Effects of feeding different probiotic types on metabolic, performance, and carcass responses of Bos indicus feedlot cattle offered a high-concentrate diet.

Two experiments were designed to evaluate the effects of different probiotic combinations on rumen fermentation characteristics, performance, and carcass characteristics of feedlot Bos indicus beef bulls offered a high-concentrate diet. In experiment 1, 30 rumen-fistulated Nellore steers were blocked by initial body weight (BW = 350 ± 35.0 kg) and within blocks (n = 10), animals were randomly assigned to receive: 1) high-concentrate diet without probiotic supplementation (n = 10; CONT), 2) CONT plus 1 g per head of a probiotic mixture containing three strains of Enterococcus faecium and one strain of Saccharomyces cerevisiae (3.5 × 109 CFU/g; n = 10; EFSC), and 3) CONT plus 2 g per head of a probiotic mixture containing Bacillus licheniformis and Bacillus subtilis (3.2 × 109 CFU/g; n = 10; BLBS). The experimental period lasted 35 d, being 28 d of adaptation and 7 d of sampling. From day 34 to day 35 of the experimental period, ruminal fluid and fecal samples were collected every 3 h, starting immediately before feeding (0 h) for rumen fermentation characteristics and apparent nutrient digestibility analysis, respectively. In experiment 2, 240 Nellore bulls were ranked by initial shrunk BW (375 ± 35.1 kg), assigned to pens (n = 4 bulls per pen), and pens randomly assigned to receive the same treatments as in experiment 1 (n = 20 pens per treatment). Regardless of treatment, all bulls received the same step-up and finishing diets throughout the experimental period, which lasted 115 d. In both experiments, data were analyzed as orthogonal contrasts to partition-specific treatment effects: 1) probiotic effect: CONT vs. PROB and 2) probiotic type: EFSC vs. BLBS (SAS Software Inc.). In experiment 1, no contrast effects were observed on nutrient intake, overall nutrient digestibility, and rumen fermentation analyses (P ≥ 0.13). Nonetheless, supplementation of probiotics, regardless of type (P = 0.59), reduced mean acetate:propionate ratio and rumen ammonia-N concentration vs. CONT (P ≤ 0.05). In experiment 2, no significant effects were observed for final BW and dry matter intake (P ≥ 0.12), but average daily gain and feed efficiency tended to improve (P ≤ 0.10) when probiotics were offered to the animals. Probiotic supplementation or type of probiotic did not affect carcass traits (P ≥ 0.22). In summary, supplementation of probiotics containing a mixture of E. faecium and S. cerevisiae or a mixture of B. licheniformis and B. subtilis reduced rumen acetate:propionate ratio and rumen ammonia-N levels and tended to improve the performance of feedlot cattle offered a high-concentrate diet.

Two experiments were designed to evaluate the effects of different probiotic combinations on rumen fermentation characteristics, performance, and carcass characteristics of feedlot Bos indicus beef bulls offered a high-concentrate diet. The two probiotics consisted of a mixture containing three strains of Enterococcus faecium and one strain of Saccharomyces cerevisiae or a mixture of Bacillus licheniformis and Bacillus subtilis. Supplementation of probiotics, regardless of type, reduced acetate:propionate ratio, and mean rumen ammonia-N concentration and tended to improve the performance of feedlot cattle offered a high-concentrate diet, demonstrating the potential of this technology to be used as a feed additive for beef cattle.

Effects of Megasphaera elsdenii administration on performance and carcass traits of finishing Bos indicus feedlot cattle.

