Hand hygiene compliance in nursing home wards: the effect of increased accessibility of alcohol-based hand rub.

BACKGROUND: Elderly nursing home residents are vulnerable to infection from micro-organisms. Hand hygiene is considered one of the most important measures to prevent transmission. AIM: To determine the effect of increased accessibility to alcohol-based hand rub (ABHR) in nursing home wards by monitoring hand hygiene compliance (HHC) among healthcare workers (HCWs). METHODS: An 11-month intervention study was conducted in a Danish six-ward nursing home. Data were collected using an automatic hand hygiene monitoring system (AHHMS). After a baseline period, one extra ABHR dispenser was placed in each of the 150 apartments. Baseline HHC was compared with the HHC during an immediate intervention period and a long-term intervention period. FINDINGS: A total of 159 HCWs were included. The AHHMS registered 341,078 hand hygiene opportunities. Overall baseline HHC was 31% (95% confidence interval: 30-32). A significant +18% absolute immediate effect (first five months) (95% CI: 17-19; P < 0.0001) and +13 percentage points (95% CI: 11-14; P < 0.0001) long-term effect (another four months) were recorded. HCWs working day shifts and short-term employees had a higher baseline HHC than HCWs working evening/night shifts. However, HCWs working night shifts achieved the greatest long-term effect with a mean +27 percentage point difference (P < 0.0001). CONCLUSION: Placing an additional ABHR dispenser strategically within staff workflow significantly increased HHC among HCWs, demonstrating a noteworthy effect. The study is the first to report the effect on nursing home dispenser accessibility as a single intervention and to show a significant unmet potential.

Fungal Planet description sheets: 154-213.

Novel species of microfungi described in the present study include the following from South Africa: Camarosporium aloes, Phaeococcomyces aloes and Phoma aloes from Aloe, C. psoraleae, Diaporthe psoraleae and D. psoraleae-pinnatae from Psoralea, Colletotrichum euphorbiae from Euphorbia, Coniothyrium prosopidis and Peyronellaea prosopidis from Prosopis, Diaporthe cassines from Cassine, D. diospyricola from Diospyros, Diaporthe maytenicola from Maytenus, Harknessia proteae from Protea, Neofusicoccum ursorum and N. cryptoaustrale from Eucalyptus, Ochrocladosporium adansoniae from Adansonia, Pilidium pseudoconcavum from Greyia radlkoferi, Stagonospora pseudopaludosa from Phragmites and Toxicocladosporium ficiniae from Ficinia. Several species were also described from Thailand, namely: Chaetopsina pini and C. pinicola from Pinus spp., Myrmecridium thailandicum from reed litter, Passalora pseudotithoniae from Tithonia, Pallidocercospora ventilago from Ventilago, Pyricularia bothriochloae from Bothriochloa and Sphaerulina rhododendricola from Rhododendron. Novelties from Spain include Cladophialophora multiseptata, Knufia tsunedae and Pleuroascus rectipilus from soil and Cyphellophora catalaunica from river sediments. Species from the USA include Bipolaris drechsleri from Microstegium, Calonectria blephiliae from Blephilia, Kellermania macrospora (epitype) and K. pseudoyuccigena from Yucca. Three new species are described from Mexico, namely Neophaeosphaeria agaves and K. agaves from Agave and Phytophthora ipomoeae from Ipomoea. Other African species include Calonectria mossambicensis from Eucalyptus (Mozambique), Harzia cameroonensis from an unknown creeper (Cameroon), Mastigosporella anisophylleae from Anisophyllea (Zambia) and Teratosphaeria terminaliae from Terminalia (Zimbabwe). Species from Europe include Auxarthron longisporum from forest soil (Portugal), Discosia pseudoartocreas from Tilia (Austria), Paraconiothyrium polonense and P. lycopodinum from Lycopodium (Poland) and Stachybotrys oleronensis from Iris (France). Two species of Chrysosporium are described from Antarctica, namely C. magnasporum and C. oceanitesii. Finally, Licea xanthospora is described from Australia, Hypochnicium huinayensis from Chile and Custingophora blanchettei from Uruguay. Novel genera of Ascomycetes include Neomycosphaerella from Pseudopentameris macrantha (South Africa), and Paramycosphaerella from Brachystegia sp. (Zimbabwe). Novel hyphomycete genera include Pseudocatenomycopsis from Rothmannia (Zambia), Neopseudocercospora from Terminalia (Zambia) and Neodeightoniella from Phragmites (South Africa), while Dimorphiopsis from Brachystegia (Zambia) represents a novel coelomycetous genus. Furthermore, Alanphillipsia is introduced as a new genus in the Botryosphaeriaceae with four species, A. aloes, A. aloeigena and A. aloetica from Aloe spp. and A. euphorbiae from Euphorbia sp. (South Africa). A new combination is also proposed for Brachysporium torulosum (Deightoniella black tip of banana) as Corynespora torulosa. Morphological and culture characteristics along with ITS DNA barcodes are provided for all taxa.

