Association of acute toxic encephalopathy with litchi consumption in an outbreak in Muzaffarpur, India, 2014: a case-control study.

BACKGROUND: Outbreaks of unexplained illness frequently remain under-investigated. In India, outbreaks of an acute neurological illness with high mortality among children occur annually in Muzaffarpur, the country's largest litchi cultivation region. In 2014, we aimed to investigate the cause and risk factors for this illness. METHODS: In this hospital-based surveillance and nested age-matched case-control study, we did laboratory investigations to assess potential infectious and non-infectious causes of this acute neurological illness. Cases were children aged 15 years or younger who were admitted to two hospitals in Muzaffarpur with new-onset seizures or altered sensorium. Age-matched controls were residents of Muzaffarpur who were admitted to the same two hospitals for a non-neurologic illness within seven days of the date of admission of the case. Clinical specimens (blood, cerebrospinal fluid, and urine) and environmental specimens (litchis) were tested for evidence of infectious pathogens, pesticides, toxic metals, and other non-infectious causes, including presence of hypoglycin A or methylenecyclopropylglycine (MCPG), naturally-occurring fruit-based toxins that cause hypoglycaemia and metabolic derangement. Matched and unmatched (controlling for age) bivariate analyses were done and risk factors for illness were expressed as matched odds ratios and odds ratios (unmatched analyses). FINDINGS: Between May 26, and July 17, 2014, 390 patients meeting the case definition were admitted to the two referral hospitals in Muzaffarpur, of whom 122 (31%) died. On admission, 204 (62%) of 327 had blood glucose concentration of 70 mg/dL or less. 104 cases were compared with 104 age-matched hospital controls. Litchi consumption (matched odds ratio [mOR] 9·6 [95% CI 3·6 - 24]) and absence of an evening meal (2·2 [1·2-4·3]) in the 24 h preceding illness onset were associated with illness. The absence of an evening meal significantly modified the effect of eating litchis on illness (odds ratio [OR] 7·8 [95% CI 3·3-18·8], without evening meal; OR 3·6 [1·1-11·1] with an evening meal). Tests for infectious agents and pesticides were negative. Metabolites of hypoglycin A, MCPG, or both were detected in 48 [66%] of 73 urine specimens from case-patients and none from 15 controls; 72 (90%) of 80 case-patient specimens had abnormal plasma acylcarnitine profiles, consistent with severe disruption of fatty acid metabolism. In 36 litchi arils tested from Muzaffarpur, hypoglycin A concentrations ranged from 12·4 µg/g to 152·0 µg/g and MCPG ranged from 44·9 µg/g to 220·0 µg/g. INTERPRETATION: Our investigation suggests an outbreak of acute encephalopathy in Muzaffarpur associated with both hypoglycin A and MCPG toxicity. To prevent illness and reduce mortality in the region, we recommended minimising litchi consumption, ensuring receipt of an evening meal and implementing rapid glucose correction for suspected illness. A comprehensive investigative approach in Muzaffarpur led to timely public health recommendations, underscoring the importance of using systematic methods in other unexplained illness outbreaks. FUNDING: US Centers for Disease Control and Prevention.

Outbreaks of unexplained neurologic illness - Muzaffarpur, India, 2013-2014.

Outbreaks of an unexplained acute neurologic illness affecting young children and associated with high case-fatality rates have been reported in the Muzaffarpur district of Bihar state in India since 1995. The outbreaks generally peak in June and decline weeks later with the onset of monsoon rains. There have been multiple epidemiologic and laboratory investigations of this syndrome, leading to a wide spectrum of proposed causes for the illness, including infectious encephalitis and exposure to pesticides. An association between illness and litchi fruit has been postulated because Muzaffarpur is a litchi fruit-producing region. To better characterize clinical and epidemiologic features of the illness that might suggest its cause and how it can be prevented, the Indian National Centre for Disease Control (NCDC) and CDC investigated outbreaks in 2013 and 2014. Clinical and laboratory findings in 2013 suggested a noninflammatory encephalopathy, possibly caused by a toxin. A common laboratory finding was low blood glucose (<70 mg/dL) on admission, a finding associated with a poorer outcome; 44% of all cases were fatal. An ongoing 2014 investigation has found no evidence of any infectious etiology and supports the possibility that exposure to a toxin might be the cause. The outbreak period coincides with the month-long litchi harvesting season in Muzaffarpur. Although a specific etiology has not yet been determined, the 2014 investigation has identified the illness as a hypoglycemic encephalopathy and confirmed the importance of ongoing laboratory evaluation of environmental toxins to identify a potential causative agent, including markers for methylenecyclopropylglycine (MCPG), a compound found in litchi seeds known to cause hypoglycemia in animal studies. Current public health recommendations are focused on reducing mortality by urging affected families to seek prompt medical care, and ensuring rapid assessment and correction of hypoglycemia in ill children.

