in '54H69' and 'Choice' were similar, but pest pressure was higher at Campbell Co. than at Montgomery Co. and always exceeded the economic threshold before insecticide was applied. Densities of potato leafhopper also did not differ between '54H69' and 'Choice' in any year at the two locations. Insecticide treatment effectively reduced potato leafhopper densities in the two cultivars, although populations were below the economic threshold at both locations when the insecticides were applied. Overall, postinsecticide treatment comparisons showed that the densities of alfalfa weevil and potato leafhoppers were similar or higher in untreated '54H69' compared with insecticide-treated 'Choice'. In addition, there were no differences in seasonal dry yields between '54H69' and 'Choice' in any year at either location. Our results indicate that the glandular-haired alfalfa '54H69' does not provide acceptable resistance to potato leafhopper and also does not offer a yield advantage to growers in Virginia.     

Glandular-haired alfalfa resistance to potato leafhopper (Homoptera: Cicadellidae) and hopperburn: development of resistance indices

Eight proprietary genotypes of glandular-haired alfalfa, Medicago sativa L., supplied by two different companies, were compared for the degree and types of resistance to the potato leafhopper, Ernpoasca fabae (Harris), and hopperburn. A tube cage no-choice bioassay was developed to test leafhopper mortality, feeding, settling preferences, severity of hopperburn symptoms (in this case, defined as both yellowing and stem growth reduction), and trichome density and type on feeding sites. Leafhopper mortality was both strongly and significantly associated with feeding and leaf trichome density; decreased hopperburn symptom severity was weakly, although significantly, associated with increased mortality. To quantify hopperburn in terms of both yellowing and stem growth reduction, we developed a ranking system that reduces overall hopperburn expression to a single number that considers the varying responses to both types of symptoms. Great variability in leafhopper settling, leafhopper mortality, and stem glandular trichome density was detected among alfalfa genotypes, suggesting that genotypic differences may be based on the concentration and/or chemical constituency of the trichome exudates. We postulate that, among variably resistant genotypes of glandular-haired alfalfa, differences among leafhopper responses and hopperburn severity are linked to forced movement from the stems to the leaves as refuge feeding sites. Principal component analysis was performed to reduce the 10 variables down to five biologically significant factors. Scores for these factors were then used to develop resistance indices for potato leafhopper resistance, hopperburn resistance, and an overall glandular-haired alfalfa resistance index.     

Influence of plant ontogeny and abiotic factors on resistance of glandular-haired alfalfa to potato leafhopper (Hemiptera: Cicadellidae)

Laboratory experiments were conducted to characterize the trichome-based defense of glandular-haired alfalfa, Medicago sativa L., against the potato leafhopper, Empoasca fabae (Harris). Within-plant variability in leafhopper resistance was examined by caging adult leafhoppers to either basal or apical stem internodes of the leafhopper-resistant, glandular-haired M. sativa genotype G98A or the susceptible, nonglandular-haired M. sativa 'Ranger'. Young, actively secreting glandular trichomes are located on apical internodes of G98A, whereas senesced gland heads are found on older, basal internodes of G98A. After 96 h, the highest cumulative leafhopper mortality and lowest number of excretory droplets were associated with apical internodes of G98A. No difference was detected in mortality and feeding levels among insects caged to basal internodes of G98A and basal and apical internodes of Ranger. The influence of abiotic factors on leafhopper resistance was evaluated by caging adult leafhoppers to either G98A or Ranger under four combinations of low and high light (250 and 1,000 micromol s(-1) m(-2)) and temperature regimes (17 and 30 degrees C). After 96 h, the highest cumulative mortality was associated with leafhoppers confined to G98A under high light and high temperature conditions. Temperature level and plant type also had an effect on the production of excretory droplets, resulting in the highest number of excretory droplets being associated with Ranger under the high temperature regime. These results indicate that certain regions of M. sativa G98A are better protected against the potato leafhopper than others and that temperature influences resistance levels of glandular-haired alfalfa.     