Molecular characterization of three genes encoding aminopeptidases N in the poplar leaf beetle Chrysomela tremulae.

Three genes encoding proteins showing sequence similarity and features typical of insect APNs were characterized in C. tremulae and designed as CtAPN1, CtAPN2 and CtAPN3. Expression analysis of the three C. tremulae APN genes showed that CtAPN2 transcript is more abundant in the fat body, whereas both CtAPN1 and CtAPN3 are specifically expressed in the midgut. Despite a similar genomic organization, lepidopteran and coleopteran APNs are phylogenetically distant, suggesting that APN gene duplication events occurred after these two insect orders split. Sequence and expression comparisons of CtAPN1, CtAPN2 and CtAPN3 cDNAs in a C. tremulae Bacillus thuringiensis (Bt)-susceptible and in a Bt-resistant strain did not show any polymorphism at the amino acid level or difference at the transcription level.

First Report of Anthracnose Caused by Colletotrichum fragariae on Cyclamen in North Carolina.

In November 2009, cyclamen (Cyclamen persicum) plants with disease symptoms from a commercial greenhouse operation in the western part of North Carolina were sent to the Plant Diseases and Insect Clinic at North Carolina State University. Symptoms consisted of coalescing reddish and tan necrotic leaf spots with concentric circles. Other symptoms included darkened vascular tissue and decay of the corm, large roots, and petioles. Diseased leaves and stems were surface sterilized in 0.5% sodium hypochlorite for 3 min, air dried, and placed in petri dishes containing alkaline water agar. After 3 days of incubation at room temperature, fungal colonies were transferred to acidified potato dextrose agar. Isolation frequency after 5 days was 33% (three of nine pieces) and 16% (one of six pieces) from small leaf spots and petioles, respectively. Pure cultures of isolates were gray and black with abundant, aerial, gray whitish mycelia. Diseased plants were also incubated in a moist chamber at room temperature and sporulation was observed within 7 days. Conidia were tapered with rounded ends and produced in the acervulus and on the tips of setae, which is consistent with the morphology of described isolates of Colletotrichum fragariae. Similar setae were also observed directly on the fine roots of the original sample. The pathogenicity of single-spore cultures was tested by spraying four 2-month-old cyclamen plants with a conidial suspension (106 conidia/ml) and the plants were kept in a humid chamber for 24 h. Noninoculated controls (four plants) were sprayed with distilled water and subjected to the same conditions. The pathogenicity test was also repeated. Inoculated plants and controls were placed in a greenhouse with a temperature range from 22 to 25°C. After 7 to 10 days, symptomatic leaves and stems were observed on all the inoculated plants but not on the control plants. Fungi reisolated from 10 symptomatic leaf tissues had identical morphological features as the original isolates. Fungal DNA was extracted with DNeasy Plant Mini DNA Extraction Kits following the manufacturer's protocol (Qiagen Inc., Valencia, CA). Sequence analysis of the rRNA internal transcribed spacer (ITS) region of the cyclamen isolate (GenBank Accession No. HQ188923), based on the fragment amplified with ITS1 and ITS4 primers, showed 100% similarity to isolates of C. fragariae deposited in GenBank (Accession Nos. FJ172290 [ATCC MYA-4443 from cyclamen] and FJ810510 [ATCC MYA-4442 from silver date palm]) and Florida isolate C16 isolated from strawberry (1). In addition, the morphology and ITS sequences of the cyclamen isolate were identical to those of the C. fragariae voucher isolate from strawberry (GU174546). Results from disease symptoms, colony and spore morphology, pathogenicity tests, and ITS sequence analysis suggest that C. fragariae was the pathogen responsible for the disease symptoms on cyclamens. To our knowledge, this is the first report of a disease caused by C. fragariae on cyclamen in North Carolina and complements an earlier report from Florida (1). Reference: (1) S. J. MacKenzie et al. Plant Dis. 92:1432, 2008.