Microevolution of highly pathogenic avian influenza A(H5N1) viruses isolated from humans, Egypt, 2007-2011.

We analyzed highly pathogenic avian influenza A(H5N1) viruses isolated from humans infected in Egypt during 2007-2011. All analyzed viruses evolved from the lineage of subtype H5N1 viruses introduced into Egypt in 2006; we found minimal evidence of reassortment and no exotic introductions. The hemagglutinin genes of the viruses from 2011 formed a monophyletic group within clade 2.2.1 that also included human viruses from 2009 and 2010 and contemporary viruses from poultry; this finding is consistent with zoonotic transmission. Although molecular markers suggestive of decreased susceptibility to antiviral drugs were detected sporadically in the neuraminidase and matrix 2 proteins, functional neuraminidase inhibition assays did not identify resistant viruses. No other mutations suggesting a change in the threat to public health were detected in the viral proteomes. However, a comparison of representative subtype H5N1 viruses from 2011 with older subtype H5N1 viruses from Egypt revealed substantial antigenic drift.

Pandemic (H1N1) 2009 and Hajj Pilgrims who received Predeparture Vaccination, Egypt.

In Egypt, vaccination against pandemic (H1N1) 2009 virus was required of pilgrims departing for the 2009 Hajj. A survey of 551 pilgrims as they returned to Egypt found 542 (98.1% [weighted]) reported receiving the vaccine; 6 (1.0% [weighted]) were infected with influenza virus A (H3N2) but none with pandemic (H1N1) 2009 virus.

Virus isolations and high population density implicate Culex antennatus (Becker) (Diptera: Culicidae) as a vector of Rift Valley Fever virus during an outbreak in the Nile Delta of Egypt.

In June, 2003, Egypt's hospital-based electronic disease surveillance system began to record increased cases of acute febrile illness from governorates in the Nile Delta. In response to a request for assistance from the Egyptian Ministry of Health and the World Health Organization (WHO), the U.S. Naval Medical Research Unit No. 3 (NAMRU-3) provided assistance in identifying the cause and extent of this outbreak. Testing of human clinical samples (n=375) from nine governorates in Egypt identified 29 cases of RVF viremia that spanned the period of June to October, and a particular focus of disease in Kafr el Sheikh governorate (7.7% RVF infection rate). Veterinary samples (n=101) collected during this time in Kafr el Sheikh and screened by immunoassay for RVFV-specific IgM identified probable recent infections in cattle (10.4%) and sheep (5%). Entomologic investigations that focused in rural, rice growing villages in the Sidi Salim District of Kafr el Sheikh during August-September, 2003, collected, identified, and tested host-seeking female mosquitoes for the presence of pathogenic viruses. Three isolates of RVF virus (RVFV) were obtained from 297 tested pools of female mosquitoes and all three RVFV isolates came from Cx. antennatus (Becker). While Cx. pipiens has been considered the primary vector of RVF virus in Egypt and is often the most common man-biting species found, Cx. antennatus was the dominant species captured at the 2003 outbreak location in Kafr el Sheikh governorate. This is the first time that Cx. antennatus has been found naturally infected with RVFV in Egypt.

Capacity-building efforts by the AFHSC-GEIS program.