This study evaluated the effects of Megasphaera elsdenii administration at the beginning of the feedlot period on performance of Bos taurus indicus bulls. On d 0, 383 Nellore bulls (initial shrunk body weight 384 ± 29.2 kg; initial age = 24 ± 2 mo) were assigned to treatments in a randomized complete block design. Treatments consisted of 1) 14 d adaptation diet and transition to a finishing diet (CONT), 2) CONT plus oral administration of 20 mL of Lactipro-NXT (M. elsdenii) on d 0 of the study (MEG-14), 3) CONT diet, consisting of 6 d of adaptation diet plus oral administration of 20 mL of Lactipro-NXT on d 0 of the study (MEG-6), and 4) No adaptation diet and oral administration of 20 mL of Lactipro-NXT on d 0 of the study (MEG-0). Experimental period lasted 119 d. No treatment effects were observed for any of the performance parameters evaluated herein (P ≥ 0.15). Nonetheless, a treatment × wk interaction was observed for DM, NEm, and NEg intakes (P < 0.0001). For all these parameters, MEG-0 and MEG-6 had a reduced intake vs. MEG-14 and CONT in the first wk of the study (P ≤ 0.05). For the carcass traits, no effects were observed for HCW (P ≥ 0.24), whereas MEG-6 had a greater REA when compared with MEG-0 and MEG-14 (quadratic effect; P = 0.04) and MEG-administered bulls tended to have a greater BFT vs. CONT (P = 0.08). In summary, M. elsdenii administration at the beginning of the feedlot period did not improve performance, whereas reducing the length of the adaptation period for 6 d improved REA of finishing Bos taurus indicus bulls.

Habitat filtering drives the local distribution of congeneric species in a Brazilian white-sand flooded tropical forest.

The investigation of ecological processes that maintain species coexistence is revealing in naturally disturbed environments such as the white-sand tropical forest, which is subject to periodic flooding that might pose strong habitat filtering to tree species. Congeneric species are a good model to investigate the relative importance of ecological processes that maintain high species diversity because they tend to exploit the same limiting resources and/or have similar tolerance limits to the same environmental conditions due to their close phylogenetic relationship. We aim to find evidence for the action and relative importance of different processes hypothesized to maintain species coexistence in a white-sand flooded forest in Brazil, taking advantage of data on the detailed spatial structure of populations of congeneric species. Individuals of three Myrcia species were tagged, mapped, and measured for diameter at soil height in a 1-ha plot. We also sampled seven environmental variables in the plot. We employed several spatial point process models to investigate the possible action of habitat filtering, interspecific competition, and dispersal limitation. Habitat filtering was the most important process driving the local distribution of the three Myrcia species, as they showed associations, albeit of different strength, to environmental variables related to flooding. We did not detect spatial patterns, such as spatial segregation and smaller size of nearby neighbors, that would be consistent with interspecific competition among the three congeneric species and other co-occurring species. Even though congeners were spatially independent, they responded to differences in the environment. Last, dispersal limitation only led to spatial associations of different size classes for one of the species. Given that white-sand flooded forests are highly threatened in Brazil, the preservation of their different habitats is of utmost importance to the maintenance of high species richness, as flooding drives the distribution of species in the community.

Effects of dietary roughage neutral detergent fiber levels and flint corn processing method on growth performance, carcass characteristics, feeding behavior, and rumen morphometrics of Bos indicus cattle1.

Flint corn processing method [coarse ground corn (CGC; 3.2 mm average particle size) or steam-flaked corn (SFC; 0.360 kg/L flake density)] was evaluated in conjunction with 4 levels of NDF from sugarcane bagasse (SCB) as roughage source (RNDF; 4%, 7%, 10%, and 13%; DM basis) to determine impact on growth performance, carcass characteristics, starch utilization, feeding behavior, and rumen morphometrics of Bos indicus beef cattle. Two hundred and forty Nellore bulls were blocked by initial BW (350 ± 37 kg), assigned to 32 feedlot pens and pens within weight block were randomly assigned, in a 2 × 4 factorial arrangement (2 corn processing and 4 levels of RNDF) to treatments. Effects of corn grain processing × RNDF level were not detected (P ≥ 0.14) for growth performance, dietary net energy concentration, carcass traits, rumen morphometrics, and feeding behavior, except for time spent ruminating and time spent resting (P ≤ 0.04), and a tendency for papillae width (P ≤ 0.09). Bulls fed SFC-based diets consumed 7% less (P = 0.001), had 10.6% greater carcass-adjusted ADG (P < 0.001) and 19% greater carcass-adjusted feed efficiency (P < 0.001) compared with bulls fed CGC-based diets. Observed net energy for maintenance and gain values were 14.9% and 19.4% greater (P < 0.001), respectively, for SFC than for CGC-based diets. Fecal starch concentration was less (P < 0.001) for bulls fed SFC compared with those fed CGC. No grain processing effects were detected (P = 0.51) for rumenitis score; however, cattle fed SFC presented smaller ruminal absorptive surface area (P = 0.03). Dry matter intake increased linearly (P = 0.02) and carcass-adjusted feed efficiency tended (P = 0.06) to decrease linearly as RNDF increased. Dietary RNDF concentration did not affect carcass characteristics (P ≥ 0.19), except for dressing percentage, which tended to decrease linearly (P = 0.06) as RNDF in finishing diets increased. Increasing RNDF in finishing diets had no effect (P = 0.26) on time spent eating, but time spent ruminating and resting increased linearly (min/d; P < 0.001) with increased dietary RNDF. Steam flaking markedly increased flint corn energy value, net energy of diets, and animal growth performance, and led to improvements on feed efficiency when compared with grinding, regardless of RNDF content of diets. Increasing dietary RNDF compromised feedlot cattle feed efficiency and carcass dressing.