[Determination of hyperregeneratory esophagopathy in dogs with clinical signs attributable to esophageal disease]. / Bestimmung der hyperregeneratorischen Ösophagopathie bei Hunden mit klinischen Anzeichen einer Speiseröhrenerkrankung.

OBJECTIVE: It was hypothesized that typical characteristics of hyperregeneratory esophagopathy (HRE) in humans such as basal cell hyperplasia and elongation of stromal papillae are also histologically detectable in canine esophageal epithelium, and that these changes are associated with clinical signs and endoscopic findings suggesting gastroesophageal reflux (GER). MATERIAL AND METHODS: Sixty-five adult dogs with clinical signs attributable to esophageal disease underwent esophagoscopy and biopsy. Clinical signs suggesting GER (regurgitation, ptyalism, painful discomfort) were prospectively evaluated through a questionnaire. Endoscopic mucosal alterations suggesting GER such as minimal endoscopic changes and obvious mucosal defects were assessed via video endoscopy. Biopsy specimens obtained from the esophageal squamous epithelium were evaluated histologically. The squamous epithelium's substructures of esophageal biopsies were quantitatively assessed through microscopic morphometry. RESULTS: Esophageal squamous epithelium was considered normal in 48 dogs, and HRE was detected histologically in 17 dogs; both pathognomonic changes (basal cell hyperplasia, elongation of stromal papillae) were consistently present. Morphometrically assessed stromal papillary length and basal cell layer thickness was significantly (each, p < 0.0001) higher in the 17 dogs with HRE than in the 48 dogs without HRE, respectively. Overall, clinical signs suggesting GER were significantly (p = 0.02) more frequently encountered and regurgitation was significantly (p = 0.009) more common in the 17 dogs with HRE than in the 48 dogs without HRE. Similarly, endoscopic changes were significantly (p = 0.002) more frequently observed and minimal endoscopic changes suggesting GER were significantly (p = 0.004) more common in 17 dogs with HRE than in the 48 dogs without HRE. CONCLUSION AND CLINICAL RELEVANCE: Typical characteristics of hyperregeneratory esophagopathy in humans are also histologically detectable in canine esophageal epithelium. Histological changes are associated with clinical signs and endoscopic findings suggesting GER.

Economic and environmental optimization of waste treatment.

This article presents the new systems engineering optimization model, OptiWaste, which incorporates a life cycle assessment (LCA) methodology and captures important characteristics of waste management systems. As part of the optimization, the model identifies the most attractive waste management options. The model renders it possible to apply different optimization objectives such as minimizing costs or greenhouse gas emissions or to prioritize several objectives given different weights. A simple illustrative case is analysed, covering alternative treatments of one tonne of residual household waste: incineration of the full amount or sorting out organic waste for biogas production for either combined heat and power generation or as fuel in vehicles. The case study illustrates that the optimal solution depends on the objective and assumptions regarding the background system--illustrated with different assumptions regarding displaced electricity production. The article shows that it is feasible to combine LCA methodology with optimization. Furthermore, it highlights the need for including the integrated waste and energy system into the model.

Development of an internal restriction control in the PCR detection of the methylenetetrahydrofolate reductase (MTHFR) C677T mutation.

BACKGROUND: Detection of the presence of 677C/T allele in the human MTHFR gene is easily achieved by amplification using primers designed to span this region. Hinf I digestion, occurring only in the 677T allele, subsequently discriminates between the two alleles. Existing methods, however, do not control for failed restriction endonuclease digestion. METHODS AND RESULTS: A new forward, modified primer was designed and placed further upstream to create a Hinf I site, which, because it is present in both alleles, would serve as an internal restriction control. CONCLUSIONS: By allowing an additional restriction to take place in both the normal and mutated alleles, the use of the new primer provided for an internal restriction control.

Development of a simple multiplex polymerase chain reaction for the simultaneous detection of factor V Leiden and prothrombin 20210A mutations.