Capacity-building initiatives related to public health are defined as developing laboratory infrastructure, strengthening host-country disease surveillance initiatives, transferring technical expertise and training personnel. These initiatives represented a major piece of the Armed Forces Health Surveillance Center, Division of Global Emerging Infections Surveillance and Response System (AFHSC-GEIS) contributions to worldwide emerging infectious disease (EID) surveillance and response. Capacity-building initiatives were undertaken with over 80 local and regional Ministries of Health, Agriculture and Defense, as well as other government entities and institutions worldwide. The efforts supported at least 52 national influenza centers and other country-specific influenza, regional and U.S.-based EID reference laboratories (44 civilian, eight military) in 46 countries worldwide. Equally important, reference testing, laboratory infrastructure and equipment support was provided to over 500 field sites in 74 countries worldwide from October 2008 to September 2009. These activities allowed countries to better meet the milestones of implementation of the 2005 International Health Regulations and complemented many initiatives undertaken by other U.S. government agencies, such as the U.S. Department of Health and Human Services, the U.S. Agency for International Development and the U.S. Department of State.

Dengue hemorrhagic fever outbreak in children in Port Sudan.

Dengue fever (DF)/dengue hemorrhagic fever (DHF) has emerged as a global public health problem with countries in Asia and the Pacific sharing more than 70% of the disease burden. In 2004-2005 a total of 312 cases admitted to Pediatric and Sea Port Hospitals in Port Sudan were clinically diagnosed as DHF. The mortality rate recorded was 3.8% (n=12). Of the cases 73.4% were patients 5-15 years of age. A total of 91.2% of cases were admitted during May and June 2005 with 49.4% residing in the eastern region of Port Sudan. Dengue shock syndrome was observed in 37 of 312 (11.9%). All patients had thrombocytopenia with platelets count ranged from <100,000 to <150,000 cell/mm³. Of the 40 sera tested using RAPID-cassette test in the Khartoum Central Public Health Lab, 36 (90%) were dengue IgM positive. A subset of these sera (n=23) were sent to NAMRU-3 and confirmed by IgM-capture ELISA; 9 of 23 were PCR positive for dengue serotype 3.

Studies on West Nile virus infection in Egypt.

We conducted a prospective cohort study to determine prevalence and incidence of West Nile virus (WNV) in Egypt. Cohorts were established in Upper (UE), Middle (ME), and Lower (LE) Egypt. Additionally, a cross-sectional serosurvey was performed in the North (NS) and South (SS) Sinai. Cohorts were bled initially and 1 year later. Sera were tested for WNV-IgG by ELISA and positive sera were confirmed by plaque reduction neutralization test (PRNT). Sentinel chicken flocks placed in the above sites were bled monthly for virus isolation and serology. Mosquitoes were collected monthly from the above sites and tested for WNV. Human seroprevalence rates were 35%, 27%, 14%, 1% and 7% in UE, ME, LE, NS and SS, respectively. Seroconversion rates were 18%, 17% and 7% in UE, ME and LE, respectively; 49% of the seroconverters reported undiagnosed febrile illness. Sentinel chickens showed seroconversion in all study sites. WNV was isolated from both sentinel chickens and mosquitoes in cohort sites. This study demonstrates that WNV was actively circulating during the study period in different areas in Egypt and causing febrile illness in a considerable proportion of individuals in the study sites.

Zoonotic transmission of avian influenza virus (H5N1), Egypt, 2006-2009.

During March 2006-March 2009, a total of 6,355 suspected cases of avian influenza (H5N1) were reported to the Ministry of Health in Egypt. Sixty-three (1%) patients had confirmed infections; 24 (38%) died. Risk factors for death included female sex, age > or = 15 years, and receiving the first dose of oseltamivir >2 days after illness onset. All but 2 case-patients reported exposure to domestic poultry probably infected with avian influenza virus (H5N1). No cases of human-to-human transmission were found. Greatest risks for infection and death were reported among women > or = 15 years of age, who accounted for 38% of infections and 83% of deaths. The lower case-fatality rate in Egypt could be caused by a less virulent virus clade. However, the lower mortality rate seems to be caused by the large number of infected children who were identified early, received prompt treatment, and had less severe clinical disease.

