A bacteriophage reagent for Salmonella: molecular studies on Felix 01

Felix 01 (F01) is a bacteriophage originally isolated by Felix and Callow which lyses almost all Salmonella strains and has been widely used as a diagnostic test for this genus. Molecular information about this phage is entirely lacking. In the present study, the DNA of the phage was found to be a double-stranded linear molecule of about 80 kb. 11.5 kb has been sequenced and in this region A + T content is 60%. There are relatively few restriction endonuclease cleavage sites in the native genome and clones show this is due to their absence rather than modification. A restriction map of the genome has been constructed. The ends of the molecule cannot be ligated although they contain 5' phosphates. At least 60% of the genome must encode proteins. In the sequenced portion, many open reading frames exist and these are tightly packed together. These have been examined for homology to published proteins but only 1 to 17 shows similarity to known proteins. F01 is therefore the prototype of a new phage family. On the basis of restriction sites, codon usage and the distribution of nonsense codons in the unused reading frames, a strong case can be made for natural selection that reacts to mRNA structure and function.     

Estrogen-induced alterations in amh and dmrt1 expression signal for disruption in male sexual development in the zebrafish

Dmrt1 and amh are genes involved in vertebrate sex differentiation. In this study, we cloned dmrt1 and amh cDNAs in zebrafish (Danio rerio) and investigated the effects of exposure to 17a-ethinylestradiol (EE2), during early life on their patterns of expression and impact on the subsequent gonadal phenotype. Expression of both amh and dmrt1 in embryos was detected as early as at 1 day post fertilization (dpf) and enhanced expression of amh from 25 dpf was associated with the period of early gonadal differentiation. Sex-dependent differences in enhanced green fluorescent protein transgene expression driven by the promoter of the germ cell-specific vas gene were exploited to show that at 28dpf and 56dpf both amh and dmrt1 mRNA were overexpressed in males compared with females. Exposure during early life to environmentally relevant concentrations of EE2 had a suppressive effect on the expression of both amh and dmrt1 mRNAs and this was associated with a cessation/retardation in male gonadal sex development. Our findings indicate that estrogen-induced suppression in expression of dmrt1 and amh during early life correlate with subsequent disruptive effects on the sexual phenotype in males.     

A hierarchical modeling approach for estimating national distributions of chemicals in public drinking water systems

Water quality studies often include the analytical challenge of incorporating censored data and quantifying error of estimation. Many analytical methods exist for estimating distribution parameters when censored data are present. This paper presents a Bayesian-based hierarchical model for estimating the national distribution of the mean concentrations of chemicals occurring in U.S. public drinking water systems using fluoride and thallium as examples. The data used are Safe Drinking Water Act compliance monitoring data (with a significant proportion of left-censored data). The model, which assumes log-normality, was evaluated using simulated data sets generated from a series of Weibull distributions to illustrate the robustness of the model. The hierarchical model is easily implemented using the Markov chain Monte Carlo simulation method. In addition, the Bayesian method is able to quantify the uncertainty in the estimated cumulative density function. The estimated fluoride and thallium national distributions are presented. Results from this study can be used to develop prior distributions for future U.S. drinking water regulatory studies of contaminant occurrence.     

Furosine is a useful indicator in pre‐baked breads

There is increasing demand for pre‐baked bread. Bakers can meet consumer requirements for breads of specific flavour and colour by pre‐baking an amount of bread in the morning and completing the baking process in the afternoon. This type of product is of special interest to sandwich bars, restaurants and large communities. In order to obtain an indicator of utility to monitor the processing of pre‐baked bread, the browning indicators furosine, hydroxymethylfurfural (HMF) and glucosylisomaltol were analysed in two independent assays of bread pre‐baked at 175 °C for between 5 and 30 min. No furosine was detected in raw or fermented dough. In the pre‐baking stage, furosine increased from 1.5‐ to 5‐fold between 5 and 30 min. The furosine values obtained in the two independent experiments were similar. HMF and glucosylisomaltol were only detected after 15 min, which is the time period commonly used for processing by the industry. Thus, among these indicators, furosine is the only one with utility for monitoring the pre‐baking process. On the other hand, a study of the baking of pre‐baked bread at 220 °C showed that HMF and glucosylisomaltol can be used to monitor the latter process. Copyright © 2004 Society of Chemical Industry     

Impact of intervention strategies on Listeria contamination patterns in crawfish processing plants: a longitudinal study

Two ready-to-eat crawfish processing plants were monitored for 2 years to study the impact of Listeria control strategies, including employee training and targeted sanitation procedures, on Listeria contamination. Environmental, raw material, and finished product samples were collected weekly during the main processing months (April to June) and tested for Listeria spp. and Listeria monocytogenes. Before implementation of control strategies (year 1), the two processing plants showed Listeria spp. prevalences of 29.5% (n = 78) in raw, whole crawfish, 5.2% (n = 155) in the processing plant environment, and 0% (n = 78) in finished products. In year 2, after plant-specific Listeria control strategies were implemented, Listeria spp. prevalence increased in raw crawfish (57.5%, n = 101), in the processing plant environment (10.8%, n = 204), and in the finished product (1.0%, n = 102). Statistical analysis showed a significant increase in Listeria spp. prevalence (P < 0.0001) and a borderline nonsignificant increase in L. monocytogenes prevalence (P = 0.097) on raw material in year 2. Borderline nonsignificant increases were also observed for Listeria spp. prevalence in environmental samples (P = 0.082). Our data showed that Listeria spp. prevalence in raw crawfish can vary significantly among seasons. However, the increased contamination prevalence for raw materials only resulted in a limited Listeria prevalence increase for the processing plant environment with extremely low levels of finished product contamination. Heat treatment of raw materials combined with Listeria control strategies to prevent cross-contamination thus appears to be effective in achieving low levels of finished product contamination, even with Listeria spp. prevalences for raw crawfish of more than 50%.     

