West Nile virus (WNV) is a neurotropic, mosquito-borne flavivirus that can cause lethal meningoencephalitis. Type I interferon (IFN) plays a critical role in controlling WNV replication, spread, and tropism. In this study, we begin to examine the effector mechanisms by which type I IFN inhibits WNV infection. Mice lacking both the interferon-induced, double-stranded-RNA-activated protein kinase (PKR) and the endoribonuclease of the 2',5'-oligoadenylate synthetase-RNase L system (PKR(-/-) x RL(-/-)) were highly susceptible to subcutaneous WNV infection, with a 90% mortality rate compared to the 30% mortality rate observed in congenic wild-type mice. PKR(-/-) x RL(-/-) mice had increased viral loads in their draining lymph nodes, sera, and spleens, which led to early viral entry into the central nervous system (CNS) and higher viral burden in neuronal tissues. Although mice lacking RNase L showed a higher CNS viral burden and an increased mortality, they were less susceptible than the PKR(-/-) x RL(-/-) mice; thus, we also infer an antiviral role for PKR in the control of WNV infection. Notably, a deficiency in both PKR and RNase L resulted in a decreased ability of type I IFN to inhibit WNV in primary macrophages and cortical neurons. In contrast, the peripheral neurons of the superior cervical ganglia of PKR(-/-) x RL(-/-) mice showed no deficiency in the IFN-mediated inhibition of WNV. Our data suggest that PKR and RNase L contribute to IFN-mediated protection in a cell-restricted manner and control WNV infection in peripheral tissues and some neuronal subtypes.

West Nile virus (WNV) is a mosquito-borne flavivirus that is neurotropic in humans, birds, and other animals. While adaptive immunity plays an important role in preventing WNV spread to the central nervous system (CNS), little is known about how alpha/beta interferon (IFN-alpha/beta) protects against peripheral and CNS infection. In this study, we examine the virulence and tropism of WNV in IFN-alpha/beta receptor-deficient (IFN- alpha/betaR-/-) mice and primary neuronal cultures. IFN-alpha/betaR-/- mice were acutely susceptible to WNV infection through subcutaneous inoculation, with 100% mortality and a mean time to death (MTD) of 4.6 +/- 0.7 and 3.8+/- 0.5 days after infection with 10(0) and 10(2) PFU, respectively. In contrast, congenic wild-type 129Sv/Ev mice infected with 10(2) PFU showed 62% mortality and a MTD of 11.9 +/- 1.9 days. IFN-alpha/betaR-/- mice developed high viral loads by day 3 after infection in nearly all tissues assayed, including many that were not infected in wild-type mice. IFN-alpha/betaR-/- mice also demonstrated altered cellular tropism, with increased infection in macrophages, B cells, and T cells in the spleen. Additionally, treatment of primary wild-type neurons in vitro with IFN-beta either before or after infection increased neuronal survival independent of its effect on WNV replication. Collectively, our data suggest that IFN-alpha/beta controls WNV infection by restricting tropism and viral burden and by preventing death of infected neurons.

OBJECTIVE: The objective of this study was to determine the etiology and risk factors for acute histoplasmosis in two outbreaks in Illinois among laborers at a landfill in 2001 and at a bridge reconstruction site in 2003. DESIGN: We performed environmental investigations during both outbreaks and also performed an analytic cohort study among bridge workers. PARTICIPANTS: Workers at the landfill during May 2001 and those at the bridge site during August 2003 participated in the study. At the landfill, workers moved topsoil from an area that previously housed a barn; at the bridge, workers observed bat guano on bridge beams. EVALUATIONS/MEASUREMENTS: We defined a case by positive immunodiffusion serology, a > or = 4-fold titer rise in complement fixation between acute and convalescent sera, or positive urinary Histoplasma capsulatum (HC) antigen. Relative risks (RR) for disease among bridge workers were calculated using bivariate analysis. RESULTS: Eight of 11 landfill workers (73%) and 6 of 12 bridge workers (50%) were laboratory-confirmed histoplasmosis cases. Three bridge workers had positive urinary HC antigen. At the bridge, seeing or having contact with bats [RR = 7.0; 95% confidence interval (CI), 1.1-43.0], jack-hammering (RR = 4.0; 95% CI, 1.2-13.3), and waste disposal (RR = 4.0; 95% CI, 1.2-13.3) were the most significant job-related risk factors for acquiring histoplasmosis. CONCLUSIONS: Workers performing activities that aerosolized topsoil and dust were at increased risk for acquiring histoplasmosis. Relevance to professional and clinical practice: Employees should wear personal protective equipment and use dust-suppression techniques when working in areas potentially contaminated with bird or bat droppings. Urinary HC antigen testing was important in rapidly identifying disease in the 2003 outbreak.