Antigenic and genetic diversity of highly pathogenic avian influenza A (H5N1) viruses isolated in Egypt.

Highly pathogenic avian influenza A virus (H5N1) has diverged antigenically and genetically since its initial detection in Asia in 1997. Viruses belonging to clade 2.2 in particular have been reported in numerous countries with the majority occurring in Egypt. Previous reports identified antigenic similarities between viruses belonging to clade 2.2. However, poultry and human viruses isolated in northern Egypt during 2007 and 2008 were found to be antigenically distinct from other clade 2.2 viruses from this country. Genetic analysis of the hemagglutinin revealed a high degree of nucleotide and amino acid divergence. The antigenic changes in Egyptian viruses isolated during 2007-08 necessitated that two of these strains be considered as potential H5N1 pre-pandemic vaccine candidates.

Effects of duration of injection drug use and age at first injection on HCV among IDU in Kabul, Afghanistan.

BACKGROUND: Hepatitis C virus (HCV) prevalence is high among injection drug users (IDUs) in Afghanistan. Duration of injection and young age at first injection are common risk factors for HCV in IDU populations. The association of HCV with these time factors was analyzed. METHODS: Socio-demographic and drug use behavior information were collected. Participants had rapid testing for HCV with recombinant immunoblot assay confirmation. Modeling of non-linear associations was performed using fractional polynomial logistic regression. RESULTS: Among 459 male IDUs, age at first injection had a constant HCV risk (odds ratio (OR): 1.01 per year; 95% confidence interval (CI): 0.98-1.03), while each additional year of injection drug use had a significantly increased risk (OR: 4.72 per year, 95% CI: 2.92-7.66). HCV risk increased significantly with each additional year of injecting drug use by groups of injectors: young (< or =22 years, OR: 1.97; 95% CI: 1.27-3.07), middle (23-28 years, OR: 1.76; 95% CI: 1.28-2.43) and older (> or =29 years, OR: 7.56; 95% CI: 3.15-18.14). CONCLUSION: The probability of HCV infection increased markedly by duration of injection drug use and varied according to age at first injection. Drug counseling and educational efforts should be directed to older drug users who have not yet initiated injecting and to young IDUs to avert infection and reduce risky drug use behaviors.

Immunization of chickens with Newcastle disease virus expressing H5 hemagglutinin protects against highly pathogenic H5N1 avian influenza viruses.

BACKGROUND: Highly-pathogenic avian influenza virus (HPAIV) and Newcastle disease virus (NDV) are the two most important poultry viruses in the world. Natural low-virulence NDV strains have been used as vaccines over the past 70 years with proven track records. We have previously developed a reverse genetics system to produce low-virulent NDV vaccine strain LaSota from cloned cDNA. This system allows us to use NDV as a vaccine vector for other avian pathogens. METHODOLOGY/PRINCIPAL FINDING: Here, we constructed two recombinant NDVs (rNDVs) each of which expresses the hemagglutinin (HA) gene of HPAIV H5N1 strain A/Vietnam/1203/2004 from an added gene. In one, rNDV (rNDV-HA), the open reading frame (ORF) of HA gene was expressed without modification. In the second, rNDV (rNDV-HAF), the ORF was modified so that the transmembrane and cytoplasmic domains of the encoded HA gene were replaced with those of the NDV F protein. The insertion of either version of the HA ORF did not increase the virulence of the rNDV vector. The HA protein was found to be incorporated into the envelopes of both rNDV-HA and rNDV-HAF. However, there was an enhanced incorporation of the HA protein in rNDV-HAF. Chickens immunized with a single dose of either rNDV-HA or rNDV-HAF induced a high titer of HPAIV H5-specific antibodies and were completely protected against challenge with NDV as well as lethal challenges of both homologous and heterologous HPAIV H5N1. CONCLUSION AND SIGNIFICANCE: Our results suggest that these chimeric viruses have potential as safe and effective bivalent vaccines against NDV and. HPAIV. These vaccines will be convenient and affordable, which will be highly beneficial to the poultry industry. Furthermore, immunization with these vaccines will permit serological differentiation of vaccinated and avian influenza field virus infected animals.

