Histologic case series of human acellular dermal matrix in superior capsule reconstruction.

BACKGROUND: Acellular dermal matrix (ADM) allografts are commonly used in the surgical treatment of complex and irreparable rotator cuff tears. Multiple studies report that superior capsule reconstruction (SCR) using ADM has resulted in short-term clinical success as assessed via radiographic and patient-reported outcomes. However, limited information is available regarding the biologic fate of these grafts in human subjects. This case series describes histologic results from 8 patients who had reoperations, during which the previously implanted ADMs were removed. These explanted ADMs were subjected to histologic analysis with the hypothesis that they would have evidence of recellularization, revascularization, and active remodeling. METHODS: Eight patients, 38-82 years old, underwent reoperation 6-38 months after undergoing SCR. ADM explants were voluntarily shipped to the manufacturer for histologic analysis. Each graft's structure and composition were qualitatively evaluated by 1 or more of the following histologic stains: hematoxylin and eosin, safranin O, and Russell-Movat pentachrome. Pan-muscle actin staining also assessed the level of neovascularization, potential myoblast or myocyte infiltration, and muscle tissue development in the graft, and was analyzed to determine the proportion of graft that had been recellularized in situ. RESULTS: Grafts showed varying levels of gross and microscopic incorporation with the host. An uneven, but high, overall degree of recellularization, revascularization, and active remodeling was observed. The degree of remodeling correlated with implant duration. These results are consistent with successful biologic reconstruction of the superior shoulder capsule. CONCLUSIONS: The present histologic analysis suggests that ADMs used in SCR undergo active recellularization, revascularization, and remodeling as early as 6 months after implantation, and that graft recellularization positively correlates with duration of implantation. These results represent a significant advancement in our knowledge regarding biologic incorporation of ADMs used in SCR.

Non-destructive Phenotyping to Identify Brachiaria Hybrids Tolerant to Waterlogging Stress under Field Conditions.

Brachiaria grasses are sown in tropical regions around the world, especially in the Neotropics, to improve livestock production. Waterlogging is a major constraint to the productivity and persistence of Brachiaria grasses during the rainy season. While some Brachiaria cultivars are moderately tolerant to seasonal waterlogging, none of the commercial cultivars combines superior yield potential and nutritional quality with a high level of waterlogging tolerance. The Brachiaria breeding program at the International Center for Tropical Agriculture, has been using recurrent selection for the past two decades to combine forage yield with resistance to biotic and abiotic stress factors. The main objective of this study was to test the suitability of normalized difference vegetation index (NDVI) and image-based phenotyping as non-destructive approaches to identify Brachiaria hybrids tolerant to waterlogging stress under field conditions. Nineteen promising hybrid selections from the breeding program and three commercial checks were evaluated for their tolerance to waterlogging under field conditions. The waterlogging treatment was imposed by applying and maintaining water to 3 cm above soil surface. Plant performance was determined non-destructively using proximal sensing and image-based phenotyping and also destructively via harvesting for comparison. Image analysis of projected green and dead areas, NDVI and shoot biomass were positively correlated (r ≥ 0.8). Our results indicate that image analysis and NDVI can serve as non-destructive screening approaches for the identification of Brachiaria hybrids tolerant to waterlogging stress.

Phenotyping Brachiaria Genotypes to Assess Rhizoctonia Resistance by Comparing Three Inoculum Types.

Rhizoctonia foliar blight, caused by Rhizoctonia solani, is an important disease of Brachiaria spp. in tropical America. Host-plant resistance is an attractive option for disease management. In this study, we evaluated three inoculum types (mycelium-infected agar disc, microdiscs suspensions, and microencapsulated-mycelium suspensions) in order to identify a rapid and accurate method for mass screening of Brachiaria genotypes for resistance to Rhizoctonia spp. in greenhouse trials. Visual damage score, area under the disease progress curve, and percent chlorophyll loss were estimated to determine the most accurate and precise method for evaluating Rhizoctonia resistance. The microencapsulated-mycelium solution (0.75 g/ml in potato dextrose broth sprayed on plants 30 days after planting) caused greater foliar damage than the other inoculum types and allowed effective discrimination between resistant and susceptible genotypes. The effectiveness of spray-applied, microencapsulated-mycelium was further corroborated by the evaluation of 350 genotypes not previously selected for resistance to Rhizoctonia spp., which varied significantly in their reaction to R. solani. The microencapsulated-mycelium methodology has several advantages over existing methods, including its high-throughput capacity, efficient use of time and space, ease of quantification of inoculum, and consistent results over replicate trials. This methodology could be applied to assess resistance to Rhizoctonia spp. in other crops.

