Development of sandwich enzyme-linked immunosorbent assay for the detection of Cronobacter muytjensii (formerly called Enterobacter sakazakii)

This study aimed to produce a polyclonal antibody against Cronobacter muytjensii (C. muytjensii, formerly called Enterobacter sakazakii) and to develop an immunoassay for its detection. The optimum production of rabbit anti-C. muytjensii immunoglobulin G (IgG) and chicken anti-C. muytjensii IgY was reached in weeks 8 and 9, respectively. Purification of rabbit anti-C. muytjensii IgG from immunized rabbit sera was accomplished using the caprylic acid and ammonium sulfate precipitation method. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 25 and 50 kDa, corresponding to a light and a heavy chain, respectively. The optimized conditions for sandwich enzyme-linked immunosorbent assay were using rabbit anti-C. muytjensii IgG (1 μg/mL) as a detection antibody and chicken anti-C. muytjensii IgY (10 μg/mL) as a capture antibody. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria tested, which included Escherichia coli O157:H7, Salmonella typhimurium, Staphylococcus aureus, Bacillus cereus and Listeria monocytogenes. The developed assay did not show cross-reactivity with other tested species of Cronobacter and Enterobacter genera such as C. turicensis, C. sakazakii, E. aerogenes, E. pulveris and E. helveticus. The detection limit of sandwich ELISA for C. muytjensii was found to be 2.0 × 10(4) colony forming units (CFU)/mL. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of sandwich ELISA for C. muytjensii, the detection limit being found to be 6.3 × 10(4) CFU/mL. These findings demonstrate that the developed method is able to detect all strains of C. muytjensii. Hence, this ELISA technique has potent application for the rapid and accurate detection of C. muytjensii in dietary foods.     

Occurrence of Cronobacter spp. in retail foods

Aims: To study the occurrence of Cronobacter spp. in foods and to investigate the phenotypic properties of the strains isolated. Methods and Results: A total of 53 strains of Cronobacter spp. isolated from 399 food samples were identified using conventional biochemical methods and MALDI‐TOF mass spectrometry. Foods of plant origin were the most frequently contaminated samples. No Cronobacter spp. were found in infant milk formula, wheat‐based infant food, pasteurized and raw cow milk, mincemeat, chicken, chickpea and potato dumpling powder. The individual species were identified as Cronobacter sakazakii (54·7%), Cronobacter malonaticus (28·4%), Cronobacter dublinensis (7·5%), Cronobacter muytjensii (7·5%) and Cronobacter turicensis (1·9%). Cronobacter sakazakii and C. malonaticus belong to biotype 1, 2, 2a, 3, 4 and 5, 5a, respectively. Cronobacter dublinensis strains were subdivided into biotypes 6 and 12. All strains were resistant to erythromycin and two of them were resistant to both erythromycin and tetracycline. Conclusions: Cronobacter spp. were isolated from various food samples pre‐eminently of plant origin and dried food ingredients. Significance and Impact of the Study: These findings will increase and detail our knowledge of the presence and diversity of Cronobacter spp. in foods.     

On the optimal path in the dynamic pool model for a fishery

Ecosystem-based fishery management (EBFM) is a new direction for fishery management, essentially reversing the order of management priorities to start with the ecosystem rather than the target species. This concept of management is a direct extension of the concept of a holistic approach incorporating interspecific interactions and physical environmental influences. However, because of the limited understanding of the complexity of marine ecosystems, few fisheries are actually managed on a multispecies basis. Even now, in order to specify a practical fishing policy we need a single-species model and utilize it by partially taking account of the effects of other factors mentioned above on the target species biomass. In fact, it is contended that in systems with moderate amounts of data, EBFM could be characterized by effective single-species management with the addition of precautionary set-asides for unknown ecosystem components. Hence, it is still necessary to examine a single-species model so as to clarify the extent of its applicability. The model investigated in this paper is what is called the dynamic pool model, which was proposed by C.W. Clark in the mid-1970s as a dynamic optimization of the classic Beverton and Holt static model for a fishery, in an attempt to make the process of growth and aging inherent in each of the creature resources reflect directly into the economic process. This dynamic model has been applied to a wide variety of commercial fish species. However, the applications have been largely confined to computer simulations using the discrete-time stand-by of the original Clark continuous-time model. This situation is caused mainly by the complexity of the mathematical structure of the Clark model. In this paper, we first specify the material related to the complexity. Subsequently, we provide a rigorous proof for the long-standing conjecture due to Clark concerning the optimal path or harvesting schedule. In addition, two derivative cases are examined: one is the case in which a year-class of fish leaves a given fishing sea area permanently before its natural biomass peaks, the other is the case in which the escapement of a year-class is required to be more than a given minimum level.     

The sampling theory of neutral alleles and an urn model in population genetics

The behaviour of a Pólya-like urn which generates Ewens' sampling formula in population genetics is investigated. Connections are made with work of Watterson and Kingman and to the Poisson-Dirichlet distribution. The order in which novel types occur in the urn is shown to parallel the age distribution of the infinitely many alleles diffusion model and consequences of this property are explored. Finally the urn process is related to Kingman's coalescent with mutation to provide a rigorous basis for this parallel.This research was partially supported by the Sloan Foundation under Grant 85-6-14 and by the National Science Foundation     

Supremum weighted log-rank test and sample size for comparing two-stage adaptive treatment strategies

In two-stage adaptive treatment strategies, patients receivean induction treatment followed by a maintenance therapy, giventhat the patient responded to the induction treatment they received.To test for a difference in the effects of different inductionand maintenance treatment combinations, a modified supremumweighted log-rank test is proposed. The test is applied to adataset from a two-stage randomized trial and the results arecompared to those obtained using a standard weighted log-ranktest. A sample-size formula is proposed based on the limitingdistribution of the supremum weighted log-rank statistic. Thesample-size formula reduces to Eng and Kosorok's sample-sizeformula for a two-sample supremum log-rank test when there isno second randomization. Monte Carlo studies show that the proposedtest provides sample sizes that are close to those obtainedby standard weighted log-rank test under a proportional hazardsalternative. However, the proposed test is more powerful thanthe standard weighted log-rank test under non-proportional hazardsalternatives.     