Role of the glandular trichomes in resistance of perennial alfalfa to the potato leafhopper (Homoptera: Cicadellidae)

Experiments were performed to elucidate resistance of glandular-haired alfalfa, Medicago sativa L., to the potato leafhopper, Empoasca fabae (Harris). The primary objective was to examine the potential role of the glandular trichomes and stem lignification for imparting resistance to this pest. During free-choice foliar discoloration experiments, the resistant alfalfa clone FGplh13 expressed lower levels of injury than the susceptible P5373 after 13 d. No-choice tests performed using nymphal potato leafhoppers showed higher levels of mortality associated with the resistant glandular-haired clone FGplh13, and no nymphs survived after 48 h. Additional no-choice experiments using FGplh13 and P5373 alfalfa with the glandular and nonglandular trichomes intact or removed show that mortality of nymphs and adults decreased after removal of the glandular trichomes from FGplh13. Nymphal and adult mortality and nymphal development time did not differ on FGplh13 with the trichomes removed and P5373 with the trichomes intact or removed. Firstinstar, and possibly second-instar, potato leafhoppers were entrapped by an exudate produced by the glandular trichomes on the FGplh13 alfalfa. Stem lignification and the number of vascular bundles did not differ between FGplh13 and P5373 alfalfas. The glandular trichomes on FGplh13 alfalfa appeared to provide the major host resistance factor, with resistance to adults being chemically based and resistance to nymphs being chemically and mechanically based. To maintain levels of potato leafhopper resistance, breeders would appear to benefit by continuing to select for the expression of the glandular trichome phenotype.     

Distribution of alfalfa weevil (Coleoptera: Curculionidae) larvae among postcutting locations

A field study was conducted in 1996 and 1997 to determine survival and spatial patterns of alfalfa weevil, Hypera postica (Gyllenhal), larvae within and between windrows of swathed alfalfa, Medicago sativa L., forage as dry matter concentration increased. After harvest, the percentage of live weevil larvae residing in swathed forage decreased as swath dry matter percentage increased. Conversely, larval populations in stubble between windrows increased with increasing swath dry matter. Larval populations in stubble under windrowed forage increased slightly as windrowed forage dry matter increased. Applications of these results for the development of alfalfa harvest systems for improved cultural control of alfalfa weevil are discussed.     

Physiological response of glandular-haired alfalfa to potato leafhopper (Hemiptera: Cicadellidae) injury

Plant tolerance to herbivory is a key approach for managing pests. In alfalfa, Medicago sativa, the potato leafhopper, Empoasca fabae, is a major pest as a result of the cascade of plant responses to piercing-sucking injury. To identify tolerance to its injury based on alfalfa physiology, experiments were conducted in the field and greenhouse. In our comparison of the response of field-grown alfalfa cultivars to standardized leafhopper densities, net photosynthesis and transpiration rates of 'Geneva' leaves were reduced by 18 and 21%, respectively, by leafhopper presence compared with a rate change of <1% of resistant 'EverGreen' leaves. Under greenhouse conditions, alfalfa clones varied in their level of gas exchange (net photosynthesis and transpiration) and stem elongation responses to leafhopper injury. For example, in the comparison of seven clones, net photosynthesis declined an average of 40.7% with leafhopper injury, although individual clones varied from 26.6 to 74.3% reduction. Internode elongation after 2 d was 60.3% less on injured stems compared with healthy stems, but again, the individual clones varied from 17.3 to 91.9%. In a time-course study of selected clones, clones varied in their level of injury just after and 3 d after insect removal. Gas exchange responses of all clones recovered by 7 d after cessation of injury. In a choice test, leafhoppers spent similar amounts of time on the susceptible clone and the most tolerant clone; however, their precise feeding behaviors were not measured. Thus, the variable response of clones to injury may be either true physiological tolerance or antixenosis from a change in feeding behavior. This study showed putative tolerance to leafhopper injury among alfalfa genotypes, suggesting that tolerance could be the basis for crop protection in alfalfa from potato leafhopper injury.     