Root adaptations to soils with low fertility and aluminium toxicity.

Background Plants depend on their root systems to acquire the water and nutrients necessary for their survival in nature, and for their yield and nutritional quality in agriculture. Root systems are complex and a variety of root phenes have been identified as contributors to adaptation to soils with low fertility and aluminium (Al) toxicity. Phenotypic characterization of root adaptations to infertile soils is enabling plant breeders to develop improved cultivars that not only yield more, but also contribute to yield stability and nutritional security in the face of climate variability. Scope In this review the adaptive responses of root systems to soils with low fertility and Al toxicity are described. After a brief introduction, the purpose and focus of the review are outlined. This is followed by a description of the adaptive responses of roots to low supply of mineral nutrients [with an emphasis on low availability of nitrogen (N) and phosphorus (P) and on toxic levels of Al]. We describe progress in developing germplasm adapted to soils with low fertility or Al toxicity using selected examples from ongoing breeding programmes on food (maize, common bean) and forage/feed (Brachiaria spp.) crops. A number of root architectural, morphological, anatomical and metabolic phenes contribute to the superior performance and yield on soils with low fertility and Al toxicity. Major advances have been made in identifying root phenes in improving adaptation to low N (maize), low P (common bean) or high Al [maize, common bean, species and hybrids of brachiariagrass, bulbous canarygrass (Phalaris aquatica) and lucerne (Medicago sativa)]. Conclusions Advanced root phenotyping tools will allow dissection of root responses into specific root phenes that will aid both conventional and molecular breeders to develop superior cultivars. These new cultivars will play a key role in sustainable intensification of crop-livestock systems, particularly in smallholder systems of the tropics. Development of these new cultivars adapted to soils with low fertility and Al toxicity is needed to improve global food and nutritional security and environmental sustainability.

Assessing the Resistance of Brachiaria Hybrids to Pathogenic Rhizoctonia.

Rhizoctonia foliar blight, caused by Rhizoctonia solani anastomosis group 1, is an economically important fungal disease found throughout the world. The fungus attacks numerous crops, including cereals, roots and tubers, legumes, and cruciferous, horticultural, and ornamental plants. In tropical America, this invasive and destructive disease also attacks most Brachiaria spp. used as forages in the ranching industry, especially in the production of cattle. Research to solve this constraint has been ongoing at the International Center for Tropical Agriculture and has generated new Brachiaria hybrids with excellent agronomic performance, tolerance to poor soils, and, particularly, high resistance to biotic factors such as Rhizoctonia foliar blight. These hybrids belong to lines obtained from Brachiaria humidicola, B. brizantha, and B. decumbens. To identify resistance among Brachiaria hybrid genotypes, the hybrid clones were evaluated for their variability in resistance, and their disease reaction was also determined and characterized. Results led to the identification of hybrids that not only were highly resistant to the blight but also had excellent agronomic characteristics.

Characterization of resistance to adult spittlebugs (Hemiptera: Cercopidae) in Brachiaria spp.

Nymphs and adults of several spittlebug (Hemiptera: Cercopidae) species are key pests of forage brachiariagrasses (Brachiaria spp.) in tropical America. To support current breeding programs, a series of experiments aimed at characterizing the mechanisms of resistance to adult feeding damage were carried out. Five genotypes were used: two susceptible checks (CIAT 0606 and CIAT 0654) and three nymph-resistant genotypes (CIAT 36087, CIAT 6294, and SX01NO/0102). Test insects were Aeneolamia varia (F.), A. reducta (Lallemand), and Zulia carbonaria (Lallemand). The nymph-resistant genotypes showed tolerance to all spittlebug species tested. Tolerance in these genotypes can be classified as only moderate given the extent of losses (60-80%) caused by both female and male adults. None of the nymph-resistant genotypes had antibiotic effects on adults feeding on foliage. The results also indicated that antixenosis for feeding is not a plausible explanation for lower damage scores and less biomass losses in resistant genotypes. The fact that adult longevity (usually 8 d) was not affected when the adults were forced to feed on roots of a genotype with strong antibiotic resistance to nymphs is regarded as additional evidence that resistances to nymphs and to adults in Brachiaria are largely independent.

Diversity of Rhizoctonia spp. Causing Foliar Blight on Brachiaria in Colombia and Evaluation of Brachiaria Genotypes for Foliar Blight Resistance.