We assessed the structure and latitudinal selection that might result in sensitivities to critical day-lengths that trigger diapause between Culex pipiens populations distributed along North-South and East-West axes in eastern North America. Strong population structure between Cx. p. pipiens and Cx. p. quinquefasciatus existed. Among Cx. p. pipiens, a 100-km increase in the latitudinal change resulted in an increased square root of F(ST) by 0.002. A 100-km increase in the longitudinal change caused an increased square root of F(ST) by 0.035. A lack of latitudinal influence on the structure between Cx. p. pipiens populations suggests a uniform signal using the 12 microsatellite markers, which might increase the risk of West Nile virus (WNV) transmission toward northern areas because of longer breeding season, extend host-seeking period, and larger population size. Northern Cx. p. pipiens may have undergone additional generations before diapause is triggered, magnifying population size when WNV amplification is peaking.

BACKGROUND: Triple-reassortant swine influenza A (H1) viruses - containing genes from avian, human, and swine influenza viruses - emerged and became enzootic among pig herds in North America during the late 1990s. METHODS: We report the clinical features of the first 11 sporadic cases of infection of humans with triple-reassortant swine influenza A (H1) viruses, occurring from December 2005 through February 2009, until just before the current epidemic of swine-origin influenza A (H1N1) among humans. These data were obtained from routine national influenza surveillance reports and from joint case investigations by public and animal health agencies. RESULTS: The median age of the 11 patients was 10 years (range, 16 months to 48 years), and 4 had underlying health conditions. Nine of the patients had had exposure to pigs, five through direct contact and four through visits to a location where pigs were present but without contact. In another patient, human-to-human transmission was suspected. The range of the incubation period, from the last known exposure to the onset of symptoms, was 3 to 9 days. Among the 10 patients with known clinical symptoms, symptoms included fever (in 90%), cough (in 100%), headache (in 60%), and diarrhea (in 30%). Complete blood counts were available for four patients, revealing leukopenia in two, lymphopenia in one, and thrombocytopenia in another. Four patients were hospitalized, two of whom underwent invasive mechanical ventilation. Four patients received oseltamivir, and all 11 recovered from their illness. CONCLUSIONS: From December 2005 until just before the current human epidemic of swine-origin influenza viruses, there was sporadic infection with triple-reassortant swine influenza A (H1) viruses in persons with exposure to pigs in the United States. Although all the patients recovered, severe illness of the lower respiratory tract and unusual influenza signs such as diarrhea were observed in some patients, including those who had been previously healthy. Copyright 2009 Massachusetts Medical Society.

