Genetic-diversity assessed by microsatellites in tropical maize populations submitted to a high-intensity reciprocal recurrent selection

BR-105 and BR-106 are important tropical maize populations, which were submitted to a high-intensity reciprocal recurrent selection, generating the IG-3 and IG-4synthetics. Using 30-microsatellite loci,we measured and compared the genetic diversity of these populations and their synthetics. The populations did not differ significantly regarding the amount of genetic diversity. As a consequence of selection, genetic variability losses, in terms of mean number of alleles per locus,proportion of polymorphic loci, and gene diversity did occur and were greater in the synthetic IG-3 than IG-4. In the synthetics, the number of loci in adherence to Hardy-Weinberg proportions was superior to that observed in the populations. The Wright's mean fixation index was higher than the mean value expected for outcrossing species (5%) indicating as light excess of homozygotic individuals in both populations. The genetic distances confirmed the favourable effects of one cycle of recurrent selection, as the synthetics became more isolated in comparison to the original populations. This revised version was published online in July 2006 with corrections to the Cover Date.     

Herbage utilisation efficiency of continuously stocked pastures during periods of restricted pasture growth

ABSTRACT

The aim of this study was to evaluate the defoliation patterns of individual tillers, efficiency of herbage utilisation, and forage production in continuously stocked Brachiaria humidicola cv. Comum swards during periods of the year of restricted pasture growth. The experiment was conducted at the Embrapa, Campo Grande-MS, Brazil, from April to October 2012. Treatments consisted of two grazing management strategies, defined by two ranges of sward height (10–15 and 20–25 cm) managed under continuous and variable stocking rates. The experiment was conducted as a completely randomised block design with three replications. During periods of resource constraints, grazing management strategies based on ‘steady state’ sward heights did not alter defoliation patterns, herbage utilisation efficiency (HUE), and forage production in Brachiaria humidicola swards under a continuous stocking method. A direct implication of our results is that grazing management strategies during the driest and coldest periods of the year in the tropics should be developed based on the conditions most suitable for better promoting faster pasture recovery and a return to production in the following spring; it is unlikely that any grazing management strategy would be successful in increasing herbage production and/or HUE in periods of restricted pasture growth.     

Nitrogen-fixing legume tree species for the reclamation of severely degraded lands in Brazil

The main challenges faced in the reclamation of severely degraded lands are in the management of the systems and finding plant species that will grow under the harsh conditions common in degraded soils. This is especially important in extremely adverse situations found in some substrates from mining activities or soils that have lost their upper horizons. Under these conditions, recolonization of the area by native vegetation through natural succession processes may be extremely limited. Once the main physical and chemical factors restrictive to plant growth are corrected or attenuated, the introduction of leguminous trees able to form symbioses with nodulating N?-fixing bacteria and arbuscular mycorrhizal fungi constitutes an efficient strategy to accelerate soil reclamation and initiate natural succession. These symbioses give the legume species a superior capacity to grow quickly in poor substrates and to withstand the harsh conditions presented in degraded soils. In this article we describe several successful results in Brazil using N?-fixing legume tree species for reclamation of areas degraded by soil erosion, construction and mining activities, emphasizing the potential of the technique to recover soil organic matter levels and restore ecosystem biodiversity and other environmental functions.     

Direct measurement of the current-phase relation of a superfluid 3He-B weak link

Direct measurements of the current-phase relation, I versus Deltaphi, for a weak link coupling two reservoirs of B-phase superfluid helium-3 (3He-B) were made over a wide range of temperatures. The weak link consists of a square array of 100-nanometer-diameter apertures. For temperatures T such that T/Tc >/= 0.6 (where Tc is the superfluid transition temperature), I approximately sin(Deltaphi). At lower temperatures, I(Deltaphi) approaches a straight line. Several remarkable phenomena heretofore inaccessible to superconducting Josephson junctions, including direct observation of quantum oscillations and continuous knowledge of Deltaphi, were also observed.     