BACKGROUND: The demand for thrombophilia testing at the molecular level is increasing, and consequently, the work load of the routine molecular laboratory is also increasing. Efforts to lighten the work load, economize on time, and strive for reduced costs while still maintaining quality assurance are thus necessary. METHODS AND RESULTS: A multiplex polymerase chain reaction (PCR) for the detection of factor V Leiden and prothrombin 20210A mutations was designed that enables the use of the same inexpensive restriction enzyme, controls for the digestion, and produces easily interpretable results. CONCLUSION: The use of this new multiplex PCR and digestion analysis enabled us to simultaneously perform a routine screen for factor V Leiden and prothrombin 20210A mutations.

Nitrification and autotrophic denitrification of source-separated urine.

In laboratory experiments, source-separated urine was stabilised with nitrification and denitrified via nitritation and anaerobic ammonium oxidation. The highest total ammonia concentration in the influent was 7,300 gN/m3, the maximum pH 9.2. In a moving bed biofilm reactor (MBBR) with Kaldnes biofilm carriers, we stabilised urine as a 1:1 ammonium nitrate solution. The maximum nitrification rate was 380 gN/m3/d corresponding to 1.7 gN/m2(biofilm)/d. Nitrite ammonium solutions were produced in a continuous flow stirred tank reactor (CSTR) with 4.8 days sludge retention time (SRT) at 30 degrees C and in a sequencing batch reactor (SBR) with more than 30 days SRT. Nitrate build-up was negligible in both reactors. Nitritation rates were 780 gN/m3/d in the CSTR and 280 gN/m3/d in the SBR, respectively. However, shortening the cycles would increase nitritation in the SBR. High concentrations of nitrous acid, salts, and presumably hydroxylamine suppressed nitrite oxidation in the nitritation reactors. In all three nitrification reactors, maximally 50% of the influent total ammonia was oxidised without pH control. None of the common inhibition or limitation approaches could explain why ammonia oxidation always stopped at pH values around 6. In a batch experiment, we showed that source-separated urine can be denitrified autotrophically by anammox bacteria.

Future waste treatment and energy systems--examples of joint scenarios.

Development and use of scenarios for large interdisciplinary projects is a complicated task. This article provides practical examples of how it has been carried out in two projects addressing waste management and energy issues respectively. Based on experiences from the two projects, recommendations are made for an approach concerning development of scenarios in projects dealing with both waste management and energy issues. Recommendations are given to develop and use overall scenarios for the project and leave room for sub-scenarios in parts of the project. Combining different types of scenarios is recommended, too, in order to adapt to the methods and tools of different disciplines, such as developing predictive scenarios with general equilibrium tools and analysing explorative scenarios with energy system analysis tools. Furthermore, as marginals identified in differing future background systems determine the outcomes of consequential life cycle assessments (LCAs), it is considered advisable to develop and use explorative external scenarios based on possible marginals as a framework for consequential LCAs. This approach is illustrated using an on-going Danish research project.

Evaluation of the quality of endoscopically obtained esophageal biopsies in the dog.

OBJECTIVE: An important premise for obtaining diagnostically relevant histology specimens is an appropriate biopsy technique. Goal of this study was to determine if biopsies of adequate quality can be obtained from the canine esophagus at the gastro-esophageal junction (GEJ) during routine upper gastrointestinal endoscopy. MATERIAL AND METHODS: Over the course of one year, 58 dogs undergoing upper gastrointestinal endoscopy because of the presence of esophageal (n = 22) or gastrointestinal (n = 36) clinical signs were prospectively included. Five biopsies were repeatedly collected from the same dorsal and ventral locations of the GEJ, fixated individually in 4% neutral buffered formaldehyde, and evaluated histopathologically after standard preparation and haematoxylin and eosin staining. The presence of esophageal squamous epithelium with a basal cell layer and lamina propria mucosae in conjunction with foveolar columnar epithelium and cardiac glands, and the absence of fundic glands in one specimen, respectively, was judged as an adequately sampled biopsy. RESULTS: Adequately sampled biopsies were reported in 45 out of 58 dogs, with 31 samples originating from the dorsal GEJ, 36 samples originating from the ventral GEJ, and with 22 samples originating from both sites, respectively. The incidence of adequately sampled biopsies increased significantly over time (r = 0,22; p < 0,05), with these biopsies being reported significantly more often during the last 6 months compared to the first 6 months of the study (p = 0,03). Histopathological evaluation of the esophageal squamous epithelium showed fibrosis, inflammation, elongation of the stromal papillae, and increased thickness of the basal cell layers in 14 out of 58 dogs. Stromal papillae of the ventral esophageal epithelium were significantly elongated in dogs with esophageal clinical signs compared to dogs with gastrointestinal clinical signs (p = 0,03). CONCLUSION AND CLINICAL RELEVANCE: After an initial learning phase adequate esophageal biopsies from the GEJ can be obtained in canine patients undergoing upper gastrointestinal endoscopy, and histological lesions can be found in these biopsies.