Feeding the combination of essential oils and exogenous α-amylase increases performance and carcass production of finishing beef cattle.

Two experiments were conducted to evaluate the performance responses of finishing feedlot cattle to dietary addition of essential oils and exogenous enzymes. The treatments in each experiment consisted of (DM basis): MON-sodium monensin (26 mg/kg); BEO-a blend of essential oils (90 mg/kg); BEO+MON-a blend of essential oils plus monensin (90 mg/kg + 26 mg/kg, respectively); BEO+AM-a blend of essential oils plus exogenous α-amylase (90 mg/kg + 560 mg/kg, respectively); and BEO+AM+PRO-a blend of essential oils plus exogenous α-amylase and exogenous protease (90 mg/kg + 560 mg/kg + 840 mg/kg, respectively). Exp. 1 consisted of a 93-d finishing period using 300 Nellore bulls in a randomized complete block design. Animals fed BEO had higher DMI (P < 0.001) but similar feed efficiency to animals fed MON (P ≥ 0.98). Compared with MON, the combination of BEO+AM resulted in 810 g greater DMI (P = 0.001), 190 g greater average daily gain (P = 0.04), 18 kg heavier final body weight (P = 0.04), and 12 kg heavier hot carcass weight (P = 0.02), although feed efficiency was not significantly different between BEO+AM and MON (P = 0.89). Combining BEO+MON tended to decrease hot carcass weight compared with BEO alone (P = 0.08) but not compared with MON (P = 0.98). Treatments did not impact observed dietary net energy values (P ≥ 0.74) or the observed:expected net energy ratio (P ≥ 0.11). In Exp. 2, five ruminally cannulated Nellore steers were used to evaluate intake, apparent total tract digestibility of nutrients, and ruminal parameters in a 5 × 5 Latin square design. Feeding BEO increased the total tract digestibility of CP compared to MON (P = 0.03). Compared to MON, feeding the combination of BEO+MON increased the intake of CP (P = 0.04) and NDF (P = 0.05), with no effects on total tract digestibility of nutrients (P ≥ 0.56), except for a tendency (P = 0.09) to increase CP digestibility. Intakes of all nutrients measured, except for ether extract (P = 0.16) were greater in animals fed BEO+AM when compared with MON (P ≤ 0.03), with no differences on total tract nutrient digestibilities (P ≥ 0.11) between these two treatments. In summary, diets containing the BEO used herein enhanced DMI of growing-finishing feedlot cattle compared with a basal diet containing MON without impair feed efficiency. A synergism between BEO and AM was detected, further increasing cattle performance and carcass production compared to MON.

Influence of light and plant size on the reproduction and growth of small palm tree species: Comparing two methods for measuring canopy openness.