Up to 50% of Brachiaria production in the tropics is affected by foliar blight caused by Rhizoctonia spp. Monothallic isolates of Rhizoctonia (n = 147) were cultured from different Brachiaria genotypes in Colombia and morphologically characterized and evaluated in pathogenicity trials in the greenhouse. Based on restriction fragment length polymorphism of the internal transcribed spacer region, 101 of the isolates were identified as Rhizoctonia solani anastomosis group (AG)-1 IA and were multinucleated, with high growth rate, brown mycelium, and high virulence; and 46 isolates were identified as Rhizoctonia sp. AG-D and were binucleated, with low growth rate, white mycelium, and lower virulence on the Brachiaria genotypes tested. The Rhizoctonia isolates also showed variation according to geographic origin, with R. solani AG-1 IA prevalent in warm lowland areas and Rhizoctonia sp. AG-D occurring in cooler areas. Ten Brachiaria genotypes were challenged with different Rhizoctonia isolates and resistant reactions were seen in three of these genotypes, including Brachiaria hybrid (International Center for Tropical Agriculture [CIAT] 36062), Brachiaria brizantha 'Marandú' (CIAT 6294), and Brachiaria hybrid 'Mulato II' (CIAT 36087). These results will contribute to a greater understanding of the interaction of diverse Rhizoctonia isolates on different Brachiaria genotypes, supporting improvement of Brachiaria spp. for disease resistance.

Independence of resistance in Brachiaria spp. to nymphs or to adult spittlebugs (Hemiptera: Cercopidae): implications for breeding for resistance.

Both nymphal and adult spittlebugs (Hemiptera: Cercopidae) cause serious economic damage to susceptible brachiariagrass [genus Brachiaria (Trin.) Griseb], pastures in tropical America. Both life stages are xylem feeders: nymphs feed primarily on roots and stems, whereas the adults feed mainly on foliage. Numerous interspecific brachiariagrass hybrids with high levels of antibiosis resistance to nymphs of several important spittlebug species have been obtained. Recent studies revealed major inconsistencies between reaction to nymphs and reaction to adults on the same host genotype. Because both insect life stages can cause severe economic damage on susceptible brachiariagrass pastures, a cultivar development strategy must take into account resistance to both life stages. To assess the degree of association between resistance to spittlebug nymphs and to adult feeding, we tested 164 hybrids and six check genotypes for resistance to both life stages of three spittlebug species: Aeneolamia varia (F.), Aeneolamia reducta (Lallemand), and Zulia carbonaria (Lallemand). Most hybrids tested were classified as resistant to nymphs. On the contrary, for all three species, the overall mean damage score of the 164 hybrids did not differ from the mean score of the susceptible checks. None of the hybrids was classified as resistant to adult feeding damage. Correlations between percentage nymph survival and adult damage scores were consistently low (r = 0.0104-0.0191). Correlations between nymphal and adult damage scores were also low (0.109-0.271), suggesting that resistances to the different life stages are largely independent. Chi-square analyses comparing frequency distributions of responses of the 164 breeding hybrids to nymphs or adults confirmed essential genetic independence of these two traits. We conclude that attention to improving genetic resistance specifically to adult feeding damage is warranted.

Screening for resistance to adult spittlebugs (Hemiptera: Cercopidae) in Brachiaria spp.: methods and categories of resistance.

Both nymphal and adult stages of several species of spittlebugs (Hemiptera: Cercopidae) are key economic pests of brachiariagrasses (Brachiaria spp.) in tropical America. Progress has been made in the characterization and development of antibiosis resistance to nymphs in brachiariagrasses. Essentially no attention has been given to screening germplasm for resistance to adults. To support current breeding programs, a series of experiments was conducted to develop a methodology to screen for adult damage and to study categories of resistance to adult feeding damage. Six host brachiariagrass genotypes were used: two susceptible checks (CIAT 0606 and CIAT 0654) and four nymph-resistant genotypes (CIAT 6294, CIAT 36062, CIAT 36087, and SX01NO/0102). Test insects were Aeneolamia varia (F.) and Zulia carbonaria (Lallemand). None of the nymph-resistant genotypes was antibiotic to adults. All four nymph-resistant genotypes showed tolerance to A. varia and Z. carbonaria feeding damage. The levels of tolerance to adults of Z. carbonaria, a larger, more aggressive species, were lower. Of the four nymph-resistant genotypes, only CIAT 6294 and CIAT 36087 showed some tolerance to Z. carbonaria expressed as lower leaf damage scores, less chlorophyll loss, and lower functional plant loss indices. The fact that a genotype like SX01NO/0102, which is highly antibiotic to nymphs, is susceptible to adult damage suggests that mechanisms of resistance to the two spittlebug life stages may be independent. Results of these studies suggest a need to incorporate routine screening for tolerance to adult feeding damage as an additional selection criterion in the breeding scheme.

Sublethal effects of antibiosis resistance on the reproductive biology of two spittlebug (Hemiptera: Cercopidae) species affecting Brachiaria spp.