Challenges when performing economic optimization of waste treatment: a review.

Strategic and operational decisions in waste management, in particular with respect to investments in new treatment facilities, are needed due to a number of factors, including continuously increasing amounts of waste, political demands for efficient utilization of waste resources, and the decommissioning of existing waste treatment facilities. Optimization models can assist in ensuring that these investment strategies are economically feasible. Various economic optimization models for waste treatment have been developed which focus on different parameters. Models focusing on transport are one example, but models focusing on energy production have also been developed, as well as models which take into account a plant's economies of scale, environmental impact, material recovery and social costs. Finally, models combining different criteria for the selection of waste treatment methods in multi-criteria analysis have been developed. A thorough updated review of the existing models is presented, and the main challenges and crucial parameters that need to be taken into account when assessing the economic performance of waste treatment alternatives are identified. The review article will assist both policy-makers and model-developers involved in assessing the economic performance of waste treatment alternatives.

Idiopathic esophagopathies resembling gastroesophageal reflux disease in dogs.

OBJECTIVE: Pathologic gastroesophageal reflux (GER) has been demonstrated experimentally in dogs, and it is suspected to occur naturally in dogs, yet its clinical significance is unknown. The aim of the study was to demonstrate clinical indicators of pathologic GER in dogs with idiopathic esophagopathies. MATERIALS AND METHODS: Dogs with clinical signs suggestive for esophageal disease (regurgitation, ptyalism, or dysphagia) and where extraesophageal and specific esophageal diseases had been ruled out, were retrospectively diagnosed with idiopathic esophagopathies. History, physical examination findings, clinicopathologic, radiographic, and endoscopic data, and treatment results were obtained from medical records, reviewed and evaluated. RESULTS: Out of 67 dogs with anamnestic esophageal signs, 12 (17.4%) dogs were identified as having idiopathic esophagopathies and were included in the study. Median age was 3.0 years (range 1.0-11.0), and median bodyweight was 28.2 kg (range 8.2-44.0). The most frequent anamnestic esophageal signs were ptyalism (10/12 dogs), regurgitation (8/12 dogs), signs of discomfort, pain (8/12 dogs), and cough (5/12 dogs). The most common radiographic abnormality was segmental esophageal dilation (8/12 dogs). Esophagoscopy revealed single mucosal surface defects at the gastroesophageal junction in 3/12 dogs. In dogs with altered esophageal motility, cytological and microbiological examinations of bronchial aspirates showed goblet cell hyperplasia (8/8 dogs), neutrophilic infiltration (5/8 dogs) and culturable bacteria (4/8 dogs), respectively. All dogs were treated with omeprazole (median 0.7 mg/kg once per day, range 0.5-1.2). Reported median treatment duration until remission of the main clinical signs was 20.0 days (range 8.0-54.0 days). This endpoint was reached in 11/12 dogs. CONCLUSION AND CLINICAL RELEVANCE: Results suggest that in some dogs with esophageal clinical signs, and where no primary disease could be identified, clinical indicators of pathologic GER such as pain, mucosal lesions and motility disturbances of the esophagus, respiratory complications, and response to therapy can be observed.

Assessment of the histological quality of endoscopic biopsies obtained from the canine gastro-esophageal junction.