Nutritional quality and yield of seedling alfalfa established with a barley companion crop and weeds

Alfalfa (Medicago sativa L.) was established with barley (Hordeum vulgare L.) and weeds to determine the effect of weed management treatments on yield and nutritional quality of alfalfa and companion crop and weeds. Alfalfa+barley and alfalfa+wild oat (Avena fatua L.) had much higher yields in August of the first year than alfalfa alone. Total yields for the first three harvests were similar for alfalfa established weed free, with broad leaf weeds, wild oat or green foxtail (Setaria viridis L.). Protein contents in forage from the August harvest were 228, 196, 117 and 181 g/kg dry matter (DM) in weed free alfalfa, alfalfa plus broadleaf weeds, alfalfa plus wild oat, and alfalfa plus green foxtail, respectively. Alfalfa+barley had higher total in vitro gas production than pure alfalfa, which is correlated with the digestibility of the forage. Total in vitro gas production indicated that alfalfa+barley+weeds had slightly lower digestibility than weed free alfalfa+barley. However, alfalfa with a high percentage of redroot pigweed (Amaranthus retroflexus L.), wild oat or green foxtail had similar or higher in vitro gas production than pure alfalfa. The results from this experiment support a previous economic study that indicated herbicide application for weed control in seedling alfalfa was not necessary.     

Multiplex CRISPR/Cas9-mediated mutagenesis of alfalfa FLOWERING LOCUS Ta1 (MsFTa1) leads to delayed flowering time with improved forage biomass yield and quality

Alfalfa (Medicago sativa L.) is a perennial flowering plant in the legume family that is widely cultivated as a forage crop for its high yield, forage quality and related agricultural and economic benefits. Alfalfa is a photoperiod sensitive long-day (LD) plant that can accomplish its vegetative and reproductive phases in a short period of time. However, rapid flowering can compromise forage biomass yield and quality. Here, we attempted to delay flowering in alfalfa using multiplex CRISPR/Cas9-mediated mutagenesis of FLOWERING LOCUS Ta1 (MsFTa1), a key floral integrator and activator gene. Four guide RNAs (gRNAs) were designed and clustered in a polycistronic tRNA–gRNA system and introduced into alfalfa by Agrobacterium-mediated transformation. Ninety-six putative mutant lines were identified by gene sequencing and characterized for delayed flowering time and related desirable agronomic traits. Phenotype assessment of flowering time under LD conditions identified 22 independent mutant lines with delayed flowering compared to the control. Six independent Msfta1 lines containing mutations in all four copies of MsFTa1 accumulated significantly higher forage biomass yield, with increases of up to 78% in fresh weight and 76% in dry weight compared to controls. Depending on the harvesting schemes, many of these lines also had reduced lignin, acid detergent fibre (ADF) and neutral detergent fibre (NDF) content and significantly higher crude protein (CP) and mineral contents compared to control plants, especially in the stems. These CRISPR/Cas9-edited Msfta1 mutants could be introduced in alfalfa breeding programmes to generate elite transgene-free alfalfa cultivars with improved forage biomass yield and quality.     

Potato leafhopper (Homoptera: Cicadellidae) injury disrupts basal transport of 14C-labelled photoassimilates in alfalfa

The potato leafhopper, Empoasca fabae (Harris), is a key pest of alfalfa, Medicago sativa L., in part because of the leafhopper's ability to disrupt upward translocation within phloem tissues. To determine if leafhopper injury also disrupts basal translocation necessary for regrowth and perenniality of alfalfa, we used radiolabeled 14CO2 to measure the basal transport of photoassimilates in injured and healthy plants. In one experiment, less 14C was transported to lower stem tissue of leafhopper-injured plants in comparison to the same tissue of healthy plants in early vegetative and early reproductive stages of alfalfa development. In a second experiment, less 14C was transported to lower stem, crown, and root tissues of injured plants in comparison to the same tissues of healthy, early reproductive plants. The disruption of basal transport caused by potato leafhopper may impact carbon storage and mobilization subsequent to defoliation, winter survival, and nitrogen fixation.     