Background. Human adenovirus type 3 (HAdV-3) causes severe respiratory illness in children, but outbreaks in long-term care facilities have not been frequently reported. We describe an outbreak of HAdV-3 infection in a long-term care facility for children with severe neurologic impairment, where only 3 of 63 residents were ambulatory.Methods. A clinical case of HAdV-3 was defined as fever (temperature,>/=38.0 degrees C) and either a worsening of respiratory symptoms or conjunctivitis in a resident, with illness onset from June through August 2005. We reviewed medical records; conducted surveillance for fever, conjunctivitis, and respiratory symptoms; and collected nasopharyngeal and conjunctival specimens from symptomatic residents. Specimens were cultured in HAdV-permissive cell lines or were analyzed by HAdV-specific polymerase chain reaction assay.Results. Thirty-five (56%) of 63 residents had illnesses that met the case definition; 17 patients (49%) were admitted to intensive care units, and 2 (6%) died. Patients were hospitalized in the intensive care unit for a total of 233 patient-days. Illness onset dates ranged from 1 June through 24 August 2005. Thirty-two patients (91%) had respiratory infection, and 3 (9%) had conjunctivitis. HAdV was identified by culture or PCR in 20 patients. Nine isolates were characterized as HAdV-3 genome type a2.Conclusions. Considering the limited mobility of residents and their reliance on respiratory care, transmission of HAdV-3 infection during this outbreak likely occurred through respiratory care provided by staff. In environments where patients with susceptible underlying conditions reside, HAdV infection should be considered when patients are identified with worsening respiratory disease, and rapid diagnostic tests for HAdV infection should be readily available to help identify and curtail the spread of this pathogen.

Background Molds are a rare cause of disseminated infection among dialysis patients.Objective. We evaluated a cluster of intravascular infections with the mold Phialemonium among patients receiving hemodialysis at the same facility in order to identify possible environmental sources and prevent further infection.Design. Environmental assessment and case-control study.Setting. A hemodialysis center affiliated with a tertiary care hospital.Methods. We reviewed surveillance and clinical microbiology records and performed a blood culture survey for all patients. The following data for case patients were compared with those for control patients: underlying illness, dialysis characteristics, medications, and other possible exposure for 120 days prior to infection. Environmental assessment of water treatment, dialysis facilities, and heating, ventilation, and air-conditioning (HVAC) systems of the current and previous locations of the dialysis center was performed. Samples were cultured for fungus; Phialemonium isolates were confirmed by sequencing of DNA. Investigators observed dialysis access site disinfection technique.Results. Four patients were confirmed as case patients, defined as a patient having intravascular infection with Phialemonium species; 3 presented with fungemia, and 1 presented with an intravascular graft infection. All case patients used a fistula or graft for dialysis access, as did 12 (75%) of 16 of control patients (P=.54). Case and control patients did not differ in other dialysis characteristics, medications received, physiologic findings, or demographic factors. Phialemonium species were not recovered from samples of water or dialysis machines, but were recovered from the condensation drip pans under the blowers of the HVAC system that supplied air to the dialysis center. Observational study of 21 patients detected suboptimal contact time with antiseptic agents used to prepare dialysis access sites.Conclusion. The report of this outbreak adds to previous published reports of Phialemonium infection occurring in immunocompromised patients who likely acquired infection in the healthcare setting. Recovery of this mold from blood culture should be considered indicative of infection until proven otherwise. Furthermore, an investigation into possible healthcare-related environmental reservoirs should be considered.

Temporal changes in the abundance Culex restuans and Culex pipiens were monitored in east-central Illinois for over a decade using infusion-baited oviposition traps. The 2 species typically exhibited a seasonal shift in relative abundance with a mean crossover date (when the proportion of egg rafts from both species is equal) of August 10 or 11, depending on leap year, with a 95% confidence interval of +/- 10.7 days. The date of crossover was linearly related to the date of last spring frost and occurred on average about 123 days after the last spring frost. Despite the predictability of crossover, the weekly pattern in the proportion of Cx. pipiens before and after crossover varied considerably, even between years with similar crossover dates. After West Nile virus became established in our area, we found that transmission based on Culex from gravid traps did not increase until Cx. pipiens abundance increased in oviposition traps. Infection rates peaked within the half-month period after crossover. The peak in Cx. pipiens abundance in oviposition traps during this 3-year period was between the 2nd half of August and the end of September. A higher magnitude of transmission in 2002 coincided with warmer temperatures during July and August and an extended period in which the 2 Culex species were in relatively equal abundance.