PREMISE OF THE STUDY: The reproduction and growth of large palms are influenced by many factors, including light and plant size, but few studies have examined smaller species (up to 2 m tall). We examined the effect of incident light and individual size on growth rates, the probability of reproduction, and the number of inflorescences of three small palm species and compared two methods for estimating canopy openness. METHODS: We measured canopy openness above the crowns of 132 adult plants and in the centers of 72 subplots (10 × 10 m) where individuals were sampled. We also recorded individual size and the number of leaves and inflorescences produced in two years. KEY RESULTS: Reproductive individuals of Butia paraguayensis tend to occur in illuminated microhabitats. Reproduction of Acrocomia emensis was correlated with stem diameter, but not with light. Reproduction was inversely related to individual size and light in Syagrus petraea, probably because this clonal palm invests heavily in sexual reproduction during its younger stages and clonal activity in older stages and may be adapted to the low-light conditions of the woodland understory. Growth was not predicted by light or individual size. Stronger correlations were found when incident light was measured directly above the crown, as opposed to the subplot center. CONCLUSIONS: The influences of light on reproduction are dependent on plant life histories, even among related and sympatric species. Light measurements directly above individual crowns provide better understanding of the reproductive effort rather than in subplot center.

Multiresidue determination of fluoroquinolones in poultry muscle and kidney according to the regulation 2002/657/EC. A systematic comparison of two different approaches: Liquid chromatography coupled to high-resolution mass spectrometry or tandem mass spectrometry.

This work involved the optimization and validation of two methods according to the Commission Decision 2002/657/EC directives for determining fluoroquinolones residues in samples of poultry muscle and kidney: ciprofloxacin, danofloxacin, difloxacin, enrofloxacin, flumequine, marbofloxacin, nalidixic acid, norfloxacin, ofloxacin, oxolinic acid, pipemidic acid and sarafloxacin. The extraction procedure was based on a QuEChERS approach, whose optimization employed a Box-Behnken 3(3) factorial design. A liquid chromatography-tandem mass spectrometry method (LC-MS/MS) was developed for determining the twelve analytes using the multiple reaction monitoring mode (MRM). Accuracy, evaluated by recovery studies, varied from 88.8 to 112.2% for the selected levels with RSD values lower than 12.3%. The second validated method employed high resolution mass spectrometry (HRMS) performed in the single ion monitoring mode (SIM), determining nine among twelve analytes. The validation parameters were evaluated as satisfactory, with recoveries from 82.5 to 114.4% and RSD lower than 8.7%. Decision limits and detection capabilities for both methods were reported. The two methods were statistically compared using the Student's t test, at 95% confidence level, resulting in no significant difference.

Characterization of 10 microsatellite loci for Bathysa australis (Rubiaceae).

PREMISE OF THE STUDY: Bathysa australis is a common subcanopy tree from the Atlantic Forest that is pollinated by bees and wasps and produces autochoric seeds. This species exhibits great phenotypic plasticity along the elevational gradient of Serra do Mar in southeastern Brazil. We expect to assess the genetic diversity and gene flow between populations of this species along the elevational gradient. • METHODS AND RESULTS: We developed a microsatellite-enriched genomic library for B. australis, and 10 microsatellite loci were successfully amplified, ranging from one to 13 alleles per locus. The observed and expected heterozygosities ranged from 0.333 to 0.900 (average: 0.629) and 0.564 to 0.900 (average: 0.742), respectively. • CONCLUSIONS: These are the first microsatellite markers developed for the genus Bathysa and may be useful in other species of the Condamineeae tribe. These primers will be an important tool for studies of population ecology and conservation genetics.

Cytogenetics and characterization of microsatellite loci for a South American pioneer tree species, Croton floribundus.

Despite the recent advances in plant population genetic studies, the lack of information regarding pedigree, ploidy level, or mode of inheritance for many polyploids can compromise the analysis of the molecular data produced. The aim of this study was to examine both microsatellite and cytogenetic characteristics of the pioneer tree Croton floribundus Spreng. (Euphorbiaceae) to test for the occurrence of polyploidy in the species and to evaluate its implications for the appropriate use of SSR markers. Seven microsatellite markers were developed and screened for 62 individuals from a semi-deciduous tropical forest in Brazil. Chromosome number, meiotic behavior, and pollen viability were evaluated from male flower buds. All SSR loci were highly polymorphic. The number of bivalents observed in meiosis n = 56 (2n = 8× = 112) and the maximum number of alleles per individual (Ni = 8) demonstrated the occurrence of polyploidy in C. floribundus. The normal meiotic pairing and the high pollen viability suggested that C. floribundus is a regular and stable polyploid, most likely an allopolyploid. The combined SSR and cytogenetic data provided new evidence on the origin and evolution of the species as well as assured the accurate use of SSR loci for population genetic studies of the polyploid pioneer species.