PCB loading from sediment in the Hudson River: congener signature analysis of pathways

The upper Hudson River (NY) was subjected to massive PCB contamination over a period of three decades. A large inventory of PCBs remains in contaminated sediments of the river, most notably in the Thompson Island Pool. During the summer, flow crossing the Thompson Island Pool exhibits a large and consistent PCB load gain. This load gain is not associated with scouring flows and is not accompanied by an increase in suspended solids. A variety of hypotheses have been proposed to explain this load gain, including flux of contaminated porewater and dissolution of unverified reservoirs of pure PCBs. A wealth of congener-specific PCB data is available for the site throughout the 1990s. Interpretation of the Thompson Island Pool load gain is facilitated by examination of the PCB congener signature of the gain and comparison to the signature of potential sources. This examination suggests that neither the flux of porewater nor the dissolution of unaltered Aroclors are the predominant source of the load gain. Instead, the congener signature is consistent with a mixed source consisting of porewater flux and non-scour flux of contaminated sediments. The non-scour sediment flux, which reaches a maximum in the beginning of the summer growing season, is likely driven by a variety of biological and anthropogenic processes, including bioturbation by benthic organisms, bioturbation by demersal fish, scour by propwash, mechanical scour by boats and floating debris in nearshore areas, and uprooting of macrophytes.     

Sodium caseinate–maltodextrin conjugate stabilized double emulsions: Encapsulation and stability

The potential food applications of water-in-oil-in-water (W₁/O/W₂) double emulsions are great, including the encapsulation of flavours or active ingredients. However, the stability of these emulsions restricts their applications in food systems. Sodium caseinate (NaCN)–maltodextrin (Md40 or Md100) conjugates were investigated for their potential to improve the stability of W₁/O/W₂ double emulsions compared to NaCN. NaCN–Md40 and NaCN–Md100 conjugates were prepared by a Maillard-type reaction by dry heat treatment of mixtures of NaCN–Md40 or NaCN–Md100 at 60 °C and 79% relative humidity for 4 days. Water-in-oil-in-water (W₁/O/W₂) double emulsions with NaCN, NaCN–Md40 or NaCN–Md100 as outer aqueous phase containing emulsifier were prepared using a two-step emulsification process. General emulsion stability was characterised by determining the droplet size distribution, viscosity characteristics and by confocal microscopy of the W₁/O/W₂ double emulsions on formation and after their storage under accelerated shelf life testing conditions at 45 °C for up to 7 days. Inner phase encapsulation and stability were characterised by monitoring the level of entrapped Vitamin B₁₂ in the inner aqueous phase on formation of the double emulsions and after storage at 45 °C for up to 7 days. Conjugate stabilized emulsions were more generally stable than NaCN stabilized emulsions. In comparison to NaCN stabilized emulsions, conjugate stabilized emulsions showed improved Vitamin B₁₂ encapsulation efficiency in the inner aqueous phase on emulsion formation and improved encapsulation stability following storage of the emulsions.     

Non-thermal pasteurization of fruit juices by combining high-intensity pulsed electric fields with natural antimicrobials

The effect of high-intensity pulsed electric fields (HIPEF) on the Salmonella Enteritidis and Escherichia coli O157:H7 populations inoculated in apple, pear, orange and strawberry juices as influenced by treatment time and pulse frequency was investigated. Combinations of HIPEF (35 kV/cm, 4 μs pulse length in bipolar mode without exceeding 40 °C) with citric acid or cinnamon bark oil against these pathogenic microorganisms in fruit juices were also evaluated. Treatment time was the more influential factor on the microbial reduction in all the fruit juices analyzed. S. Enteritidis and E. coli O157:H7 were reduced by more than 5.0 log₁₀ units in orange juice treated by only HIPEF; whereas strawberry, apple and pear juices were pasteurized when HIPEF was combined with citric acid at 0.5, 1.5, 1.5%, respectively, or cinnamon bark oil at 0.05, 0.1 and 0.1%, respectively. Synergistic and additive killing effects against S. Enteritidis and E. coli O157:H7 in fruit juices by combining treatments were observed.

Industrial relevance

The use of high-intensity pulsed electric fields treatment as a non-thermal pasteurization method in combination with organic acids or essential oils is an effective process for eliminating S. Enteritidis and E. coli O157:H7 populations in fruit juices upper 5.0 log₁₀ reductions. Therefore, combinations of those treatments may help to ensure the microbiological safety in juice products, and to reduce the risk of food-borne illness caused by the consumption of these kinds of foods.