Persistence of Influenza A (H1N1) Virus on Stainless Steel Surfaces.

UNLABELLED: As annual influenza epidemics continue to cause significant morbidity and economic burden, an understanding of viral persistence and transmission is critical for public health officials and health care workers to better protect patients and their family members from infection. The infectivity and persistence of two influenza A (H1N1) virus strains (A/New Caledonia/20/1999 and A/Brisbane/59/2007) on stainless steel (SS) surfaces were evaluated using three different surface matrices (2% fetal bovine serum, 5 mg/ml mucin, and viral medium) under various absolute humidity conditions (4.1 × 10(5) mPa, 6.5 × 10(5) mPa, 7.1 × 10(5) mPa, 11.4 × 10(5) mPa, 11.2 × 10(5) mPa, and 17.9 × 10(5) mPa) for up to 7 days. Influenza A virus was deposited onto SS coupons (7.07 cm(2)) and recovered by agitation and sonication in viral medium. Viral persistence was quantified using a tissue culture-based enzyme-linked immunosorbent assay (ELISA) to determine the median (50%) tissue culture infective dose (TCID50) of infectious virus per coupon. Overall, both strains of influenza A virus remained infectious on SS coupons, with an approximate 2 log10 loss over 7 days. Factors that influenced viral persistence included absolute humidity, strain-absolute humidity interaction, and time (P ≤ 0.01). Further studies on the transfer of influenza A virus from fomites by hand and the impact of inanimate surface contamination on transmission should be performed, as this study demonstrates prolonged persistence on nonporous surfaces. IMPORTANCE: This study tested the ability of two influenza A (H1N1) virus strains to persist and remain infectious on stainless steel surfaces under various environmental conditions. It demonstrated that influenza A (H1N1) viruses can persist and remain infectious on stainless steel surfaces for 7 days. Additional studies should be conducted to assess the role played by contaminated surfaces in the transmission of influenza A virus.

Multi-environment Evaluation of Maize Inbred Lines for Resistance to Fusarium Ear Rot and Fumonisins.

Fusarium verticillioides causes Fusarium ear rot (FER) of maize and produces fumonisins, which affects grain quality. Host-plant resistance can reduce both FER and fumonisins in maize. In this study, 18 maize inbred lines were evaluated for resistance to F. verticillioides and fumonisin accumulation at five localities in South Africa. Additive main effects and multiplicative interaction analyses revealed significant environment × genotype interactions, with inbred lines CML 390, US 2540W, RO 424W, and VO 617y-2 consistently exhibiting low FER severity (≤5.4%), fungal target DNA (≤0.1 ng µl-1), and fumonisin levels (≤5.6 ppm). Genotype main effect and genotype × environment biplots showed that inbred lines CML 390, US 2540W, and RO 424W were most resistant to FER, fungal colonization, and fumonisin accumulation, respectively, while inbred line RO 424W was most stable in its resistance response over environments. These inbred lines also demonstrated broad adaptability by consistently exhibiting resistance to FER, fungal colonization, and fumonisins across localities. The identified lines could serve as valuable sources of resistance against F. verticillioides and its fumonisins in local breeding programs.

Inactivation of bacterial biothreat agents in water, a review.

Water supplies and water distribution systems have been identified as potential targets for contamination by bacterial biothreat agents. Since the 2001 Bacillus anthracis bioterrorist attacks, additional efforts have been aimed at research to characterize biothreat organisms in regards to their susceptibility to disinfectants and technologies currently in use for potable water. Here, we present a review of research relevant to disinfection of bacteria with the potential to pose a severe threat to public health and safety, and their potential surrogates. The efficacy of chlorine, monochloramine, chlorine dioxide, and ultraviolet light to inactivate each organism in suspension is described. The complexities of disinfection under varying water conditions and when the organisms are associated with biofilms in distribution systems are discussed.

Survival and persistence of nonspore-forming biothreat agents in water.