Isolation of cryptococcus, Candida, Aspergillus, Rhodotorula and nocardia from meningitis patients in egypt.

Meningitis occurs throughout Egypt and is largely attributed to bacterial pathogens, but there is little information on fungal etiologies of meningitis. We, therefore, investigated fungal infections among Egyptian patients with acute and subacute meningitis who tested negative for bacterial and viral agents. A total of 1000 cerebrospinal fluid (CSF) samples collected from nine governorates of Egypt during 1998-2002 were initially stained with Gram's, India ink, and lacto-phenol cotton-blue stains, and examined under light microscope to detect fungal elements. All CSF samples were cultured on brain heart infusion, Wickerham and Staib agar media for fungus isolation. CSF with suspected Cryptococcus neoformans infections were also tested by latex agglutination test for antigen detection. Species identification of selected isolates was carried out at the Mycotic Diseases Branch, CDC, Atlanta, Georgia, USA. Fungal agents were detected microscopically and by culture in 17 of 1000 (1.7%) CSF samples tested. Ten of 17 were identified as C. neoformans var grubii (serotype A), 4 as Candida albicans, and one each of Aspergillus candidus, Rhodotorula mucilaginosa (rubra) and Nocardia spp (actinomycetes). Out of the 17 cases with fungal CSF infection, 8 died (Cryptococcus-3, Candida-2, Aspergillus, Rhodotorula and Nocardia) and 2 suffered neurological sequelae. Of the 10 cryptococcal meningitis patients, 4 were HIV positive and one was diagnosed with lymphoma. To our knowledge, this is the first study on isolation of fungi other than Cryptococcus from CSF of Egyptian patients with acute/subacute meningitis. Consideration must now be given to cryptococcosis and candidiasis as potential etiologies of meningitis in Egypt.

Risk factors for brucellosis in Samarqand Oblast, Uzbekistan.

OBJECTIVES: This study was conducted to identify the potential risk factors for human brucellosis infection in Samarqand, Uzbekistan. METHODS: Clinically identified cases admitted to different hospitals during 2004-2006 (N=144), and age-, sex- and residence-matched control patients (N=288) with other unrelated conditions, were included in this study. Structured questionnaires were completed and consent forms signed. Patients and controls were tested on site for Brucella infection by standard tube agglutination test and culture. Statistical analyses were performed with Stata software for univariate and multivariate analysis. RESULTS: Among the 144 patients with confirmed brucellosis, 137 (95.1%) owned farm animals, 135 (93.8%) were from rural areas, and 119 (82.6%) were enrolled during the animal breeding season. Multivariate analysis indicated that brucellosis was highly associated with contact with aborted animals (adjusted matched odds ratio (AMOR) 87.19, 95% confidence interval (CI) 9.36-911.85; p<0.001), slaughtering/butchering animals (AMOR 35.35, 95% CI 6.25-199.77; p<0.001) in the household, consumption of raw milk (AMOR 54.13, 95% CI 1.98-1476.13; p=0.018), and being in a family that had brucellosis sharing the same exposure (AMOR 15.93, 95% CI 1.37-184.97; p=0.027). CONCLUSIONS: To reduce the burden of brucellosis in Samarqand Oblast, veterinary services should be improved. Also public health education programs should be increased. Implementing these measures will minimize exposure to infected farm animals and reduce the risk of infection.

A cross-sectional assessment of utilization of addiction treatment among injection drug users in Kabul, Afghanistan.