Validation of an LC-MS/MS method for malachite green (MG), leucomalachite green (LMG), crystal violet (CV) and leucocrystal violet (LCV) residues in fish and shrimp.

A quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous analyses of malachite green (MG), crystal violet (CV) and its major metabolites, leucomalachite green (LMG) and leucocrystal violet (LCV) residues in fish and shrimp samples has been validated. Fish and shrimp samples were extracted with citrate buffer/acetonitrile, and the extracts were purified on strong cation-exchange (SCX) solid-phase extraction (SPE) cartridge. After conversion of LMG into MG using a post column oxidation reactor containing lead (IV) oxide (PbO(2)), the effluents were analysed. Residues were analysed using positive-ion electrospray ionisation (ESI). Identification and quantification of analytes were based on the ion transitions monitored by multiple reaction monitoring (MRM). Validation of the method was carried out in accordance with the Decision 2002/657/EC, which establishes criteria and procedures for the validation of methods. The following parameters were determined: decision limit (CCα), detection capability (CCß), linearity, accuracy, precision, selectivity, specificity and matrix effect. The decision limits (CCα) for MG, LMG, CV and LCV were 0.164, 0.161, 0.248 and 0.860 µg kg(-1). The respective detection capabilities (CCß) were 0.222, 0.218, 0.355 and 1.162 µg kg(-1). Typical recoveries (intermediate precision) in shrimp, for MG, CV, LMG and LCV for 2.0 µg kg(-1) level fortified samples using the optimised procedure were in the range 69%, 97%, 80.3% and 71.8%, respectively. The findings demonstrate the suitability of the method to detect simultaneously MG, CV and its metabolite (LMG and LCV) in fish and shrimp.

Seed banks on Attalea phalerata (Arecaceae) stems in the Pantanal wetland, Brazil.

BACKGROUND AND AIMS: Seeds can accumulate in the soil or elsewhere, such as on the stems of palms when these are covered by persistent sheaths. These sheaths could act as a safe site for some species. Here, we studied whether persistent sheaths of the palm Attalea phalerata (Arecaceae) are available sites for seed accumulation in the Pantanal wetland of Brazil. We also investigated whether the composition, richness and diversity of species of seeds in the persistent sheaths are determined by habitat (riparian forest and forest patches) and/or season (wet and dry). METHODS: All accumulated material was collected from ten persistent sheaths along the stems of 64 A. phalerata individuals (16 per habitat and 16 per season). The material was then individually inspected under a stereomicroscope to record seed species and number. KEY RESULTS: Of the 640 sheaths sampled, 65 % contained seeds (n = 3468). This seed bank included 75 species belonging to 12 families, and was primarily composed of small, endozoochoric seeds, with a few abundant species (Cecropia pachystachya and Ficus pertusa). Moraceae was the richest family (four species) and Urticaceae the most abundant (1594 seeds). Stems of A. phalerata in the riparian forest had 1·8 times more seeds and 1·3 times more species than those in forest patches. In the wet season we sampled 4·1 times more seeds and 2·2 more species on palm stems than in the dry season. Richness did not differ between habitats, but was higher in the wet season. Abundance was higher in forest patches and in the wet season. CONCLUSIONS: Attalea phalerata stems contain a rich seed bank, comparable to soil seed banks of tropical forests. As most of these seeds are not adapted to grow in flooding conditions, palm stems might be regarded as safe sites for seeds (and seedlings) to escape from the seasonal flooding of the Pantanal.

Characterization of microsatellite loci in Himatanthus drasticus (Apocynaceae), a medicinal plant from the Brazilian savanna.

PREMISE OF THE STUDY: We developed a new set of microsatellite markers for studying the genome of the janaguba tree, Himatanthus drasticus (Mart.) Plumel, which is used in folk medicine in northeastern Brazil. These novel markers are being used to evaluate the effect of harvesting on the genetic structure and diversity of natural populations of this species. • METHODS AND RESULTS: Microsatellite loci were isolated from an enriched H. drasticus genomic library. Nine primer pairs successfully amplified polymorphic microsatellite regions, with an average of 8.5 alleles per locus. The average values of observed and expected heterozygosity were 0.456 and 0.601, respectively. • CONCLUSIONS: The microsatellite markers described here are valuable tools for population genetics studies of H. drasticus. The majority of the primers also amplified sequences in the genome of another species of the same genus. This new set of markers may be useful in designing a genetic conservation strategy and a sustainable management plan for the species.