OBJECTIVE: In the dog biopsy samples from the gastro-esophageal junction (GEJ) are rarely obtained during routine gastroscopy. The aim of this pilot study was to assess the histological quality of endoscopic biopsies sampled from the canine esophagus and cardia. It was hypothesised that it is possible to sample adequate specimens from these sites. MATERIALS AND METHODS: For this purpose 10 dogs with an indication for gastroscopy were enrolled in a prospective study. Biopsy samples were obtained with standard biopsy forceps for single use exactly from the GEJ thus containing preferably columnar epithelium from the cardia and squamous epithelium from the esophagus, respectively. In every dog the specimens were examined for size, layers and site, respectively. Study endpoint was reached when specimens originated from cardia and esophagus, showing at least epithelium and lamina propria mucosae, and a diameter >2mm on the slide, respectively. RESULTS: 72 biopsy specimens (median 7, range 5-10) obtained from the GEJ were examined in 10 dogs. Specimens from the esophagus containing squamous epithelium with lamina propria mucosae were found in 5 of 10 (50.0%) dogs. Specimens from the cardia containing columnar epithelium with lamina propria mucosae were found in 10 of 10 (100.0%) dogs. Four of 10 (40.0%), and 10 of 10 (100.0%) dogs showed at least one specimen >2mm on the slide originating from the esophagus, and from the cardia, respectively. Histological quality was found to be adequate in 4 of 10 (40.0%) dogs, showing specimens of adequate size, originating from both esophagus and cardia, and containing at least epithelium and lamina propria mucosae. CONCLUSION AND CLINICAL RELEVANCE: The pilot study provides evidence that during routine gastroscopy it is possible to sample endoscopic biopsies from the cardia and with limitations from the esophagus showing a quality adequate for histological examination of the epithelium and the lamina propria mucosae.

Long-term affected energy production of waste to energy technologies identified by use of energy system analysis.

Affected energy production is often decisive for the outcome of consequential life-cycle assessments when comparing the potential environmental impact of products or services. Affected energy production is however difficult to determine. In this article the future long-term affected energy production is identified by use of energy system analysis. The focus is on different uses of waste for energy production. The Waste-to-Energy technologies analysed include co-combustion of coal and waste, anaerobic digestion and thermal gasification. The analysis is based on optimization of both investments and production of electricity, district heating and bio-fuel in a future possible energy system in 2025 in the countries of the Northern European electricity market (Denmark, Norway, Sweden, Finland and Germany). Scenarios with different CO(2) quota costs are analysed. It is demonstrated that the waste incineration continues to treat the largest amount of waste. Investments in new waste incineration capacity may, however, be superseded by investments in new Waste-to-Energy technologies, particularly those utilising sorted fractions such as organic waste and refuse derived fuel. The changed use of waste proves to always affect a combination of technologies. What is affected varies among the different Waste-to-Energy technologies and is furthermore dependent on the CO(2) quota costs and on the geographical scope. The necessity for investments in flexibility measures varies with the different technologies such as storage of heat and waste as well as expansion of district heating networks. Finally, inflexible technologies such as nuclear power plants are shown to be affected.

Altered gene expression in Choristoneura fumiferana and Manduca sexta in response to sublethal intoxication by Bacillus thuringiensis Cry1Ab toxin.

In order to understand how lepidopteran insects react physiologically to Bacillus thuringiensis crystal toxin ingestion, transcriptional profiling of Choristoneura fumiferana larvae exposed to sublethal doses of Cry1Ab protoxin were monitored using a C. fumiferana-specific cDNA microarray derived from a protoxin-specific subtractive library. Differential gene expression occurred primarily between 2 and 5 h postingestion. Metabolic enzymes such as lipases and proteases were generally repressed, whereas genes involved in detoxification, immune system regulation or general stress response were upregulated. A similar protoxin-specific transcriptional pattern was also observed with Manduca sexta larvae, using three upregulated genes (serpin, cytochrome P450 and carboxyl/cholinesterase) and one downregulated gene (beta-glucosidase), suggesting that a susceptible larval response to Cry toxin exposure might be universal among lepidopterous insects.

Transcriptional response of Choristoneura fumiferana to sublethal exposure of Cry1Ab protoxin from Bacillus thuringiensis.

Bacillus thuringiensis is a microbial control agent active against Choristoneura fumiferana, a lepidopteran defoliator of North American forests. Although the B. thuringiensis insecticidal crystal protoxins have a relatively narrow host range, there is concern about their impact on non-target species where intoxication effects may not be overt. Larval toxicity effects can be assessed at the molecular level by determining altered transcriptional profiles in response to sublethal protoxin exposure in sensitive insects. Subtraction hybridization libraries were created using two larval populations, control and protoxin-fed and were characterized by sequencing 1091 clones. Differential mRNA expression of selected clones, as measured by quantitative polymerase chain reaction, identified a number of metabolic and stress-related genes that were either transcriptionally enhanced or repressed after protoxin exposure.