After a severe outbreak of West Nile virus (WNV) in Cook County, Illinois, in 2002, detections of WNV in mosquitoes were frequent across the state in the following years despite small numbers of human cases. We conducted a spatio-temporal analysis of Culex (subgenus Culex) mosquitoes collected in 2004 in three mosquito abatement districts (MAD) in Cook County by calculating monthly estimates of mosquito density, prevalence of infected mosquitoes, and exposure intensity, which in turn is a product of mosquito density and infection rates. Mosquito infections were detected early at three sites in late May and were widely detected throughout the three MADs in the summer with infection rates as high as 13 per 1000 in August. Exposure intensities were higher at sites adjacent to the Des Plaines River, especially in August and September. The aggregated pattern of WNV transmission along the river might be related to the existence of substantial forest preserves and wetlands that might produce ecological conditions favorable for mosquito proliferation and interactions between mosquitoes and birds.

BACKGROUND: Pontiac fever (PF), a legionellosis with influenza-like symptoms and high attack rates, is rarely reported. Travel-related outbreaks can elude detection because infected persons are often widely removed geographically from the transmission source before illness onset. Thirty-one persons staying at an Illinois hotel during August 9 to 11, 2002, reported influenza-like symptoms to local health departments within 24 to 48 hours of checkout. We investigated to identify the cause and source of illness to guide control measures. METHODS: Hotel water samples were collected for culture. A telephone questionnaire detailing illness symptoms and exposures was administered to all who were guests at the hotel from August 9 to 15 (n = 380). A case was defined as onset of fever, headache, and myalgia in a guest in the 14 days following the hotel stay. Patient sera were tested by hemagglutination assay for antibodies to Legionella species. RESULTS: Among 204 questionnaire respondents from 15 states and Canada, 50 met the case definition. Among persons exposed to the swimming pool/whirlpool spa area, 63%(47 of 75) became ill versus 3%(3 of 110) of unexposed persons (relative risk 23.0, 95% CI 7.4-71.1). Illness risk increased with increasing time exposed to the pool/spa. Approximately 95 to 115 bathers per day, two to three times above the usual number, used the spa during August 9 to 11. Three Legionella species, L. dumoffii, L. maceachernii, and L. micdadei, were isolated from spa filter backwash cultures. Two of 15 ill persons with acute- and convalescent-phase sera had a greater than fourfold rise in antibody titer to L. micdadei. CONCLUSIONS: PF was associated with exposure to a hotel pool/spa area. Heavy bather usage likely contributed to a decreased effectiveness of the disinfectant in the whirlpool spa, possibly promoting bacterial aerosolization. Linking case information from many states is essential in identifying and eliminating the source of disease transmission in travel-related outbreaks of PF. Clinicians should be aware of PF in the differential diagnosis of patients with influenza-like symptoms following recent travel, particularly with exposure to a communal-use whirlpool spa.

Although foodborne outbreaks of illness are relatively common, they are rarely caused by chemical agents. An outbreak of gastrointestinal illness occurred among students at two schools shortly after lunch was served. A cohort study, an environmental investigation, and microbiological and toxicological laboratory testing of food samples were performed. A case was defined as a student or teacher who ate food prepared in the kitchen at school A on 25 November 2002 (and served at schools A and B) and who later developed headache or symptoms of gastrointestinal tract irritation, with onset within 180 min of eating lunch. Among 312 persons interviewed, 157 persons became ill (attack rate = 49%; attack rate 41% for school A, 11% for school B). Onset of illness occurred within 60 min for 81% of cases; 91% of students reported that their chicken tenders smelled unusual. Eating chicken tenders that smelled unusual was associated with being a case (relative risk 9.2, 95% confidence interval 1.4 to 62.6, P < 0.05). Ammonia was detected in uncooked chicken tenders at levels as high as 2,468 ppm. The chicken had been contaminated during a warehouse leak of ammonia refrigerant. This outbreak of ammonia poisoning is only the second reported in food, and the first in a solid food. Heated chicken tenders contaminated with ammonia can cause acute illness within a short period of time.