AIMS: To determine whether nonspore-forming biothreat agents can survive and persist in potable water that does not contain a disinfectant. METHODS AND RESULTS: Autoclaved, de-chlorinated Atlanta municipal water was inoculated with eight isolates of bacterial biothreat agents (106 CFU ml⁻¹). The inoculated water samples were incubated at 5, 8 (Francisella tularensis only) or 25°C and assayed for viability by culture and by the presence of metabolic activity as measured by esterase activity (ScanRDI, AES Chemunex). Viability as determined by culture varied from 1 to 30 days, depending upon the organism and the temperature of the water. All organisms were determined viable as measured by esterase activity for the entire 30 days, regardless of the incubation temperature. CONCLUSION: Francisella tularensis was culturable for at least 21 days if held at 8°C. The remaining nonspore-forming bacterial biothreat agents were found to be metabolically active for at least 30 days in water held at 5 or 25°C. SIGNIFICANCE AND IMPACT OF THE STUDY: The data can assist public health officials to determine the safety of drinking water after contamination with a biothreat agent.

Chlorine dioxide inactivation of bacterial threat agents.

AIMS: To evaluate the efficacy of chlorine dioxide (ClO(2)) against seven species of bacterial threat (BT) agents in water. METHODS AND RESULTS: Two strains of Bacillus anthracis spores, Yersinia pestis, Francisella tularensis, Burkholderia pseudomallei, Burkholderia mallei and Brucella species were each inoculated into a ClO(2) solution with an initial concentration of 2.0 (spores only) and 0.25 mg l(-1) (all other bacteria) at pH 7 or 8, 5 or 25°C. At 0.25 mg l(-1) in potable water, six species were inactivated by at least three orders of magnitude within 10 min. Bacillus anthracis spores required up to 7 h at 5°C for the same inactivation with 2.0 mg l(-1) ClO(2). CONCLUSIONS: Typical ClO(2) doses used in water treatment facilities would be effective against all bacteria tested except B. anthracis spores that would require up to 7 h with the largest allowable dose of 2 mg l(-1) ClO(2). Other water treatment processes may be required in addition to ClO(2) disinfection for effective spore removal or inactivation. SIGNIFICANCE AND IMPACT OF STUDY: The data obtained from this study provide valuable information for water treatment facilities and public health officials in the event that a potable water supply is contaminated with these BT agents.

Chlorine disinfection of Francisella tularensis.

AIMS: To determine the range of free available chlorine (FAC) required for disinfection of the live vaccine strain (LVS) and wild-type strains of Francisella tularensis. METHODS AND RESULTS: Seven strains of planktonic F. tularensis were exposed to 0·5 mg·l(-1) FAC for two pH values, 7 and 8, at 5 and 25°C. LVS was inactivated 2 to 4 times more quickly than any of the wild-type F. tularensis strains at pH 8 and 5°C. CONCLUSIONS: Free available chlorine residual concentrations routinely maintained in drinking water distribution systems would require up to two hours to reduce all F. tularensis strains by 4 log10. LVS was inactivated most quickly of the tested strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides contact time (CT) values that are useful for drinking water risk assessment and also suggests that LVS may not be a good surrogate in disinfection studies.

Most-probable-number rapid viability PCR method to detect viable spores of Bacillus anthracis in swab samples.

A comparison of Most-Probable-Number Rapid Viability (MPN RV) PCR and traditional culture methods for the quantification of Bacillus anthracis Sterne spores in macrofoam swabs from a multi-center validation study was performed. The purpose of the study was to compare environmental swab processing methods for recovery, detection, and quantification of viable B. anthracis spores from surfaces. Results show that spore numbers provided by the MPN RV-PCR method were typically within 1-log of the values from a plate count method for all three levels of spores tested (3.1x10(4), 400, and 40 spores sampled from surfaces with swabs) even in the presence of debris. The MPN method tended to overestimate the expected result, especially at lower spore levels. Blind negative samples were correctly identified using both methods showing a lack of cross contamination. In addition to detecting low levels of spores in environmental conditions, the MPN RV-PCR method is specific, and compatible with automated high-throughput sample processing and analysis protocols, enhancing its utility for characterization and clearance following a biothreat agent release.

UV light inactivation of bacterial biothreat agents.

Seven species of bacterial biothreat agents were tested for susceptibility to UV light (254 nm). All gram-negative organisms tested required <12 mJ/cm(2) for a 4-log(10) reduction in viability (inactivation). Tailing off of the B. anthracis spore inactivation curves began close to the 2-log(10) inactivation point, with a fluence of approximately 40 mJ/cm(2), and 3-log(10) inactivation still was not achieved with a fluence of 120 mJ/cm(2).

Evaluation of a macrofoam swab protocol for the recovery of Bacillus anthracis spores from a steel surface.