Several greenhouse experiments were used to measure how high levels of antibiosis resistance to nymphs in two interspecific Brachiaria (brachiariagrass) hybrids affect life history parameters of the spittlebugs Aeneolamia varia (F.) and Zulia carbonaria (Lallemand), two of the most important spittlebug (Hemiptera: Cercopidae) species affecting Brachiaria production in Colombia. The A. varia-resistant hybrid CIAT 36062, the Z. carbonaria-resistant hybrid SX01NO/0102, and the susceptible accession CIAT 0654 were used to compare the effect of all possible combinations of food sources for nymphs and adults. Calculation of growth indexes showed a significant impact of antibiosis resistance on the biology of immature stages of both species. Median survival times of adults feeding on resistant genotypes did not differ from those recorded on the susceptible genotype, suggesting that factors responsible for high mortality of nymphs in the resistant hybrids did not affect adult survival. Rearing nymphs of A. varia on CIAT 36062 and of Z. carbonaria on SX01NO/0102 had deleterious sublethal effects on the reproductive biology of resulting adult females. It is concluded that high nymphal mortality and subsequent sublethal effects of nymphal antibiosis on adults should have a major impact on the demography of the two spittlebug species studied.

Response of resistant and susceptible Brachiaria spp. genotypes to simultaneous infestation with multiple species of spittlebugs (Hemiptera: Cercopidae).

The response of one susceptible and three resistant Brachiaria spp. (Hemiptera: Cercopidae) genotypes to individual or combined attacks by nymphs of Aeneolamia varia (F.), Aeneolamia reducta (Lallemand), Zulia carbonaria (Lallemand), and Zulia pubescens (F.) was studied. We assessed the effect of infesting plants of the susceptible check BRX 44-02 and of the A. varia-resistant genotypes CIAT 6294 and CIAT 36062 with A. varia, Z. carbonaria, or Z. pubescens either alone or in two-species combinations. In a second trial, we studied the performance of BRX 44-02, CIAT 6294, and the multiple resistant clone SX01NO/0102 exposed to individual or combined attack by A. reducta and Z. carbonaria. In a third trial, we compared the response of BRX 44-02, CIAT 6294, and CIAT 36062 to individual A. varia, Z. carbonaria, or Z. pubescens attack as opposed to a combined three-species attack. Plant damage scores and percentage of nymphal survival were recorded in all three trials. Data on percentage of survival indicated that competition between and among spittlebug species occurs. However, we found no evidence of interaction between species competition and different levels of resistance to spittlebug. Rather, host genotype reactions conformed to previously known categories of resistance regardless of the presence of more than one spittlebug species. Resistance rather than competition seems to have been the overriding factor determining nymph survival and resistance expression (damage scores) in these experiments. Our results corroborate the need to develop brachiariagrass genotypes with multiple resistance to spittlebugs.

Antibiosis and tolerance to five species of spittlebug (Homoptera: Cercopidae) in Brachiaria spp.: implications for breeding for resistance.

Several genera and species of spittlebugs (Homoptera: Cercopidae) are economic pests of Brachiaria spp. grasses in tropical America. To support current breeding programs aimed at obtaining multiple spittlebug resistance, we undertook a series of studies on antibiosis and tolerance as possible mechanisms of resistance to five major spittlebug species affecting Brachiaria spp. in Colombia: Aeneolamia varia (F.), Aeneolamia reducta (Lallemand), Zulia carbonaria (Lallemand), Zulia pubescens (F.), and Mahanarva trifissa (Jacobi). Four host genotypes, well known for their reaction to A. varia attack, were used to compare their resistance to other spittlebug species: CIAT 0654 and CIAT 0606 (susceptible) and CIAT 6294 and CIAT 36062 (resistant). CIAT 0654 and CIAT 36062 were used in antibiosis studies. Tolerance studies were conducted with CIAT 0654, CIAT 6294, and CIAT 36062. Sixty-five hybrid-derived clones were used to identify levels of multiple resistance to three spittlebug species. The levels of antibiosis resistance in CIAT 36062 clearly differed by spittlebug species and were classified as follows: very high for M. trifissa, high for A. varia and A. reducta, moderate for Z. pubescens, and absent for Z. carbonaria. Our results suggest the presence of true tolerance to Z. carbonaria in CIAT 6294 and CIAT 36062, true tolerance to Z. pubescens in CIAT 6294 and a combination of tolerance and antibiosis as mechanisms of resistance to Z. pubescens in CIAT 36062. Of the 65 hybrid clones tested with A. varia, A. reducta, and Z. carbonaria, 15 combined resistance to two species and three showed antibiosis resistance to all three spittlebug species.