Stocks of carbon and nitrogen and partitioning between above- and belowground pools in the Brazilian coastal Atlantic Forest elevation range.

We estimated carbon and nitrogen stocks in aboveground biomass (AGB) and belowground biomass (BGB) along an elevation range in forest sites located on the steep slopes of the Serra do Mar on the north coast of the State of São Paulo, southeast Brazil. In elevations of 100 m (lowland), 400 m (submontane), and 1000 m (montane) four 1-ha plots were established, and above- (live and dead) and belowground (live and dead) biomass were determined. Carbon and nitrogen concentrations in each compartment were determined and used to convert biomass into carbon and nitrogen stocks. The carbon aboveground stock (C(AGB)) varied along the elevation range from approximately 110 to 150 Mg·ha(-1), and nitrogen aboveground stock (N(AGB)), varied from approximately 1.0 to 1.9 Mg·ha(-1). The carbon belowground stock (C(BGB)) and the nitrogen belowground stock (N(BGB)) were significantly higher than the AGB and varied along the elevation range from approximately 200-300 Mg·ha(-1), and from 14 to 20 Mg·ha(-1), respectively. Finally, the total carbon stock (C(TOTAL)) varied from approximately 320 to 460 Mg·ha(-1), and the nitrogen total stock (N(TOTAL)) from approximately 15 to 22 Mg·ha(-1). Most of the carbon and nitrogen stocks were found belowground and not aboveground as normally found in lowland tropical forests. The above- and belowground stocks, and consequently, the total stocks of carbon and nitrogen increased significantly with elevation. As the soil and air temperature also decreased significantly with elevation, we found a significantly inverse relationship between carbon and nitrogen stocks and temperature. Using this inverse relationship, we made a first approach estimate that an increase of 1°C in soil temperature would decrease the carbon and nitrogen stocks in approximately 17 Mg·ha(-1) and 1 Mg·ha(-1) of carbon and nitrogen, respectively.

Mating systems of Psychotria tenuinervis (Rubiaceae): distance from anthropogenic and natural edges of Atlantic forest fragment.

The aim of this study was to determine and compare the mating systems among Psychotria tenuinervis populations at anthropogenic edges, natural edges, and the forest interior using allozyme electrophoresis of naturally pollinated progeny arrays. P. tenuinervis showed low outcrossing rates, varying from 37% to 50% of the mating attributable to outcrossing and 50% to 63% attributable to self-fertilization, in the three habitats. The forest interior had the highest outcrossing rate (t(m) = 0.50 and t(s) = 0.43) among the three habitats. However, there were no differences in either multilocus or single-locus rates among the three habitats, indicating that the contribution of biparental inbreeding to the apparent selfing rate in these populations was very low. The multilocus (t(m)) and single-locus (t(s)) outcrossing rates for the P. tenuinervis in the sample plots within each habitat showed great heterogeneity. In conclusion, edge creation seems not to influence its mating systems. Additionally, although P. tenuinervis is a distylous species, the population's inbreeding can be attributed almost entirely to self-fertilization.

Quality of seeds produced by Psychotria tenuinervis (Rubiaceae): distance from anthropogenic and natural edges of Atlantic forest fragment.

The aim of this study was to investigate whether there were differences in the genetic variability and rate and velocity of the seed germination produced by Psychotria tenuinervis located at anthropogenic edges, natural edges, and in the forest interior. The populations of P. tenuinervis showed no differences in genetic variability or structure among the three habitats. There was, however, an indication of inbreeding, which was significantly higher in natural edges than in anthropogenic edges and the forest interior. Within-habitat variation was considerable, but there were no differences in seed mass or rate and velocity of germination among the three habitats. These results suggest that seed characteristics were not influenced by the genetic pattern of P. tenuinervis and that other characteristics of the forest fragment, such as gaps, edge age, and type of matrix exert more influence on seed mass and germination than the distance from the edges.