A protocol to recover Bacillus anthracis spores from a steel surface using macrofoam swabs was evaluated for its accuracy, precision, reproducibility, and limit of detection. Macrofoam swabs recovered 31.7 to 49.1% of spores from 10-cm2 steel surfaces with a < or =32.7% coefficient of variation in sampling precision and reproducibility for inocula of > or =38 spores.

Inactivation of spores of Bacillus anthracis Sterne, Bacillus cereus, and Bacillus thuringiensis subsp. israelensis by chlorination.

Three species of Bacillus were evaluated as potential surrogates for Bacillus anthracis for determining the sporicidal activity of chlorination as commonly used in drinking water treatment. Spores of Bacillus thuringiensis subsp. israelensis were found to be an appropriate surrogate for spores of B. anthracis for use in chlorine inactivation studies.

Preliminary report on reduction of esophagitis by amifostine in patients with non-small-cell lung cancer treated with chemoradiotherapy.

Esophagitis is a major toxicity of chemoradiotherapy for lung cancer. Twenty-four patients with non-small-cell lung cancer received induction chemotherapy (paclitaxel/carboplatin) followed by concurrent thoracic irradiation (RT) and weekly paclitaxel. Acute esophagitis was scored weekly. Since a high rate of grade 3 esophagitis was noted in the initial group of 12 patients, amifostine (AMI) 500 mg intravenously twice weekly was added to the regimen in the subsequent 12 patients. Esophagitis Index (EI) was calculated as an area under the curve reflecting esophagitis grade over time. Median number of AMI doses was 12 per patient. AMI was well tolerated. Two patients were not evaluable for esophagitis. The incidence of grade 3 esophagitis was 18% in the initial 11 patients versus 9% in the AMI-treated patients (P = not significant). Mean EI was numerically lower in the AMI-treated patients than in the initial group (5.1 vs. 11.6, P = 0.14). The product of RT dose and length of esophagus in the RT field was larger in the AMI group (934 vs. 761, P = 0.035). Median survival time for all patients was 12.4 months. Esophagitis Index, a novel measure of the severity and duration of acute esophagitis, may be reduced in lung cancer patients receiving twice-weekly AMI with thoracic RT and paclitaxel. Twice weekly AMI did not eliminate grade 3 esophagitis; therefore, dose escalation of AMI is planned. The effect of AMI was not due to the shorter irradiated esophageal length. A phase III randomized trial is now open to assess AMI's effect on esophagitis.

Volatile organic compounds associated with microbial growth in automobile air conditioning systems.

Volatile organic compounds from Penicillium viridicatum and Methylobacterium mesophilicum growing on laboratory media and on component materials of automobile air conditioners were analyzed with gas chromatography and mass spectrometry. P. viridicatum produced compounds such as 4-methyl thiazole, terpenes and alcohols, whereas M. mesophilicum produced dimethyl disulfide, dimethyl trisulfide, and chlorophenol with growth on laboratory media. In comparison with laboratory media, fewer volatiles were detected from colonized foam insulation materials. Biofilms of M. mesophilicum on aluminum evaporator components produced mainly dimethyl disulfide. These biofilms, after inoculation with P. viridicatum, produced offensive smelling alcohols and esters such as 2-methyl propanol, 3-penten-2-ol, and the ethyl ester of butanoic acid. The moisture and substrates innate to the automobile air conditioning systems provided an environment suitable for microbial biofilm development and odor production. Reduction of retained moisture in the air conditioning system coupled with use of less susceptible or antimicrobial substrates are advised for remediation of the noxious odors.

Effects of hydrogel/silver coatings on in vitro adhesion to catheters of bacteria associated with urinary tract infections.

Sections of sterile all-silicone-, hydrogel/silver-all-silicone-, and hydrogel/silver-latex-Foley urinary catheters were exposed to suspensions of bacteria and Candida albicans associated with urinary tract infections. The adhesion of these microorganisms to the catheters was determined with a radiolabel-cell procedure and scanning electron microscopy. Anomalous data with the radiolabel procedure were produced with the hydrogel/silver-latex catheters for certain species. These aberrant data were related to adhesion on the untreated cut ends of the latex catheter. Radiolabel-cell-adhesion procedures that involve sections of coated materials may need to be supplemented with additional procedures such as scanning electron microscopy for valid interpretations of the data. Adhesion to the hydrogel/silver catheters by both Gram-positive- and Gram-negative bacteria most commonly associated with nosocomial urinary tract infections, including a strain of Pseudomonas aeruginosa noted for its superior adhesion capacity, was significantly lower than the adhesion to the control all-silicone catheter.