The purpose of this study was to describe prior use of detoxification and addiction-treatment programs among injection drug users (IDUs) in Kabul, Afghanistan. From 2005-2006, IDUs (n = 464) recruited into this cross-sectional study completed an interviewer-administered questionnaire and whole blood rapid testing with fingerstick samples for HIV, syphilis, and hepatitis C antibody and B surface antigen testing. Participants were predominantly male (99.8%), Afghan (98.9%), and had little formal education. Correlates of detoxification and addiction treatment were identified with logistic regression. The majority (94.0%, n = 435) felt great/urgent need for treatment, of whom 56.3% (n = 245) reported inability to access treatment. Prior detoxification was associated with new needle use with each injection (AOR = 1.91, 95% CI: 1.12-3.26) and prior incarceration (AOR = 1.81, 95% CI: 1.04-3.13). The study's limitations are noted. Rapid scale-up and subsidy of needle and syringe programs and opioid agonist treatment is urgently needed in Kabul.

Oseltamivir resistance mutation N294S in human influenza A(H5N1) virus in Egypt.

In December 2006, three human specimens were received that were suspected positive for influenza A(H5N1). The specimens were tested using real time PCR. And the presence of A(H5N1) virus was confirmed in 2 patients (16F and 26M), The NA sequence from A(H5N1) positive specimens collected before and after antiviral therapy revealed a mutation (N294S) (N295S according to N1 numbering), previously associated with resistance to oseltamivir. When tested with NA inhibition assays, the two N294S viruses from Egypt exhibited from 57 to 138-fold reduction in susceptibility to oseltamivir, depending on the assay. To our knowledge, this is the first time oseltamivir resistance has been detected in A(H5N1) infecting a human prior to treatment.

Outbreak of Q fever among US military in western Iraq, June-July 2005.

An outbreak of Q fever occurred in 22 (58%) of 38 Marines deployed to Iraq in 2005. Fever (in 100% of patients), respiratory symptoms (76%), and gastrointestinal symptoms (53%) were common. Possible risk factors included dust and exposure to animals and ticks.

Correlates of receptive and distributive needle sharing among injection drug users in Kabul, Afghanistan.

We describe receptive and distributive needle/syringe sharing among injection drug users (IDUs) in Kabul, Afghanistan. In this cross-sectional study, IDUs completed an interviewer-administered questionnaire. Logistic regression identified correlates of needle sharing in the last six months. Receptive and distributive sharing in the last six months were reported by 28.2% and 28.7% of participants, respectively, and were both independently associated with reported difficulty obtaining new syringes (Receptive sharing: AOR = 2.60, 95% CI: 1.66-4.06; Distributive: AOR = 1.56, 95% CI: 1.02-2.39). Receptive and distributive sharing are common among IDU in Kabul; scaling up availability of sterile, no-cost injecting equipment is urgently needed.

Epidemiology and risk factors for endemic typhoid fever in Uzbekistan.

BACKGROUND: To investigate the risk factors for infection with endemic typhoid fever in the Samarkand region of Uzbekistan. METHODS: Case-control study of culture-confirmed bloodstream infection with Salmonella Typhi. Patients were compared to age-matched community controls. Salmonella Typhi isolates were tested for antimicrobial susceptibility. RESULTS: We enrolled 97 patients and 192 controls. The median age of patients was 19 years. In a conditional regression model, consumption of unboiled surface water outside the home [adjusted odds ratio (aOR)=3.0, 95% confidence interval (CI)=1.1-8.2], use of antimicrobials in the 2 weeks preceding onset of symptoms (aOR=12.2, 95% CI 4.0-37.0), and being a student (aOR=4.0, 95% CI 1.4-11.3) were independently associated with typhoid fever. Routinely washing vegetables (aOR 0.06, 95% CI 0.02-0.2) and dining at a tea-house (aOR 0.4, 95% CI 0.2-1.0) were associated with protection against illness. Salmonella Typhi resistant to ampicillin, chloramphenicol, and trimethoprim-sulfamethoxazole was identified in 6 (15%) of 41 isolates tested. CONCLUSIONS: Endemic typhoid fever in Uzbekistan is transmitted by contaminated water. Recent use of antimicrobials also increased risk of infection. Targeted efforts at improving drinking water quality, especially for students and young adults, are likely to decrease transmission of typhoid fever. Measures to decrease the unnecessary use of antimicrobials would be expected to reduce the risk of typhoid fever and decrease the spread of multiple drug-resistant Salmonella Typhi.