A.G. Freifeld, J. Meza, B. Schweitzer, L. Shafer, A.C. Kalil. A.R. Sambol. Seroprevalence of West Nile virus infection in solid organ transplant recipients. Transpl Infect Dis 2009. All rights reserved Background. Of people infected with mosquito-borne West Nile virus (WNV),<1% develop neuroinvasive disease (NID). Population studies suggest that people older than 65 years may be at higher risk for neurologic symptoms. It has been suggested that solid organ transplant (SOT) recipients are also at higher risk for WNV NID, but definitive serologic and epidemiologic data are lacking. Methods. A serologic screening survey, using a US Food & Drug Administration-approved enzyme-linked immunosorbant assay to detect WNV immunoglobulin-G (IgG) antibody responses in cohorts of SOT recipients and non-immunocompromised controls, was undertaken at a large Midwestern university organ transplant center in the aftermath of the summer 2003 WNV regional outbreak. Hemagglutination-inhibition testing was used to confirm WNV IgG-positive results and differentiate them from positive results caused by Saint Louis encephalitis virus, another flavivirus that is endemic in the Midwestern US. Findings. The rate of WNV IgG-seropositive responses did not differ between SOT recipients and non-immunocompromised controls, and were 12% and 10%, respectively. Retrospective chart review showed no documented WNV NID in the seropositive SOT recipients, suggesting an incidence of WNV NID may be as low as 0.7% in this population. Interpretation. Asymptomatic WNV infection is common among immunocompromised SOT patients, occurring as often as it does in non-immunocompromised controls. Our data indicated that severe WNV NID is less frequent in SOT patients, contrary to what has been suggested in other studies.

Neurotrophic West Nile virus (WNV) disease is a severe arbovirus infection in which neuronal loss is the likely anatomical substrate for the high morbidity and mortality. We investigated whether cerebrospinal fluid (CSF) protein biomarkers were elevated in vivo and related to disease severity in patients with WNV infection. This exploratory study included 114 patients (24 acute WNV, 77 noninflammatory controls, six peripheral neuropathies, seven aseptic meningoencephalitis). CSF levels of neuronal (neurofilaments, NfH-SMI35) and glial (glial fibrillary acidic protein, GFAP, S100B) biomarkers were measured by enzyme-linked immunosorbent assay (ELISA). Immunocytochemistry was performed in two fatal WNV cases. A significant proportion of patients with WNV had pathological CSF levels for NfH-SMI35 (58%, median concentration 1.01 ng/mL), GFAP (58%, 10 pg/mL), and S100B (90%, 1.29 ng/mL). The results were consistent with postmortem evidence for neuronal death and astrogliosis. Surprisingly, CSF protein biomarker levels were also found to be pathological in a considerable proportion of patients who presented with WNV fever only (100% for GFAP and S100B and 43% for NfH-SMI35). Elevated CSF protein biomarker levels are suggestive of neuronal death and glial pathology in human WNV infection. The results indicate the presence of neuroinvasive disease across the spectrum of WNV disease, including WNV fever. Muscle Nerve, 2009.

In Guadeloupe, West Nile virus (WNV) activity was first observed in equids in 2002, and a high seroprevalence was found in 2003. The objective of our study was to determine individual and environmental factors associated with the risk of WNV seropositivity during 2002-2003. Fieldwork was conducted to retrospectively determine the location of equids at the time of virus circulation and to collect information regarding environmental and individual variables. Sera were collected from 369 equids out of an estimated total population of less than 500. Thirty-four environmental and individual variables were investigated. Equids had a higher risk (p<0.001) for WNV seropositivity if they lived within the proximity "distance less than 1.5km" of marshes or swamp forests "a large freshwater formation behind mangroves" or if they remained outside after dusk. Equids living within the proximity of ouassous shrimp (Macrobrachium rosenbergii) basins or sugar cane fields had a lower risk (p<0.001) for WNV seropositivity. These results confirm that WNV circulation is more likely in the humid coastal areas of Guadeloupe. The identification of risk factors is useful for predicting future emergence sites of WNV in the archipelago and other Neotropical islands, and to better target sentinel surveillance in the region.