Controlling microbial contamination on beef and lamb meat during processing

SUMMARY The microbiological quality of carcases, meat and environmental surfaces was evaluated in commercial boning rooms processing beef and lamb. There was considerable variation in the level of microbial contamination on both carcases and meat, with counts ranging from less than 20 to 10⁸/cm² on carcases and to 2 times 10⁷/cm² on meat. The level of microbial contamination on meat was influenced by the level of carcase contamination at boning and by the boning process itself. Carcase contamination was the major determinant of microbiological quality, as more than 70% of carcases had microbial counts greater than 10³/cm². Cutting boards were a major source for microbial dissemination during boning, particularly when carcase counts were less than 10³/cm². If carcases were heavily contaminated, the contamination of processing surfaces was irrelevant in determining microbial loads on meat. Where carcase contamination was at low to moderate levels, the contribution of the boning process to the contamination on meat assumed increased significance. Under these conditions, improved sanitation of cutting surfaces in the boning room resulted in a significant reduction in microbial contamination on the surface of meat. These results can form the basis for ensuring that improvements made in carcase management before boning, to improve microbiological quality, will be preserved through attention to cutting board hygiene during boning.     

Effectiveness of passive gas-scavenging canisters attached to isoflurane anesthesia systems under standard-use conditions in a laboratory animal facility

Chronic exposure to trace levels of waste anesthetic gases has been linked to higher incidences of neurologic and reproductive dysfunction, hepatic and renal toxicity, and neoplasia in health care professionals. We have shown that low levels of isoflurane emissions are likely in conventional laboratory animal treatment rooms during the use of standard anesthesia delivery systems equipped with activated charcoal canisters for passive gas scavenging. In the present study, we surveyed the effectiveness of canisters (attached to well‐maintained precision isoflurane vaporizers) in current use throughout our AAALAC‐accredited laboratory animal facility. Canisters (Omnicon f/air) had been weighed prior to use and then attached to dual‐loop systems (face mask and induction box circuits) from 1 week to 6 months of service. Isoflurane emissions were measured using a pre‐calibrated, portable infrared spectrophotometer by attaching each canister to the face‐mask circuit, occluding the face mask and closing the stopcock to the induction circuit, and running the system at uniform isoflurane concentration (2%) and oxygen flow rate (1 L minute^?1). Samples were taken in animal procedure rooms (size range, 45–80 m³) in which the air turnover rate ranged between 20 and 30 nonrecirculating changes per hour. Nine of the 60 canisters (15%) in current use were found to have exceeded the manufacturer's recommended use‐life (defined as a weight increase of 50 g). Of these nine, seven canisters did not scavenge isoflurane at all (indicated by emissions greatly exceeding 100 ppm). Isoflurane was not detected in the operator's breathing zone under normal use conditions (i.e. gas directed to both circuits at once). Of the 51 canisters that had not exceeded their rated use‐life, 12 (23.5%) exhausted isoflurane at >2 ppm. Our data show that (i) the potential for exposure to waste isoflurane emissions will depend on the configuration of the delivery system and that (ii) enhanced attention to canister surveillance may be warranted even in well‐run facilities.     

The isolation of salmonella from kangaroos and feral pigs processed for human consumption

A total of 154 feral pig carcases and 81 kangaroo carcases were examined for the presence of Salmonella, coliforms and total aerobic counts. Approximately 34% of pig carcases yielded one or more serotypes of Salmonella, while about 11% of kangaroo carcases were contaminated with salmonella. The results differed widely between sampling occasions. A total of 13 serotypes were isolated from feral pigs with S. anatum (31 isolates) and S. typhimurium (9 isolates) being the predominant serotypes. Coliforms were isolated from approximately 90% of carcases. The mean log10 coliform count on feral pigs was 4.39 +/- 1.45/g and the mean log10 total count was 6.15 +/- 1.15/g. About 21% of carcases were contaminated with more than 100,000 coliforms/g. A total 3 serotypes were isolated from kangaroos (S. bahrenfeld, S. binza, and S. onderstepoort). The mean log10 coliform count on kangaroos was 3.54 +/- 1.04. More than 50% of kangaroo carcases were contaminated with less than 100 coliforms/g. About 15% of carcases were contaminated with more than 10,000 coliforms/g. The mean log10 total count was 5.2 +/- 1.01/g.     

Comparison of inoculation regimes for the experimental production of swine erysipelas arthritis: II Serological findings in a gel diffusion precipitin test and enzyme-linked immunosorbent assay