Host suitability and gas exchange response of grapevines to potato leafhopper (Hemiptera: Cicadellidae)

Although potato leafhopper, Empoasca fabae (Harris) (Hemiptera: Cicadellidae), is highly polyphagous, classic host studies do not recognize grapevines (Vitis spp.), as suitable hosts. Recently, injury has been reported and reproduction documented within grape vineyards, suggesting a host expansion for the leafhopper. To document this apparent expansion in host use, we determined whether grape plants were suitable hosts for potato leafhopper reproduction, measured the consequence of feeding injury on gas exchange rates of grape leaves, and compared the susceptibility to feeding injury among cultivars. We found that potato leafhopper adults survived equally well on grape (Vitis vinifera L.), alfalfa (Medicago sativa L.), and fava bean (Vicia faba L.). The total number of offspring was greater on fava bean but did not differ between alfalfa and grape. Injury to grapevines was assessed by measuring gas exchange responses of leaves in field cages and in greenhouse tests. We found marginally significant declines in photosynthesis and transpiration rates in the field (9.6 and 13.2%, respectively), and much stronger effects in greenhouse tests (ranging between 22 and 52%). Our results verify that Vitis is a suitable host, and that potato leafhopper is capable of injuring its gas exchange physiology. We discuss possible explanations for the host expansion, and its potential to damage commercial grapevines.     

Growth of potato cultivars in response to application of phosphate fertiliser

Three experiments examined the response of potato cultivars from different maturity groups to fertiliser phosphorus application under conditions of relatively low residual soil P. Initial expansion of the leaf canopy was much more rapid following P application and maximum cover was attained sooner. In the absence of applied P, maximum ground cover was reduced substantially in early-maturing cultivars but not in maincrop cultivars. At early harvests, all cultivars showed very large growth responses to P application, but cv. Rocket grown from well-sprouted seed showed a significantly larger response than cv. Desiree. In the absence of applied P, ground cover was in some cases higher at later growth stages allowing a substantial degree of growth compensation so that final harvests showed only small effects of P application on dry matter yields. At final harvests, no significant interactions between cultivar and phosphorus fertiliser level were found for either dry matter production or P uptake despite some apparent effects. Phosphorus application increased dry matter production through increased interception of solar radiation with no effect on efficiency of utilisation.     

Effect of wheel traffic and green manure treatments on forage yield and crown rot in alfalfa (Medicago sativa)

Harvesting alfalfa damages crowns and increases the opportunity for entry of pathogens. Incorporation of green manures into soil increases population density of streptomycetes with broad pathogen antagonist activity. This study aimed to measure the impact of wheel traffic on forage yield and plant health and the effect of green manures to reduce disease. Buckwheat and sorghum-sundangrass were incorporated into soil 3 weeks before seeding alfalfa. Total bacteria, streptomycete, and pathogen antagonist densities were measured prior to planting green manures and alfalfa. Wheel traffic was applied 2 days after each forage harvest. Wheel traffic reduced forage yield 12 % to 17 % depending on year and location, significantly reduced plant counts, and increased crown rot compared to the no traffic control. Cultivar had a significant effect on yield, plant counts, and crown rot. Streptomycete density and pathogen antagonists increased when fall-sown green manure crops were incorporated in spring. Forage yields were significantly higher in plots with greater antagonist density when traffic was applied. Green manure treatments did not affect plant counts or crown rot. Mechanical wheel traffic reduces forage yield and increases disease. Green manure crops may provide benefits in alfalfa production systems by increasing pathogen antagonists.     

Impact of Bemisia whiteflies (Homoptera: Aleyrodidae) on alfalfa growth, forage yield, and quality