In September 2008, CDC, the Food and Drug Administration (FDA), and state health departments began a nationwide investigation into an increase in false-positive test results obtained with a commercially available West Nile virus (WNV) immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA). The investigation revealed that, in the United States, one lot of the commercially available test kits was the source of the false-positive results. That lot was recalled, and a second lot distributed outside the United States also was recalled. During July 1--September 30, 2008, the kit lot implicated in the United States resulted in positive tests on 568 specimens collected from 518 patients in 42 states and the District of Columbia (DC). A total of 166 (29%) specimens were retested at CDC, and 119 (72%) had false-positive results. A higher false-positive percentage were found among patients without evidence of neuroinvasive disease (77%) than patients with evidence of neuroinvasive disease (47%). Of the 518 patients, 249 (48%) had been reported to CDC as persons with WNV disease; however, only 45 (18%) had confirmatory testing that supported their inclusion in national surveillance data. Commercially available WNV test kits should be used to determine a presumptive diagnosis of WNV neuroinvasive disease. These kits should not be used to test specimens from persons without compatible illness, and any positive result should be confirmed by additional testing at a state health department or CDC.

West Nile disease in humans has been detected for the first time in Italy in two regions, Emilia-Romagna and Veneto. We conclude that also West Nile fever cases should be specifically targeted by surveillance.

On 19 September the Hungarian reference laboratory for viral zoonoses reported the first two cases of West Nile virus (WNV) neuroinvasive infection in Hungary in 2008. As of 31 October a total of 14 confirmed cases were identified.

In 2003, the occurrence and location of horses with clinical signs of West Nile virus infection were identified in the southern portion of Saskatchewan with the help of veterinarians, owners, and the regional laboratory. A total of 133 clinical cases were reported between July 30 and September 19, 2003; however, postseason surveillance suggests that the number of cases was underestimated. The case fatality rate was 43.8%(95% CI 35.2, 52.4). Factors associated with fatality in clinical cases included sex, week of onset of clinical signs, and coat color. Reported clinical cases clustered within regional health authority districts, suggesting regional differences in geographic factors, potentially including climate and mosquito control, that could contribute to the risk of disease. However, most of the variation in the risk of fatality in clinical cases is explained at the individual level rather than the Regional Health Authority level, which suggests the outcome of clinical disease is primarily determined by characteristics of, or management factors affecting, the individual horse.

This report summarizes 2007 West Nile virus (WNV) surveillance data reported to CDC through ArboNET as of 3 a.m. Mountain Standard Time, November 13, 2007. A total of 43 states had reported 3,304 cases of human WNV illness to CDC. A total of 1,803 (55%) cases for which such data were available occurred in males; median age of patients was 51 years (range: 1 month--97 years). Dates of illness onset ranged from January 8 to November 6; a total of 93 cases were fatal.

Background: West Nile virus (WNV) was first identified in the United States in 1999. In addition to a spectrum of systemic manifestations, several ocular conditions secondary to the virus have been reported, including chorioretinitis, uveitis and optic neuritis. Age and diabetes mellitus (DM) have been reported to be associated risk factors for the more severe forms of the systemic disease. Only seven cases of occlusive retinal vasculitis have been reported in patients with WNV infection. Case history: A 60-year-old Asian male presented with complaints of decreased vision in his left eye. He had been hospitalised approximately seven weeks earlier with meningo-encephalitis secondary to presumed WNV infection, at which time he was also diagnosed with DM. The visual loss coincided with the manifestation of systemic WNV infection. Old peripheral chorioretinal lesions without active inflammation in both eyes were consistent with WNV infection. In addition, retinal haemorrhage and cotton wool spots were noted in the posterior pole of both eyes with severe macular ischaemia in the left eye. Conclusion: Occlusive retinal vasculitis is an uncommon ocular manifestation of WNV, which should be suspected in patients with meningitis or encephalitis who reside in endemic areas with ocular findings of the disease.