Incidence of Endemic Entomopathogenic Nematodes Following Application of Steinernema riobrave for Control of Diaprepes abbreviatus

Control of Diaprepes abbreviatus by endemic and exotic entomopathogenic nematodes (EPN) was monitored during 2000-2001 in two citrus orchards in central Florida (Bartow and Poinciana). Caged sentinel insect larvae were buried beneath citrus trees for 7 days at 1 to 2-month intervals from April to October each year. At Bartow, the survey occurred in experimental plots that were (i) not treated with commercial EPN, (ii) treated twice annually since 1998 with commercially formulated Steinernema riobrave, or (iii) treated twice annually with S. riobrave and liquid fertilization (15 times/year) occurred in place of dry fertilizer (3 times/year) used in the other treatments. Four endemic EPN species, in addition to S. riobrave, were recovered from the sandy soil at Bartow: S. diaprepesi, Heterorhabditis zealandica, H. indica, and H. bacteriophora. Mean insect mortality in control plots was 39.4% (range = 13% to 74%), with seasonal maxima in May to July each year. Endemic EPN were recovered from 55% (range = 22% to 81%) of the cadavers each month. Total numbers of endemic EPN recovered in all plots during 2 years were directly related to the numbers of adult weevils (D. abbreviatus and Pachnaeus litus) captured in modified Tedder''s traps and inversely related to recovery of S. riobrave. Insect mortality was higher and cadavers containing endemic EPN were more numerous in untreated control plots than in S. riobrave-treated plots, except during months in which S. riobrave was applied. In treated plots, endemic EPN were recovered from cadavers at twice the rate of S. riobrave. Suppression of endemic EPN in plots treated with S. riobrave, combined with inferior persistence by the introduced species, may have attenuated the net efficacy of S. riobrave against D. abbreviatus. In contrast, H. indica was the only endemic nematode recovered from the sandy clay loam soil at Poinciana, where the average mortality of D. abbreviatus was 12% (range 3% to 20%) and incidence of H. indica did not exceed 8%. Results of these surveys suggest that the regional patterns in the abundance and damage to citrus caused by D. abbreviatus in Florida are regulated by endemic EPN and other soilborne enemies of the weevil.     

施肥对降香黄檀营养生长和生殖生长的影响

研究施肥对降香黄檀（Dalbergia odorifera T.Chen.）营养生长和生殖生长的影响,为不同经营目标的降香黄檀人工林培育提供技术支撑。本研究以8年生降香黄檀为对象,设置N（185.6 g N/株）、P（120 g P₂O₅/株）、K（120 g K₂O/株）、PK（120 g P₂O₅+120 g K₂O/株）、NPK（185.6 g N+120 g P₂O₅+120 g K₂O/株）以及不施肥（对照）等六个处理,调查施肥后盛花期内一年生新梢、叶、花的生长情况。结果表明：6个施肥处理间降香黄檀营养生长和生殖生长均差异显著（P<0.05）。N处理的营养枝率、营养枝复叶数和小叶宽分别比对照提高40.25%、21.75%和9.52%,花序直径和花序生物量则降低12.75%和48.63%,显示N肥能促进降香黄檀营养生长,抑制生殖生长,有利于大径材培育;P、PK处理的营养枝率较对照显著降低47.96%和46.84%,表明P肥和K肥能促进生殖生长,有利于良种选育;NPK处理能同时显著促进营养生长和生殖生长,其营养枝率、营养枝长度、营养枝复叶数、营养枝小叶长、宽和枝生物量比对照提高26.04%、68.16%、32.98%、15.20%、11.40%和83.60%,花序数量和花序生物量亦提高54.20%和49.84%。因此,在降香黄檀人工林培育实践中,可通过调整肥料的NPK配比（偏向N或PK）以实现营养生长或生殖生长调控之目的。     

Codon evolution is governed by linear formulas

When nucleotide (G, C, T and A) contents were plotted against each nucleotide, their relationships were clearly expressed by a linear formula, y = αx + β in the coding and non-coding regions. This linear relationship was obtained from the complete single-stranded DNA. Similarly, nucleotide contents at all three codon positions were expressed by linear regression lines based on the content of each nucleotide. In addition, 64 codon usages were also expressed by linear formulas against nucleotide content. Thus, the nucleotide content not only in coding sequence but also in non-coding sequence can be expressed by a linear formula, y = αx + β, in 145 organisms (112 bacteria, 15 archaea and 18 eukaryotes). Based on these results, the ratio of C/T, G/T, C/A or G/A one can essentially estimate all four nucleotide contents in the complete single-stranded DNA, and the determination of any ratio of two kinds of nucleotides can essentially estimate four nucleotide contents, nucleotide contents at the three different codon positions and codon distributions at 64 codons in the coding region. The maximum and minimum values of G content were ∼0.35 and ∼0.15, respectively, among various organisms examined. Codon evolution occurs according to linear formulas between these two values. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.