A 3-yr project was initiated in 1993 to examine the effects of insecticides and sustained whitefly, Bemisia argentifolii Bellows & Perring [aka. B tabaci Gennadius (Strain B)], feeding on alfalfa plant growth and vigor in greenhouse cage studies, and to determine the impact of natural Bemisia whitefly populations on alfalfa forage yields and quality in a large-plot field experiment. Alfalfa plant growth and vigor after exposure to imidacloprid and a mixture of fenpropathrin and acephate insecticides did not differ from untreated plants in the greenhouse. Consequently, foliar and soil applied insecticides were used to manipulate whitefly densities on alfalfa plants to measure whitefly feeding effects on plant growth and forage yield. Heavy whitefly densities on untreated alfalfa plants in the greenhouse resulted in significant reductions in relative growth rates and net assimilation rates as compared with imidacloprid-treated plants that were maintained relatively whitefly-free. Reductions in alfalfa plant growth measured between infested and treated plants were proportional to whitefly densities. Field plot results derived from three crop seasons were relatively consistent with our greenhouse trials. Both experimental approaches clearly showed that alfalfa plants exposed to high densities of whitefly immatures and adults grew at a significantly slower rate and produced less foliage. As a result of reduced growth rates, alfalfa maturity in the naturally infested plots was estimated to be approximately 7-10 d behind managed plots. Delays in maturity resulted in significant reductions in forage yields of 13-18% during August-September harvests when whitefly populations reached peak abundance. Whitefly feeding stresses also effected hay quality through the reduction of crude protein content and contamination of foliage with honeydew and sooty mold. The status of the Bemisia whiteflies as an economic pest to alfalfa is clearly evident from these studies, but the damage potential of whiteflies in the southwestern United States appears to be restricted to one or two harvest periods during the summer coinciding with peak adult populations and their dispersal from alternate host crops.     

Influence of alfalfa cultivar on suitability of Acyrthosiphon kondoi (Homoptera: Aphididae) for survival and development of hippodamia convergens and Coccinella septempunctata (Coleoptera: Coccinellidae)

Hippodamia convergens Guérin-Méneville and Coccinella septempunctata L. (Coleoptera: Coccinellidae) larvae were supplied daily with 1, 2, 4, or 16 mg of Acyrthosiphon kondoi Shinji (Homoptera: Aphididae) reared on one of two susceptible ('OK08' or 'CUF-101') or one resistant ('54H55') alfalfa cultivar (IMedicago sativa L.) . Hippodamia convergens survived to the adult stage when supplied with > or = 1 mg of A. kondoi per day from both susceptible and aphid-resistant cultivars, whereas C. septempunctata required > or = 2 mg of A. kondoi per day (from each cultivar) for survival to the adult stage. For both H. convergens and C. septempunctata, no consistent differences in survivorship or developmental times were observed between predator larvae supplied with increasing daily levels of A. kondoi from susceptible (OKO8 orCUF-101) versus resistant (54H55) cultivars. Additionally, alfalfa cultivar had no indirect influence on adult weight of H. convergens or C. septempunctata. Results from our study suggest that the resistant alfalfa cultivar (54H55) would have little to no effect on the nutritional value of A. kondoi for ladybeetle predators.     

Suppression of the root-lesion nematode (Pratylenchus penetrans) in alfalfa (Medicago sativa) by Streptomyces spp.

Strains of Streptomyces were tested for their ability to reduce population densities of the root-lesion nematode (RLN), Pratylenchus penetrans, in roots of alfalfa (Medicago sativa) in growth chamber assays. Previously, these strains were shown to suppress potato scab disease, caused by Streptomyces scabies, in field experiments and to inhibit in vitrogrowth of a wide range of plant-pathogenic fungi and bacteria. Inoculation with Streptomyces at planting significantly reduced RLN population densities in roots of both susceptible and resistant alfalfa varieties grown in either heat-treated or untreated soil. Reductions in RLN population densities were observed 6 weeks after nematode inoculation. Shoot dry matter was not affected by any treatment; root dry weight was reduced in Streptomycesplus nematode treatments compared to the nematode inoculation alone in some experiments but was not affected by Streptomyces when RLN was absent. Mutant strains not producing antibiotics in vitro also reduced RLN population densities in alfalfa roots and all strains maintained high population densities after inoculation into heat-treated soil and on alfalfa roots. These strains may be useful in multi-crop, multi-pathogen management programs to augment genetic resistance to plant diseases.     