Managed care companies and the practice of medicine.

One of the major issues pertaining to the pending legislation for Patient's Bill of Rights is the potential of liability health care plans, particularly when they decline coverage they consider not medically necessary. We call these contracts "managed health care" plans. But, realistically, when is it managing? When is it medicine? When, if at all, does management undermine medicine? And if it does, should managed care organizations--and their representatives--be held legally liable for medical decisions that go wrong? A panel of seven experts examines these questions from medical, payment, patient, legal, insurance, and governing viewpoints.

The occurrence and persistence of mixed biofilms in automobile air conditioning systems.

Twelve automobile air conditioner systems from six manufacturers and three countries, selected mostly because of complaints of unpleasant odors in the passenger compartment, were examined for microbial growth by direct microscopy and enrichment culture. Mixed populations of fungi and bacteria (with occasional protozoa) were observed in biofilms in at least some of the components from all used units. The aluminum heat exchanger fins from ten evaporators demonstrated bacterial biofilms that yielded Methylobacterium mesophilicum. Penicillium viridicatum colonized components from four units. These bacteria and fungi were recoverable repeatedly from these units during 'dry' storage of up to 27 months. This report associates a bacterial-fungal community with disagreeable air quality in some automobiles.

Neoadjuvant and adjuvant therapy of non-small cell lung cancer.

Present treatment options for non-small cell lung cancer in the adjuvant and neoadjuvant settings are far from ideal. The bottom line of improvement in overall survival by any treatment modality used has yet to be achieved. For this to happen, better drug therapy will be required. Future trials should be carefully designed with intraoperative staging and stratification of patients by histology, stage, and potentially immunologic markers, ploidy, and molecular biologic markers such as growth factors and their receptors. Combination of individual therapies to maximal toxicity in an attempt to overcome inherent tumor resistance will be key. The use of colony-stimulating factors, with high-dose chemotherapy integrated with hyperfractionated radiotherapy, may be seen in future studies. Combining standard chemotherapy and radiotherapy with cytokines or immunotherapy using interleukin-1 and -2, or tumor necrosis factor or monoclonal antibodies to growth factors may be seen in innovative trials. By such meticulously and intelligently designed trials, progress in adjuvant and neoadjuvant treatment of the important group of patients with locally advanced non-small cell lung cancer will occur.

Diffuse contralateral pulmonary metastases in malignant mesothelioma. An unusual radiographic presentation.

We describe a patient with malignant pleural mesothelioma involving the lung parenchyma bilaterally in a diffuse nodular fashion. This pattern of metastasis is seldom reported for this tumor.

Occult prostatic carcinoma metastatic to the medial canthal area. Diagnosis by immunohistochemistry.

A 91-year-old man developed a mass in the right medial canthal and lacrimal sac region, which was found histopathologically to be a poorly differentiated neoplasm. The results of a complete systemic evaluation, including urologic consultation, rectal examination, bone scan, and determination of serum acid phosphatase level, were normal. In spite of the negative evaluation for prostatic carcinoma, immunohistochemical studies using immunoperoxidase stains for prostatic-specific antigen demonstrated that the lacrimal sac tumor was metastatic carcinoma from an occult primary neoplasm in the prostate gland. The presence of tumor in the prostate gland was then confirmed by a needle biopsy. Modern immunohistochemical techniques are particularly valuable in the diagnosis of selected ophthalmic tumors in which the diagnosis cannot be determined by routine diagnostic techniques.

Hepatic granulomatosis and sarcoidosis. Further observations.

The etiology of hepatic granulomatosis discovered in patients with normal chest roentgenograms is controversial. Among 18 such patients studied in recent years, sarcoidosis was identified as the cause in 15 by demonstration of granulomatous inflammation in extrahepatic tissues, employing a variety of biopsy methods. Serum angiotensin levels were raised in all 10 patients tested and gallium scans were helpful in four of six cases. Prolonged fever was the most common symptom, but three patients had severe right upper quadrant pain, a manifestation of hepatic sarcoidosis not previously described. Improvement on corticosteroids was observed in the 14 treated patients, but incomplete response led to use of cytotoxic drugs in three cases. Sarcoidosis presents with normal chest roentgenograms more often than is appreciated and appears to be a common cause of unexplained persistent hepatic granulomatosis. A firm diagnosis should be established since treatment is long and frustrating. The prognosis is good but treatment for years is required in most cases.