The serological response of pigs to Erysipelothrix rhusiopathiae inoculation was monitored by a gel diffusion precipitin test (GDPT) using a crude, serotype-specific, autoclaved antigen and an enzyme-linked immunosorbent assay (ELISA) using a heat-extracted, alcohol precipitated and molecular seived antigen previously shown to react with serum from pigs infected with serotypes 1 or 2. All pigs receiving 3 or 5 weekly intravenous inoculations of either a highly virulent (VRS 229) or a lowly virulent isolate (VRS 252) produced GDPT-reactive antibody within 3 weeks, but only 44% were still reactive at 8 to 9.5 weeks. The ELISA response was significantly higher in pigs inoculated with the highly virulent strain, and was similar in pigs receiving 3 or 5 doses of either strain. In a dose-response trial, after 3 doses of VRS 229, GDPT reactivity occurred earlier and was stronger in pigs given higher doses of E. rhusiopathiae, but the response peaked 3 to 5 weeks after the start of challenge and was short lived. GDPT reactivity correlated with dose, but not with the severity of arthritis. The ELISA demonstrated specific IgG antibody was present by 2 weeks, and persisted to at least 11 weeks. The ELISA reactivity was significantly higher in pigs with arthritis than in pigs that received low doses and were not arthritic. Within groups of pigs with arthritis a significant, dose dependent, linear ELISA response developed but did not correlate with the presence or degree of arthritis at slaughter. Non-arthritic pigs had similar low ELISA responses to uninoculated controls.     

Biofilm-forming ability profiling of Staphylococcus aureus and Staphylococcus epidermidis mastitis isolates

Biofilm-forming ability has been increasingly recognized as an important virulence factor in Staphylococci, facilitating their persistence in the host, evading its defences and allowing bacterial survival at high antimicrobial concentrations. Staphylococcus aureus remains a major pathogen of chronic mastitis, but in the last years Staphylococcus epidermidis has emerged as a relevant mastitis pathogen. The present work aimed at the evaluation of the biofilm-forming ability of Staphylococci field isolates from bovine subclinical mastitis and at the development of a fluorescent in situ hybridisation (FISH) protocol that would allow the direct observation of biofilm formation in milk samples. The analysis of phenotypic expression in Congo Red Agar (CRA) and by FISH, showed that 37.5% of the S. aureus isolates produced biofilm, while by optical density measurement only 18.75% isolates revealed this phenotype. The results showed a fair agreement according to the kappa coefficient test (kappa = 0.259). Regarding S. epidermidis mastitis isolates, 37.5% revealed the ability to produce biofilm, but only four isolates were positive by all methods. This agreement was moderate (kappa = 0.467). The application of FISH to artificially contaminated milk samples allowed the direct observation of biofilm production by 37.5% isolates, showing total agreement with the CRA results. This method better mimics the in vivo conditions, especially in terms of the presence of calcium and iron, which in high concentrations, respectively, are known to inhibit or induce biofilm production.     

Biometrical, biological, biochemical and molecular characteristics of Meloidogyne incognita isolates and related species

Meloidogyne incognita is one of the most polyphagous species of root-knot nematodes occurring in Brazil and worldwide. Eight M. incognita isolates were studied, representing two enzymatic phenotypes (esterase and malate desydrogenase: I1/N1, I2/N1) and four cryptic Meloidogyne sp.1 (S2/N1) isolates, representing one cytological type (3n?=?40–46). Three M. hispanica isolates (Hi3/N1, 2n?=?32–36) and two of an atypical Meloidogyne sp.2 (S2a/N3, 3n?=?40–44) were included in this study for comparison. All isolates were tested with three M. incognita-specific molecular markers. The primer pairs B06F/R, miF/R and incK14F/R amplified three species-specific fragments of 1,200?bp, 955?bp and 399?bp, respectively for M. incognita and Meloidogyne sp.1 isolates. No amplification occurred in the M. hispanica and Meloidogyne sp.2 isolates, except with primers miF/R (1,650?bp). The genetic variability of the Meloidogyne spp. isolates was evaluated, using RAPD and ISSR markers. The phylogenetic analyses revealed two strongly supported monophyletic clades: clade I, consisting of M. hispanica and the atypical Meloidogyne sp.2 isolates, and clade II, clustering together all M. incognita and the Meloidogyne sp.1 isolates. Considering the biometrical, cytological and molecular approaches, it was possible to conclude that the isolates with three enzymatic phenotypes (I1/N1, I2/N1 and S2/N1) presented the characteristics described for M. incognita. Some correlations were detected between the isozymatic phenotypes and the tree topology (S2a/N3, Hi3/N1, I1/N1, S2/N1), but no strict correlation could be observed for the phenotype I2/N1 and one isolate of S2/N1. Morphologically, the Msp.2 isolates differ from M. incognita and M. hispanica by the female stylet features presenting straight cone tip and round pear shaped knobs, posteriorly sloping. The results of this study suggested that the Msp.2 isolates with phenotypes S2aN3 belong to a new or an unidentified species closely related to M. hispanica.