The detection of viable vegetative cells of Bacillus sporothermodurans using propidium monoazide with semi-nested PCR

AbstractBacillus sporothermodurans produces highly heat-resistant spores that can survive ultra-high temperature (UHT) treatment in milk. Therefore, we developed a rapid, specific and sensitive semi-nested touchdown PCR assay combined with propidium monoazide (PMA) treatment for the detection of viable B. sporothermodurans vegetative cells. The semi-nested touchdown PCR alone proved to be specific for B. sporothermodurans, and the achieved detection limit was 4 CFU/mL from bacterial culture and artificially contaminated UHT milk. This method combined with PMA treatment was shown to amplify DNA specifically from viable cells and presented a detection limit of 102 CFU/mL in UHT milk. The developed PMA-PCR assay shows applicability for the specific detection of viable cells of B. sporothermodurans from UHT milk. This method is of special significance for applications in the food industry by reducing the time required for the analysis of milk and dairy products for the presence of this microorganism

Ammonia levels on in vitro degradation of fibrous carbohydrates from buffel grass

This study was carried out to examine the degradation dynamics of neutral detergent fibre (NDF) and the profile of volatile fatty acids that originate from the fermentation of buffel grass (Cenchrus ciliaris, L.) with various levels of ammonia in the growth medium. The treatments consisted of six levels of ammonia in the growth medium (0, 5, 10, 15, 20 and 30 mg/dL), which were obtained by adding urea. These in vitro incubation times were evaluated in three replicates per time for 0, 3, 6, 9, 12, 24, 36, 48, 72 and 96 hours. Rumen concentrations of acetate and propionate responded quadratically to the ammonia levels. The treatment with 15 mg/dL of ammonia nitrogen in the rumen fluid provided mean acetate and propionate contents of 57.6 mM and 23.1 mM, respectively. Urea addition elevated the degradation rate of potentially degradable fraction of NDF (pdNDF) from 2.5% to 20.1% in comparison with the treatment without urea and to a reduction in estimated discrete lag time from 0.34 to 2.31 hours. Urea supplementation increased the specific microorganism growth rate from 2.6% to 20.1%. At the end of the incubation trial, NDF degradation showed a quadratic response, with maximum value obtained at 17.76 mg/dL of ammonia in the rumen fluid. Urea improves the degradation dynamics of NDF from deferred buffel grass and increases the concentrations of acetate and propionate.Keywords: Cenchrus ciliaris, degradation kinetics, non-protein nitrogen, ure

Local search heuristics for the multidimensional assignment problem

The Multidimensional Assignment Problem (MAP) (abbreviated s-AP in the case of s dimensions) is an extension of the well-known assignment problem. The most studied case of MAP is 3-AP, though the problems with larger values of s also have a large number of applications. We consider several known neighborhoods, generalize them and propose some new ones. The heuristics are evaluated both theoretically and experimentally and dominating algorithms are selected. We also demonstrate that a combination of two neighborhoods may yield a heuristics which is superior to both of its components

Particle swarm optimization for the Steiner tree in graph and delay-constrained multicast routing problems

This paper presents the first investigation on applying a particle swarm optimization (PSO) algorithm to both the Steiner tree problem and the delay constrained multicast routing problem. Steiner tree problems, being the underlining models of many applications, have received significant research attention within the meta-heuristics community. The literature on the application of meta-heuristics to multicast routing problems is less extensive but includes several promising approaches. Many interesting research issues still remain to be investigated, for example, the inclusion of different constraints, such as delay bounds, when finding multicast trees with minimum cost. In this paper, we develop a novel PSO algorithm based on the jumping PSO (JPSO) algorithm recently developed by Moreno-Perez et al. (Proc. of the 7th Metaheuristics International Conference, 2007), and also propose two novel local search heuristics within our JPSO framework. A path replacement operator has been used in particle moves to improve the positions of the particle with regard to the structure of the tree. We test the performance of our JPSO algorithm, and the effect of the integrated local search heuristics by an extensive set of experiments on multicast routing benchmark problems and Steiner tree problems from the OR library. The experimental results show the superior performance of the proposed JPSO algorithm over a number of other state-of-the-art approaches