Glandular trichomes on perennial alfalfa affect host-selection behavior of Empoasca fabae

Selected behavior of Empoasca fabae (Harris) (Homoptera: Cicadellidae) was examined to elucidate resistance of commercially-available glandular-haired alfalfa to this key forage pest. The overall objective was to assess the effects of the glandular trichomes on the behavior of nymphs and adults. Studies of host-plant acceptance by E. fabae nymphs found first and third instars to show a higher degree of change in settling location on the glandular-haired FGplh13 alfalfa than on the nonglandular P5373 alfalfa. Nymphs also cleaned their tarsi more frequently when in contact with the glandular trichomes on FGplh13 alfalfa, and in addition a larger number of nymphs jumped/fell off the surface of FGplh13 alfalfa. The glandular trichomes on FGplh13 also impeded nymphal mobility more effectively than the nonglandular trichomes on P5373. No choice, time-course analysis of adult host-plant acceptance behavior found that adults settled less frequently on FGplh13 alfalfa with the glandular trichomes intact, than on either FGplh13 with the glandular trichomes removed, or P5373 alfalfa with the nonglandular trichomes intact or removed. Free choice, time-course analysis of adult host-preference behavior determined that at each observation, stems of the nonglandular P5373 alfalfa were preferred over FGplh13 alfalfa. Similarly, at each observation, stems of FGplh13 alfalfa with the glandular trichomes removed were preferred over FGplh13 with the glandular trichomes intact. These data provide additional evidence for the localization of a resistance factor in the glandular trichomes of FGplh13 alfalfa. An antixenotic resistance mechanism also appears to be present, which may function, in part, through a tactile avenue.     

Mechanical and salivary aspects of potato leafhopper probing in alfalfa stems

Studies were conducted to separate the salivary and mechanical aspects of probing damage by the potato leafhopper,Empoasca fabae (Harris) (Homoptera: Cicadellidae), to stem vascular tissue of alfalfa,Medicago sativa L. Implantation of salivary gland tissue or fed-upon artificial diet under the stem epidermis yielded no evidence, three days later, of hopperburn-associated, anatomical changes. Mechanical puncturing of stems with implements approximating the size and shape of leafhopper stylets caused some anatomical changes, three days later, similar to those underlying hopperburn, i.e. tracks of necrosis, chlorosis, cell enlargement, and cell division. These changes, however, were much less severe than those observed in tissues three days after potato leafhopper probing. In contrast, puncturing through salivary gland or Malpighian tubule tissues produced extreme hyperplasia and other symptoms of wounding in cells near the puncture. This was similar to but more severe than effects from leafhopper probing, and was probably caused by leafhopper structural tissues or larger amounts of saliva being conveyed into the plant than normally occurs during leafhopper probing. We conclude that both salivation and mechanical wounding by leafhopper stylets are probably necessary to cause hopperburn-associated anatomical changes to vascular tissue in stems of alfalfa. This conclusion supports our hypothesis that hopperburn is a saliva-enhanced wound response.     

Increase in forage yield in narrowleaf birdsfoot trefoil (Lotus tenuis Waldst & Kit ex Willd) in a permanent pasture with foliar applied gibberellic acid (GA3), and phosphorus

Lotus tenuis is a perennial legume with a good adaptation to infertile, heavy and waterlogging soils. It can replace alfalfa in these sites with a similar feeding value. An important constraint is its weak competitive capacity with other graminae and weed species in permanent pastures, having consequently a poor forage yield. The objective of the present research is to overcome this disadvantage, enhancing its competitive ability with foliar applications of GA3 (GA) and phosphorus (P), increasing L. tenuis forage yield. Field experiments were conducted during 1994 with foliar application of GA (50 mg.l-1) and during 1995 with foliar application of GA (25 and 50 mg.l-1), phosphorus (8 kg.ha-1, as P2O5) and their combinations, in permanent pastures with L. tenuis and other companion grasses. In 1994 GA 50 increased significantly L. tenuis dry matter (DML) in 64.6% but not the dry matter of graminae fraction (DMG) and in consequence the total dry matter of the pasture (TDMP) was increased. In 1995 all GA treatments and their combinations with phosphorus enhanced DML but not DMG. In this sense GA 25 + P was the most effective treatment with a 151% increment of DML. Consequently TDMP was significantly increased due to a larger participation of L. tenuis in the forage yield. This increase was achieved due to a greater length and diameter of L. tenuis branches, with a logical modification in leaf:stem ratio. Moreover GA treatments reduced L. tenuis flower number. Phosphorus treatment, applied alone, showed an increase in the DML. GA treatments did not modify the feeding value of the forage in L. tenuis and graminae fractions, except GA 50 and GA 50 + P in acid detergent fiber (ADF), neutral detergent fiber (NDF) and crude protein (CP), respect to the control. The total crude protein (CP.m-2) was enhanced in all GA and GA + P treatments. Foliar GA3 and phosphorus spray applications increased the competitiveness of trefoil for light, on account of morphological changes in the spatial disposition of L. tenuis stems reaching faster the top of the pasture canopy. This practice can be an adequate alternative to increase the forage yield and total crude protein in permanent and cultivated pastures with a low cost-benefit ratio.     

Fiber digestibility in juvenile Galapagos tortoises (Geochelone nigra) and implications for the development of captive animals

Digestive strategies have been recognized to be a key factor for healthy growth in juvenile Galapagos giant tortoises (Geochelone nigra). The aim of present study was to investigate digestive coefficients with special regard to fiber fractions. Four captive bred Galapagos giant tortoises 4–5 years of age were fed a controlled diet for 32 days. The diet consisted of 77% hay, 15% tortoise pellets, and 8% apples on a dry matter basis. On a dry matter basis diet analysis showed: 95.7% organic matter, 11.3% crude protein, 20.5% crude fiber, 22.6% acid detergent fiber, 5.0% acid detergent lignin, and 17.6% cellulose. Based on total fecal collection during 7 days average dry matter digestibilities were calculated: 65% for dry matter, 67% for organic matter, 63% for crude protein, 55% for crude fiber, 49% for acid detergent fiber, 41% for acid detergent lignin, 54% for cellulose. An increase in crude fiber content resulted in a reduced digestibility in comparative evaluations of data for different tortoise species, and in a comparison of tortoises and mammalian hindgut fermenters. Compared to some mammalian hindgut‐fermenting herbivore species (domestic horses, Asian elephants, Indian rhinoceroses) on a diet of hay and concentrates, the juvenile Galapagos giant tortoises showed a digestion of similar efficiency. If a reduction in dietary digestibility is warranted in juvenile Galapagos giant tortoises, it is concluded that dietary fiber levels should be increased and it is proposed that crude fiber levels of 30–40% on a dry matter basis should be achieved. Zoo Biol 24:185–191, 2005. © 2005 Wiley‐Liss, Inc.     

The digestibility and nutritive value for dairy heifers of rations containing cellulose fibre

Cellulose residue (cellufiber) from an ammonium base acid sulphite paper process was accepted by dairy heifers to the extent of 40% of the dry matter of an all-roughage ration when mixed with maize (Zea mays) silage at time of feeding. No adverse physiological effects were observed during digestion trials lasting 80 days in which cattle were given successively diets containing 10, 20, 30 or 40% of the ration dry matter as cellufiber. Apparent digestibility of crude protein decreased and digestibility of crude fibre and acid detergent fibre increased as the cellufiber level in the ration increased. Digestibility coefficients of dry matter, nitrogen-free extract and gross energy were unaffected.Uniformity of values for total digestible nutrients, digestible energy and metabolizable energy and, also, of the body weight gains by the test animals indicated only small differences in nutritive value between rations. However, estimated nutritive values of ration components indicated appreciable differences. The TDN of cellufiber was 56% which was approximately equal to that of alfalfa (Medicago sativa) hay and was 82% of the value of maize silage. However, the digestible energy value of cellufiber (1.93 Mcal/kg DM) was only 77% of alfalfa hay and 69% of maize silage. Similarly, its metabolizable energe value of 1.41 Mcal/kg DM was 69% of that of alfalfa hay and 61% of that of maize silage.